The role of short journey transportation in the spreading of swine pathogens and antimicrobial-resistant bacteria

The role of short journey transportation in the spreading of swine pathogens and antimicrobial-resistant bacteria | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article The role of short journey transportation in the spreading of swine pathogens and antimicrobial-resistant bacteria Marta Masserdotti, Nicoletta Formenti, Anna Donneschi, Flavia Guarneri, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4251132/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: The transport of live pigs poses a risk to on-farm biosecurity. Trucks can carry pathogens with significant economic and health impacts, including antimicrobial-resistant (AMR) bacteria. This study aimed to investigate the microbiological contamination of trucks before and after loading, focusing on AMR bacteria and other major pathogens transmissible through faeces. Samples were collected by swabbing the internal surface of disinfected empty trucks at farm entry (‘clean’) and after loading (‘dirty’), and were tested for total plate count (TPC), specific bacteria and viruses. Escherichia coli isolates were also phenotypically and molecularly tested for the presence of extended-spectrum β-lactamase (ESBL), other β-lactamases (AmpC) and carbapenemase. Results: Bacterial counts (both TPC and Enterobacteriaceae count) and the probability of testing positive for E. coli , ESBL/AmpC-producing E. coli and Rotavirus A varied significantly depending on the truck condition, being significantly higher in “dirty” than in “clean” trucks. Despite a non-significant difference, positivity to Rotavirus B showed the same tendency. Conversely, the truck condition had no effect on Rotavirus C. Salmonella spp., PRRSV, and carbapenemase-producing E. coli were detected only in samples collected on “dirty” trucks. Conclusions: Although the prevalence of most agents in ‘clean’ samples was close to zero, the relatively frequent occurrence of E. coli and some rotaviruses highlights the importance of improving sanitisation procedures. The detection of ESBL/AmpC- and carbapenemase-producing E. coli was of particular concern. These findings confirm the role of trucks in spreading pathogens of concern and AMR, highlighting the importance of effective monitoring and proper sanitisation procedures. biosecurity AMR clean and dirty trucks swabs carbapenemase-producing E. coli Rotavirus C ESBL/AmpC Figures Figure 1 Figure 2 Background The approach of veterinary medicine to animal health has transformed considerably over time, especially in the livestock field, turning more and more to preventing diseases than to treating them ( 1 ). The key element to achieve effective prevention is biosecurity, a system of practices and measures aimed at minimising the risk of introduction, establishment and spread of animal infections within and among animal populations. According to the World Organization for Animal Health (WOAH)( 2 ), implementing biosecurity plans is thus pivotal for effective disease prevention within individual farms as well as in whole regions. A low risk of pathogen introduction and/or spread within the herd leads in turn to a reduction in the use of drugs, especially antimicrobials ( 3 ). Specific to pig farming, a clear link between the implementation of biosecurity measures and improvements in parameters related to antimicrobial use and production has been demonstrated and quantified ( 4 , 5 ). A key measure to prevent the spread of diseases within a farm, is to avoid mixing pigs of different ages ( 6 , 7 ). This can be achieved either by establishing a rigorous workflow and separation of the groups within the farm or by allocating the groups to different farming sites. Especially in high-income countries characterized by more intensive pig production and bigger herd sizes, the second solution is generally preferred, and pigs typically follow a path that begins on the birth farm and then transitions to a fattening farm, sometimes including a nursery farm dedicated to caring for weaned piglets. Consequently, transporting animals to and from the farm is a common occurrence ( 8 ) and is also one of the most critical hazards for farm biosecurity, as it involves breaching the biosecurity interface and potentially having animals, as well as trucks or even drivers, carry around pathogens of concern for both animal health (e.g. Porcine reproductive and respiratory syndrome virus (PRRSV) ( 9 ), Porcine epidemic diarrhoea virus (PEDV) ( 10 – 12 ), African swine fever virus (ASFV) ( 13 )) and public health (e.g. Salmonella spp. ( 14 ), AMR pathogens ( 15 – 17 )). Efficient truck cleaning and disinfection procedures are thus crucial to prevent the spread of pathogens from one farm to another, to the abattoir, and to operators. Nevertheless, trucks often show residues of contamination when entering the farm ( 16 ). This is due either to improper execution of the cleaning and disinfection procedures, or to protocols that target only some specific pathogens and are thus partially effective. Traditionally, legislative and research efforts in the animal transportation field have been focused mostly on improving welfare conditions rather than sanitary ones, and studies on the role of trucks as a potential threat to on-farm biosecurity are scarce. Furthermore, the lack of studies and data is even more limited when specifically considering the spread of AMR pathogens, as recently reported by the European Food Safety Authority (EFSA) ( 18 ). The World Health Organization recently published a list of bacteria for which new antimicrobials are urgently needed, particularly highlighting the threat posed by bacteria relevant at the nosocomial level that have become resistant to carbapenems and third-generation cephalosporins ( 19 ). Among these bacteria are Enterobacteriaceae , which are showing rising levels of AMR ( 20 , 21 ). Extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae or carbapenemase-producing Enterobacteriaceae are a significant threat to public health due to their association with increased health costs, hospitalisation, and mortality rate ( 22 ). Specifically, E. coli is frequently included in AMR monitoring programs worldwide ( 23 ) as it is a cross-species pathogen with a great capacity to accumulate resistance genes, mostly through horizontal gene transfer ( 24 ) ( 25 ). There is, therefore, an urgent need to investigate and quantify the impact of animal transportation ( 18 ) in the transmission and diffusion of AMR, assessing the role of different risk factors such as the hygiene of the transport vehicle. Based on the above considerations, Enterobacteriaceae are considered a sensitive indicator of ineffective cleaning ( 26 , 27 ) and can also be used to test for the presence of AMR. In the present work, we investigated the contamination of pig transport vehicles before and after animal loading procedures, to assess the effectiveness of cleaning and disinfection measures to which transport vehicles are commonly subjected. We focused on the main swine pathogens transmissible through faeces due to their permanence in the organic matrix and consequent feasibility of sampling: Enterobacteriaceae ( E. coli , Salmonella spp.), Brachyspira spp., Lawsonia intracellularis , PRRSV, PEDV, and Rotavirus A, B, C, H. Additionally, we investigated ESBL/AmpC- and carbapenemase-producing E. coli pathogens in order to evaluate the general risk of AMR spread associated with animal transportation. Results A total of 84 swabs collected from 42 trucks (42 “dirty” and 42 “clean” swabs) were tested for specific pathogens and bacterial counts. The prevalences of the pathogens considered and their distribution between swabs from “clean” and “dirty” trucks are reported in Table 1 and Fig. 1 . Table 1 Overall prevalence of the investigated pathogens and percentage of positive swabs sampled in “clean” and “dirty” pig transport trucks. Microorganism Prevalence (%) % on “clean” trucks % on “dirty” trucks E. coli 40.47% (34 / 84) 14.71% (5 / 34) 85.29% (29 / 34) ESBL/AmpC producing E. coli 10.71% (9 / 84) 11.11% (1 / 9) 88.89% (8 / 9) Carbapenemase producing E. coli 11.90% (10 / 84) 0% 100% (10 / 10) Salmonella spp. 3.85% (3 / 84) 0% 100% (3 / 3) Brachyspira spp. 0% 0% 0% Lawsonia intracellularis 5.13% (4 / 78) 25% (1 / 4) 75% (3 / 4) PRRSV 10.26% (8 / 78) 0% 100% (8 / 8) PEDV 0% 0% 0% Rotavirus A 16.67% (13 / 78) 15.38% (2 / 13) 84.62% (11 / 13) Rotavirus B 33.33% (26 / 78) 34.62% (9 / 78) 65.38% (17 / 78) Rotavirus C 56.41% (44 / 78) 52.27% (23 / 78) 47.73% (21 / 78) Rotavirus H 5.13% (4 / 78) 75% (3 / 4) 25% (1 / 4) For most of the examined pathogens, at least one positive swab was found in both “clean” and “dirty” trucks, with the exception of PRRSV, carbapenemase-producing E. coli and Salmonella spp., which were detected only in “dirty” samples. Regarding the latter, the three isolates were serotyped, resulting in one Salmonella Typhimurium monophasic variant and two Salmonella Choleraesuis . No samples positive to Brachyspira spp. or PEDV were detected. Regarding those pathogens detected in both conditions and with an overall prevalence higher than 10%, the probability of testing positive was significantly higher in “dirty” samples than in “clean” ones for E. coli (p < 0.0001) , ESBL/AmpC-producing E. coli (p = 0.026) and Rotavirus A (p = 0.011). Despite a non-significant difference (p = 0.0504), positivity to Rotavirus B showed the same tendency. The truck condition had conversely no effect on Rotavirus C presence (p = 0.10), which varied instead depending on the production stage (p = 0.0002), with swabs from weaning and growing farms being more likely to be positive than the ones from fattening farms. Similarly, ESBL/AmpC-producing E. coli were more likely to be found in swabs collected from weaning than fattening farms (p = 0.046). Bacterial counts varied significantly depending on the truck condition as well: both TPC (p < 0.0001) and Enterobacteriaceae counts (p < 0.0001) were significantly higher in “dirty” samples than in “clean” ones (Fig. 2 ). Complete results of all models, including parameter estimates, are reported in Supplementary Table 1. Regarding AMR bacteria, the proportion of isolates resistant to the different molecules examined, and their distribution between “clean” and “dirty” condition are shown in Table 2 for E. coli isolates and in Table 3 for ESBL/AmpC-producing E. coli and carbapenemase-producing E. coli. Table 2 Percentage of resistance of E. coli isolates (n = 34) for each molecule analyzed, distribution of these percentages between swabs taken on the “clean” and the “dirty” trucks. The epidemiological cut-off is available online (ECOFFs, EUCAST, www.eucast.org , last accessed on 28th February 2024). Molecule (Tentative) Epidemiological cut-off values (T)ECOFF % Resistance % on “dirty” trucks % on “clean” trucks Ampicillin 8 64.71% (22 / 34) 90.91% (20 / 22) 9.09% (2 / 22) Tetracycline 8 61.76% (21 / 34) 85.71% (18 / 21) 14,29% (3 / 21) Florfenicol 16 55.88% (19 / 34) 89.47% (17 / 19) 10.53% (2 / 19) Trimetoprim + Sulfametoxazole 0,5 50% (17 / 34) 88.23% (15 / 17) 11.76% (2 / 17) Enrofloxacin 0,125 32.35% (11 / 34) 90.91% (10 / 11) 9.09% (1 / 11) Gentamicin 2 26.47% (9 / 34) 100% (9 / 9) 0% Amoxicillin + Clavulanic acid ( 8 ) 23.53% (8 / 34) 100% (8 / 8) 0% Flumequina 2 23.53% (8 / 34) 100% (8 / 8) 0% Cefazolin 4 20.59% (7 / 34) 100% (7 / 7) 0% Kanamycin ( 16 ) 17.64% (6 / 34) 100% (6 / 6) 0% Colistin 2 2.94% (1 / 34) 100% (1 / 1) 0% 67.65% (23/34) of the tested E. coli samples were resistant to sulfisoxazole. The majority of these (20/23) were collected on “dirty” trucks. 