Phenotypic and Molecular Characterization of Aerobic Bacteria from the Vagina of Apparently Healthy and Infertile Bitches in Zaria, Nigeria

Phenotypic and Molecular Characterization of Aerobic Bacteria from the Vagina of Apparently Healthy and Infertile Bitches in Zaria, Nigeria | 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 Phenotypic and Molecular Characterization of Aerobic Bacteria from the Vagina of Apparently Healthy and Infertile Bitches in Zaria, Nigeria Paul Habila Mamman, Bala Ningi Umar, Abubakar Nafi'u Kakudi, Akeem Olayiwola Ahmed, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-1580205/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 Infertility is a major constrain to a successful dog breeding in many developing countries like Nigeria. The aim of this study therefore, was to establish the aerobic microflora of the vagina of Nigerian bitches. Bacteria were isolated and characterized including susceptibility to commonly used antimicrobials. A total of 169 bacteria were isolated; of which 103 (60.9%) were from apparently healthy bitches and 66 (39.1%) from bitches with reproductive anomalies. These include Staphylococcus saprophyticus (27.2%), Streptococcus species (14.8%), and Corynebacterium species (13.6%). Also, Gemella palaticanis (1.8%), and Schaalia canis (1.3%) which are new to the study area were isolated for the first time in Nigeria. Nearly all the infertile bitches (90.1%) had mixed infection which might be responsible for their illness. Gentamicin was the most effective drug. Micrococcus species and Escherichia coli exhibited multiple drug resistance. Thus, screening of dogs for pathogenic bacteria before breeding is recommended. Aerobes Bitches Gemella palaticanis Schaalia canis 16S rRNA sequencing Figures Figure 1 Introduction Reproduction in dogs is a complex biological process that is governed by various external and internal parameters having different influencing factors. Similarly, reproductive disorders in bitches are usually multifactorial in origin, involving multiple predisposing factors (de Paula Antunes et al., 2016 ; Hollinshead and Hanlon, 2017 ). Failure to detect and correct these anomalies associated factors which predispose bitches to infertility may lead to irreversible syndromes of the reproductive tract (Gavrilovic et al., 2008 ; Runcan and da Silva, 2018 ). Infertility is one of the common reproductive disorders affecting dogs; which comprise a long list of anatomical, hormonal, nutritional, genetic, management and infectious causes (Fontbonne, 2011 ; Zubair et al., 2014 ; Hollinshead and Hanlon, 2017 ; Anon, 2021). Generally, these are broadly categorized as infectious and non-infectious causes. Bacteria are the leading cause of infectious canine infertility (Zubair et al., 2014 ; Hollinshead and Hanlon, 2017 ).Infertility of infectious origin poses a serious threat to the breeding and management of bitches; inflammatory diseases associated with the reproductive system of bitches are significant causes of such reproductive failure. Consequently, male dog contract infection from the infected female and rapidly spread it to other females through coitus. These infections are therefore of vital interest for economic and public health reasons and may create serious problems for the commercial litter producers; hence the need for an in-depth knowledge about the bacterial flora colonizing the reproductive tract of bitch. Globally, efforts have been made over the last 30 years to address the genital flora of normal fertile dogs as well as those with reproductive tract diseases (Olson and Mather, 1978 ; Allen and Dagnall, 1982 ; Groppetti et al., 2012 ; Golińska et al., 2021 ), however, results might differ due to the animals’ immune status, nutrition, age, breed, and geographical location. It has been documented that the species of bacteria isolated from cranial and caudal vagina of bitchesare similar