Pasteurella multocida from deep nasal swabs and tracheobronchial lavage in calves

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To guide practitioners, knowledge of the bacteria involved in an outbreak and their antibiotic susceptibility is warranted. To this end, samples from the upper or lower respiratory tract of calves can be submitted for bacteriological culture and susceptibility testing of relevant isolates. However, it is debated whether isolates from the upper respiratory tract are representative of bacteria causing infections in the lower tract. In this study, we used MALDI-TOF MS, multilocus sequence typing (MLST) and core-genome multilocus sequence typing (cgMLST) to compare isolates from deep nasal swabs (DNS) and from tracheobronchial lavage (TBL) in 219 calves from 25 farms. Results: The predominant bacterial pathogen in this study was Pasteurella multocida , which was isolated from 37.4% of DNS and 22.4% of TBL. There was no statistically significant difference in isolation frequency between healthy and diseased calves for DNS (p = 0.778) or TBL (p = 0.410). At least one P. multocida isolate was obtained from 83.3% of the calf groups. At the 49 samplings where P. multocida was isolated from TBL, the same species was isolated from DNS at 29 samplings (59.2%). From 28 of these samplings, isolates were evaluated by MLST and at 24 samplings (86.0%) P. multocida from DNS and TBL were of the same sequence type (ST). Moreover, at 21 of the samplings (75.0%), cgMLST showed that the genetic distance within isolate pairs was less than two alleles, and DNS and TBL isolates were considered identical. At seven samplings (25%), the genetic distance within isolate pairs was greater, and DNS and TBL isolates were considered nonidentical. Within farms and groups of calves, P. multocida of several different STs were isolated. Conclusions: Pasteurella multocida was readily isolated from DNS and in calves where this species was isolated also from TBL, isolates from DNS and TBL were identical at 75% of the samplings. This suggests that during an outbreak of BRD, submission of DNS samples from 4–6 calves could be a convenient approach for practitioners seeking guidance on bacterial pathogens present and their antibiotic susceptibility. Bovine respiratory diseases cattle cgMLST MSLT sampling. Figures Figure 1 Background Bovine respiratory disease (BRD) is a common and costly problem in intensively raised cattle worldwide [ 1 – 5 ]. The background of BRD is multifactorial and includes infectious agents and predisposing factors related to the immunological and general status of the animals and to management and housing [ 6 – 8 ]. Due to its multifactorial background, the pathogenesis of BRD varies, but in general, viral infections of the upper respiratory tract precede bacterial infections of the lower respiratory tract [ 9 ]. The bacteria commonly involved often reside in the upper airways of healthy calves and include Pasteurella multocida , Mannheimia haemolytica , Histophilus somni and Mycoplasma bovis [ 8 – 10 ]. The control of BRD is complex and involves measures to hinder the spread of infectious agents and to mitigate predisposing factors [ 7 , 11 – 13 ]. Therapeutic, prophylactic or metaphylactic antibiotic treatments are also used to control BRD, but routine use of antibiotics should be avoided due to the risk of emerging antibiotic resistance [ 6 , 11 , 12 , 14 ]. For reasons of animal welfare and to minimise economic losses, however, it is often necessary to use therapeutic antibiotic treatments when BRD manifests in a group of animals [ 11 ]. Considering the increased occurrence of acquired antibiotic resistance, it is important that treatment is guided by knowledge of the antibiotic susceptibility of the relevant bacteria [ 5 , 15 – 18 ]. Information on the antibiotic susceptibility of bacteria involved in BRD is available from national monitoring programs, e.g., [ 19 – 24 ], and from other scientific literature, e.g., [ 16 , 25 – 27 ]. However, such compiled data are derived from sources with varying study designs and from different farm types and might not be representative of the situation on a specific farm [ 18 ]. It is therefore preferable if information is available for relevant bacteria isolated from acutely affected and untreated animals from the farm where treatments are to be instituted [ 18 ]. This is even more relevant for farms experiencing treatment failures where the presence of antibiotic resistance could cause re-evaluation of treatment protocols [ 28 ]. Moreover, in some countries, susceptibility testing of isolates from animals on a farm is mandatory for the use of specific antibiotics on that farm [ 16 , 29 ]. To obtain isolates of bacterial respiratory pathogens for susceptibility testing, it is convenient and simple for practitioners to collect samples for bacteriological culture from the upper respiratory tract using nasal swabs (NS), deep nasal swabs (DNS) or nasopharyngeal swabs (NPS) [ 5 , 28 ]. Samples from the lower respiratory tract can also be collected in a clinical setting. Several different techniques are available, including bronchoalveolar lavage (BAL), tracheobronchial lavage (TBL), transtracheal wash (TTW) or transtracheal swab (TTW) [ 5 , 28 , 30 ]. These methods are, however, more complicated, and invasive than NS, DNS or NPS and are considered more stressful for the animals [ 15 , 25 , 31 ]. To collect samples from the upper respiratory tract is convenient, but it is debated whether isolates from NS, DNS or NPS are representative of respiratory pathogens in the lower respiratory tract, which are the targets for antibiotic therapy [ 5 , 18 , 32 ]. However, in studies that have compared paired isolates of Pasteurellacae from the upper and lower respiratory tracts of calves, there is generally moderate to almost perfect agreement at the species level [ 15 , 28 , 29 , 33 – 35 ]. For M. haemolytica , there was also a high agreement between paired isolates from the upper and lower respiratory tracts when molecular methods were used to compare isolates [ 15 , 35 , 36 ]. However paired isolates of P. multocida from the upper and lower respiratory tracts of calves have not been compared by molecular methods. The aim of this study was to evaluate whether samples from the upper respiratory tract collected by deep nasal swabs (DNS) from calves provide comparable information regarding bacterial pathogens as if the lower respiratory tract of the same calves were sampled by tracheobronchial lavage (TBL). Our hypothesis is that bacterial pathogens in the lower respiratory tract of a calf can generally also be cultured from DNS from the same calf or from other calves in the same group. To test this hypothesis, we used data from field investigations and reanalysed a collection of strains from the investigations using matrix-assisted laser-desorption ionisation-time-of-flight (MALDI-TOF MS), multilocus sequence typing (MLST) and core-genome multilocus sequence typing (cgMLST). MLST is a method, first described and evaluated in Neisseria meningitidis , in which the gene sequences of, usually, seven housekeeping genes are compared [ 37 ]. The number of identical gene variants in an isolate-to-isolate comparison makes it possible to calculate the evolutionary relationships between isolates of the same species. The core-genome multilocus sequence typing method also compares gene variants obtained from genome analysis. The genome sequences enable the comparison of 1609 genes instead of 7; hence, cgMLST can show relatedness between isolates with considerably higher detail than MLST. Methods Field investigations The data and bacterial isolates in this study are from field investigations from 1997–2000 on Swedish cattle farms experiencing outbreaks of BRD and affiliated with a health control program through the Swedish Animal Health Service [ 38 ]. To manage the outbreaks, the calves were examined clinically, and those with a rectal temperature > 39.5°C and at least one of the clinical signs of nasal discharge, cough or abnormal respiratory sounds on auscultation were considered BRD cases. To assess antibiotic susceptibility of possible bacterial respiratory pathogens, 4–6 calves from a group with clinical signs of BRD were sampled at each visit. When only a few diseased calves were available, healthy calves from the group were sampled instead. Samples were collected from the posterior nasal cavity (DNS) using cotton swabs (Copan, Amies charcoal medium). The swab was entered into the nasal cavity to its full length (≈ 12 cm) after dry cleaning of the outer nares. Bacteriological samples were also collected from the lower respiratory tract by tracheobronchial lavage (TBL). The catheter used for TBL was designed to protect the sample from contamination when it was passed through the nasal cavity [ 30 ]. In brief, a catheter made of Silastic tubing (diameter 5 mm, length 50 cm) with an inner lining of Teflon tubing was passed through the nasal cavity into the trachea. A silicon rubber stopper 10 cm from the upper end of the catheter hindered passage more than 40 cm into the airways. When located in the trachea, a 100 cm flexible Teflon tube was passed through the catheter and through a slit in a silicon tip sealing the catheter. The Teflon tube was then passed down to the region of the tracheal bifurcation where 20 ml of isotonic saline was instilled and immediately aspirated. The mean volume of fluid retracted as a sample was 4.5 ml (range 0.5–15 ml). In all, 37 different farms were visited, 400 calves were examined and sampled, and 80 of the calves were examined and sampled again within 3–9 weeks. The DNS and TBL samples collected were kept at + 8°C and transported within 24 h to the National Veterinary Agency (SVA), where they were cultivated for respiratory bacterial pathogens on blood agar, blue agar and hematin agar. Colonies with a macroscopic appearance in agreement with Pasteurellacae were subcultured and identified by biochemical tests. The culture results were registered as “Pasteurella-like bacteria” (PLB), “Other bacteria” (OB) or “Negative culture” (NEG). Descriptive data for farms and individual calves from the field investigations were stored together with laboratory results. Isolates identified as PLB were stored at -80°C, and isolates identified as OB were not stored. As the calves were examined and sampled in the context of the routines for managing clinical outbreaks of BRD, no ethical approval was needed. Selection of calves from the field investigations For the present study, calves from the field investigations were selected using the following criteria: 1 both DNS and TBL should have been collected; 2 no antibiotic treatment within 3 days prior to sampling; 3 data should be available for at least 3 calves in the calf group; 4 isolates identified as PLB should be available in the strain collection. Selection of isolates for species identification by MALDI-TOF To refine the identification of bacterial species, isolates identified as PLB in the field investigations were analysed by MALDI-TOF MS. From the calves selected for this study, 85 isolates from DNS and 51 from TBL were available in the strain collection. These isolates were analysed by the MALDI Biotyper system (Bruker Daltonics, Bremen, Germany) to identify the species. Material from a single colony from the agar plate was spotted on a MALDI plate without pretreatment. The spots were covered with 1 µl of matrix solution consisting of α-cyano-4-hydroxycinnamic acid (HCCA), air-dried at room temperature, and introduced into the MALDI-TOF mass spectrometer for analysis. The spectra of all the isolates were compared to the spectra in the database, and identification was provided with a reliability score. A score ≥ 2.0 was considered reliable for species identification. Whole-genome Sequencing and Genome Assembly At 29 samplings, P. multocida was isolated from both DNS and TBL on the same calf. The genomes of 28 of these isolate pairs (56 isolates from 25 different calves) were sequenced and compared (one isolate pair was not available). Colony material was collected from blood agar plates for DNA extraction using a Qiagen EZ1 DNA Tissue Kit (Qiagen, Halden, Germany). Nextera Library preparation (Illumina, Foster City, United States) and paired-end sequencing (2 x 150bp) was performed at Clinical Genomics Stockholm, SciLifeLab (Solna, Sweden) using an Illumina NovaSeq 6000 instrument. The raw reads for each sample were quality checked using FastQC vo.11.9 [ 39 ], trimmed using Trimmomatic v0.39 [ 40 ] and assembled using SPAdes v3.14.0 [ 41 ]. The assemblies were error-corrected using Pilon v1.23 [ 42 ]. Details on the analysis parameters and FastQC data can be found in Additional file 1, Supplementary Table 1. Multilocus Sequence Typing For P. multocida , there are two public MLST schemes [ 43 , 44 ] available at PubMLST [ 45 ], of which we chose to use the multihost MLST scheme of Davies et al. [ 43 ]. Core-Genome Multilocus Sequence Typing and Minimum Spanning Tree Construction There were, at the time of the writing of this publication and to our knowledge, no cgMLST schemes for P. multocida suitable for global nomenclature. Therefore, we constructed an ad hoc cgMLST scheme from the project data suitable for analysis of this dataset alone using the cgMLST Target Definer v.1.5 function in SeqSphere + version 6.0.2 (Ridom, Würzburg, Germany). We used the annotated genome with the GenBank accession number NZ_CP008918.1 as the seed genome and the genomes NZ_CP015569.1, NZ_CP037861.1 and NZ_CP037865.1 as