14.71% (5/34) of the E. coli samples were resistant to amminosidine, all collected on “dirty” trucks. Table 3 Percentage of resistance of ESBL/AmpC-producing E. coli isolates (n = 9) and carbapenemase-producing E. coli isolates (n = 10) for each molecule analyzed, distribution of these percentages between swabs taken on the “clean” and the “dirty” trucks. The epidemiological cut-off is available online (ECOFFs, EUCAST, www.eucast.org , last accessed on 28th February 2024). Molecule (Tentative) Epidemiological cut-off values (T)ECOFF ESBL/AmpC – producing E. coli Carbapenemase – producing E. coli % Resistance % on “dirty” trucks % on “clean” trucks % Resistance % on “dirty” trucks % on “clean” trucks Cefotaxime 0,25 100% (9 / 9) 88.89% (8 / 9) 11,11% (1 / 9) 80% (8 / 10) 100% (8 / 8) 0% (0 / 8) Cefepime 0,125 88.89% (8 / 9) 87.50% (7 / 8) 12.5% (1 / 8) 50% (5 / 10) 100% (5 / 5) 0% (0 / 5) Ceftazidime 1 55.56% (5 / 9) 100% (5 / 5) 0% (0 / 5) 10% (1 / 10) 100% (1 / 1) 0% (0 / 1) Cefotaxime + Clavulanic acid 0,25 33.33% (3 / 9) 100% (3 / 3) 0% (0 / 3) 60% (6 / 10)* 100% (6 / 6) 0% (0 / 6) Cefoxitin 16 33.33% (3 / 9) 100% (3 / 3) 0% (0 / 3) 10% (1 / 10)** 100% (1 / 1) 0% (0 / 1) Ertapenem (0,03) 33.33% (3 / 9) 100% (3 / 3) 0% (0 / 3) 100% (10 / 10) 100% (10 / 10) 0% (0 / 10) Ceftazidime + Clavulanic acid 1 33.33% (3 / 9) 100% (3 / 3) 0% (0 / 3) 10% (1 / 10) 100% (1 / 1) 0% (0 / 1) Temocillin 16 11.11% (1 / 9)* 100% (1 / 1) 0% (0 / 1) 100% (10 / 10) 100% (10 / 10) 0% (0 / 10) Imipenem 0,5 0% 0% 0% 20% (2 / 10)** 100% (2 / 2) 0% (0 / 2) Meropenem 0,06 0% 0% 0% 80% (8 / 10)* 100% (8 / 8) 0% (0 / 8) The 7 ESBL/AmpC-producing E. coli isolates that were subjected to molecular investigation by PCR were found to be positive for bla CTX-M, bla TEM and bla CMY (alone or in combination), and none was positive for bla SHV . One isolate was negative to all the sought genes (Table 4 ). All of the carbapenemase-producing E. coli isolates that were subjected to molecular investigation by PCR were found to be positive only for the bla OXA-48 gene (Table 4 ). Table 4 Distribution of prevalence of resistance genes found, either singly or in combination, in ESBL/AmpC- and carbapenemase-producing E. coli isolates. bla CTX−M bla CTX−M+TEM bla CMY+TEM bla SHV Prevalence of resistance genes found, either singly or in combination, in ESBL/AmpC-producing E. coli isolates 42.86% (3 / 7) 28.67% (2 / 7) 14.29% (1 / 7) 0% (0 / 7) bla OXA−48 bla NDM bla KPC bla VIM Prevalence of resistance genes found in carbapenemase-producing E. coli isolates 100% (9 / 9) 0% 0% 0% Discussion We investigated the potential role played by vehicles for transporting live pigs in the spread of a comprehensive panel of pathogens transmissible through faeces, including antibiotic-resistant pathogens, an aspect that has gaps in knowledge and requires dedicated investigations, as recently reported by EFSA ( 28 ). Overall, we found significantly lower bacterial counts and pathogen occurrence in sanitized trucks entering the farms, compared to loaded trucks. In particular, several pathogens of zoo-economic and zoonotic importance, namely carbapenemase-producing E. coli , Salmonella spp. and PRRSV were never detected on “clean” trucks entering the farm. These are encouraging results regarding the effectiveness of cleaning and disinfection procedures. However, residual contamination by E. coli , L. intarcellularis , and rotaviruses was found in sanitised trucks at farm entry. In particular, Rotaviruses B, C and H were frequently detected on “clean” trucks, with Rotavirus C occurrence being equally likely on “clean” and “dirty” trucks, suggesting that current sanitization protocols might be ineffective against them. In general, Rotaviruses are known for their high environmental resistance: drying does not inactivate all viromes ( 29 ), and while for Rotavirus A the effectiveness of phenolic disinfectants regardless of the concentration of organic substance has been demonstrated, glutaraldehyde-based disinfectants or peroxide compounds have proved to be effective only if preceded by careful cleaning and removal of the organic matrix ( 30 ). Additionally, there is a lack of information regarding the effectiveness of disinfection procedures against Rotavirus C and H, whose potential pathogenicity has only recently been highlighted ( 7 , 31 ). Therefore, it is necessary to deepen the knowledge of the epidemiology and the environmental resistance of these pathogens, to develop a protocol that can be effective also towards them. Studies on the role of trucks as a potential threat to on-farm biosecurity are usually mainly related to specific outbreak episodes ( 9 , 11 , 12 ), and are thus mostly focused on a single pathogen, not assessing the sanitary risk from a broader epidemiological perspective. For instance, two studies were conducted in our same intensive pig production area in Northern Italy to investigate the role of vehicles as a source of PEDV transmission following the occurrence of an epidemic wave ( 10 ), and to assess the propagation of Brachyspira hyodysenteriae by vehicles for transport to slaughterhouse ( 32 ). Both studies reported relatively high levels of truck contamination, but interestingly, we didn’t find any of the two pathogens in our investigation. Our results on the prevalence of AMR bacteria are consistent with findings from previous studies carried at environmental level ( 33 , 34 ) or by faecal samplings ( 35 – 37 ). Our results highlight a relatively common occurrence of resistance against common antimicrobials ( 38 ), such as tetracycline, chloramphenicol, and trimethoprim/sulfamethoxazole, while resistance to last-resort antimicrobials, such as colistin, broad-spectrum cephalosporins and carbapenems is less frequent. Notably, AbuOun et al. (2020) did not detect any carbapenem-resistant strains, which emerged instead from the present study and from the one conducted by Peng et al. (2022). With regard to ESBL/AmpC-producing strains, our findings are consistent with recent studies that highlight bla CTX−M as the most common gene conferring resistance to beta-lactams, detected ubiquitously in community and healthcare contexts, in the environment, in food, and in animal species ( 39 ). For carbapenemase-producing E. coli strains, only bla OXA−48 -encoding strains were found in this study. This gene was reported for the first time in E. coli isolated from pigs in 2019 in Germany ( 40 ), and it is speculated that transmission occurred from humans, in which this resistance gene was already circulating. Data on the dissemination of carbapenem-resistant Enterobacteriaceae (CRE) in animal species and their potential for transmission to humans are still scarce, and in Europe its prevalence in livestock seems to be low, unlike in Asia and North Africa ( 41 ). However, considering the importance of carbapenems as last-resort antimicrobials for humans, an integrated surveillance approach of CRE emergence is paramount. When considering the prevalence of AMR pathogens across pig production phases, the trend that is observed in the present study is consistent with other studies, where they were more common in early production stages ( 42 ). Overall, the data about AMR pathogens obtained from trucks in this study are comparable with those available in literature, collected at the farm level or by animal sampling. This suggests that animal transport vehicles may represent an alternative epidemiological indicator of AMR pathogens, other than a means to assess the risk related to livestock movements in the spread of AMR microorganisms ( 15 , 16 ). In general, the high pathogen contamination levels detected on outgoing trucks suggest that loaded trucks may provide a potential indicator of the microorganisms circulating in the farm, and above all highlight that ineffective cleaning and disinfection procedures may represent a breach in the biosecurity of the farms visited afterwards. However, it must be noted that our study presents some limitations: the limited number of farms which were enrolled based on opportunistic criteria related to logistic constraints, and farmers’ agreement to participate. Such convenience sampling implies that our conclusions might not be extended to the whole population. However, we believe that this might make our results even more conservative, as the farmers who volunteered for the study are probably among those with the best biosecurity procedures. On the whole, the lack of provisions, guidelines or regulations regulating precisely the hygiene of means of transport should be highlighted: for instance in the European Union, the Article 125 of Regulation (EU) 2016/429 on the prevention of transport-related diseases merely recommends that all equipment and means of transport must be subjected to thorough cleaning, disinfection, and pest control measures to eliminate any potential biohazard, and that other biosecurity measures must be taken depending on the risks associated with the transport operations concerned. This regulation has been supplemented, but does not contain any specific provisions yet. Despite the present study was not focused on the loading procedures, observing the loading of pigs has uncovered critical biosecurity weaknesses that require training and awareness of transporters and breeders, as recently prescribed by Articles 11 and 13 of the Animal Health Law (Reg. CE 2016/429). To achieve the highest biosecurity standards, trucks should be allowed to stand for a period after passing through disinfection arches, transportation staff should not be involved in animal handling or entering animal housings, disposable personal protective equipment should always be readily available, and transporters should not be allowed to enter the truck’s cockpit after donning personal protective equipment. Conclusion To the best of our knowledge, this study is among the firsts to investigate trucks as mechanical vectors of pathogens relevant to pig breeding and public health, including AMR bacteria. Microbiological investigations carried out on disinfected trucks dedicated to animal transport at farm entry highlight the risk of introducing pathogens, showing that the cleaning and disinfection protocols currently in use are only partially effective, particularly against Rotavirus C. On the other hand, the generally lower bacterial counts and pathogen prevalence on sanitized trucks, and in particular the rare occurrence of AMR bacteria, are an encouraging result. Further studies will have to evaluate the effectiveness of these protocols on farms and especially in farrowing and weaning units. Overall, these findings suggest that there is a fair focus on cleaning and disinfection procedures in animal transport, but at the same time highlight the need for further studies assessing the efficacy of protocols against specific pathogens, and for the definition of guidelines to allow greater uniformity and greater awareness of transporters and operators about such procedures, so that the protocols are carried out with the appropriate attention and therefore greater effectiveness. Methods Sampling To investigate the contamination of animal transport vehicles and assess the efficacy of standard cleaning and disinfection procedures, from March to July 2023 we surveyed trucks used for animal transport in 11 pig farms (3 weaning farms, 4 growing farms, 3 fattening farms, and 1 farm that loaded animals both at growing and at fattening stage) located in Lombardy, Northern Italy (farms’ size are listed in Suppl. Table 2). The trucks were employed for short journeys (less than 8 hours) to transport pigs from farm to farm or from farm to slaughterhouse. The farms were enrolled in the study based on opportunistic criteria related to logistic constraints, feasibility and voluntary acceptance to participate by farmers. For each truck, two samples were collected in duplicate, for a total of four samples: two before the start of loading procedures, at the time of arrival of the vehicle on the farm and after it had been subjected to the disinfection measures adopted by the company (hereafter, "clean” samples); and two at the end of loading procedures before the truck left the farm (hereafter, "dirty” samples). Sterile gauze swabs were rubbed on the floor and on the lower portion of the interior walls of the truck and/or trailer to cover a total area of approximately 100 cm 2 , avoiding those areas that were still wet from the residual disinfectant from the biosecurity procedures at the entrance of the farm. The disinfectant is usually applied on the external surfaces of the clean truck entering the farm either manually, by an operator, or automatically, by a disinfection arch located at the entrance. The swabs were then placed in pairs in sterile sampling bags, stored at room temperature, within ≤ 6 hours delivered to the laboratory where swabs were stored at + 4°C until further analyses. A total number of 84 samples (buffer pairs) were collected (Table 5 ). Of the 84 swabs taken, 78 were subjected to both microbiological and molecular investigations, while the remaining 6 could