between normal individuals and those with reproductive anomalies (Doig et al., 1981 ; Hutchins et al., 2014 ). Likewise, the type of bacteria isolated do not seem to vary with the stages of estrus cycle, though an increased number of organisms appear to be present during pro-estrus and estrus (Allen and Dagnall, 1982 ; Golińska et al., 2021 ). Notwithstanding, in spite of the vast advances in the knowledge of infectious agents affecting fertility in bitches, the relationship between the presence of bacteria in the reproductive tract and infertility in the bitch remains poorly understood (Golińska et al., 2021 ). However, a heavy growth of a uniform bacterium or potentially fatal organisms like Brucella canis needs to be investigated as similarly reported by Freshman ( 1991 ). Likewise, culturing the vaginal swabs during an overt disease process is valuable in choosing an appropriate therapy. The culture results can be combined with a thorough history, physical examination and vaginal cytology, to adequately assess clinical significance. In addition, some clinicians advocate the employment of systemic antibiotic therapy in bitches when a heavy growth of bacterium is identified at the anterior vagina (Johnston et al., 2001 ). Moreover, in Nigeria, dog breeders often treat bitches with antibiotics at the time of mating to guard against genital infections, and this indiscriminate use may result in development of partial or complete antimicrobial resistance. Hence, antibiogram studies on bacterial isolates assume great importance, and such studies are essential prerequisites for rational use of antimicrobial agents. This study therefore, was aimed at determining the presence and antimicrobial profile of vaginal aerobic bacterial flora from both apparently healthy and infertile bitches in Zaria, Nigeria. Materials And Methods Experimental Dogs The study was carried out on both clinical and non-clinical cases (presented for vaccination) of bitches presented to the Small Animal Clinic of the Veterinary Teaching Hospital, Ahmadu Bello University (VTH-ABU), Zaria; as well as bitches at their homes within Zaria metropolis. Medical history of each bitch was taken. Bitches were termed healthy or infertile based on the history of previous mating and outcomes. They were considered infertile when presented with a history of having been mated three or more times during previous estrous cycles and had not conceived to a healthy male dog that had sired pups before. Based on these inclusion criteria, available 70 healthy, and 30 infertile bitches were evaluated. Sample Collection A total of 100 samples were obtained from the anterior vagina of the bitches using sterile disposable swabs. Each bitch was restrained in a standing position on the examination table and the tail deflected laterally to expose the peri-vulvar area. The peri-vulvar area was initially cleaned with 5% povidone iodine antiseptic solution (WOSAN®). An eight-inch sterile swab was then carefully introduced, directing the swab initially dorso-cranially to avoid the clitoral fossa and urethral orifice and then longitudinally as previously described (Olson, 1975 ). The tip of the swab was dipped into the anterior vagina then softly rolled around the vaginal wall. Thereafter, the swab was withdrawn gently into its labeled sterile container. The samples collected were packed in a cool box containing ice pack and transported to the Department of Veterinary Microbiology laboratory, ABU Zaria for analyses. Bacterial Isolation and Phenotypic Identification Samples collected were inoculated on prepared plates of blood agar base (Oxoid, Hampshire, UK) enriched with 10% sheep blood for primary isolation. The inoculated plates were incubated aerobically at 37 0 C for 24 hours, and the isolated colonies were Gram stained for preliminary identification. Gram negative organisms from the primary culture were sub-cultured on