the penetration genomes and excluded hits found in the plasmid sequence NC_017035.1, gene duplicates and truncated genes. This resulted in a cgMLST scheme consisting of 1611 targets, as documented in Supplementary Table 1. Using the results from 1609 of the cgMLST targets, the phylogenetic distance was calculated using the Minimum Spanning Tree method (Kruskal JB 1956) using GrapeTree v.1.5.0 [ 46 ]. Statistical calculations Differences in culture results between DNS and TBL were evaluated by Fischer’s exact test, with P ≤ 0.05 considered evidence of a significant difference. Agreement between culture results for DNS and TBL for P. multocida was evaluated by Kappa statistics and McNemar’s test [ 47 ]. Kappa values (ƙ) were interpreted to indicate the strength of agreement as follows: 0.8, almost perfect. The exact McNemar significance probability test was used to determine the potential for bias between DNS and TBL, and values ≤ 0.05 were considered evidence of bias. Statistical calculations were made using the GraphPad software. Results Bacteriological culture Using the specified criteria, 219 samplings of 171 different calves were selected for this study, 48 of which were examined and sampled twice (Table 1 ). The calves were from 30 different groups on 25 farms, and 12 calf groups were sampled twice (Table 1 ). Ninety of the samplings were of calves with clinical signs of BRD, and 129 were from healthy calves (Table 1 ). Among the 219 DNS samples, P. multocida was isolated from 82 (37.4%), M. haemolytica from one (0.5%), Moraxella bovoculi from two (0.9%) and OB from 143 samples (65.3%) (Table 2 ). There was no difference in the isolation frequency of P. multocida between healthy (38.0%) and diseased calves (36.7%) (P = 0.778). Among the 219 TBL samples, P. multocida was isolated from 49 (22.4%), M. bovoculi from two (0.9%), and OB from 33 (15.1%), whereas most samples, 135 (61.6%), yielded no growth. There was no difference in the isolation frequency of P. multocida between healthy (20.2%) and diseased calves (25.6%) (P = 0.410), but the proportion of negative cultures was greater in healthy (68.2%) than in diseased calves (52.2%) (P = 0.023). Table 1 Descriptive data from field investigations from 1997–2000 for the 25 farms and 171 calves (48 calves sampled twice) selected for the present study. Farm ID Type a Calf group Sampling 1 Interval S1 - S2 (days) Sampling 2 Total number of samplings Number of calves b Age (days) Mean (range) Number of calves Age (days) Mean (range) 1 Meat A 6 (1) 61 (19–171) 6 2 Meat A 4 (2) NA c 33 4 (3) 8 B 4 (4) NA 4 C 6 (4) NA 43 4 (1) 10 5 Meat A 6 (3) 80 (63–130) 6 6 Meat A 5 (2) 85 (46–129) 35 4 (1) 127 (81–164) 9 B 6 (0) 95 (76–131) 6 7 Meat A 6 (3) 98 (84–113) 29 4 (0) 131 (123–143) 10 9 Meat A 3 (0) 118 (94–154) 59 3 (1) NA 6 10 Meat A 4 (1) 88 (58–115) 49 5 (3) 126 (107–159) 9 B 6 (2) NA 49 5 (0) NA 11 11 Meat A 4 (2) 55 (49–58) 34 4 (2) 89 (84–93) 8 B 5 (3) 51 (43–59) 5 13 Meat A 6 (3) NA 33 6 (0) NA 12 15 Meat A 6 (3) NA 6 16 Meat A 6 (4) 102 (79–128) 6 17 Dairy A 6 (4) NA 6 18 Meat A 5 (4) 92 (72–112) 5 22 Meat A 5 (1) 99 (83–124 32 6 (1) 139 (123–164) 11 25 Meat A 6 (2) 103 (92–130) 6 28 Meat A 5 (3) 98 (86–113) 5 29 Dairy A 5 (2) 55 (20–79) 5 31 Meat A 6 (4) 67 (44–116) 6 33 Meat A 5 (3) 78 (62–91) 5 34 Meat A 6 (4) 102 (77–134) 6 35 Meat A 6 (6) NA 6 36 Meat A 6 (2) 60 (43–97) 6 40 Dairy A 6 (1) 52 (19–73) 29 6 (2) 81 (48–102) 12 44 Meat A 6 (1) 77 (73–86) 6 46 Dairy A 6 (2) 60 (25–83) 20 6 (0) 80 (45–103) 12 Total 162 (76) 80 (19–171) 57 (14 ) 108 (45–164) 219 a Meat = farms specialised in raising calves for meat production, Dairy = farms with milk production; b Number of calves with clinical signs of BRD in parenthesis; c Data not available. HERE Table 1 Crosstabulation of culture results from DNS and TBL showed that when P. multocida or M. bovoculi was isolated from TBL, the same species was isolated from DNS at 60.8% (31/51) of the samplings (Table 2 ). At the remaining 20 samplings where P. multocida was isolated from TBL, M. haemolytica was isolated from DNS at one sampling and OB at 19 samplings. Conversely, when P. multocida or M. bovoculi were isolated from DNS, the same species was isolated from TBL at,36.9% (31/84) of the samplings (Table 2 ). At the remaining 53 samplings where P. multocida was isolated from DNS, the TBL samples were negative at 46 samplings and yielded OB at seven samplings. The agreement between DNS and TBL culture results for P. multocida was slight for healthy calves, with a possible bias in the dataset (ƙ 0,14 ± 0,16, McNemar P = 0.001), and fair for calves with BRD, with no bias in the dataset (ƙ 0,34 ± 0,20, McNemar P = 0.078). Table 2 Crosstabulation of cultivation results for deep nasal swabs (DNS) and tracheobronchial lavage fluid (TBL) from 219 samplings of 171 calves and 48 calves were sampled twice. TBL P. multocida M. bovoculi Other bacteria Negative culture Total DNS P. multocida 29 (13.2%) 7 (3.2%) 46 (21.0%) 82 (37.4%) M. haemolytica 1 (0.5%) 1 (0.5%) M. bovoculi 2 (0.9%) 2 (0.9%) Other bacteria 19 (11.9%) 26 (11.9%) 89 (40.6%) 134 (61.2%) Total 49 (22.4%) 2 (0.9%) 33 (15.1%) 135 (61.6%) 219 (100%) HERE Table 2 Pasteurella multocida was isolated from DNS in at least one calf in 35 (83.3%) of the 42 calf groups sampled (Table 3 ). The median isolation frequency of P. multocida in a group was 33% (range: 0-100%). From TBL, P. multocida was isolated from at least one calf in a group at 31 (73.8%) of the groups sampled and the median isolation frequency was 23% (range: 0–75%) (Table 3 ). Table 3 Cross tabulation within calf groups of cultivation results for deep nasal swabs (DNS) and tracheobronchial lavage fluid (TBL) from 42 sampling occasions of 171 calves (48 sampled twice) in 30 groups (12 sampled twice). Farm ID Calf group Sampling number TBL Frequency of P. multocida in calf group DNS P. multocida M. bovoculi Other Negative Total DNS TBL 1 A 1 P. multocida 1 1 17% 17% Other 1 1 3 5 2 A 1 P. multocida 1 1 25% 25% M. bovoculi 2 2 Other 1 1 2 P. multocida 1 1 2 50% 25% Other 2 2 B 1 P. multocida 1 1 2 50% 25% Other 2 2 C 1 P. multocida 1 1 2 4 67% 17% Other 2 2 2 P. multocida 1 1 25% 25% Other 1 2 3 5 A 1 P. multocida 3 3 50% 50% Other 1 2 3 6 A 1 P. multocida 2 1 3 60% 60% M. haemolytica 1 1 Other 1 1 B 1 P. multocida 1 2 3 75% 25% Other 1 1 2 P. multocida 1 1 17% 0% Other 5 5 7 A 1 P. multocida 2 2 4 67% 33% Other 2 2 2 P. multocida 1 2 3 75% 25% Other 1 1 9 A 1 P. multocida 1 1 33% 0% Other 2 2 2 P. multocida 1 1 2 67% 33% Other 1 1 10 A 1 P. multocida 1 3 4 100% 25% 2 Other 1 1 3 5 0% 20% B 1 P. multocida 1 1 2 33% 33% Other 1 2 1 4 2 P. multocida 1 1 2 40% 40% Other 1 2 3 11 A 1 P. multocida 2 1 3 75% 75% Other 1 1 2 P. multocida 2 2 4 100% 50% B 1 P. multocida 2 1 3 60% 40% Other 1 1 2 13 A 1 P. multocida 1 1 17% 50% Other 3 1 1 5 2 P. multocida 1 1 17% 0% Other 2 3 5 15 A 1 Other 1 2 3 6 0% 17% 16 A 1 Other 1 5 6 0% 0% 17 A 1 Other 2 1 3 6 0% 33% 18 A 1 Other 2 3 5 0% 0% 22 A 1 P. multocida 2 2 40% 0% Other 3 3 2 P. multocida 1 1 17% 17% Other 1 2 2 5 25 A 1 P. multocida 1 1 17% 0% Other 2 3 5 28 A 1 P. multocida 3 1 4 80% 60% Other 1 1 29 A 1 P. multocida 1 1 20% 0% Other 1 3 4 31 A 1 Other 1 5 6 0% 17% 33 A 1 P. multocida 1 1 20% 0% Other 4 4 34 A 1 P. multocida 2 2 33% 17% Other 1 3 4 35 A 1 P. multocida 2 2 33% 33% Other 4 4 36 A 1 P. multocida 2 2 33% 17% Other 1 3 4 40 A 1 P. multocida 1 1 2 4 67% 17% Other 2 2 2 P. multocida 1 3 4 67% 0% Other 1 1 2 44 A 1 Other 6 6 0% 0% 46 A 1 P. multocida 4 4 67% 17% Other 1 1 2 2 P. multocida 3 3 50% 33% Other 2 1 3 Total 49 2 33 135 219 37% 22% HERE Table 3 Genotyping Among the 56 isolates from 28 samplings where P. multocida was isolated in both DNS and TBL, four different STs were found (Table 4 , Fig. 1 A). Twenty-five isolates were ST-68, 21 were ST-3, and seven were ST-19. Three isolates were of a new sequence type which was reported to the Public databases for molecular typing and microbial genome diversity (PubMLST) and named ST-202. At 24 of the 28 samplings (85.7%), P. multocida from DNS and TBL had the same STs and were closely related, whereas at four samplings (14.3%), the STs of the DNS and TBL isolates differed (Table 4 , Fig. 1 B). In the three calves that were sampled twice, P. multocida of the same ST was isolated on both occasions from one calf, whereas there was a shift in STs between samplings in two calves (Table 4 ). Table 4 Sequence types (STs) of P. multocida from 28 samplings of 25 calves (3 sampled twice) where isolates were available from both nasal swabs (NS) and tracheo-bronchial lavage fluid (TBL). Farm ID Calf group Calf ID Sequence types Sampling 1 Sampling 2 DNS TBL DNS TBL 2 A 950 ST-3 ST-3 951 ST-68 ST-68 B 960 ST-3 ST-3 C 35 ST-68 ST-68 5 A 1737 ST-68 ST-68 1739 ST-68 ST-68 1749 ST-3 ST-19 6 A 428 ST-68 ST-68 444 ST-68 ST-68 ST-3 ST-3 7 A 1016 ST-19 ST-3 2039 ST-19 ST-202 ST-202 ST-202 9 A 5062 ST-3 ST-3 10 A 9205 ST-68 ST-68 B 9725 ST-68 ST-68 ST-68 ST-68 11 A 72 ST-19 ST-19 91 ST-19 ST-19 1011 ST-68 ST-68 6051 ST-68 ST-68 B 828 ST-3 ST-3 28 A 1077 ST-68 ST-3 5079 ST-3 ST-3 7067 ST-68 ST-68 35 A 25 ST-3 ST-3 30 ST-3 ST-3 40 A 482 ST-3 ST-3 HERE Table 4 In groups of calves and within farms, P. multocida of more than one ST was often isolated. Thus, of the seven sampling occasions where two calves from a group were sampled, P. multocida of a single ST was isolated from both calves on four occasions (Farm 6, Group A, Sampling 1; Farm 11, Group A Sampling 1; Farm 11, Group A, Sampling 2; Farm 35, Group A, Sampling 1), whereas the isolates were of two or more STs on three occasions (Table 4 ). Similarly, on the three farms where more than one calf group was sampled, isolates of a single ST were found only on Farm 10 (Table 4 ). The minimum spanning tree (MST) from the cgMLST data (Fig. 1 A) showed that the P. multocida isolates clustered into three groups with closely related isolates. The genetic distance between DNS and TBL isolates from the same calf on the same sampling occasion was 0 alleles in 17 isolate pairs and 1 allele in 4 isolate pairs. We interpret this as indicating that the P. multocida isolates within these pairs were identical in 21 of the 28 isolate pairs (75%) (Fig. 1 B). In 3 isolate pairs, the genetic distance was between 26 and 74 alleles, and in 4 isolate pairs, it was between 1041 and 1443 alleles (Fig. 1 B). The isolates within these 7 pairs were considered nonidentical. The data for this study have been deposited in the European Nucleotide Archive (ENA) at EMBL-EBI under accession number PRJEB73847 [ 48 ]. HERE FIGURE 1 Discussion In this study, 37.9% DNS yielded relevant bacterial pathogens ( P. multocida and M. haemolytica ). Pasteurella multocida was highly predominant and was isolated from DNS in at least one calf in 35 of the 42 calf groups sampled. This confirms that this species can be readily isolated from the upper airways of both healthy and diseased calves [ 2 ]. A wide range of isolation frequencies for P. multocida (20–70%) have previously been reported for calves sampled by DNS or NPS [ 15 , 17 , 25 , 28 , 29 , 31 , 33 , 36 , 49 – 54 ]. The variation between studies is likely due to differences in age and disease status of the sampled animals, farm type, sample type and testing method [ 55 ]. In agreement with this, the isolation frequency varied substantially between farms and calf groups in our study, as previously observed by others [ 29 , 31 , 54 ]. This could be due to differences in disease status between farms and calf groups, as discussed above, but in contrast to previous reports [ 29 , 33 ], there was no statistically significant difference in isolation frequency between healthy and diseased calves in our study. Overgrowth of contaminants and mixed cultures from DNS were common in our study (data not shown) and made it difficult to recognise relevant bacterial pathogens in some samples. This has also been identified as problematic by others [ 29 ] and is challenging when the aim is to obtain relevant bacterial isolates for susceptibility testing. It has been suggested that this could be partly overcome by cleaning the calves’ nares prior to sampling but also by efforts in the laboratory to identify relevant pathogens in mixed cultures [ 28 ]. On submission of samples, it might therefore be important to inform the laboratory that the main aim is not to identify specific infections in individual animals but to obtain relevant isolates for susceptibility testing representative of the group or farm. Most TBL samples (61.6%) yielded no bacterial growth, but P. multocida was isolated from 22.4% of the samples and from at least one calf in 31 (73.8%) of the 42 calf groups. This is within the range of results (5–44%) reported for healthy calves sampled by BAL or TTW [ 29 , 33 , 56 , 57 ]. Our data are also in agreement with the isolation frequencies for calves with BRD sampled with the same TBL technique as in our study (16–34%) [ 59 , 59 ] and in studies using BAL (21–30%) [ 29 , 36 ]. Higher isolation frequencies in diseased calves (40–80%) were obtained in other studies using TTW or BAL [ 28 , 33 , 52 , 56 ]. Isolation frequencies in samples from the lower respiratory tract are likely to be influenced by the same factors as those discussed above for samples from the upper respiratory tract, but it is probable that the disease status of the calves is of greater importance. We found no statistically significant difference in isolation frequency between healthy and diseased calves, but several studies have reported that P. multocida is more often isolated from the lower respiratory