only be subjected to bacteriological tests due to a sampling error. Table 5 Number of collected samples by rearing stage. A farm (*) was sampled both when loading growing pigs and when loading fattening pigs. Rearing stage Number of farms Number of samples Number of trucks Weaning piglets transported to a nursery or to a wean-to-finish barn 3 22 11 Growing pigs transported from a nursery to a finishing barn 5* 22 11 Finishing pigs transported to slaughterhouse 4* 40 20 Total 11 84 42 Microbiological analyses The Total Plate Count (TPC) was performed manually after incubation at 37°C for 48h on PCA medium of the − 3 dilution, according to internal testing method ( 43 – 45 ). Such testing method has a superior detection limit of 1,5 x 10 6 CFU/cm 2 . The quantification of Enterobacteriaceae contamination was performed according to ISO 21528-2 procedures, and the biochemical confirmation was obtained through the oxidase test. The detection and identification of Salmonella spp. strains was performed according to ISO 6579-1:2017/Amd1:2020 and ISO/TR 6579-3:2014 procedures. Concerning E. coli , bacteriological examinations were performed by using standard bacteriological cultures ( 46 ), while for ESBL/AmpC- and carbapenemase-producing E. coli the following method was applied: after a pre-enriching phase with BPW (1:10 ratio) at 37°C for 18–24 hours, a first selective incubation at 37 ºC for 18 hours was performed for detecting ESBL/AmpC- and carbapenemase-producing E. coli using a solid medium (respectively, McConkey + cefotaxime 1 mg/L at 41 ± 2°C and CHROMID® CARBA SMART at 37°C, both for 18–24 hours). At the end of this first incubation, the suspected colonies were transplanted on solid McConkey soil and incubated at 37°C for 18–24 hours. A single bacterial colony from each phenotype-positive sample was resuspended in 250 µL of DNase-Rnase free water and DNA was extracted by lysis-boiling (98°C for 10 min) for further molecular characterization. After molecular confirmation, the plates were subjected to antimicrobial susceptibility testing (the panels of tested antimicrobials are listed in Suppl. Table 3). E. coli isolates were also tested for aminosidine and sulfisoxazole, and the classification of the resistance profile was based on the clinical cut-off of the internal test method ( 47 – 52 ). Minimum inhibitory concentrations (MICs) were determined by broth microdilution, and the strains were classified as “resistant” or “susceptible” according to the Epidemiological cut-off values (ECOFFs) recommended by the European Committee on Antimicrobial Susceptibility Testing (EUCAST, www.eucast.org ). When unavailable, clinical breakpoints were used ( 47 – 52 ). Molecular analyses The screening for Brachyspira spp., Lawsonia intracellularis , PRRSV, PEDV, and Rotavirus A, B, C, H was performed by molecular analysis. In particular, PRRSV screening was performed using a commercial kit (virotype PRRSV RT-PCR Kit, Indical Bioscience) that allows the detection of PRRS virus and its discrimination in European, American and highly pathogenic American strain. All the other pathogens were searched for by means of in-house methods. In detail, Brachyspira hyodysenteriae, B. pilosicoli , and Lawsonia intracellularis were searched by real-Time PCR using the kit QuantiNova Probe PCR mastermix (Qiagen) and targeting the nox gene region in Brachyspira spp. and the aspA gene in L. intracellularis ( 53 , 54 ). The reaction for the identification of PEDV allows for the detection of the presence of a nucleic acid region of 111 bp of the S1 gene (Spike, region 1) ( 55 , 56 ). The performed Rotavirus Real-time RT-PCR method allows the detection of an RNA portion of the VP6 region specific to the A, B, C, H groups of Rotavirus in pigs. The method is based on the use of two multiplex reactions one-step Real-Time RT-PCR: in the first multiplex reaction specific regions of groups A (98 bp) and C (86 bp) are amplified, and in the second multiplex reaction the specific regions of groups B (89 bp) and H (93 bp) are amplified ( 31 ). The primers sequences used to detect the presence of Brachyspira hyodisenteriae and B. pilosicoli , Lawsonia intracellularis , PRRSV, PEDV, and Rotavirus A, B, C, H are listed in Suppl. Table 4. Finally, a multiplex-PCR was used to search for the genes coding for ESBL/AmpC ( bla CTX−M 593 bp, bla SHV 237 bp, bla TEM 445 bp, bla CMY 820 bp) ( 57 , 58 ) and carbapenemases ( bla VIM 390 bp, bla NDM 621 bp, bla KPC 798 bp, blaOXA-48 438 bp) ( 59 ). We investigated 7 out of the 9 ESBL/AmpC-producing E. coli isolates and 9 out of the 10 carbapenemase-producing E. coli isolates that were detected on plate. The impossibility of carrying out the molecular investigation in the remaining isolates is linked to the absence of regrowth of the strain during the processing phases or to pollution of the strain. Statistical analysis To assess the effectiveness of cleaning and disinfection protocols, the effect of the independent variable “condition” (i.e., "clean” or ”dirty") on: i) the presence/absence of pathogens with overall prevalence greater than 10% (i.e., E. coli , ESBL/AmpC-producing E. coli , and Rotaviruses A, B, C), and on ii) TPC and Enterobacteriaceae counts (ln-transformed values [ln(x + 1]) were analyzed through mixed logistic regressions and linear mixed models, respectively. In all models, the productive category (i.e., weaning, growing or fattening) was included as a covariate and farm IDs were included as random intercepts to account for different samplings within the same farm. When significant, factors with more than two levels (namely, productive category) were tested post hoc through t-tests on differences of least-square means, applying Holm correction for multiple comparisons. Results were considered statistically significant when p < 0.05. All the analyses were carried out using the packages lme4 and emmeans in R (R Core Team 2021) version 4.3.1 (2023-06-16). Abbreviations AMR Antimicrobial-Resistant/Antimicrobial Resistance ASF(V) African Swine Fever (Virus) CRE Carbapenem-Resistant Enterobacteriaceae EC Enterobacteriaceae count (T)ECOFF (Tentative) Epidemiological cut-off values ESBL Extended-Spectrum β-lactamase EUCAST European Committee on Antimicrobial Susceptibility Testing EFSA European Food Safety Authority PEDV Porcine Epidemic Diarrhoea Virus PRRS(V) Porcine Respiratory and Reproductive Syndrome Virus TPC Total Plate Count WOAH World Organisation for Animal Health Declarations Ethics approval and consent to participate Not applicable Consent for publication Not applicable Availability of data and materials All data are available upon reasonable request from the authors. Competing interests The authors declare that they have no competing interests. Funding This work was supported by Italian Ministry of Health (Agreement ClassyFarm 2021-2023). The Funder had no role in the design, analysis and reporting of the study. Authors' contributions Conceived and designed the experiments: N.F., C.R., G.L.A., C.L.. Performed sampling: M.M., A.D., E.G.. Performed laboratory experiments: F.G., C.B., M.B.B.. Analyzed the data: M.M., N.F., C.R., F.S., C.B., M.B.B.. Contributed reagents/materials/analysis tools: M.B.B., G.L.A.. Writing—original draft preparation: M.M., N.F., C.R., G.L.A., C.L.. Writing—review and editing: M.M., N.F., A.D., F.G., F.S., C.R., E.G., C.B., M.B.B., G.L.A., C.L.. Supervision: M.B.B., G.L.A., C.L.. Funding acquisition: G.L.A.. All authors read and approved the final manuscript. Acknowledgements We thank Laura Birbes, Paola Giangrossi, Fabiana Gatti, Gloria Garbin, Claudia Alberti and Chiara Boifava for their help with the project activities. References European Commission. Animal Health Strategy for the European Union. (2007–2013) where ‘Prevention is better than cure’ [Internet]. 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Koutsoumanis K, Allende A, Álvarez-Ordóñez A, Bolton D, Bover‐Cid S, Chemaly M et al. Transmission of antimicrobial resistance (AMR) during animal transport. EFSA J. 2022;20(10). Ward RL, Bernstein DI, Knowlton DR, Sherwood JR, Young EC, Cusack TM et al. Prevention of surface-to-human transmission of rotaviruses by treatment with disinfectant spray. J Clin Microbiol. 1991;29(9). Chandler-Bostock R, Mellits KH. Efficacy of disinfectants against porcine rotavirus in the presence and absence of organic matter. Lett Appl Microbiol. 2015;61(6). Ferrari E, Salogni C, Martella V, Alborali GL, Scaburri A, Boniotti MB. Assessing the Epidemiology of Rotavirus A, B, C and H in Diarrheic Pigs of Different Ages in Northern Italy. Pathogens. 2022;11(4). Giacomini E, Gasparrini S, Lazzaro M, Scali F, Boniotti MB, Corradi A, et al. The role of transportation in the spread of Brachyspira hyodysenteriae in fattening farms. BMC Vet Res. 2018;14(1):10. Peng Z, Hu Z, Li Z, Zhang X, Jia C, Li T et al. Antimicrobial resistance and population genomics of multidrug-resistant Escherichia coli in pig farms in mainland China. [cited 2023 Sep 26]; https://doi.org/10.1038/s41467-022-28750-6 . AbuOun M, O’Connor HM, Stubberfield EJ, Nunez-Garcia J, Sayers E, Crook DW, et al. Characterizing Antimicrobial Resistant Escherichia coli and Associated Risk Factors in a Cross-Sectional Study of Pig Farms in Great Britain. Front Microbiol. 2020;11:861. Zhang P, Shen Z, Zhang C, Song L, Wang B, Shang J et al. Surveillance of antimicrobial resistance among Escherichia coli from chicken and swine, China, 2008–2015. Vet Microbiol [Internet]. 2017 May 1 [cited 2024 Mar 27];203:49–55. https://pubmed.ncbi.nlm.nih.gov/28619166/ . Burow E, Rostalski A, Harlizius J, Gangl A, Simoneit C, Grobbel M et al. Antibiotic resistance in Escherichia coli from pigs from birth to slaughter and its association with antibiotic treatment. Prev Vet Med [Internet]. 2019 [cited 2024 Mar 27];165:52–62. http://creativecommons.org/licenses/BY-NC-ND/4.0/ . Liu X, Liu Q, Cheng Y, Liu R, Zhao R, Wang J, et al. Effect of Bacterial Resistance of Escherichia coli From Swine in Large-Scale Pig Farms in Beijing. Front Microbiol. 2022;13:820833. Lekagul A, Tangcharoensathien V, Yeung S. Patterns of antibiotic use in global pig production: A systematic review. 2019 [cited 2024 Mar 27]; https://doi.org/10.1016/j.vas.2019.100058 . Castanheira M, Simner PJ, Bradford PA. Extended-spectrum β-lactamases: An update on their characteristics, epidemiology and detection. Volume 3. JAC-Antimicrobial Resistance; 2021. Irrgang A, Pauly N, Tenhagen BA, Grobbel M, Kaesbohrer A, Hammerl JA. Spill-over from public health? First detection of an OXA-48-producing Escherichia coli in a German pig farm. Microorganisms. 2020;8(6). Köck R, Daniels-Haardt I, Becker K, Mellmann A, Friedrich AW, Mevius D et al. Carbapenem-resistant Enterobacteriaceae in wildlife, food-producing, and companion animals: a systematic review. Vol. 24, Clinical Microbiology and Infection. 2018. Pholwat S, Pongpan T, Chinli R, Rogawski McQuade ET, Thaipisuttikul I, Ratanakorn P et al. Antimicrobial Resistance in Swine Fecal Specimens Across Different Farm Management Systems. Front Microbiol. 2020;11. Tiecco G. Microbiologia degli alimenti di origine animale. Seconda Edizione. Bologna: Edagricole; 1992. pp. 15–7. ICMSF. Microorganisms in Foods 1: Their Significance and Methods of Enumeration. 2nd Edition. Toronto: University of Toronto Press. 1988. 112–124 p. Hayes PR. Food Microbiology and Hygiene. London: Elsevier Applied Science; 1985. pp. 144–57. BSOP 54 National Standard Method Inoculation of Culture Media. issued by Standards Unit, Evaluations and Standards Laboratory, Specialist and Reference Microbiology Division, Issue No. 4, Issue Date 03.05.05 . The European Committee on Antimicrobial Susceptibility Testing. Breakpoint tables for interpretation of MICs and zone diameters. 2021;11.0. http://www.eucast.org . Lubbers BV, Miller C, Papich MG. Clinical and Laboratory Standards Institute. Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals. p. 170. Société Française de Microbiologie. Comité de l’antibiogramme de la Société Française de Microbiologie RECOMMANDATIONS. 2014. Jean-Yves Coordonnateur M, Jean-Winoc DECOUSSER, Hôpital Henri Mondor L, Nicolas Hôpital Bicêtre FORTINEAU, Kremlin-Bicêtre C, Marisa Anses HAENNI, Eric Anses LJOUY, Isabelle Anses LKEMPF et al. P, Comité de l’antibiogramme de la Société Française de Microbiologie RECOMMANDATIONS VÉTÉRINAIRES. 2020. 