MacConkey agar (Oxoid, Hampshire, UK) for differentiation based on their lactose fermentation properties (Cottral, 1978 ). The resulting lactose and non-lactose fermenters were sub-cultured on eosin methylene blue agar (Oxoid, Hampshire, UK) and xylose lysine deoxycholate agar (Oxoid, Hampshire, UK) for probable identification of Escherichia coli and Salmonella species respectively. The isolates were then subjected to biochemical characterization, including sugar fermentation, oxidase, triple sugar iron, urease, citrate, methyl red, indole, Voges–Proskauer, phenylalanine deaminase, catalase and motility tests as previously described (Cowan and Steel, 2002 ). Genetic Identification of Some Isolates Five of the bacterial isolates (1ZN2019–5ZN2019) not clearly identified phenotypically were characterized by molecular means. DNA extraction was carried out in accordance with manufacturer’s instruction using ZR fungal/bacterial DNA miniPrep™ as previously described (Ahmed et al., 2017 ); and conventional polymerase chain reactions (PCR) assay targeting the 16S rRNA gene was done as par standard protocol described by Wagner et al. ( 2008 ) using Ribose 1: 5’-GGACTACAGGGTATCTAAT-3’, and Ribose 2: 5’-AGAGTTTGATCCTGG-3’ primers. Partial genome sequencing of the amplified product was done at Inqaba laboratory (South Africa). The obtained 16S rDNA sequences were compared with sequences available in GenBank database of National Center for Biotechnology Information (NCBI) using the algorithm BLASTn program (Wagner et al., 2008 ; Thomas et al., 2014 ). Sequences obtained were edited using BioEdit version 10.2. Alignment of the sequences was done using Clustal W; while the phylogenetic analyses was done using MEGA version 6.0. The resulting phylogenetic tree was an Unrooted Neighbour-Joining Tree built with Kimura 2 Parameter with Gama distribution (K 2-G) (Kimura, 1980 ). Boostrap replicate of 1000 was used to test the robustness of the tree. Seven hundred and eight (708) nucleotide regions were used for construction of the phylogenetic tree. Antimicrobial Susceptibility Test The pure bacterial isolates were subjected to antimicrobial sensitivity test using the Kirby-Bauer disk diffusion method (Bauer et al., 1966 ), then interpreted based on Clinical Laboratory Standard Institute Guidelines (CLSI, 2016). The drug impregnated disks (Oxoid, Hampshire, UK) included: tetracycline (30µg), gentamicin (10µg), streptomycin (10µg), amoxycilin/clavulanic acid (30µg), ampicillin (10µg), penicillin (10 units), oxacillin (1µg), cefoxitin (30µg), vancomycin (30µg), erythromycin (15µg), and ciprofloxacin (5µg). Data Analyses Descriptive statistics was applied to calculate frequencies, and percentages of bacterial occurrence rates using IBM SPSS software version 26 (NY, USA). Statistical analysis was performed to determine significant differences in the occurrence rate of each microbial species within the tested groups using Fisher’s exact test. P-values less than 0.05 were considered statistically significant. Results A total of 169 bacterial isolates were identified from the entire samples analyzed, out of which 103 were obtained from apparently healthy bitches, while 66from infertile ones. Staphylococcus saprophyticus had the highest occurrence rate of 27.2%, followed by Streptococcus species (14.8%), and Corynebacterium species (13.6%). While the lowest occurrence rates were the molecularly identified Schaalia canis (1.3%) and Gemella palaticanis (1.8%) both of which were isolated from infertile bitches only (Table 1 ). Broadly, 15 different categories of bacteria were isolated with an overall occurrence rate of 11.3%. Samples from the healthy bitches yielded 9.8% isolation rate while 14.7% was recorded from the infertile bitches. Therefore, infertile bitches had significantly higher (P = 0.007) bacterial occurrence rate than the healthy bitches. Among the different bacteria isolated, only