tract of diseased calves [ 29 , 33 , 57 , 59 ], although others have found no difference [ 56 ]. It is also likely that the sampling method, specifically the volume of fluid instilled, influences the isolation frequency, as previously suggested [ 58 ]. A large, instilled volume probably increases the likelihood of isolating bacteria in the lower respiratory tract. This is corroborated by a generally higher isolation frequency in studies using TTW or BAL, where volumes of 50–180 mL fluid were instilled [ 28 , 33 , 52 , 56 ], than in studies where smaller volumes were used [ 29 , 58 , 59 ]. In our study, we instilled 20 ml of isotonic saline with a mean retraction volume of 4.5 ml, and it is likely that the number of positive TBL samples would have been greater if a larger volume had been used. There was a strong predominance of P. multocida in both DNS and TBL in our study. In TBL, this was the only species of Pasteurellacae isolated, whereas M. haemolytica was isolated from DNS in one calf. Histophilus somni was isolated neither in DNS nor in TBL. A strong predominance of P. multocida in samples from the respiratory tract of calves was also found in most other studies [ 28 , 29 , 31 , 36 , 49 , 50 , 52 , 58 , 59 ], although a more balanced panorama of Pasteurellacae [ 56 ] or a predominance of M. haemolytica [ 15 , 35 ] has also been reported. The main aim of this study was to evaluate whether isolates from DNS are representative of the bacterial flora of the lower respiratory tract of calves. Since P. multocida was the only relevant pathogen isolated from both DNS and TBL, only this species could be evaluated. At 53 samplings, P. multocida was isolated only from DNS, at 20 only from TBL and at 29 samplings from both DNS and TBL. At the species level, the agreement between DNS and TBL in healthy calves was slight (ƙ 0.14), and the McNemar test (P = 0.001) indicated that the data were biased. This finding is similar to what has been previously reported for P. multocida from the upper and lower respiratory tract of healthy calves (fair, ƙ 0.28) [ 29 ]. A greater agreement can be expected for diseased calves since P. multocida is considered a commensal in calves [ 2 , 25 , 53 ], and its presence in the upper respiratory tract does not imply infection in the lower respiratory tract [ 2 ]. For diseased calves in our study, the agreement was fair (ƙ 0,34), and the McNemar test indicated an unbiased dataset (P = 0.078). Previously, agreements ranging from moderate to almost perfect (ƙ 0.48->0.8) have been reported for calves with BRD [ 28 , 29 , 36 ] and for a mixture of healthy and diseased calves [ 33 ]. In these studies, moderate to almost perfect agreement was also reported for M. haemolytica , and slight to substantial agreement was reported for H. somni . The lesser agreement for P. multocida between DNS and TBL in our study could be a consequence of underdiagnosis of relevant pathogens due to mixed cultures in DNS and the small fluid volume used in TBL, as discussed above. Although an agreement at the species level between DNS and TBL suggests that the same bacterial strain was isolated from the upper and lower respiratory tract, this is not necessarily true. Moreover, neither does a comparison of STs validate the presence of the same strain. At the 28 samplings where P. multocida were subtyped by MLST, isolates from DNS and TBL had the same STs at 24 samplings, but at four samplings they differed, indicating that different strains were isolated. However, when cgMLST data were used to compare the 1609 alleles of the isolates, there was no difference between the isolate pairs at 17 samplings, and there was a difference of one allele at four samplings. We considered this to confirm that the same strain of P. multocida was present in DNS and TBL in 75% of the sample pairs. The isolates in the three DNS/TBL pairs with differences ranging from 26 to 74 alleles are most likely also closely related. However, in the in four pairs where DNS and TBL isolates differed by 1041 to 1443 alleles, it is evident that different strains of P. multocida were isolated in DNS and TBL. There are no previous reports evaluating genotypes of P. multocida from the upper and lower respiratory tract, but our findings agree with previous reports on M. haemolytica evaluated by molecular methods [ 15 , 35 , 36 ]. Thus, the same P. multocida strain was isolated from the upper and lower respiratory tract at 75% of the samplings where isolates were available from both DNS and TBL. This suggests that isolates from DNS generally provide relevant results for individual calves. However, within calf groups and farms, MLST showed that often P. multocida of more than one ST were isolated and cgMLST revealed that isolates of the same ST were not always identical. Apparently, more than one strain of P. multocida can be present in individual animals as well as in a group of calves and the panorama of strains on a farm is variable. This agrees with other studies where MLST of P. multocida has shown that isolates with different STs can be present in a group of calves, although often one or two types predominate on a farm, [ 52 , 54 , 56 , 60 ] and it is proposed that the movement of animals may cause greater variability [ 60 ]. In our study, ST-68 was the most common P. multocida ST. To our knowledge, this is the first findings of ST-68 in farm animals. The second most common ST was ST-3, which was previously found in pigs from China [ 61 ] and in association with porcine pneumonia in Spain together with ST-19 [ 62 ], the third most common ST in our collection. Thus, the epidemiology of bacterial respiratory pathogens within a farm is complex [ 63 ], but for practitioners seeking guidance on antibiotic therapy, it is neither possible nor relevant to identify all bacterial pathogens and strains present. For guidance on antibiotic therapy in a group of calves, it is most likely sufficient to obtain a few representative isolates of the bacterial respiratory pathogens present and test them for antibiotic susceptibility. To this end, the practitioner has two options, either to sample the upper respiratory tract or the lower respiratory tract of a reasonable number of calves [ 36 ]. Sampling the lower respiratory tract has been considered more cost effective and appropriate in practice because samples are less contaminated, and the interpretation of culture results is therefore more straightforward [ 29 ]. Additionally, isolates from the lower respiratory tract are considered more relevant for identifying the pathogen causing infection in individual calves [ 5 , 18 , 32 ]. However, for practitioners, it is undoubtedly more convenient to collect samples from the upper respiratory tract, which is also considered less harmful for the animals [ 25 , 31 ]. The risk of failure to isolate relevant pathogens from the upper respiratory tract in a group of calves can be minimised by sampling a larger number of animals, as previously suggested [ 15 ]. Additionally, our study showed that P. multocida isolated from upper respiratory tract samples are generally representative of isolates obtained from the lower respiratory tract of the same calf. This finding agrees with several other studies [ 15 , 28 , 33 , 35 , 36 ], and susceptibility testing of isolates collected by NPS or DNS from diseased calves was considered to yield relevant results for individual animals or calf groups [ 33 , 35 , 64 ]. Neither sampling the upper nor the lower respiratory tract would overcome the possibility that there can be several different strains on a farm, as discussed above. Regardless of the sampling site, the culture results should therefore be interpreted with caution [ 36 ]. To reduce the risk of missing strains of the same bacterial species with different antibiotic susceptibility, several isolates could be selected on culture, but this approach might not be possible or cost effective in a clinical context. Our study has several limitations. First, the isolates were collected between 1997 and 2000, and data regarding the STs of P. multocida and the occurrence of specific respiratory pathogens could be irrelevant for the current situation in Sweden. Moreover, the data and bacterial isolates in this study are from clinical investigations on farms with BRD outbreaks, and sampling and bacteriological cultures were performed to guide practitioners in managing the outbreaks and not in the context of an experimental study. Therefore, the processing of samples in the laboratory was performed as part of the routine work and focused on identifying Pasteurellacae . More elaborate efforts to identify isolates in the samples were not made, and only one isolate from each sample was further evaluated and saved. This probably led to underdiagnosis of bacterial pathogens in both DNS and TBL. Another limitation is that Mycoplasma spp. were not cultured because the methodology was not available at the laboratory at the time of the field investigations. Despite these limitations, we believe that the evaluation of DNS in relation to TBL and conclusions regarding the sampling of calves are relevant. Conclusions In this study, P. multocida was readily isolated from the upper airways of calves. Although there was a large variation in isolation frequency between farms and calf groups, at least one P. multocida isolate was obtained from DNS in 83.3% of the calf groups. In 75% of the calves, where P. multocida was isolated from both DNS and TBL, isolates from DNS and TBL were identical. During an outbreak of BRD, a pragmatic and convenient approach for practitioners to gain insight into the bacterial respiratory pathogens present and their antibiotic susceptibility could be to collect and submit samples from the upper respiratory tract of 4–6 calves. Declarations Authors’ contributions BB participated in the field investigations on which the data and isolates of this study were based and designed the present study; BH performed the bacteriological analyses; MM performed the molecular analyses; BB performed the statistical analyses; BB and MM interpreted the results; BB, MM and MP drafted the manuscript; and all the authors commented and refined the draft. All authors have read and approved the final version of the manuscript. Acknowledgements The contributions of VMD Sven Viring and VMD Charina Gånheim to the field investigations on which this study is based are gratefully acknowledged. Likewise, VMD Helene Wahlström is acknowledged for constructive criticism of this manuscript. Competing interests The authors declare that they have no competing interests. Availability of data and materials The datasets used and analysed during the current study are available from the corresponding author on reasonable request. Consent for publication Not applicable. Ethics approval The data and isolates on which this study is based were collected from calves at farm visits made by the Swedish animal health services in 1997–2000. The farm visits, including sampling of calves, were part of routine work in managing outbreaks of BRD on farms affiliated with the organisation. Hence, this is not an experimental study and ethics approval and consent to participate are therefore not relevant. Prior publication Data have not been published previously. Funding The processing of data, analysis of isolates from the strain collection and elaboration of the manuscript were conducted within the SvarmPat programme. This programme aims to counteract emergence and spread of antibiotic resistance in farm animal bacterial pathogens and is financed by the Swedish Board of Agriculture. SvarmPat is run in cooperation between the Department of Animal Health and Antimicrobial Strategies at the Swedish Veterinary Agency and the Swedish consultancy company Farm and Animal Health. References Gay E, Barnouin J. A nation-wide epidemiological study of acute bovine respiratory disease in France. Prev Vet Med. 2009; DOI: 10.1016/j.prevetmed.2009.02.013. Griffin D, Chengappa MM, Kuszak J, McVey DS. Bacterial pathogens of the bovine respiratory disease complex. Vet Clin North Am Food Anim Pract. 2010; DOI: 10.1016/j.cvfa.2010.04.004. Hay KE, Morton JM, Mahony TJ, Clements AC, Barnes TS. 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Details on the analysis parameters and FastQC data for whole-genome sequencing and genome assembly of 56 isolates of Pasteurella multocida from deep nasal swabs and tracheobronchial lavage from calves. Colony material was collected from blood agar plates for DNA extraction using a Qiagen EZ1 DNA Tissue Kit (Qiagen, Halden, Germany). Nextera Library preparation (Illumina, Foster City, United States) and paired-end sequencing (2 x 150bp) was performed at Clinical Genomics Stockholm, SciLifeLab (Solna, Sweden) using an Illumina NovaSeq 6000 instrument. The raw reads for each sample were quality checked using FastQC vo.11.9, trimmed using Trimmomatic v0.39 and assembled using SPAdes v3.14.0. The assemblies were error-corrected using Pilon v1.23. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4162560","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":292211016,"identity":"c2dd4a5f-a753-4776-a054-f361047e49c5","order_by":0,"name":"Mattias Myrenås","email":"","orcid":"","institution":"Department of Animal Health and Antimicrobial Strategies, Swedish Veterinary Agency (SVA)","correspondingAuthor":false,"prefix":"","firstName":"Mattias","middleName":"","lastName":"Myrenås","suffix":""},{"id":292211017,"identity":"a9e1d8fc-f5d1-44e7-ae2e-a35816ff6cab","order_by":1,"name":"Märit Pringle","email":"","orcid":"","institution":"Department od Animal Health and Antimicrobial Strategies, Swedish Veterinary Agency (SVA)","correspondingAuthor":false,"prefix":"","firstName":"Märit","middleName":"","lastName":"Pringle","suffix":""},{"id":292211018,"identity":"eba5feeb-c80e-4ba9-9494-9f4687bc62cc","order_by":2,"name":"Boel Harbom","email":"","orcid":"","institution":"Department of Animal Health and Antimicrobial Strategies, Swedish Veterinary Agency (SVA)","correspondingAuthor":false,"prefix":"","firstName":"Boel","middleName":"","lastName":"Harbom","suffix":""},{"id":292211019,"identity":"8409d383-fd74-4e50-81dc-e996b2504ff1","order_by":3,"name":"Björn Bengtsson","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0002-2012-0515","institution":"Department of Animal Health and Antimicrobial Strategies, Swedish Veterinary Agency (SVA)","correspondingAuthor":true,"prefix":"","firstName":"Björn","middleName":"","lastName":"Bengtsson","suffix":""}],"badges":[],"createdAt":"2024-03-25 10:27:04","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4162560/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4162560/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s13028-024-00781-7","type":"published","date":"2024-11-05T15:57:07+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":55066730,"identity":"ea3afdcd-d1d5-4728-a9f1-54dcf0b15727","added_by":"auto","created_at":"2024-04-22 04:39:36","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":348008,"visible":true,"origin":"","legend":"\u003cp\u003eMinimum spanning tree constructed from core genome multilocus sequence typing data consisting of 1609 loci showing 56 \u003cem\u003eP. multocida\u003c/em\u003e isolates sampled from the nasal cavity (DNS) or tracheobronchial lavage (TBL). Panel A: ST-3 is shown in light blue, ST-19 in dark orange, ST-68 in dark blue and ST-202 in light orange. Panel B: cgMLST differences between DNS and TBL sampled on the same occasion. Pairs of identical isolates (0 or 1 allele difference from 1609) are coloured yellow, related isolates (26 to 74 alleles) are coloured magenta, and nonrelated isolates (1041 to 1443 allele difference) are coloured cyan. The circle size indicates the number of isolates. Branches are dotted when there are differences in alleles greater than 200.\u003c/p\u003e","description":"","filename":"Figure1P.mfromDNSandTBLincalves22mars2024.png","url":"https://assets-eu.researchsquare.com/files/rs-4162560/v1/80bb70bf805ad787a5e664cb.png"},{"id":68749820,"identity":"eabef82e-48c3-42d7-b8e6-5511412c22d6","added_by":"auto","created_at":"2024-11-11 16:05:44","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1611735,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4162560/v1/b67e1a20-3568-473c-9a1d-1d585fc9a305.pdf"},{"id":55066731,"identity":"ec90b337-a2d9-49ea-b8e8-0d94fe06aa81","added_by":"auto","created_at":"2024-04-22 04:39:36","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":17760,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eADDITIONAL FILES\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAdditional file 1\u003c/strong\u003e (.xls)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSupplementary Table 1\u003c/strong\u003e. Details on the analysis parameters and FastQC data for whole-genome sequencing and genome assembly of 56 isolates of \u003cem\u003ePasteurella multocida\u003c/em\u003e from deep nasal swabs and tracheobronchial lavage from calves. Colony material was collected from blood agar plates for DNA extraction using a Qiagen EZ1 DNA Tissue Kit (Qiagen, Halden, Germany). Nextera Library preparation (Illumina, Foster City, United States) and paired-end sequencing (2 x 150bp) was performed at Clinical Genomics Stockholm, SciLifeLab (Solna, Sweden) using an Illumina NovaSeq 6000 instrument. The raw reads for each sample were quality checked using FastQC vo.11.9, trimmed using Trimmomatic v0.39 and assembled using SPAdes v3.14.0. The assemblies were error-corrected using Pilon v1.23.\u003c/p\u003e","description":"","filename":"Additionalfile1SupplementaryTable1.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4162560/v1/08ebf383c81220de77a5137c.xlsx"}],"financialInterests":"","formattedTitle":"Pasteurella multocida from deep nasal swabs and tracheobronchial lavage in calves","fulltext":[{"header":"Background","content":"\u003cp\u003eBovine respiratory disease (BRD) is a common and costly problem in intensively raised cattle worldwide [\u003cspan additionalcitationids=\"CR2 CR3 CR4\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. The background of BRD is multifactorial and includes infectious agents and predisposing factors related to the immunological and general status of the animals and to management and housing [\u003cspan additionalcitationids=\"CR7\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Due to its multifactorial background, the pathogenesis of BRD varies, but in general, viral infections of the upper respiratory tract precede bacterial infections of the lower respiratory tract [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. The bacteria commonly involved often reside in the upper airways of healthy calves and include \u003cem\u003ePasteurella multocida\u003c/em\u003e, \u003cem\u003eMannheimia haemolytica\u003c/em\u003e, \u003cem\u003eHistophilus somni\u003c/em\u003e and \u003cem\u003eMycoplasma bovis\u003c/em\u003e [\u003cspan additionalcitationids=\"CR9\" citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe control of BRD is complex and involves measures to hinder the spread of infectious agents and to mitigate predisposing factors [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan additionalcitationids=\"CR12\" citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Therapeutic, prophylactic or metaphylactic antibiotic treatments are also used to control BRD, but routine use of antibiotics should be avoided due to the risk of emerging antibiotic resistance [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. For reasons of animal welfare and to minimise economic losses, however, it is often necessary to use therapeutic antibiotic treatments when BRD manifests in a group of animals [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eConsidering the increased occurrence of acquired antibiotic resistance, it is important that treatment is guided by knowledge of the antibiotic susceptibility of the relevant bacteria [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan additionalcitationids=\"CR16 CR17\" citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Information on the antibiotic susceptibility of bacteria involved in BRD is available from national monitoring programs, e.g., [\u003cspan additionalcitationids=\"CR20 CR21 CR22 CR23\" citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e], and from other scientific literature, e.g., [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan additionalcitationids=\"CR26\" citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. However, such compiled data are derived from sources with varying study designs and from different farm types and might not be representative of the situation on a specific farm [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. It is therefore preferable if information is available for relevant bacteria isolated from acutely affected and untreated animals from the farm where treatments are to be instituted [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. This is even more relevant for farms experiencing treatment failures where the presence of antibiotic resistance could cause re-evaluation of treatment protocols [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Moreover, in some countries, susceptibility testing of isolates from animals on a farm is mandatory for the use of specific antibiotics on that farm [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTo obtain isolates of bacterial respiratory pathogens for susceptibility testing, it is convenient and simple for practitioners to collect samples for bacteriological culture from the upper respiratory tract using nasal swabs (NS), deep nasal swabs (DNS) or nasopharyngeal swabs (NPS) [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Samples from the lower respiratory tract can also be collected in a clinical setting. Several different techniques are available, including bronchoalveolar lavage (BAL), tracheobronchial lavage (TBL), transtracheal wash (TTW) or transtracheal swab (TTW) [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. These methods are, however, more complicated, and invasive than NS, DNS or NPS and are considered more stressful for the animals [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTo collect samples from the upper respiratory tract is convenient, but it is debated whether isolates from NS, DNS or NPS are representative of respiratory pathogens in the lower respiratory tract, which are the targets for antibiotic therapy [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. However, in studies that have compared paired isolates of \u003cem\u003ePasteurellacae\u003c/em\u003e from the upper and lower respiratory tracts of calves, there is generally moderate to almost perfect agreement at the species level [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan additionalcitationids=\"CR34\" citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. For \u003cem\u003eM. haemolytica\u003c/em\u003e, there was also a high agreement between paired isolates from the upper and lower respiratory tracts when molecular methods were used to compare isolates [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. However paired isolates of \u003cem\u003eP. multocida\u003c/em\u003e from the upper and lower respiratory tracts of calves have not been compared by molecular methods.\u003c/p\u003e \u003cp\u003eThe aim of this study was to evaluate whether samples from the upper respiratory tract collected by deep nasal swabs (DNS) from calves provide comparable information regarding bacterial pathogens as if the lower respiratory tract of the same calves were sampled by tracheobronchial lavage (TBL). Our hypothesis is that bacterial pathogens in the lower respiratory tract of a calf can generally also be cultured from DNS from the same calf or from other calves in the same group. To test this hypothesis, we used data from field investigations and reanalysed a collection of strains from the investigations using matrix-assisted laser-desorption ionisation-time-of-flight (MALDI-TOF MS), multilocus sequence typing (MLST) and core-genome multilocus sequence typing (cgMLST). MLST is a method, first described and evaluated in \u003cem\u003eNeisseria meningitidis\u003c/em\u003e, in which the gene sequences of, usually, seven housekeeping genes are compared [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. The number of identical gene variants in an isolate-to-isolate comparison makes it possible to calculate the evolutionary relationships between isolates of the same species. The core-genome multilocus sequence typing method also compares gene variants obtained from genome analysis. The genome sequences enable the comparison of 1609 genes instead of 7; hence, cgMLST can show relatedness between isolates with considerably higher detail than MLST.\u003c/p\u003e"},{"header":"Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eField investigations\u003c/h2\u003e \u003cp\u003eThe data and bacterial isolates in this study are from field investigations from 1997\u0026ndash;2000 on Swedish cattle farms experiencing outbreaks of BRD and affiliated with a health control program through the Swedish Animal Health Service [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. To manage the outbreaks, the calves were examined clinically, and those with a rectal temperature\u0026thinsp;\u0026gt;\u0026thinsp;39.5\u0026deg;C and at least one of the clinical signs of nasal discharge, cough or abnormal respiratory sounds on auscultation were considered BRD cases. To assess antibiotic susceptibility of possible bacterial respiratory pathogens, 4\u0026ndash;6 calves from a group with clinical signs of BRD were sampled at each visit. When only a few diseased calves were available, healthy calves from the group were sampled instead. Samples were collected from the posterior nasal cavity (DNS) using cotton swabs (Copan, Amies charcoal medium). The swab was entered into the nasal cavity to its full length (\u0026asymp;\u0026thinsp;12 cm) after dry cleaning of the outer nares. Bacteriological samples were also collected from the lower respiratory tract by tracheobronchial lavage (TBL). The catheter used for TBL was designed to protect the sample from contamination when it was passed through the nasal cavity [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. In brief, a catheter made of Silastic tubing (diameter 5 mm, length 50 cm) with an inner lining of Teflon tubing was passed through the nasal cavity into the trachea. A silicon rubber stopper 10 cm from the upper end of the catheter hindered passage more than 40 cm into the airways. When located in the trachea, a 100 cm flexible Teflon tube was passed through the catheter and through a slit in a silicon tip sealing the catheter. The Teflon tube was then passed down to the region of the tracheal bifurcation where 20 ml of isotonic saline was instilled and immediately aspirated. The mean volume of fluid retracted as a sample was 4.5 ml (range 0.5\u0026ndash;15 ml). In all, 37 different farms were visited, 400 calves were examined and sampled, and 80 of the calves were examined and sampled again within 3\u0026ndash;9 weeks.