2020. Wayne PA. Methods for Antimicrobial Susceptibility Testing of Infrequently Isolated or Fastidious Bacteria Isolated From Animals. A CLSI supplement for global application. 1st ed CLSI supplement VET06 [Internet]. 2017; . Weinstein MP. Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing. 282 p. Willems H, Reiner G. A multiplex real-time PCR for the simultaneous detection and quantitation of Brachyspira hyodysenteriae, Brachyspira pilosicoli and Lawsonia intracellularis in pig faeces. Berl Munch Tierarztl Wochenschr. 2010;123(5–6):205–9. Gasparrini S, Alborali GL, Pitozzi A, Guarneri F, Giacomini E, Baldo V, et al. Characterization of Brachyspira hyodysenteriae isolates from Italy by multilocus sequence typing and multiple locus variable number tandem repeat analysis. J Appl Microbiol. 2017;123(2):340–51. Li Z, Chen F, Yuan Y, Zeng X, Wei Z, Zhu L et al. Sequence and phylogenetic analysis of nucleocapsid genes of porcine epidemic diarrhea virus (PEDV) strains in China. Arch Virol. 2013;158(6). Li ZL, Zhu L, Ma JY, Zhou QF, Song YH, Sun BL et al. Molecular characterization and phylogenetic analysis of porcine epidemic diarrhea virus (PEDV) field strains in south China. Virus Genes. 2012;45(1). Fang H, Ataker F, Hedin G, Dornbusch K. Molecular epidemiology of extended-spectrum β-lactamases among Escherichia coli isolates collected in a Swedish hospital and its associated health care facilities from 2001 to 2006. J Clin Microbiol. 2008;46(2). Rehman MA, Hasted TL, Persaud-Lachhman MG, Yin X, Carrillo C, Diarra MS. Genome Analysis and Multiplex PCR Method for the Molecular Detection of Coresistance to Cephalosporins and Fosfomycin in Salmonella enterica Serovar Heidelberg. J Food Prot. 2019;82(11). Poirel L, Walsh TR, Cuvillier V, Nordmann P. Multiplex PCR for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis. 2011;70(1). Additional Declarations No competing interests reported. Supplementary Files Supplementarymaterial.docx Additional Files Legends The Additional File (.wrd) contains Suppl. Table 1 Results of mixed logistic regressions and linear mixed models explaining variation in pathogen presence and bacterial counts detected in samples from pig transport trucks before and after loading procedures. Suppl. Table 2 Consistency of the enrolled farms Suppl. Table 3 Panel of antimicrobials that were tested for each pathogen Suppl. Table 4 Primer and sequences to detect Brachyspira spp., Lawsonia intracellularis , Porcine Epidemic Diarrhoea Coronavirus and Rotavirus A, B, C, H. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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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-4251132","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":293027505,"identity":"9b898988-6941-42b6-8587-f18507e3f6d4","order_by":0,"name":"Marta Masserdotti","email":"","orcid":"","institution":"Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna \"Bruno Ubertini\"","correspondingAuthor":false,"prefix":"","firstName":"Marta","middleName":"","lastName":"Masserdotti","suffix":""},{"id":293027506,"identity":"39d79d4a-95d2-4f2f-b64b-f8b0d8bee950","order_by":1,"name":"Nicoletta 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08:28:18","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4251132/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4251132/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":55087897,"identity":"683da50b-5029-4210-88d2-98a9f40aea2b","added_by":"auto","created_at":"2024-04-22 11:48:06","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":46146,"visible":true,"origin":"","legend":"\u003cp\u003eTotal plate counts (a) and Enterobacteriaceae counts (b) from swabs sampled on the internal surfaces of pig transport trucks at farm entry (“clean” condition, white boxplot) and after loading (“dirty condition”, grey boxplot). Statistics were performed by linear mixed modelling. The superior detection limit of the TPC method is of 1,5 x 10\u003csup\u003e6 \u003c/sup\u003eCFU/cm\u003csup\u003e2\u003c/sup\u003e.\u003c/p\u003e","description":"","filename":"Figure1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4251132/v1/988828b07f77d4de1367c009.jpg"},{"id":55087430,"identity":"1d33884e-1094-49fd-ab20-0f86209ccf2a","added_by":"auto","created_at":"2024-04-22 11:40:06","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":53399,"visible":true,"origin":"","legend":"\u003cp\u003ePrevalence of the examined pathogens in swabs sampled from “clean” (white bars) and “dirty” pig transport trucks (grey bars). Bars indicate 95% confidence limits. When indicated, statistics were performed by linear mixed modelling. In all the other cases, statistical analysis was not performed due to low prevalence or exclusive occurrence in one condition. \u003cem\u003eBrachyspira\u003c/em\u003e spp., Coronaviruses and Influenza A were never detected.\u003c/p\u003e","description":"","filename":"Figure2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4251132/v1/f5c1f28305d9b58db06af810.jpg"},{"id":71719441,"identity":"2d9372a0-62d1-4928-a8c6-3b7a5e8a2d95","added_by":"auto","created_at":"2024-12-18 04:46:55","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":916543,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4251132/v1/5e4b0b14-345a-4c2c-ae66-50272a4a079a.pdf"},{"id":55087428,"identity":"8af7bbb6-558f-4faf-b4a0-8f7584fca7a7","added_by":"auto","created_at":"2024-04-22 11:40:06","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":19006,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAdditional Files Legends\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe Additional File (.wrd) contains\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSuppl. Table 1\u003c/strong\u003e Results of mixed logistic regressions and linear mixed models explaining variation in pathogen presence and bacterial counts detected in samples from pig transport trucks before and after loading procedures.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSuppl. Table 2\u003c/strong\u003e Consistency of the enrolled farms\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSuppl. Table 3\u003c/strong\u003e Panel of antimicrobials that were tested for each pathogen\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSuppl. Table 4\u003c/strong\u003e Primer and sequences to detect \u003cem\u003eBrachyspira\u003c/em\u003espp., \u003cem\u003eLawsonia intracellularis\u003c/em\u003e, Porcine Epidemic Diarrhoea Coronavirus\u003cstrong\u003e \u003c/strong\u003eand Rotavirus A, B, C, H.\u003c/p\u003e","description":"","filename":"Supplementarymaterial.docx","url":"https://assets-eu.researchsquare.com/files/rs-4251132/v1/fadd3167196e2b4b66e01f08.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"The role of short journey transportation in the spreading of swine pathogens and antimicrobial-resistant bacteria","fulltext":[{"header":"Background","content":"\u003cp\u003eThe approach of veterinary medicine to animal health has transformed considerably over time, especially in the livestock field, turning more and more to preventing diseases than to treating them (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e). The key element to achieve effective prevention is biosecurity, a system of practices and measures aimed at minimising the risk of introduction, establishment and spread of animal infections within and among animal populations. According to the World Organization for Animal Health (WOAH)(\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e), implementing biosecurity plans is thus pivotal for effective disease prevention within individual farms as well as in whole regions. A low risk of pathogen introduction and/or spread within the herd leads in turn to a reduction in the use of drugs, especially antimicrobials (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e). Specific to pig farming, a clear link between the implementation of biosecurity measures and improvements in parameters related to antimicrobial use and production has been demonstrated and quantified (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eA key measure to prevent the spread of diseases within a farm, is to avoid mixing pigs of different ages (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e). This can be achieved either by establishing a rigorous workflow and separation of the groups within the farm or by allocating the groups to different farming sites. Especially in high-income countries characterized by more intensive pig production and bigger herd sizes, the second solution is generally preferred, and pigs typically follow a path that begins on the birth farm and then transitions to a fattening farm, sometimes including a nursery farm dedicated to caring for weaned piglets. Consequently, transporting animals to and from the farm is a common occurrence (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e) and is also one of the most critical hazards for farm biosecurity, as it involves breaching the biosecurity interface and potentially having animals, as well as trucks or even drivers, carry around pathogens of concern for both animal health (e.g. Porcine reproductive and respiratory syndrome virus (PRRSV) (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e), Porcine epidemic diarrhoea virus (PEDV) (\u003cspan additionalcitationids=\"CR11\" citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e), African swine fever virus (ASFV) (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e)) and public health (e.g. \u003cem\u003eSalmonella\u003c/em\u003e spp. (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e), AMR pathogens (\u003cspan additionalcitationids=\"CR16\" citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e)). Efficient truck cleaning and disinfection procedures are thus crucial to prevent the spread of pathogens from one farm to another, to the abattoir, and to operators. Nevertheless, trucks often show residues of contamination when entering the farm (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e). This is due either to improper execution of the cleaning and disinfection procedures, or to protocols that target only some specific pathogens and are thus partially effective. Traditionally, legislative and research efforts in the animal transportation field have been focused mostly on improving welfare conditions rather than sanitary ones, and studies on the role of trucks as a potential threat to on-farm biosecurity are scarce. Furthermore, the lack of studies and data is even more limited when specifically considering the spread of AMR pathogens, as recently reported by the European Food Safety Authority (EFSA) (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe World Health Organization recently published a list of bacteria for which new antimicrobials are urgently needed, particularly highlighting the threat posed by bacteria relevant at the nosocomial level that have become resistant to carbapenems and third-generation cephalosporins (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e). Among these bacteria are \u003cem\u003eEnterobacteriaceae\u003c/em\u003e, which are showing rising levels of AMR (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e). Extended-spectrum β-lactamase (ESBL)-producing \u003cem\u003eEnterobacteriaceae\u003c/em\u003e or carbapenemase-producing \u003cem\u003eEnterobacteriaceae\u003c/em\u003e are a significant threat to public health due to their association with increased health costs, hospitalisation, and mortality rate (\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e). Specifically, \u003cem\u003eE. coli\u003c/em\u003e is frequently included in AMR monitoring programs worldwide (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e) as it is a cross-species pathogen with a great capacity to accumulate resistance genes, mostly through horizontal gene transfer (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e) (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e). There is, therefore, an urgent need to investigate and quantify the impact of animal transportation (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e) in the transmission and diffusion of AMR, assessing the role of different risk factors such as the hygiene of the transport vehicle. Based on the above considerations, \u003cem\u003eEnterobacteriaceae\u003c/em\u003e are considered a sensitive indicator of ineffective cleaning (\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e) and can also be used to test for the presence of AMR.