G. palaticanis and Streptococcus species (other than Strept . hyovaginalis ) differ significantly (P ˂ 0.05) between the infertile and healthy bitches (Table 1 ). Similarly, majority of the isolates were Gram positive with only Escherichia coli and Proteus vulgaris beingGram negative (Tables 2 and 3 ). Based on the sequence results of the16S rRNA gene, isolates 1ZN2019 with 762bp sequence size was identified as Enterococcus dispar (Accession No: MN585805); 2ZN2019 with 739bp sequence size as Schaalia canis (Accession No: MN585806); and 3ZN2019 with 761bp sequence size as Streptococcus hyovaginalis (Accession No: MN585807). Isolates 4ZN2019 which has 768bp sequence, and 5ZN2019 with 757bp sequence sizes were respectively identified as Enterococcus faecium (Accession No: MN585808) and Gemella palaticanis (Accession No: MN585809). The obtained bacterial sequences were deposited in the GeneBank ( https://submit.ncbi.nlm.nih.gov/subs/?search=SUB6437530 ). The dendogram of the phylogenetic tree drawn from the five identified bacterial isolates and other related organisms earlier deposited from different parts of the world is presented as Fig. 1 . The bacteria isolated in the present study exhibited varying degree of sensitivity to different antimicrobials. Among all the 11 antimicrobials tested, gentamicin proved to be the most effective as 89.25% of all the isolates tested were sensitive to it. This is followed by a cephalosporin, cefoxitin which showed 70.00% sensitivity, while the highest rate of resistance was recorded among the penicillins (penicillin, oxacillin, amoxicillin/clavulanic acid, and ampicillin) which had about 73.00% average resistance to all the isolates tested (Table 4 ). The detailed antimicrobial resistance pattern of the isolates is shown in Table 5 . Several categories of the isolates particularly E. coli, Streptococcus species (other than Strep . hyovaginalis ), and Enterococcus species (other than E. faecium ) exhibited multidrug resistance (MDR), resisting at leastthree different classes of antimicrobials.Similarly, Micrococcus species showed an extended spectrum of drug resistance (XDR) as they were resistant to at least one agent in nearly all the classes of antimicrobial tested. Table 1 Occurrence of aerobic bacteria recovered from the vagina of apparently healthy and infertile bitches in Zaria, Nigeria S/No Name of isolate Healthy Bitches (n = 70) Infertile Bitches (n = 30) P-values Frequency Percentage (%) Frequency Percentage (%) 1 Staphylococcus saprophyticus 32 45.7 14 46.7 1.000 2 Staphylococcus aureus 10 14.3 6 20.0 0.554 3 Staphylococcus schleiferi 6 8.6 0 0.0 0.174 4 Streptococcus hyovaginalis 0 0.0 1 3.3 0.300 5 Other Streptococcus species 12 17.1 12 40.0 0.021٭ 6 Corynebacterium species 14 20.0 9 30.0 0.306 7 Enterococcus faecium 0 0.0 1 3.3 0.300 8 Enterococcus dispar 0 0.0 1 3.3 0.300 9 Other Enterococcus species 8 11.4 1 3.3 0.272 10 Micrococcus species 5 7.1 6 20.0 0.082 11 Proteus vulgaris 6 8.6 5 16.7 0.298 12 Escherichia coli 7 10.0 3 10.0 1.000 13 Baccilus species 3 4.3 2 6.7 0.635 14 Gemellapalaticanis 0 0.0 3 10.0 0.025٭ 15 Schaaliacanis 0 0.0 2 6.7 0.088 Total 103 66 P-values with an asterisk٭are statistically significant (P˂0.05). Table 2 Morphological and Biochemical characteristics of some aerobic Gram-positive bacteria isolated from the vagina of apparently healthy and infertile bitches in Zaria, Nigeria S/No Colonial morphology Microscopic features Biochemical tests Organism identified 1 Small, raised, β-hemolytic colonies on blood agar Gram positive cocci in cluster Catalase, coagulase, glucose, trehalose, maltose, mannitol, and sucrose positive; while negative for xylose, and arabinose Staphylococcus aureus 2 Small, raised, β-hemolytic colonies on blood agar Gram positive cocci in cluster Catalase, glucose, trehalose, mannitol, and sucrose positive; while negative for coagulase, maltose, xylose, and arabinose