\u003c/p\u003e \u003cp\u003eThe DNS and TBL samples collected were kept at +\u0026thinsp;8\u0026deg;C and transported within 24 h to the National Veterinary Agency (SVA), where they were cultivated for respiratory bacterial pathogens on blood agar, blue agar and hematin agar. Colonies with a macroscopic appearance in agreement with \u003cem\u003ePasteurellacae\u003c/em\u003e were subcultured and identified by biochemical tests. The culture results were registered as \u0026ldquo;Pasteurella-like bacteria\u0026rdquo; (PLB), \u0026ldquo;Other bacteria\u0026rdquo; (OB) or \u0026ldquo;Negative culture\u0026rdquo; (NEG).\u003c/p\u003e \u003cp\u003eDescriptive data for farms and individual calves from the field investigations were stored together with laboratory results. Isolates identified as PLB were stored at -80\u0026deg;C, and isolates identified as OB were not stored.\u003c/p\u003e \u003cp\u003eAs the calves were examined and sampled in the context of the routines for managing clinical outbreaks of BRD, no ethical approval was needed.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eSelection of calves from the field investigations\u003c/h2\u003e \u003cp\u003eFor the present study, calves from the field investigations were selected using the following criteria: \u003csup\u003e1\u003c/sup\u003e both DNS and TBL should have been collected; \u003csup\u003e2\u003c/sup\u003e no antibiotic treatment within 3 days prior to sampling; \u003csup\u003e3\u003c/sup\u003e data should be available for at least 3 calves in the calf group; \u003csup\u003e4\u003c/sup\u003e isolates identified as PLB should be available in the strain collection.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eSelection of isolates for species identification by MALDI-TOF\u003c/h2\u003e \u003cp\u003eTo refine the identification of bacterial species, isolates identified as PLB in the field investigations were analysed by MALDI-TOF MS. From the calves selected for this study, 85 isolates from DNS and 51 from TBL were available in the strain collection. These isolates were analysed by the MALDI Biotyper system (Bruker Daltonics, Bremen, Germany) to identify the species. Material from a single colony from the agar plate was spotted on a MALDI plate without pretreatment. The spots were covered with 1 \u0026micro;l of matrix solution consisting of α-cyano-4-hydroxycinnamic acid (HCCA), air-dried at room temperature, and introduced into the MALDI-TOF mass spectrometer for analysis. The spectra of all the isolates were compared to the spectra in the database, and identification was provided with a reliability score. A score\u0026thinsp;\u0026ge;\u0026thinsp;2.0 was considered reliable for species identification.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eWhole-genome Sequencing and Genome Assembly\u003c/h2\u003e \u003cp\u003eAt 29 samplings, \u003cem\u003eP. multocida\u003c/em\u003e was isolated from both DNS and TBL on the same calf. The genomes of 28 of these isolate pairs (56 isolates from 25 different calves) were sequenced and compared (one isolate pair was not available). Colony material was collected from blood agar plates for DNA extraction using a Qiagen EZ1 DNA Tissue Kit (Qiagen, Halden, Germany). Nextera Library preparation (Illumina, Foster City, United States) and paired-end sequencing (2 x 150bp) was performed at Clinical Genomics Stockholm, SciLifeLab (Solna, Sweden) using an Illumina NovaSeq 6000 instrument. The raw reads for each sample were quality checked using FastQC vo.11.9 [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e], trimmed using Trimmomatic v0.39 [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e] and assembled using SPAdes v3.14.0 [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. The assemblies were error-corrected using Pilon v1.23 [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. Details on the analysis parameters and FastQC data can be found in Additional file 1, Supplementary Table\u0026nbsp;1.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eMultilocus Sequence Typing\u003c/h2\u003e \u003cp\u003eFor \u003cem\u003eP. multocida\u003c/em\u003e, there are two public MLST schemes [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e] available at PubMLST [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e], of which we chose to use the multihost MLST scheme of Davies et al. [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eCore-Genome Multilocus Sequence Typing and Minimum Spanning Tree Construction\u003c/h2\u003e \u003cp\u003eThere were, at the time of the writing of this publication and to our knowledge, no cgMLST schemes for \u003cem\u003eP. multocida\u003c/em\u003e suitable for global nomenclature. Therefore, we constructed an ad hoc cgMLST scheme from the project data suitable for analysis of this dataset alone using the cgMLST Target Definer v.1.5 function in SeqSphere\u0026thinsp;+\u0026thinsp;version 6.0.2 (Ridom, W\u0026uuml;rzburg, Germany). We used the annotated genome with the GenBank accession number NZ_CP008918.1 as the seed genome and the genomes NZ_CP015569.1, NZ_CP037861.1 and NZ_CP037865.1 as the penetration genomes and excluded hits found in the plasmid sequence NC_017035.1, gene duplicates and truncated genes. This resulted in a cgMLST scheme consisting of 1611 targets, as documented in Supplementary Table\u0026nbsp;1. Using the results from 1609 of the cgMLST targets, the phylogenetic distance was calculated using the Minimum Spanning Tree method (Kruskal JB 1956) using GrapeTree v.1.5.0 [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eStatistical calculations\u003c/h2\u003e \u003cp\u003eDifferences in culture results between DNS and TBL were evaluated by Fischer\u0026rsquo;s exact test, with P\u0026thinsp;\u0026le;\u0026thinsp;0.05 considered evidence of a significant difference. Agreement between culture results for DNS and TBL for \u003cem\u003eP. multocida\u003c/em\u003e was evaluated by Kappa statistics and McNemar\u0026rsquo;s test [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. Kappa values (ƙ) were interpreted to indicate the strength of agreement as follows: \u0026lt; 0.20, slight; 0.20\u0026ndash;0.40, fair; 0.41\u0026ndash;0.60, moderate; 0.61\u0026ndash;0.8, substantial; and \u0026gt;\u0026thinsp;0.8, almost perfect. The exact McNemar significance probability test was used to determine the potential for bias between DNS and TBL, and values\u0026thinsp;\u0026le;\u0026thinsp;0.05 were considered evidence of bias. Statistical calculations were made using the GraphPad software.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n \u003ch2\u003eBacteriological culture\u003c/h2\u003e\n \u003cp\u003eUsing the specified criteria, 219 samplings of 171 different calves were selected for this study, 48 of which were examined and sampled twice (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). The calves were from 30 different groups on 25 farms, and 12 calf groups were sampled twice (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). Ninety of the samplings were of calves with clinical signs of BRD, and 129 were from healthy calves (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). Among the 219 DNS samples, \u003cem\u003eP. multocida\u003c/em\u003e was isolated from 82 (37.4%), \u003cem\u003eM. haemolytica\u003c/em\u003e from one (0.5%), \u003cem\u003eMoraxella bovoculi\u003c/em\u003e from two (0.9%) and OB from 143 samples (65.3%) (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). There was no difference in the isolation frequency of \u003cem\u003eP. multocida\u003c/em\u003e between healthy (38.0%) and diseased calves (36.7%) (P\u0026thinsp;=\u0026thinsp;0.778). Among the 219 TBL samples, \u003cem\u003eP. multocida\u003c/em\u003e was isolated from 49 (22.4%), \u003cem\u003eM. bovoculi\u003c/em\u003e from two (0.9%), and OB from 33 (15.1%), whereas most samples, 135 (61.6%), yielded no growth. There was no difference in the isolation frequency of \u003cem\u003eP. multocida\u003c/em\u003e between healthy (20.2%) and diseased calves (25.6%) (P\u0026thinsp;=\u0026thinsp;0.410), but the proportion of negative cultures was greater in healthy (68.2%) than in diseased calves (52.2%) (P\u0026thinsp;=\u0026thinsp;0.023).\u003c/p\u003e\u0026nbsp;\u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eDescriptive data from field investigations from 1997\u0026ndash;2000 for the 25 farms and 171 calves (48 calves sampled twice) selected for the present study.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eFarm ID\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eType \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eCalf group\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eSampling 1\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eInterval\u003c/p\u003e\n \u003cp\u003eS1 - S2\u003c/p\u003e\n \u003cp\u003e(days)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eSampling 2\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eTotal number of samplings\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNumber of calves\u003c/strong\u003e \u003csup\u003e\u003cstrong\u003eb\u003c/strong\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eAge\u003c/strong\u003e (days)\u003c/p\u003e\n \u003cp\u003eMean (range)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNumber of calves\u003c/strong\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eAge\u003c/strong\u003e (days)\u003c/p\u003e\n \u003cp\u003eMean (range)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e61 (19\u0026ndash;171)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4 (2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNA \u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4 (3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4 (4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4 (1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e80 (63\u0026ndash;130)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5 (2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e85 (46\u0026ndash;129)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4 (1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e127 (81\u0026ndash;164)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e95 (76\u0026ndash;131)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e98 (84\u0026ndash;113)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4 (0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e131 (123\u0026ndash;143)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3 (0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e118 (94\u0026ndash;154)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3 (1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4 (1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e88 (58\u0026ndash;115)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5 (3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e126 (107\u0026ndash;159)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5 (0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4 (2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e55 (49\u0026ndash;58)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4 (2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e89 (84\u0026ndash;93)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5 (3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e51 (43\u0026ndash;59)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e102 (79\u0026ndash;128)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5 (4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e92 (72\u0026ndash;112)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5 (1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e99 (83\u0026ndash;124\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e139 (123\u0026ndash;164)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e103 (92\u0026ndash;130)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5 (3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e98 (86\u0026ndash;113)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5 (2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e55 (20\u0026ndash;79)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e67 (44\u0026ndash;116)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5 (3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e78 (62\u0026ndash;91)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e102 (77\u0026ndash;134)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (6)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e60 (43\u0026ndash;97)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e52 (19\u0026ndash;73)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e81 (48\u0026ndash;102)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMeat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e77 (73\u0026ndash;86)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e60 (25\u0026ndash;83)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6 (0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e80 (45\u0026ndash;103)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e162\u003c/strong\u003e (76)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e80\u003c/strong\u003e (19\u0026ndash;171)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e57\u003c/strong\u003e (14\u003cstrong\u003e)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e108\u003c/strong\u003e (45\u0026ndash;164)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e219\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"9\"\u003e\u003csup\u003ea\u003c/sup\u003e Meat\u0026thinsp;=\u0026thinsp;farms specialised in raising calves for meat production, Dairy\u0026thinsp;=\u0026thinsp;farms with milk production; \u003csup\u003eb\u003c/sup\u003e Number of calves with clinical signs of BRD in parenthesis; \u003csup\u003ec\u003c/sup\u003e Data not available.