\u003c/p\u003e \u003cp\u003eIn the present work, we investigated the contamination of pig transport vehicles before and after animal loading procedures, to assess the effectiveness of cleaning and disinfection measures to which transport vehicles are commonly subjected. We focused on the main swine pathogens transmissible through faeces due to their permanence in the organic matrix and consequent feasibility of sampling: Enterobacteriaceae (\u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eSalmonella\u003c/em\u003e spp.), \u003cem\u003eBrachyspira\u003c/em\u003e spp., \u003cem\u003eLawsonia intracellularis\u003c/em\u003e, PRRSV, PEDV, and Rotavirus A, B, C, H. Additionally, we investigated ESBL/AmpC- and carbapenemase-producing \u003cem\u003eE. coli\u003c/em\u003e pathogens in order to evaluate the general risk of AMR spread associated with animal transportation.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eA total of 84 swabs collected from 42 trucks (42 \u0026ldquo;dirty\u0026rdquo; and 42 \u0026ldquo;clean\u0026rdquo; swabs) were tested for specific pathogens and bacterial counts. The prevalences of the pathogens considered and their distribution between swabs from \u0026ldquo;clean\u0026rdquo; and \u0026ldquo;dirty\u0026rdquo; trucks are reported in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eOverall prevalence of the investigated pathogens and percentage of positive swabs sampled in \u0026ldquo;clean\u0026rdquo; and \u0026ldquo;dirty\u0026rdquo; pig transport trucks.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMicroorganism\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePrevalence (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e% on \u0026ldquo;clean\u0026rdquo; trucks\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e% on \u0026ldquo;dirty\u0026rdquo; trucks\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eE. coli\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e40.47%\u003c/p\u003e \u003cp\u003e(34 / 84)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e14.71%\u003c/p\u003e \u003cp\u003e(5 / 34)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e85.29%\u003c/p\u003e \u003cp\u003e(29 / 34)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eESBL/AmpC producing \u003cem\u003eE. coli\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10.71%\u003c/p\u003e \u003cp\u003e(9 / 84)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e11.11%\u003c/p\u003e \u003cp\u003e(1 / 9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e88.89%\u003c/p\u003e \u003cp\u003e(8 / 9)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCarbapenemase producing \u003cem\u003eE. coli\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11.90%\u003c/p\u003e \u003cp\u003e(10 / 84)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(10 / 10)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eSalmonella spp.\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.85%\u003c/p\u003e \u003cp\u003e(3 / 84)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(3 / 3)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eBrachyspira spp.\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eLawsonia intracellularis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.13%\u003c/p\u003e \u003cp\u003e(4 / 78)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e25%\u003c/p\u003e \u003cp\u003e(1 / 4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e75%\u003c/p\u003e \u003cp\u003e(3 / 4)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePRRSV\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10.26%\u003c/p\u003e \u003cp\u003e(8 / 78)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(8 / 8)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePEDV\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRotavirus A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e16.67%\u003c/p\u003e \u003cp\u003e(13 / 78)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15.38%\u003c/p\u003e \u003cp\u003e(2 / 13)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e84.62%\u003c/p\u003e \u003cp\u003e(11 / 13)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRotavirus B\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e33.33%\u003c/p\u003e \u003cp\u003e(26 / 78)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e34.62%\u003c/p\u003e \u003cp\u003e(9 / 78)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e65.38%\u003c/p\u003e \u003cp\u003e(17 / 78)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRotavirus C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e56.41%\u003c/p\u003e \u003cp\u003e(44 / 78)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e52.27%\u003c/p\u003e \u003cp\u003e(23 / 78)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e47.73%\u003c/p\u003e \u003cp\u003e(21 / 78)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRotavirus H\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.13%\u003c/p\u003e \u003cp\u003e(4 / 78)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e75%\u003c/p\u003e \u003cp\u003e(3 / 4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e25%\u003c/p\u003e \u003cp\u003e(1 / 4)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eFor most of the examined pathogens, at least one positive swab was found in both \u0026ldquo;clean\u0026rdquo; and \u0026ldquo;dirty\u0026rdquo; trucks, with the exception of PRRSV, carbapenemase-producing \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eSalmonella\u003c/em\u003e spp., which were detected only in \u0026ldquo;dirty\u0026rdquo; samples. Regarding the latter, the three isolates were serotyped, resulting in one \u003cem\u003eSalmonella Typhimurium\u003c/em\u003e monophasic variant and two \u003cem\u003eSalmonella Choleraesuis\u003c/em\u003e. No samples positive to \u003cem\u003eBrachyspira\u003c/em\u003e spp. or PEDV were detected.\u003c/p\u003e \u003cp\u003eRegarding those pathogens detected in both conditions and with an overall prevalence higher than 10%, the probability of testing positive was significantly higher in \u0026ldquo;dirty\u0026rdquo; samples than in \u0026ldquo;clean\u0026rdquo; ones for \u003cem\u003eE. coli (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001)\u003c/em\u003e, ESBL/AmpC-producing \u003cem\u003eE. coli\u003c/em\u003e (p\u0026thinsp;=\u0026thinsp;0.026) and Rotavirus A (p\u0026thinsp;=\u0026thinsp;0.011). Despite a non-significant difference (p\u0026thinsp;=\u0026thinsp;0.0504), positivity to Rotavirus B showed the same tendency. The truck condition had conversely no effect on Rotavirus C presence (p\u0026thinsp;=\u0026thinsp;0.10), which varied instead depending on the production stage (p\u0026thinsp;=\u0026thinsp;0.0002), with swabs from weaning and growing farms being more likely to be positive than the ones from fattening farms. Similarly, ESBL/AmpC-producing \u003cem\u003eE. coli\u003c/em\u003e were more likely to be found in swabs collected from weaning than fattening farms (p\u0026thinsp;=\u0026thinsp;0.046).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eBacterial counts varied significantly depending on the truck condition as well: both TPC (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) and \u003cem\u003eEnterobacteriaceae\u003c/em\u003e counts (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) were significantly higher in \u0026ldquo;dirty\u0026rdquo; samples than in \u0026ldquo;clean\u0026rdquo; ones (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Complete results of all models, including parameter estimates, are reported in Supplementary Table\u0026nbsp;1.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eRegarding AMR bacteria, the proportion of isolates resistant to the different molecules examined, and their distribution between \u0026ldquo;clean\u0026rdquo; and \u0026ldquo;dirty\u0026rdquo; condition are shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e for E. \u003cem\u003ecoli\u003c/em\u003e isolates and in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e for ESBL/AmpC-producing \u003cem\u003eE. coli\u003c/em\u003e and carbapenemase-producing \u003cem\u003eE. coli.\u003c/em\u003e\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePercentage of resistance of \u003cem\u003eE. coli\u003c/em\u003e isolates (n\u0026thinsp;=\u0026thinsp;34) for each molecule analyzed, distribution of these percentages between swabs taken on the \u0026ldquo;clean\u0026rdquo; and the \u0026ldquo;dirty\u0026rdquo; trucks. The epidemiological cut-off is available online (ECOFFs, EUCAST, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e\u003ca href=\"http://www.eucast.org\" target=\"_blank\"\u003ewww.eucast.org\u003c/a\u003e\u003c/span\u003e\u003cspan address=\"http://www.eucast.org\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e, last accessed on 28th February 2024).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMolecule\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(Tentative)\u003c/p\u003e \u003cp\u003eEpidemiological cut-off values\u003c/p\u003e \u003cp\u003e(T)ECOFF\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e% Resistance\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e% on \u0026ldquo;dirty\u0026rdquo; trucks\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e% on \u0026ldquo;clean\u0026rdquo; trucks\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAmpicillin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e64.71%\u003c/p\u003e \u003cp\u003e(22 / 34)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e90.91%\u003c/p\u003e \u003cp\u003e(20 / 22)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.09%\u003c/p\u003e \u003cp\u003e(2 / 22)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTetracycline\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e61.76%\u003c/p\u003e \u003cp\u003e(21 / 34)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e85.71%\u003c/p\u003e \u003cp\u003e(18 / 21)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14,29%\u003c/p\u003e \u003cp\u003e(3 / 21)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFlorfenicol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e55.88%\u003c/p\u003e \u003cp\u003e(19 / 34)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e89.47%\u003c/p\u003e \u003cp\u003e(17 / 19)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e10.53%\u003c/p\u003e \u003cp\u003e(2 / 19)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTrimetoprim\u0026thinsp;+\u0026thinsp;Sulfametoxazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e50%\u003c/p\u003e \u003cp\u003e(17 / 34)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e88.23%\u003c/p\u003e \u003cp\u003e(15 / 17)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e11.76%\u003c/p\u003e \u003cp\u003e(2 / 17)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEnrofloxacin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e32.35%\u003c/p\u003e \u003cp\u003e(11 / 34)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e90.91%\u003c/p\u003e \u003cp\u003e(10 / 11)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.09%\u003c/p\u003e \u003cp\u003e(1 / 11)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGentamicin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e26.47%\u003c/p\u003e \u003cp\u003e(9 / 34)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(9 / 9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAmoxicillin\u0026thinsp;+\u0026thinsp;Clavulanic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e23.53%\u003c/p\u003e \u003cp\u003e(8 / 34)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(8 / 8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFlumequina\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e23.53%\u003c/p\u003e \u003cp\u003e(8 / 34)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(8 / 8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCefazolin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e20.59%\u003c/p\u003e \u003cp\u003e(7 / 34)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(7 / 7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eKanamycin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17.64%\u003c/p\u003e \u003cp\u003e(6 / 34)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(6 / 6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eColistin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.94%\u003c/p\u003e \u003cp\u003e(1 / 34)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(1 / 1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e67.65% (23/34) of the tested \u003cem\u003eE. coli\u003c/em\u003e samples were resistant to sulfisoxazole. The majority of these (20/23) were collected on \u0026ldquo;dirty\u0026rdquo; trucks. 