S. saprophyticus 3 Small, raised, whitish β-hemolytic colonies Gram positive cocci in cluster Catalase, glucose, and sucrose positive; while negative for coagulase, trehalose, mannitol, maltose, xylose, and arabinose S. schleiferi 4 Glossy-dew drop-like, α, and β-hemolytic colonies Gram positive cocci in short and long chains Positive for glucose; and negative for catalase Streptococcus sppother than St. hyovaginalis 5 Glossy-dew drop-like, γ-hemolytic colonies Gram positive cocci in short chains and branches Glucose, lactose, dextrose, raffinose, arabinose, and urease positive; while negative for catalase, coagulase, trehalose, mannitol, maltose, xylose, and sucrose Enterococcus species other than E . faecium and E. dispar 6 Small, raised, β-hemolytic colonies with little yellowish pigmentation on blood agar Gram positive cocci with tetrad appearance Positive for catalase, and glucose; but negative for coagulase Micrococcus species 7 Large, flat, β-hemolytic colonies with serrated edges Gram positive rods in short chains Catalase, glucose, lactose, and urease positive; while negative for sucrose Baccilus species 8 Small, raised, moist, greyish, γ-hemolytic colonies Gram positive, pleomorphic rods with Chinese letters appearance Positive for catalase, glucose, and lactose; but negative for coagulase, sucrose, indole, and urease Corynebacterium species 9 Pin-point β-hemolytic colonies on blood agar Gram positive cocci mostly in pairs ٭٭٭ Gemellapalaticanis 10 Tiny granular-like, β-hemolytic colonies Gram positive coccobaccilli in chains ٭٭٭ Schaaliacanis Note: Organisms with٭٭٭ were identified via molecular characterization Table 3 Biochemical characteristics of some aerobic Gram-negative bacteria isolated from the vagina of apparently healthy and infertile bitches in Zaria, Nigeria S/No. Biochemical Tests Conducted Name of Bacterium CAT OF TSI NIT IND URE CIT MR VP MOT OXD GLU SUC LAC 1 + F K/A.H 2 S + + + - + - + -/+ + + - Proteus vulgaris 2 + F A/A. G + + - - + - + - + - + Escherichia coli Key: CAT = catalase, OF = oxidative/fermentative, TSI = triple sugar iron, NIT = nitrate reduction, IND = indole, URE = urease, CIT = citrate, MR = methyl red, VP = Voges Proskauer, MOT = motility, OXD = oxidase, GLU = glucose, SUC = sucrose, LAC = lactose, F = fermenter, A = acid, K = alkaline, G = gas, H 2 S = hydrogen sulphide, -=negative, and + = positive Table 4 Percentage antimicrobial susceptibility profile of aerobic bacteria isolated from the vagina of apparently healthy and infertile bitches in Zaria, Nigeria Antibiotic (µg) Staphylococcus Streptococcus Enterococcus Micrococcus Corynebacterium Schaalia Gemella Escherichia S I R S I R S I R S I R S I R S I R S I R S I R TE (30) 7 25 68 50 0 50 0 0 100 0 0 100 0 100 0 70 20 10 0 0 100 90 10 0 CN (10) 94 0 6 100 0 0 90 10 0 50 0 50 90 10 0 90 10 0 100 0 0 100 0 0 AMC (30) 5 0 95 20 20 60 0 0 100 0 0 100 0 10 90 0 10 90 80 20 0 0 0 100 FOX (30) 80 15 5 80 20 0 80 20 0 50 0 50 80 20 0 80 10 10 100 0 0 10 90 0 OX (1) 0 40 60 20 0 80 0 0 100 20 30 50 0 50 50 0 40 60 80 20 0 0 0 100 VA (30) 40 30 30 80 20 0 0 10 90 0 100 0 0 100 0 0 30 70 60 20 20 0 0 100 AMP (10) 0 20 80 0 40 60 0 20 80 0 0 100 0 10 90 0 20 80 0 40 60 0 0 100 P (10) 0 10 90 20 40 40 0 20 80 0 10 90 0 20 80 20 70 10 20 20 60 0 0 100 E (15) 50 10 40 40 20 40 80 20 0 0 50 50 50 30 20 80 10 10 80 20 0 0 0 100 S (10) 40 30 30 40 40 20 80 20 0 0 0 100 80 20 0 70 20 10 20 60 20 20 80 0 CIP (5) 30 60 10 40 40 20 0 100 0 0 0 100 50 30 20 100 0 0 40 40 20 50 50 0 Note: Staphylococcus = ( S. aureus & S. saprophyticus ), Streptococcus = ( Strep. Hyovaginalis & other species), Enterococcus = ( E. faecium, E. dispar , & other species), Micrococcus = ( Micrococcus species), Corynebacterium ( Corynebacterium species), Schaalia = ( S . canis ), Gemella = ( G . palaticanis ), Escherichia = ( E . coli ), TE = Tetracycline, CN = Gentamicin, AMC = Amoxicillin/Clavulanicacid, FOX = Cefoxitin, OX = Oxacillin, VA = Vancomycin, AMP = Ampicillin, P = Penicillin, E = Erythromycin, S = Streptomycin, CIP = Ciprofloxacin. Using the CLSI, 2016: R = Resistant, I = Intermediate, and S = Sensitive Table 5 Antimicrobial resistance patterns of some aerobic bacteria isolated from the vagina of apparently healthy and infertile bitches in Zaria, Nigeria S/No Name of Isolate Resistance Pattern Class of Antimicrobials Resisted 1 Staphylococcus saprophyticus TE-AMC-OX-AMP-P 2 2 Staphylococcus aureus AMC-AMP-P-OX 1 4 Streptococcus hyovaginalis AMC-OX-AMP-P 1 5 Other Streptococcus species TE-AMC-OX-AMP-P-E 3 6 Corynebacterium species AMC-OX-AMP-P 1 7 Enterococcus faecium AMC-OX-AMP-P 1 8 Enterococcus dispar TE-AMC-OX-VA-AMP-P-S 4 9 Other Enterococcus species TE-AMC-OX-VA-AMP-P 3 10 Micrococcus species TE-CN-AMC-FOX-OX-AMP-P-E-S-CIP 6 11 Escherichia coli AMC-OX-VA-AMP-P-E 3 12 Gemellapalaticanis TE-AMP-P 2 13 Schaaliacanis AMC-OX-VA-AMP 2 TE = Tetracycline, AMC = Amoxicillin/Clavulanicacid, OX = Oxacillin, AMP = Ampicillin, P = Penicillin, E = Erythromycin, VA = Vancomycin,S = Streptomycin, CN = Gentamicin,FOX = Cefoxitin, and CIP = Ciprofloxacin Discussion This study assessed the presence and antibiogram of aerobic bacterial populations in the vagina of apparently healthy and infertile bitches. The findings showed some level of variation in number and type of bacteria between the two different groups. The incidence of common pathogens, particularly, Staphylococcus saprophyticus, S. aureus, E. coli, Streptococcus s pecies, Bacillus species, Enterococcus , and Corynebacterium species was similar between the healthy and infertile bitches. This outcome corelates well with the results of other parallel studies (Findik et al., 2003 ; Groppetti et al., 2012 ; Antonov, 2019 ; Golińskaet al., 2021 ) which documented Staphylococcus, Streptococcus, Enterococcus, Corynebacterium, Proteus and Escherichia to be the most common genera colonizing the vagina of dogs. However, the present study established significant difference in the overall bacterial occurrence rate between healthy and infertile bitches; This contradicts the studies of Hutchins et al. ( 2014 ) and Golińska et al. ( 2021 ) who reported that the total number of bacteria in the vagina of healthy and non-healthy dogs at different stages of estrus cycle was almost the same when conducting similar studies. The exact reason for this variation is not clear, but one possible explanation is the relatively large sample size (100) of the current study as against the 44 and 39 respectively utilized by the two previous studies. This is to say that, a probable significant difference in the prevalence of specific bacteria would have been identified if much higher sample sizes were employed in those preceding research. The restricted vaginal presence of Staphylococcus schleiferi within healthy bitches could have a protective competitive role against potentially more dangerous pathogens like Gemella palaticanis and Schaalia canis which were never isolated from the healthy bitches in the present study. This statement is in line with the findings of Feldman and Nelson ( 2003 ) who documented that normal vaginal microbiota has been shown to protect genito-urinary tract against pathogenic bacteria through nutrient and epithelial cell receptors competition. Conversely, the relative higher incidence of Proteus vulgaris observed in infertile bitches agrees with the findings of Golińska et al. ( 2021 ) who reported significantly higher prevalence of Gram-negative rods (other than E. coli ) in the vagina of dogs with genital tract infections. Likewise, the significantly higher occurrence rate of Streptococcus species in infertile bitches might be responsible for the infertility in those dogs. This is because β-haemolytic Streptococci , as well as Proteus, E. coli and Enterococcus species were previously associated with infertility