\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n \u003ch2\u003eHERE Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e\u003c/h2\u003e\n \u003cp\u003eCrosstabulation of culture results from DNS and TBL showed that when \u003cem\u003eP. multocida\u003c/em\u003e or \u003cem\u003eM. bovoculi\u003c/em\u003e was isolated from TBL, the same species was isolated from DNS at 60.8% (31/51) of the samplings (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). At the remaining 20 samplings where \u003cem\u003eP. multocida\u003c/em\u003e was isolated from TBL, \u003cem\u003eM. haemolytica\u003c/em\u003e was isolated from DNS at one sampling and OB at 19 samplings. Conversely, when \u003cem\u003eP. multocida\u003c/em\u003e or \u003cem\u003eM. bovoculi\u003c/em\u003e were isolated from DNS, the same species was isolated from TBL at,36.9% (31/84) of the samplings (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). At the remaining 53 samplings where \u003cem\u003eP. multocida\u003c/em\u003e was isolated from DNS, the TBL samples were negative at 46 samplings and yielded OB at seven samplings. The agreement between DNS and TBL culture results for \u003cem\u003eP. multocida\u003c/em\u003e was slight for healthy calves, with a possible bias in the dataset (ƙ 0,14\u0026thinsp;\u0026plusmn;\u0026thinsp;0,16, McNemar P\u0026thinsp;=\u0026thinsp;0.001), and fair for calves with BRD, with no bias in the dataset (ƙ 0,34\u0026thinsp;\u0026plusmn;\u0026thinsp;0,20, McNemar P\u0026thinsp;=\u0026thinsp;0.078).\u003c/p\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003c/p\u003e\u0026nbsp;\u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eCrosstabulation of cultivation results for deep nasal swabs (DNS) and tracheobronchial lavage fluid (TBL) from 219 samplings of 171 calves and 48 calves were sampled twice.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" style=\"width: 4.707%;\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\" style=\"width: 13.8521%;\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\" style=\"width: 49.0874%;\"\u003e\n \u003cp\u003eTBL\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" style=\"width: 5.8367%;\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 4.707%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 13.8521%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 11.1623%;\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 10.6244%;\"\u003e\n \u003cp\u003e\u003cem\u003eM. bovoculi\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.6417%;\"\u003e\n \u003cp\u003eOther bacteria\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 14.659%;\"\u003e\n \u003cp\u003eNegative culture\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 11.1623%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\" style=\"width: 4.707%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDNS\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 13.8521%;\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 11.1623%;\"\u003e\n \u003cp\u003e29 (13.2%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 10.6244%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.6417%;\"\u003e\n \u003cp\u003e7 (3.2%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 14.659%;\"\u003e\n \u003cp\u003e46 (21.0%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 11.1623%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e82\u003c/strong\u003e (37.4%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 13.8521%;\"\u003e\n \u003cp\u003e\u003cem\u003eM. haemolytica\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 11.1623%;\"\u003e\n \u003cp\u003e1 (0.5%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 10.6244%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.6417%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 14.659%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 11.1623%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003c/strong\u003e (0.5%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 13.8521%;\"\u003e\n \u003cp\u003e\u003cem\u003eM. bovoculi\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 11.1623%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 10.6244%;\"\u003e\n \u003cp\u003e2 (0.9%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.6417%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 14.659%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 11.1623%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e (0.9%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 13.8521%;\"\u003e\n \u003cp\u003eOther bacteria\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 11.1623%;\"\u003e\n \u003cp\u003e19 (11.9%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 10.6244%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.6417%;\"\u003e\n \u003cp\u003e26 (11.9%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 14.659%;\"\u003e\n \u003cp\u003e89 (40.6%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 11.1623%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e134\u003c/strong\u003e (61.2%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 4.707%;\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 13.8521%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 11.1623%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e49\u003c/strong\u003e (22.4%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 10.6244%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e (0.9%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.6417%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e33\u003c/strong\u003e (15.1%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 14.659%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e135\u003c/strong\u003e (61.6%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 11.1623%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e219\u003c/strong\u003e (100%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003c/p\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n \u003ch2\u003eHERE Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e\u003c/h2\u003e\n \u003cp\u003e\u003cem\u003ePasteurella multocida\u003c/em\u003e was isolated from DNS in at least one calf in 35 (83.3%) of the 42 calf groups sampled (Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e). The median isolation frequency of \u003cem\u003eP. multocida\u003c/em\u003e in a group was 33% (range: 0-100%). From TBL, \u003cem\u003eP. multocida\u003c/em\u003e was isolated from at least one calf in a group at 31 (73.8%) of the groups sampled and the median isolation frequency was 23% (range: 0\u0026ndash;75%) (Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\u0026nbsp;\u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eCross tabulation within calf groups of cultivation results for deep nasal swabs (DNS) and tracheobronchial lavage fluid (TBL) from 42 sampling occasions of 171 calves (48 sampled twice) in 30 groups (12 sampled twice).\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eFarm\u003c/p\u003e\n \u003cp\u003eID\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eCalf\u003c/p\u003e\n \u003cp\u003egroup\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eSampling\u003c/p\u003e\n \u003cp\u003enumber\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eTBL\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eFrequency of \u003cem\u003eP. multocida\u003c/em\u003e in calf group\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDNS\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eM. bovoculi\u003c/em\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eNegative\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTotal\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDNS\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTBL\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"11\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"3\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003e25%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003e25%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eM. bovoculi\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e50%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e25%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e50%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e25%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003eC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e67%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e25%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e25%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e50%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e50%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"7\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"3\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003e60%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003e60%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eM. haemolytica\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e75%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e25%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e67%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e33%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e75%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e25%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e33%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e67%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e33%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"6\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e25%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e33%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e33%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e40%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e40%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e75%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e75%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e60%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e40%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e50%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e33%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e40%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e80%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e60%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e20%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e20%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e33%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e33%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e33%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e33%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e67%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e67%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003e46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e67%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e17%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP. multocida\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e50%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e33%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOther\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e49\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e33\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e135\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e219\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e37%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e22%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n \u003ch2\u003eHERE Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e\u003c/h2\u003e\n \u003cdiv id=\"Sec15\" class=\"Section3\"\u003e\n \u003ch2\u003eGenotyping\u003c/h2\u003e\n \u003cp\u003eAmong the 56 isolates from 28 samplings where \u003cem\u003eP. multocida\u003c/em\u003e was isolated in both DNS and TBL, four different STs were found (Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e, Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eA). Twenty-five isolates were ST-68, 21 were ST-3, and seven were ST-19. Three isolates were of a new sequence type which was reported to the Public databases for molecular typing and microbial genome diversity (PubMLST) and named ST-202. At 24 of the 28 samplings (85.7%), \u003cem\u003eP. multocida\u003c/em\u003e from DNS and TBL had the same STs and were closely related, whereas at four samplings (14.3%), the STs of the DNS and TBL isolates differed (Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e, Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eB). In the three calves that were sampled twice, \u003cem\u003eP. multocida\u003c/em\u003e of the same ST was isolated on both occasions from one calf, whereas there was a shift in STs between samplings in two calves (Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\u0026nbsp;\u003ctable id=\"Tab4\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eSequence types (STs) of \u003cem\u003eP. multocida\u003c/em\u003e from 28 samplings of 25 calves (3 sampled twice) where isolates were available from both nasal swabs (NS) and tracheo-bronchial lavage fluid (TBL).