14.71% (5/34) of the \u003cem\u003eE. coli\u003c/em\u003e samples were resistant to amminosidine, all collected on \u0026ldquo;dirty\u0026rdquo; trucks.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePercentage of resistance of ESBL/AmpC-producing \u003cem\u003eE. coli\u003c/em\u003e isolates (n\u0026thinsp;=\u0026thinsp;9) and carbapenemase-producing \u003cem\u003eE. coli\u003c/em\u003e isolates (n\u0026thinsp;=\u0026thinsp;10) for each molecule analyzed, distribution of these percentages between swabs taken on the \u0026ldquo;clean\u0026rdquo; and the \u0026ldquo;dirty\u0026rdquo; trucks. The epidemiological cut-off is available online (ECOFFs, EUCAST, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e\u003ca href=\"http://www.eucast.org\" target=\"_blank\"\u003ewww.eucast.org\u003c/a\u003e\u003c/span\u003e\u003cspan address=\"http://www.eucast.org\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e, last accessed on 28th February 2024).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eMolecule\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e(Tentative)\u003c/p\u003e \u003cp\u003eEpidemiological cut-off values\u003c/p\u003e \u003cp\u003e(T)ECOFF\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c5\" namest=\"c3\"\u003e \u003cp\u003eESBL/AmpC \u0026ndash; producing \u003cem\u003eE. coli\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c8\" namest=\"c6\"\u003e \u003cp\u003eCarbapenemase \u0026ndash; producing\u003c/p\u003e \u003cp\u003e\u003cem\u003eE. coli\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e% Resistance\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e% on \u0026ldquo;dirty\u0026rdquo; trucks\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e% on \u0026ldquo;clean\u0026rdquo; trucks\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e% Resistance\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e% on \u0026ldquo;dirty\u0026rdquo; trucks\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e% on \u0026ldquo;clean\u0026rdquo; trucks\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCefotaxime\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(9 / 9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e88.89%\u003c/p\u003e \u003cp\u003e(8 / 9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e11,11%\u003c/p\u003e \u003cp\u003e(1 / 9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e80%\u003c/p\u003e \u003cp\u003e(8 / 10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(8 / 8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 8)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCefepime\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e88.89%\u003c/p\u003e \u003cp\u003e(8 / 9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e87.50%\u003c/p\u003e \u003cp\u003e(7 / 8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e12.5%\u003c/p\u003e \u003cp\u003e(1 / 8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e50%\u003c/p\u003e \u003cp\u003e(5 / 10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(5 / 5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 5)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCeftazidime\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e55.56%\u003c/p\u003e \u003cp\u003e(5 / 9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(5 / 5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e10%\u003c/p\u003e \u003cp\u003e(1 / 10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(1 / 1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 1)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCefotaxime\u0026thinsp;+\u0026thinsp;Clavulanic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e33.33%\u003c/p\u003e \u003cp\u003e(3 / 9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(3 / 3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e60%\u003c/p\u003e \u003cp\u003e(6 / 10)*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(6 / 6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 6)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCefoxitin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e33.33%\u003c/p\u003e \u003cp\u003e(3 / 9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(3 / 3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e10%\u003c/p\u003e \u003cp\u003e(1 / 10)**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(1 / 1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 1)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eErtapenem\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(0,03)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e33.33%\u003c/p\u003e \u003cp\u003e(3 / 9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(3 / 3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(10 / 10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(10 / 10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 10)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCeftazidime\u0026thinsp;+\u0026thinsp;Clavulanic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e33.33%\u003c/p\u003e \u003cp\u003e(3 / 9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(3 / 3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e10%\u003c/p\u003e \u003cp\u003e(1 / 10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(1 / 1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 1)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTemocillin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e11.11%\u003c/p\u003e \u003cp\u003e(1 / 9)*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(1 / 1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(10 / 10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(10 / 10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 10)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eImipenem\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e20%\u003c/p\u003e \u003cp\u003e(2 / 10)**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(2 / 2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 2)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMeropenem\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e80%\u003c/p\u003e \u003cp\u003e(8 / 10)*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(8 / 8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 8)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe 7 ESBL/AmpC-producing \u003cem\u003eE. coli\u003c/em\u003e isolates that were subjected to molecular investigation by PCR were found to be positive for \u003cem\u003ebla\u003c/em\u003e\u003csub\u003eCTX-M,\u003c/sub\u003e \u003cem\u003ebla\u003c/em\u003e\u003csub\u003eTEM\u003c/sub\u003e and \u003cem\u003ebla\u003c/em\u003e\u003csub\u003eCMY\u003c/sub\u003e (alone or in combination), and none was positive for \u003cem\u003ebla\u003c/em\u003e\u003csub\u003eSHV\u003c/sub\u003e. One isolate was negative to all the sought genes (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). All of the carbapenemase-producing \u003cem\u003eE. coli\u003c/em\u003e isolates that were subjected to molecular investigation by PCR were found to be positive only for the \u003cem\u003ebla\u003c/em\u003e\u003csub\u003eOXA-48\u003c/sub\u003e gene (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDistribution of prevalence of resistance genes found, either singly or in combination, in ESBL/AmpC- and carbapenemase-producing \u003cem\u003eE. coli\u003c/em\u003e isolates.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ebla\u003c/em\u003e\u003csub\u003eCTX\u0026minus;M\u003c/sub\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e\u003cem\u003ebla\u003c/em\u003e\u003csub\u003eCTX\u0026minus;M+TEM\u003c/sub\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003ebla\u003c/em\u003e\u003csub\u003eCMY+TEM\u003c/sub\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003ebla\u003c/em\u003e\u003csub\u003eSHV\u003c/sub\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePrevalence of resistance genes found, either singly or in combination, in ESBL/AmpC-producing \u003cem\u003eE. coli\u003c/em\u003e isolates\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e42.86%\u003c/p\u003e \u003cp\u003e(3 / 7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e28.67%\u003c/p\u003e \u003cp\u003e(2 / 7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14.29%\u003c/p\u003e \u003cp\u003e(1 / 7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003cp\u003e(0 / 7)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e\u003cb\u003ebla\u003c/b\u003e\u003csub\u003e\u003cb\u003eOXA\u0026minus;48\u003c/b\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003ebla\u003c/b\u003e\u003csub\u003e\u003cb\u003eNDM\u003c/b\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003ebla\u003c/b\u003e\u003csub\u003e\u003cb\u003eKPC\u003c/b\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003ebla\u003c/b\u003e\u003csub\u003e\u003cb\u003eVIM\u003c/b\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePrevalence of resistance genes found in carbapenemase-producing \u003cem\u003eE. coli\u003c/em\u003e isolates\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003cp\u003e(9 / 9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eWe investigated the potential role played by vehicles for transporting live pigs in the spread of a comprehensive panel of pathogens transmissible through faeces, including antibiotic-resistant pathogens, an aspect that has gaps in knowledge and requires dedicated investigations, as recently reported by EFSA (\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eOverall, we found significantly lower bacterial counts and pathogen occurrence in sanitized trucks entering the farms, compared to loaded trucks. In particular, several pathogens of zoo-economic and zoonotic importance, namely carbapenemase-producing \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eSalmonella\u003c/em\u003e spp. and PRRSV were never detected on \u0026ldquo;clean\u0026rdquo; trucks entering the farm. These are encouraging results regarding the effectiveness of cleaning and disinfection procedures. However, residual contamination by \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eL. intarcellularis\u003c/em\u003e, and rotaviruses was found in sanitised trucks at farm entry. In particular, Rotaviruses B, C and H were frequently detected on \u0026ldquo;clean\u0026rdquo; trucks, with Rotavirus C occurrence being equally likely on \u0026ldquo;clean\u0026rdquo; and \u0026ldquo;dirty\u0026rdquo; trucks, suggesting that current sanitization protocols might be ineffective against them. In general, Rotaviruses are known for their high environmental resistance: drying does not inactivate all viromes (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e), and while for Rotavirus A the effectiveness of phenolic disinfectants regardless of the concentration of organic substance has been demonstrated, glutaraldehyde-based disinfectants or peroxide compounds have proved to be effective only if preceded by careful cleaning and removal of the organic matrix (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e). Additionally, there is a lack of information regarding the effectiveness of disinfection procedures against Rotavirus C and H, whose potential pathogenicity has only recently been highlighted (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e). Therefore, it is necessary to deepen the knowledge of the epidemiology and the environmental resistance of these pathogens, to develop a protocol that can be effective also towards them.\u003c/p\u003e \u003cp\u003eStudies on the role of trucks as a potential threat to on-farm biosecurity are usually mainly related to specific outbreak episodes (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e), and are thus mostly focused on a single pathogen, not assessing the sanitary risk from a broader epidemiological perspective. For instance, two studies were conducted in our same intensive pig production area in Northern Italy to investigate the role of vehicles as a source of PEDV transmission following the occurrence of an epidemic wave (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e), and to assess the propagation of \u003cem\u003eBrachyspira hyodysenteriae\u003c/em\u003e by vehicles for transport to slaughterhouse (\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e). Both studies reported relatively high levels of truck contamination, but interestingly, we didn\u0026rsquo;t find any of the two pathogens in our investigation.