in bitches (John et al., 2011 ). Similarly, the significantly higher occurrence of G . palaticanis in the anterior vagina of infertile bitches could be associated with the development of genital tract infection that might eventually result in infertility, however, a focused case control study would be required to ascertain this hypothesis. Bacteria of the Genera Pasteurella, Pseudomonas, Klebsiella , and Enterobacter which were previously reported to reside within dog’s reproductive tract (John et al., 2011 ; Antonov, 2019 ) were not identified in the present study. This variation could be due to the canine intrinsic and genetic variability, indiscriminate use of antibiotics in the study area, or inherent limitations of culture-based systems compared to molecular identification as similarly documented by Lyman et al. ( 2019 ). Another likely cause of this disparity in the type of vaginal microflora is the possible sample contamination from the vestibule or vulva especially in the case of John et al. ( 2011 ) who used vaginal discharges rather than guarded aseptic swab collection from the anterior vagina. Varying grades of sensitivity to different antimicrobials were reported in the isolates from both healthy and infertile bitches with aminoglycoside, gentamicin being the most effective as equally observed by John et al. ( 2011 ). Likewise, several isolates displayed resistance to at least two classes of antibiotics termed multidrug resistant phenotypes. In this study, the β-lactam antibiotics; ampicillin, penicillin, oxacillin, and amoxicillin/clavulanic acid which are known to be effective against many Gram-positive pathogens were surprisingly found to be resistant (with average resistance of 81.25%, 68.75%, 62.50% and 79.38% respectively). Gram-negative bacteria, (such as E. coli ) have been established to develop resistance to multiple antibiotics mostly due to their inherent resistance to macrolide antibiotics (such as erythromycin), efflux system, as well as decreased drug uptake mediated by chromosomal mutations or via acquisition of resistance genes (Lartey et al., 1994 ; Rossolini et al., 2017 ). However, the non-susceptibility of E. coli to multiple antimicrobials observed in the current study is alarming. Consequently, the extremely drug-resistant Micrococcus species most likely resulted from the acquisition of multiple resistance determinants over a long period sequel to the known indiscriminate prescription, acquisition and usage of antimicrobials in the study area (FMAEH, 2017; Ogwucheet al., 2021 ). Therefore, the multidrug resistance nature of Streptococcus species (other than Strep. hyovaginalis ), Enterococcus species (other than E. faecium & E. dispar ), as well as the extensive antimicrobial resistance of Micrococcus species recorded in the current study highlight the need for an improved fight against antimicrobial resistance in the study area. In conclusion, this study has established the most common bacteria colonizing the anterior vagina of bitches in Zaria, Nigeria. The occurrence of aerobic bacteria was significantly higher in infertile bitches compared to the healthy ones suggesting the possibility of some identified pathogens being responsible for the infertility. Furthermore, varying degree of sensitivity to antimicrobials, and development of multiple drug resistance were identified among the isolates highlighting the need for rational and judicious selection of drugs when managing infertility in dogs. We recommend dog breeders to screen their bitches and studs before breeding especially for the presence of possible multi-drug resistant bacteria which can pose a threat to pregnancy. This should be preceded by thorough clinical and vaginal-cytological examinations. Declarations Acknowledgement The authors wish to thank Dr. K. H. Ahmad for his help in reading and correcting the manuscript. Funding The authors declare that no specific