\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eFarm ID\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eCalf group\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eCalf ID\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eSequence types\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eSampling 1\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eSampling 2\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDNS\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTBL\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDNS\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTBL\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e950\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e951\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e960\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1737\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1739\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1749\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e428\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e444\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1016\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2039\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-202\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-202\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-202\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5062\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e9205\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e9725\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1011\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6051\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e828\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1077\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5079\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e7067\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e482\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eST-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n \u003ch2\u003eHERE Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e\u003c/h2\u003e\n \u003cp\u003eIn groups of calves and within farms, \u003cem\u003eP. multocida\u003c/em\u003e of more than one ST was often isolated. Thus, of the seven sampling occasions where two calves from a group were sampled, \u003cem\u003eP. multocida\u003c/em\u003e of a single ST was isolated from both calves on four occasions (Farm 6, Group A, Sampling 1; Farm 11, Group A Sampling 1; Farm 11, Group A, Sampling 2; Farm 35, Group A, Sampling 1), whereas the isolates were of two or more STs on three occasions (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). Similarly, on the three farms where more than one calf group was sampled, isolates of a single ST were found only on Farm 10 (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eThe minimum spanning tree (MST) from the cgMLST data (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eA) showed that the \u003cem\u003eP. multocida\u003c/em\u003e isolates clustered into three groups with closely related isolates. The genetic distance between DNS and TBL isolates from the same calf on the same sampling occasion was 0 alleles in 17 isolate pairs and 1 allele in 4 isolate pairs. We interpret this as indicating that the \u003cem\u003eP. multocida\u003c/em\u003e isolates within these pairs were identical in 21 of the 28 isolate pairs (75%) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eB). In 3 isolate pairs, the genetic distance was between 26 and 74 alleles, and in 4 isolate pairs, it was between 1041 and 1443 alleles (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eB). The isolates within these 7 pairs were considered nonidentical.\u003c/p\u003e\n \u003cp\u003eThe data for this study have been deposited in the European Nucleotide Archive (ENA) at EMBL-EBI under accession number PRJEB73847 [\u003cspan class=\"CitationRef\"\u003e48\u003c/span\u003e].\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\n \u003ch2\u003eHERE FIGURE \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e\u003c/h2\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn this study, 37.9% DNS yielded relevant bacterial pathogens (\u003cem\u003eP. multocida and M. haemolytica\u003c/em\u003e). \u003cem\u003ePasteurella multocida\u003c/em\u003e was highly predominant and was isolated from DNS in at least one calf in 35 of the 42 calf groups sampled. This confirms that this species can be readily isolated from the upper airways of both healthy and diseased calves [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. A wide range of isolation frequencies for \u003cem\u003eP. multocida\u003c/em\u003e (20\u0026ndash;70%) have previously been reported for calves sampled by DNS or NPS [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e, \u003cspan additionalcitationids=\"CR50 CR51 CR52 CR53\" citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. The variation between studies is likely due to differences in age and disease status of the sampled animals, farm type, sample type and testing method [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e]. In agreement with this, the isolation frequency varied substantially between farms and calf groups in our study, as previously observed by others [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e, \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. This could be due to differences in disease status between farms and calf groups, as discussed above, but in contrast to previous reports [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e], there was no statistically significant difference in isolation frequency between healthy and diseased calves in our study.\u003c/p\u003e \u003cp\u003eOvergrowth of contaminants and mixed cultures from DNS were common in our study (data not shown) and made it difficult to recognise relevant bacterial pathogens in some samples. This has also been identified as problematic by others [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e] and is challenging when the aim is to obtain relevant bacterial isolates for susceptibility testing. It has been suggested that this could be partly overcome by cleaning the calves\u0026rsquo; nares prior to sampling but also by efforts in the laboratory to identify relevant pathogens in mixed cultures [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. On submission of samples, it might therefore be important to inform the laboratory that the main aim is not to identify specific infections in individual animals but to obtain relevant isolates for susceptibility testing representative of the group or farm.\u003c/p\u003e \u003cp\u003eMost TBL samples (61.6%) yielded no bacterial growth, but \u003cem\u003eP. multocida\u003c/em\u003e was isolated from 22.4% of the samples and from at least one calf in 31 (73.8%) of the 42 calf groups. This is within the range of results (5\u0026ndash;44%) reported for healthy calves sampled by BAL or TTW [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e, \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e]. Our data are also in agreement with the isolation frequencies for calves with BRD sampled with the same TBL technique as in our study (16\u0026ndash;34%) [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e, \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e] and in studies using BAL (21\u0026ndash;30%) [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. Higher isolation frequencies in diseased calves (40\u0026ndash;80%) were obtained in other studies using TTW or BAL [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIsolation frequencies in samples from the lower respiratory tract are likely to be influenced by the same factors as those discussed above for samples from the upper respiratory tract, but it is probable that the disease status of the calves is of greater importance. We found no statistically significant difference in isolation frequency between healthy and diseased calves, but several studies have reported that \u003cem\u003eP. multocida\u003c/em\u003e is more often isolated from the lower respiratory tract of diseased calves [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e, \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e], although others have found no difference [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e]. It is also likely that the sampling method, specifically the volume of fluid instilled, influences the isolation frequency, as previously suggested [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e]. A large, instilled volume probably increases the likelihood of isolating bacteria in the lower respiratory tract. This is corroborated by a generally higher isolation frequency in studies using TTW or BAL, where volumes of 50\u0026ndash;180 mL fluid were instilled [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e], than in studies where smaller volumes were used [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e, \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e]. In our study, we instilled 20 ml of isotonic saline with a mean retraction volume of 4.5 ml, and it is likely that the number of positive TBL samples would have been greater if a larger volume had been used.\u003c/p\u003e \u003cp\u003eThere was a strong predominance of \u003cem\u003eP. multocida\u003c/em\u003e in both DNS and TBL in our study. In TBL, this was the only species of \u003cem\u003ePasteurellacae\u003c/em\u003e isolated, whereas \u003cem\u003eM. haemolytica\u003c/em\u003e was isolated from DNS in one calf. \u003cem\u003eHistophilus somni\u003c/em\u003e was isolated neither in DNS nor in TBL. A strong predominance of \u003cem\u003eP. multocida\u003c/em\u003e in samples from the respiratory tract of calves was also found in most other studies [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e, \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e, \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e], although a more balanced panorama of \u003cem\u003ePasteurellacae\u003c/em\u003e [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e] or a predominance of \u003cem\u003eM. haemolytica\u003c/em\u003e [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e] has also been reported.\u003c/p\u003e \u003cp\u003eThe main aim of this study was to evaluate whether isolates from DNS are representative of the bacterial flora of the lower respiratory tract of calves. Since \u003cem\u003eP. multocida\u003c/em\u003e was the only relevant pathogen isolated from both DNS and TBL, only this species could be evaluated. At 53 samplings, \u003cem\u003eP. multocida\u003c/em\u003e was isolated only from DNS, at 20 only from TBL and at 29 samplings from both DNS and TBL. At the species level, the agreement between DNS and TBL in healthy calves was slight (ƙ 0.14), and the McNemar test (P\u0026thinsp;=\u0026thinsp;0.001) indicated that the data were biased. This finding is similar to what has been previously reported for \u003cem\u003eP. multocida\u003c/em\u003e from the upper and lower respiratory tract of healthy calves (fair, ƙ 0.28) [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. A greater agreement can be expected for diseased calves since \u003cem\u003eP. multocida\u003c/em\u003e is considered a commensal in calves [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e], and its presence in the upper respiratory tract does not imply infection in the lower respiratory tract [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. For diseased calves in our study, the agreement was fair (ƙ 0,34), and the McNemar test indicated an unbiased dataset (P\u0026thinsp;=\u0026thinsp;0.078). Previously, agreements ranging from moderate to almost perfect (ƙ 0.48-\u0026gt;0.8) have been reported for calves with BRD [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e] and for a mixture of healthy and diseased calves [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. In these studies, moderate to almost perfect agreement was also reported for \u003cem\u003eM. haemolytica\u003c/em\u003e, and slight to substantial agreement was reported for \u003cem\u003eH. somni\u003c/em\u003e. The lesser agreement for \u003cem\u003eP. multocida\u003c/em\u003e between DNS and TBL in our study could be a consequence of underdiagnosis of relevant pathogens due to mixed cultures in DNS and the small fluid volume used in TBL, as discussed above.