\u003c/p\u003e \u003cp\u003eOur results on the prevalence of AMR bacteria are consistent with findings from previous studies carried at environmental level (\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e) or by faecal samplings (\u003cspan additionalcitationids=\"CR36\" citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e). Our results highlight a relatively common occurrence of resistance against common antimicrobials (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e), such as tetracycline, chloramphenicol, and trimethoprim/sulfamethoxazole, while resistance to last-resort antimicrobials, such as colistin, broad-spectrum cephalosporins and carbapenems is less frequent. Notably, AbuOun et al. (2020) did not detect any carbapenem-resistant strains, which emerged instead from the present study and from the one conducted by Peng et al. (2022).\u003c/p\u003e \u003cp\u003eWith regard to ESBL/AmpC-producing strains, our findings are consistent with recent studies that highlight bla\u003csub\u003eCTX\u0026minus;M\u003c/sub\u003e as the most common gene conferring resistance to beta-lactams, detected ubiquitously in community and healthcare contexts, in the environment, in food, and in animal species (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e). For carbapenemase-producing \u003cem\u003eE. coli\u003c/em\u003e strains, only bla\u003csub\u003eOXA\u0026minus;48\u003c/sub\u003e-encoding strains were found in this study. This gene was reported for the first time in \u003cem\u003eE. coli\u003c/em\u003e isolated from pigs in 2019 in Germany (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e), and it is speculated that transmission occurred from humans, in which this resistance gene was already circulating. Data on the dissemination of carbapenem-resistant \u003cem\u003eEnterobacteriaceae\u003c/em\u003e (CRE) in animal species and their potential for transmission to humans are still scarce, and in Europe its prevalence in livestock seems to be low, unlike in Asia and North Africa (\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e). However, considering the importance of carbapenems as last-resort antimicrobials for humans, an integrated surveillance approach of CRE emergence is paramount. When considering the prevalence of AMR pathogens across pig production phases, the trend that is observed in the present study is consistent with other studies, where they were more common in early production stages (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e). Overall, the data about AMR pathogens obtained from trucks in this study are comparable with those available in literature, collected at the farm level or by animal sampling. This suggests that animal transport vehicles may represent an alternative epidemiological indicator of AMR pathogens, other than a means to assess the risk related to livestock movements in the spread of AMR microorganisms (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e). In general, the high pathogen contamination levels detected on outgoing trucks suggest that loaded trucks may provide a potential indicator of the microorganisms circulating in the farm, and above all highlight that ineffective cleaning and disinfection procedures may represent a breach in the biosecurity of the farms visited afterwards.\u003c/p\u003e \u003cp\u003eHowever, it must be noted that our study presents some limitations: the limited number of farms which were enrolled based on opportunistic criteria related to logistic constraints, and farmers\u0026rsquo; agreement to participate. Such convenience sampling implies that our conclusions might not be extended to the whole population. However, we believe that this might make our results even more conservative, as the farmers who volunteered for the study are probably among those with the best biosecurity procedures.\u003c/p\u003e \u003cp\u003eOn the whole, the lack of provisions, guidelines or regulations regulating precisely the hygiene of means of transport should be highlighted: for instance in the European Union, the Article 125 of Regulation (EU) 2016/429 on the prevention of transport-related diseases merely recommends that all equipment and means of transport must be subjected to thorough cleaning, disinfection, and pest control measures to eliminate any potential biohazard, and that other biosecurity measures must be taken depending on the risks associated with the transport operations concerned. This regulation has been supplemented, but does not contain any specific provisions yet.\u003c/p\u003e \u003cp\u003eDespite the present study was not focused on the loading procedures, observing the loading of pigs has uncovered critical biosecurity weaknesses that require training and awareness of transporters and breeders, as recently prescribed by Articles 11 and 13 of the Animal Health Law (Reg. CE 2016/429). To achieve the highest biosecurity standards, trucks should be allowed to stand for a period after passing through disinfection arches, transportation staff should not be involved in animal handling or entering animal housings, disposable personal protective equipment should always be readily available, and transporters should not be allowed to enter the truck\u0026rsquo;s cockpit after donning personal protective equipment.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eTo the best of our knowledge, this study is among the firsts to investigate trucks as mechanical vectors of pathogens relevant to pig breeding and public health, including AMR bacteria. Microbiological investigations carried out on disinfected trucks dedicated to animal transport at farm entry highlight the risk of introducing pathogens, showing that the cleaning and disinfection protocols currently in use are only partially effective, particularly against Rotavirus C. On the other hand, the generally lower bacterial counts and pathogen prevalence on sanitized trucks, and in particular the rare occurrence of AMR bacteria, are an encouraging result. Further studies will have to evaluate the effectiveness of these protocols on farms and especially in farrowing and weaning units. Overall, these findings suggest that there is a fair focus on cleaning and disinfection procedures in animal transport, but at the same time highlight the need for further studies assessing the efficacy of protocols against specific pathogens, and for the definition of guidelines to allow greater uniformity and greater awareness of transporters and operators about such procedures, so that the protocols are carried out with the appropriate attention and therefore greater effectiveness.\u003c/p\u003e"},{"header":"Methods","content":"\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eSampling\u003c/h2\u003e \u003cp\u003eTo investigate the contamination of animal transport vehicles and assess the efficacy of standard cleaning and disinfection procedures, from March to July 2023 we surveyed trucks used for animal transport in 11 pig farms (3 weaning farms, 4 growing farms, 3 fattening farms, and 1 farm that loaded animals both at growing and at fattening stage) located in Lombardy, Northern Italy (farms\u0026rsquo; size are listed in Suppl. Table\u0026nbsp;2). The trucks were employed for short journeys (less than 8 hours) to transport pigs from farm to farm or from farm to slaughterhouse. The farms were enrolled in the study based on opportunistic criteria related to logistic constraints, feasibility and voluntary acceptance to participate by farmers.\u003c/p\u003e \u003cp\u003eFor each truck, two samples were collected in duplicate, for a total of four samples: two before the start of loading procedures, at the time of arrival of the vehicle on the farm and after it had been subjected to the disinfection measures adopted by the company (hereafter, \"clean\u0026rdquo; samples); and two at the end of loading procedures before the truck left the farm (hereafter, \"dirty\u0026rdquo; samples). Sterile gauze swabs were rubbed on the floor and on the lower portion of the interior walls of the truck and/or trailer to cover a total area of approximately 100 cm\u003csup\u003e2\u003c/sup\u003e, avoiding those areas that were still wet from the residual disinfectant from the biosecurity procedures at the entrance of the farm. The disinfectant is usually applied on the external surfaces of the clean truck entering the farm either manually, by an operator, or automatically, by a disinfection arch located at the entrance.\u003c/p\u003e \u003cp\u003eThe swabs were then placed in pairs in sterile sampling bags, stored at room temperature, within \u0026le;\u0026thinsp;6 hours delivered to the laboratory where swabs were stored at +\u0026thinsp;4\u0026deg;C until further analyses. A total number of 84 samples (buffer pairs) were collected (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). Of the 84 swabs taken, 78 were subjected to both microbiological and molecular investigations, while the remaining 6 could only be subjected to bacteriological tests due to a sampling error.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eNumber of collected samples by rearing stage. A farm (*) was sampled both when loading growing pigs and when loading fattening pigs.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRearing stage\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNumber of farms\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNumber of samples\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNumber of trucks\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWeaning piglets transported to a nursery or to a wean-to-finish barn\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGrowing pigs transported from a nursery to a finishing barn\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFinishing pigs transported to slaughterhouse\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTotal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e42\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eMicrobiological analyses\u003c/h3\u003e\n\u003cp\u003eThe Total Plate Count (TPC) was performed manually after incubation at 37\u0026deg;C for 48h on PCA medium of the \u0026minus;\u0026thinsp;3 dilution, according to internal testing method (\u003cspan additionalcitationids=\"CR44\" citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e). Such testing method has a superior detection limit of 1,5 x 10\u003csup\u003e6\u003c/sup\u003e CFU/cm\u003csup\u003e2\u003c/sup\u003e. The quantification of \u003cem\u003eEnterobacteriaceae\u003c/em\u003e contamination was performed according to ISO 21528-2 procedures, and the biochemical confirmation was obtained through the oxidase test. The detection and identification of Salmonella spp. strains was performed according to ISO 6579-1:2017/Amd1:2020 and ISO/TR 6579-3:2014 procedures.