\u003c/p\u003e \u003cp\u003eAlthough an agreement at the species level between DNS and TBL suggests that the same bacterial strain was isolated from the upper and lower respiratory tract, this is not necessarily true. Moreover, neither does a comparison of STs validate the presence of the same strain. At the 28 samplings where \u003cem\u003eP. multocida\u003c/em\u003e were subtyped by MLST, isolates from DNS and TBL had the same STs at 24 samplings, but at four samplings they differed, indicating that different strains were isolated. However, when cgMLST data were used to compare the 1609 alleles of the isolates, there was no difference between the isolate pairs at 17 samplings, and there was a difference of one allele at four samplings. We considered this to confirm that the same strain of \u003cem\u003eP. multocida\u003c/em\u003e was present in DNS and TBL in 75% of the sample pairs. The isolates in the three DNS/TBL pairs with differences ranging from 26 to 74 alleles are most likely also closely related. However, in the in four pairs where DNS and TBL isolates differed by 1041 to 1443 alleles, it is evident that different strains of \u003cem\u003eP. multocida\u003c/em\u003e were isolated in DNS and TBL. There are no previous reports evaluating genotypes of \u003cem\u003eP. multocida\u003c/em\u003e from the upper and lower respiratory tract, but our findings agree with previous reports on \u003cem\u003eM. haemolytica\u003c/em\u003e evaluated by molecular methods [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThus, the same \u003cem\u003eP. multocida\u003c/em\u003e strain was isolated from the upper and lower respiratory tract at 75% of the samplings where isolates were available from both DNS and TBL. This suggests that isolates from DNS generally provide relevant results for individual calves. However, within calf groups and farms, MLST showed that often \u003cem\u003eP. multocida\u003c/em\u003e of more than one ST were isolated and cgMLST revealed that isolates of the same ST were not always identical. Apparently, more than one strain of \u003cem\u003eP. multocida\u003c/em\u003e can be present in individual animals as well as in a group of calves and the panorama of strains on a farm is variable. This agrees with other studies where MLST of \u003cem\u003eP. multocida\u003c/em\u003e has shown that isolates with different STs can be present in a group of calves, although often one or two types predominate on a farm, [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e, \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e, \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e] and it is proposed that the movement of animals may cause greater variability [\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn our study, ST-68 was the most common \u003cem\u003eP. multocida\u003c/em\u003e ST. To our knowledge, this is the first findings of ST-68 in farm animals. The second most common ST was ST-3, which was previously found in pigs from China [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e] and in association with porcine pneumonia in Spain together with ST-19 [\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e], the third most common ST in our collection.\u003c/p\u003e \u003cp\u003eThus, the epidemiology of bacterial respiratory pathogens within a farm is complex [\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e], but for practitioners seeking guidance on antibiotic therapy, it is neither possible nor relevant to identify all bacterial pathogens and strains present. For guidance on antibiotic therapy in a group of calves, it is most likely sufficient to obtain a few representative isolates of the bacterial respiratory pathogens present and test them for antibiotic susceptibility. To this end, the practitioner has two options, either to sample the upper respiratory tract or the lower respiratory tract of a reasonable number of calves [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. Sampling the lower respiratory tract has been considered more cost effective and appropriate in practice because samples are less contaminated, and the interpretation of culture results is therefore more straightforward [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Additionally, isolates from the lower respiratory tract are considered more relevant for identifying the pathogen causing infection in individual calves [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. However, for practitioners, it is undoubtedly more convenient to collect samples from the upper respiratory tract, which is also considered less harmful for the animals [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. The risk of failure to isolate relevant pathogens from the upper respiratory tract in a group of calves can be minimised by sampling a larger number of animals, as previously suggested [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Additionally, our study showed that \u003cem\u003eP. multocida\u003c/em\u003e isolated from upper respiratory tract samples are generally representative of isolates obtained from the lower respiratory tract of the same calf. This finding agrees with several other studies [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e], and susceptibility testing of isolates collected by NPS or DNS from diseased calves was considered to yield relevant results for individual animals or calf groups [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eNeither sampling the upper nor the lower respiratory tract would overcome the possibility that there can be several different strains on a farm, as discussed above. Regardless of the sampling site, the culture results should therefore be interpreted with caution [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. To reduce the risk of missing strains of the same bacterial species with different antibiotic susceptibility, several isolates could be selected on culture, but this approach might not be possible or cost effective in a clinical context.\u003c/p\u003e \u003cp\u003eOur study has several limitations. First, the isolates were collected between 1997 and 2000, and data regarding the STs of \u003cem\u003eP. multocida\u003c/em\u003e and the occurrence of specific respiratory pathogens could be irrelevant for the current situation in Sweden. Moreover, the data and bacterial isolates in this study are from clinical investigations on farms with BRD outbreaks, and sampling and bacteriological cultures were performed to guide practitioners in managing the outbreaks and not in the context of an experimental study. Therefore, the processing of samples in the laboratory was performed as part of the routine work and focused on identifying \u003cem\u003ePasteurellacae\u003c/em\u003e. More elaborate efforts to identify isolates in the samples were not made, and only one isolate from each sample was further evaluated and saved. This probably led to underdiagnosis of bacterial pathogens in both DNS and TBL. Another limitation is that \u003cem\u003eMycoplasma\u003c/em\u003e spp. were not cultured because the methodology was not available at the laboratory at the time of the field investigations. Despite these limitations, we believe that the evaluation of DNS in relation to TBL and conclusions regarding the sampling of calves are relevant.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eIn this study, \u003cem\u003eP. multocida\u003c/em\u003e was readily isolated from the upper airways of calves. Although there was a large variation in isolation frequency between farms and calf groups, at least one \u003cem\u003eP. multocida\u003c/em\u003e isolate was obtained from DNS in 83.3% of the calf groups. In 75% of the calves, where \u003cem\u003eP. multocida\u003c/em\u003e was isolated from both DNS and TBL, isolates from DNS and TBL were identical. During an outbreak of BRD, a pragmatic and convenient approach for practitioners to gain insight into the bacterial respiratory pathogens present and their antibiotic susceptibility could be to collect and submit samples from the upper respiratory tract of 4\u0026ndash;6 calves.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eAuthors\u0026rsquo; contributions\u003c/p\u003e\n\u003cp\u003eBB participated in the field investigations on which the data and isolates of this study were based and designed the present study; BH performed the bacteriological analyses; MM performed the molecular analyses; BB performed the statistical analyses; BB and MM interpreted the results; BB, MM and MP drafted the manuscript; and all the authors commented and refined the draft. All authors have read and approved the final version of the manuscript.\u003c/p\u003e\n\u003cp\u003eAcknowledgements\u003c/p\u003e\n\u003cp\u003eThe contributions of VMD Sven Viring and VMD Charina G\u0026aring;nheim to the field investigations on which this study is based are gratefully acknowledged. Likewise, VMD Helene Wahlstr\u0026ouml;m is acknowledged for constructive criticism of this manuscript.\u003c/p\u003e\n\u003cp\u003eCompeting interests \u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003eAvailability of data and materials \u003c/p\u003e\n\u003cp\u003eThe datasets used and analysed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003eConsent for publication\u003c/p\u003e\n\u003cp\u003eNot applicable. \u003c/p\u003e\n\u003cp\u003eEthics approval\u003c/p\u003e\n\u003cp\u003eThe data and isolates on which this study is based were collected from calves at farm visits made by the Swedish animal health services in 1997\u0026ndash;2000. The farm visits, including sampling of calves, were part of routine work in managing outbreaks of BRD on farms affiliated with the organisation. Hence, this is not an experimental study and ethics approval and consent to participate are therefore not relevant.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePrior publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData have not been published previously.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe processing of data, analysis of isolates from the strain collection and elaboration of the manuscript were conducted within the SvarmPat programme. This programme aims to counteract emergence and spread of antibiotic resistance in farm animal bacterial pathogens and is financed by the Swedish Board of Agriculture. SvarmPat is run in cooperation between the Department of Animal Health and Antimicrobial Strategies at the Swedish Veterinary Agency and the Swedish consultancy company Farm and Animal Health.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eGay E, Barnouin J. 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BMC Microbiol. 2011; DOI: 10.1186/1471-2180-11-115.\u003c/li\u003e\n\u003cli\u003eCarter HF, Wills RW, Scott MA, Thompson AC, Singer RS, Loy JD, et al. Assessment of Diversity of Antimicrobial Resistance Phenotypes and Genotypes of \u003cem\u003eMannheimia haemolytica\u003c/em\u003e Isolates from Bovine Nasopharyngeal Swabs. Front Vet Sci. 2022; DOI: 10.3389/fvets.2022.883389.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Bovine respiratory diseases, cattle, cgMLST, MSLT, sampling.","lastPublishedDoi":"10.21203/rs.3.rs-4162560/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4162560/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eBovine respiratory disease (BRD) is common in intensively raised cattle and is often treated with antibiotics. To guide practitioners, knowledge of the bacteria involved in an outbreak and their antibiotic susceptibility is warranted. To this end, samples from the upper or lower respiratory tract of calves can be submitted for bacteriological culture and susceptibility testing of relevant isolates. However, it is debated whether isolates from the upper respiratory tract are representative of bacteria causing infections in the lower tract. In this study, we used MALDI-TOF MS, multilocus sequence typing (MLST) and core-genome multilocus sequence typing (cgMLST) to compare isolates from deep nasal swabs (DNS) and from tracheobronchial lavage (TBL) in 219 calves from 25 farms.\u003c/p\u003e\u003ch2\u003eResults:\u003c/h2\u003e \u003cp\u003eThe predominant bacterial pathogen in this study was \u003cem\u003ePasteurella multocida\u003c/em\u003e, which was isolated from 37.4% of DNS and 22.4% of TBL. There was no statistically significant difference in isolation frequency between healthy and diseased calves for DNS (p\u0026thinsp;=\u0026thinsp;0.778) or TBL (p\u0026thinsp;=\u0026thinsp;0.410). At least one \u003cem\u003eP. multocida\u003c/em\u003e isolate was obtained from 83.3% of the calf groups. At the 49 samplings where \u003cem\u003eP. multocida\u003c/em\u003e was isolated from TBL, the same species was isolated from DNS at 29 samplings (59.2%). From 28 of these samplings, isolates were evaluated by MLST and at 24 samplings (86.0%) \u003cem\u003eP. multocida\u003c/em\u003e from DNS and TBL were of the same sequence type (ST). Moreover, at 21 of the samplings (75.0%), cgMLST showed that the genetic distance within isolate pairs was less than two alleles, and DNS and TBL isolates were considered identical. At seven samplings (25%), the genetic distance within isolate pairs was greater, and DNS and TBL isolates were considered nonidentical. Within farms and groups of calves, \u003cem\u003eP. multocida\u003c/em\u003e of several different STs were isolated.\u003c/p\u003e\u003ch2\u003eConclusions:\u003c/h2\u003e \u003cp\u003e \u003cem\u003ePasteurella multocida\u003c/em\u003e was readily isolated from DNS and in calves where this species was isolated also from TBL, isolates from DNS and TBL were identical at 75% of the samplings. This suggests that during an outbreak of BRD, submission of DNS samples from 4\u0026ndash;6 calves could be a convenient approach for practitioners seeking guidance on bacterial pathogens present and their antibiotic susceptibility.\u003c/p\u003e","manuscriptTitle":"Pasteurella multocida from deep nasal swabs and tracheobronchial lavage in calves","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-04-22 04:39:32","doi":"10.21203/rs.3.rs-4162560/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"24d713bd-7285-4f40-a743-e79d1e4b35ee","owner":[],"postedDate":"April 22nd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-11-11T15:59:24+00:00","versionOfRecord":{"articleIdentity":"rs-4162560","link":"https://doi.org/10.1186/s13028-024-00781-7","journal":{"identity":"acta-veterinaria-scandinavica","isVorOnly":false,"title":"Acta Veterinaria Scandinavica"},"publishedOn":"2024-11-05 15:57:07","publishedOnDateReadable":"November 5th, 2024"},"versionCreatedAt":"2024-04-22 04:39:32","video":"","vorDoi":"10.1186/s13028-024-00781-7","vorDoiUrl":"https://doi.org/10.1186/s13028-024-00781-7","workflowStages":[]},"version":"v1","identity":"rs-4162560","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4162560","identity":"rs-4162560","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}