\u003c/p\u003e \u003cp\u003eConcerning \u003cem\u003eE. coli\u003c/em\u003e, bacteriological examinations were performed by using standard bacteriological cultures (\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e), while for ESBL/AmpC- and carbapenemase-producing \u003cem\u003eE. coli\u003c/em\u003e the following method was applied: after a pre-enriching phase with BPW (1:10 ratio) at 37\u0026deg;C for 18\u0026ndash;24 hours, a first selective incubation at 37 \u0026ordm;C for 18 hours was performed for detecting ESBL/AmpC- and carbapenemase-producing \u003cem\u003eE. coli\u003c/em\u003e using a solid medium (respectively, McConkey\u0026thinsp;+\u0026thinsp;cefotaxime 1 mg/L at 41\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C and CHROMID\u0026reg; CARBA SMART at 37\u0026deg;C, both for 18\u0026ndash;24 hours). At the end of this first incubation, the suspected colonies were transplanted on solid McConkey soil and incubated at 37\u0026deg;C for 18\u0026ndash;24 hours. A single bacterial colony from each phenotype-positive sample was resuspended in 250 \u0026micro;L of DNase-Rnase free water and DNA was extracted by lysis-boiling (98\u0026deg;C for 10 min) for further molecular characterization. After molecular confirmation, the plates were subjected to antimicrobial susceptibility testing (the panels of tested antimicrobials are listed in Suppl. Table\u0026nbsp;3). \u003cem\u003eE. coli\u003c/em\u003e isolates were also tested for aminosidine and sulfisoxazole, and the classification of the resistance profile was based on the clinical cut-off of the internal test method (\u003cspan additionalcitationids=\"CR48 CR49 CR50 CR51\" citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e). Minimum inhibitory concentrations (MICs) were determined by broth microdilution, and the strains were classified as \u0026ldquo;resistant\u0026rdquo; or \u0026ldquo;susceptible\u0026rdquo; according to the Epidemiological cut-off values (ECOFFs) recommended by the European Committee on Antimicrobial Susceptibility Testing (EUCAST, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e\u003ca href=\"http://www.eucast.org\" target=\"_blank\"\u003ewww.eucast.org\u003c/a\u003e\u003c/span\u003e\u003cspan address=\"http://www.eucast.org\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). When unavailable, clinical breakpoints were used (\u003cspan additionalcitationids=\"CR48 CR49 CR50 CR51\" citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e).\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eMolecular analyses\u003c/h2\u003e \u003cp\u003eThe screening for \u003cem\u003eBrachyspira\u003c/em\u003e spp., \u003cem\u003eLawsonia intracellularis\u003c/em\u003e, PRRSV, PEDV, and Rotavirus A, B, C, H was performed by molecular analysis. In particular, PRRSV screening was performed using a commercial kit (virotype PRRSV RT-PCR Kit, Indical Bioscience) that allows the detection of PRRS virus and its discrimination in European, American and highly pathogenic American strain. All the other pathogens were searched for by means of in-house methods. In detail, \u003cem\u003eBrachyspira hyodysenteriae, B. pilosicoli\u003c/em\u003e, and \u003cem\u003eLawsonia intracellularis\u003c/em\u003e were searched by real-Time PCR using the kit QuantiNova Probe PCR mastermix (Qiagen) and targeting the nox gene region in \u003cem\u003eBrachyspira\u003c/em\u003e spp. and the aspA gene in \u003cem\u003eL. intracellularis\u003c/em\u003e (\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e, \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e). The reaction for the identification of PEDV allows for the detection of the presence of a nucleic acid region of 111 bp of the S1 gene (Spike, region 1) (\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe performed Rotavirus Real-time RT-PCR method allows the detection of an RNA portion of the VP6 region specific to the A, B, C, H groups of Rotavirus in pigs. The method is based on the use of two multiplex reactions one-step Real-Time RT-PCR: in the first multiplex reaction specific regions of groups A (98 bp) and C (86 bp) are amplified, and in the second multiplex reaction the specific regions of groups B (89 bp) and H (93 bp) are amplified (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe primers sequences used to detect the presence of \u003cem\u003eBrachyspira hyodisenteriae\u003c/em\u003e and \u003cem\u003eB. pilosicoli\u003c/em\u003e, \u003cem\u003eLawsonia intracellularis\u003c/em\u003e, PRRSV, PEDV, and Rotavirus A, B, C, H are listed in Suppl. Table\u0026nbsp;4.\u003c/p\u003e \u003cp\u003eFinally, a multiplex-PCR was used to search for the genes coding for ESBL/AmpC (\u003cem\u003ebla\u003c/em\u003e\u003csub\u003eCTX\u0026minus;M\u003c/sub\u003e 593 bp, \u003cem\u003ebla\u003c/em\u003e\u003csub\u003eSHV\u003c/sub\u003e 237 bp, \u003cem\u003ebla\u003c/em\u003e\u003csub\u003eTEM\u003c/sub\u003e 445 bp, \u003cem\u003ebla\u003c/em\u003e\u003csub\u003eCMY\u003c/sub\u003e 820 bp) (\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e, \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e) and carbapenemases (\u003cem\u003ebla\u003c/em\u003e\u003csub\u003eVIM\u003c/sub\u003e 390 bp, \u003cem\u003ebla\u003c/em\u003e\u003csub\u003eNDM\u003c/sub\u003e 621 bp, \u003cem\u003ebla\u003c/em\u003e\u003csub\u003eKPC\u003c/sub\u003e 798 bp, blaOXA-48 438 bp) (\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e). We investigated 7 out of the 9 ESBL/AmpC-producing \u003cem\u003eE. coli\u003c/em\u003e isolates and 9 out of the 10 carbapenemase-producing \u003cem\u003eE. coli\u003c/em\u003e isolates that were detected on plate. The impossibility of carrying out the molecular investigation in the remaining isolates is linked to the absence of regrowth of the strain during the processing phases or to pollution of the strain.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eTo assess the effectiveness of cleaning and disinfection protocols, the effect of the independent variable \u0026ldquo;condition\u0026rdquo; (i.e., \"clean\u0026rdquo; or \u0026rdquo;dirty\") on: i) the presence/absence of pathogens with overall prevalence greater than 10% (i.e., \u003cem\u003eE. coli\u003c/em\u003e, ESBL/AmpC-producing \u003cem\u003eE. coli\u003c/em\u003e, and Rotaviruses A, B, C), and on ii) TPC and \u003cem\u003eEnterobacteriaceae\u003c/em\u003e counts (ln-transformed values [ln(x\u0026thinsp;+\u0026thinsp;1]) were analyzed through mixed logistic regressions and linear mixed models, respectively. In all models, the productive category (i.e., weaning, growing or fattening) was included as a covariate and farm IDs were included as random intercepts to account for different samplings within the same farm. When significant, factors with more than two levels (namely, productive category) were tested \u003cem\u003epost hoc\u003c/em\u003e through t-tests on differences of least-square means, applying Holm correction for multiple comparisons. Results were considered statistically significant when p\u0026thinsp;\u0026lt;\u0026thinsp;0.05. All the analyses were carried out using the packages \u003cem\u003elme4\u003c/em\u003e and \u003cem\u003eemmeans\u003c/em\u003e in R (R Core Team 2021) version 4.3.1 (2023-06-16).\u003c/p\u003e \u003c/div\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eAMR\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eAntimicrobial-Resistant/Antimicrobial Resistance\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eASF(V)\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eAfrican Swine Fever (Virus)\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eCRE\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eCarbapenem-Resistant \u003cem\u003eEnterobacteriaceae\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eEC\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003e \u003cem\u003eEnterobacteriaceae\u003c/em\u003e count\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003e(T)ECOFF\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003e(Tentative) Epidemiological cut-off values\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eESBL\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eExtended-Spectrum β-lactamase\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eEUCAST\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eEuropean Committee on Antimicrobial Susceptibility Testing\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eEFSA\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eEuropean Food Safety Authority\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003ePEDV\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ePorcine Epidemic Diarrhoea Virus\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003ePRRS(V)\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ePorcine Respiratory and Reproductive Syndrome Virus\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eTPC\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eTotal Plate Count\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eWOAH\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eWorld Organisation for Animal Health\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data are available upon reasonable request from the authors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by Italian Ministry of Health (Agreement ClassyFarm 2021-2023).\u0026nbsp;The Funder had no role in the design, analysis and reporting of the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eConceived and designed the experiments: N.F., C.R., G.L.A., C.L.. Performed sampling: M.M., A.D., E.G.. Performed laboratory experiments: F.G., C.B., M.B.B.. Analyzed the data: M.M., N.F., C.R., F.S., C.B., M.B.B.. Contributed reagents/materials/analysis tools: M.B.B., G.L.A.. Writing\u0026mdash;original draft preparation: M.M., N.F., C.R., G.L.A., C.L.. Writing\u0026mdash;review and editing: M.M., N.F., A.D., F.G., F.S., C.R., E.G., C.B., M.B.B., G.L.A., C.L.. Supervision: M.B.B., G.L.A., C.L.. Funding acquisition: G.L.A..\u0026nbsp;All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank Laura Birbes, Paola Giangrossi, Fabiana Gatti, Gloria Garbin, Claudia Alberti and Chiara Boifava for their help with the project activities.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eEuropean Commission. Animal Health Strategy for the European Union. 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Diagn Microbiol Infect Dis. 2011;70(1).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"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":"biosecurity, AMR, clean and dirty trucks, swabs, carbapenemase-producing E. coli, Rotavirus C, ESBL/AmpC","lastPublishedDoi":"10.21203/rs.3.rs-4251132/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4251132/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eThe transport of live pigs poses a risk to on-farm biosecurity. Trucks can carry pathogens with significant economic and health impacts, including antimicrobial-resistant (AMR) bacteria. This study aimed to investigate the microbiological contamination of trucks before and after loading, focusing on AMR bacteria and other major pathogens transmissible through faeces. Samples were collected by swabbing the internal surface of disinfected empty trucks at farm entry (‘clean’) and after loading (‘dirty’), and were tested for total plate count (TPC), specific bacteria and viruses. \u003cem\u003eEscherichia coli\u003c/em\u003e isolates were also phenotypically and molecularly tested for the presence of extended-spectrum β-lactamase (ESBL), other β-lactamases (AmpC) and carbapenemase.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eBacterial counts (both TPC and \u003cem\u003eEnterobacteriaceae\u003c/em\u003e count) and the probability of testing positive for \u003cem\u003eE. coli\u003c/em\u003e, ESBL/AmpC-producing \u003cem\u003eE. coli\u003c/em\u003e and Rotavirus A varied significantly depending on the truck condition, being significantly higher in “dirty” than in “clean” trucks. Despite a non-significant difference, positivity to Rotavirus B showed the same tendency. Conversely, the truck condition had no effect on Rotavirus C. \u003cem\u003eSalmonella\u003c/em\u003e spp., PRRSV, and carbapenemase-producing \u003cem\u003eE. coli\u003c/em\u003e were detected only in samples collected on “dirty” trucks.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions: \u003c/strong\u003eAlthough the prevalence of most agents in ‘clean’ samples was close to zero, the relatively frequent occurrence of \u003cem\u003eE. coli\u003c/em\u003e and some rotaviruses highlights the importance of improving sanitisation procedures. The detection of ESBL/AmpC- and carbapenemase-producing \u003cem\u003eE. coli\u003c/em\u003e was of particular concern. These findings confirm the role of trucks in spreading pathogens of concern and AMR, highlighting the importance of effective monitoring and proper sanitisation procedures.\u003c/p\u003e","manuscriptTitle":"The role of short journey transportation in the spreading of swine pathogens and antimicrobial-resistant bacteria","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-04-22 11:40:01","doi":"10.21203/rs.3.rs-4251132/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":"72e2b49c-d3c4-4c76-b925-be5707c9a9c2","owner":[],"postedDate":"April 22nd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-12-18T04:38:46+00:00","versionOfRecord":[],"versionCreatedAt":"2024-04-22 11:40:01","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4251132","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4251132","identity":"rs-4251132","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}