funds, grants, or other support were received from funding agencies in the public, commercial, or not-for-profit sectors during the preparation of this manuscript. Conflict of Interest The authors have no relevant financial or non-financial interests to disclose. Authors’ Contributions All the authors have contributed substantially to the concept, design, and execution of the study. Material preparation, data collection and analysis were performed by Dr. B.N. Umar, Dr. A.N. Kakudi, and Dr. A.O. Ahmed. The first draft of the manuscript was written by Dr. P.H. Mamman and all the authors reviewed the previous versions of the manuscript. All authors read and approved the final manuscript. Data Availability Statement The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request. Ethics Approval The bitches used in this study were managed in accordance with standard ethics. The study was performed in line with the principles of the declaration of Helsinki; and it was approved by the Ahmadu Bello University Committee on Animal Use and Care (ABUCAUC). Informed Consent Not applicable References Ahmed OA, Mamman PH, Raji MA, Kwanashie, CN, Raufu IA,Aremu A (2017) Distribution of Virulence Genes in Salmonella Serovars isolated from poultry farms in Kwara State, Nigeria. Ceylon Journal of Science, 46, 69–76. https://doi.org/10.4038/cjs.v46i4.7469. Allen WE,Dagnall GJR (1982) Some observations on the aerobic bacterial flora of the genital tract of the dog and bitch. Journal of Small Animal Practice, 23, 325–335. Anonymous (2021)High Street Epping Veterinary Clinic : Factors Leading to Infertility in Bitches . Retrieved on October 28, 2021 from https://www.highstreeteppingvetclinic.com/factors-leading-to-infertility-in-bitch.pml Antonov A (2019) Normal and Pathological Vaginal Microflora in the Bitch-A Review. Journal of Advances in Microbiology,15, 1-5.DOI: 10.9734/JAMB/2019/v15i130074 Bauer AW, Kirby WM, Sherris JC,Turck M (1966) Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology, 45, 493–496. CLSI. M100S27: (2016) Performance Standards for Antimicrobial Susceptibility Testing. Clinical Laboratory Standard Institute2016;M100S27 Cottral GE (1978) Manual of Standardised Methods for Veterinary Microbiology . National Research Council. Subcommittee on Standardized Methods for Veterinary Microbiology. Comelll University Press, Ithaca, New York, Pp. 731. Cowan ST, Steel KJ (2002) Manual for The Identification of Medical Bacteria. 2nd edition. Cambridge University press, United Kindom, Pp. 217. de Paula Antunes JMA, de Carvalho Freire DA, de Medeiros Oliveira IVP, Moura GHF, de Castro Demoner L, Ferreira HIP (2016) Infectious Causes of Abortion, Stillbirth and Neonatal Death in Bitches. In: Abdelhay Essayed K.H (Ed). Canine Medicine: Recent Topics and Advanced Research . Janeza Trdine 9, 5100, Rijeka, Croatia. pp.55-73. Doig PA, Ruhnke HL,Bosu WTK (1981) The Genital mycoplasma and ureaplasma flora of healthy and diseased dogs. Canadian Journal of Comparative Medicine, 45, 223–231. Federal Ministries of Agriculture, Environment and Health; (FMAEH 2017) Antimicrobial Use and Resistance in Nigeria: Situation Analysis and Recommendations. Pp. 1-105 Feldman EC, Nelson RW (2003) Canine and feline endocrinology and reproduction. 3rd Ed. Saunders, Philadelphia, USA, Pp. 1044-1104. Findik M, Maral N, Keskin O, Kalender H, Erdeger J, Aslan S (2003) The relationship between the stages of sexual cycle, the pregnancy and postpartum periods and vaginal flora in kangal breed bitches. Turkish Journal of Veterinary and Animal Science, 27, 761-765. Fontbonne A (2011) Infertility in bitches and queens: recent advances. Revista Brasileira de Reprodução Animal , 35(2), 202-209. Freshman JL (1991) Clinical approach to infertility in the cycling bitch. Veterinary Clinics of North America: Small Animal Practice, 21(3), 427-435. Gavrilovic BB, Andersson K, Forsberg C (2008) Reproductive patterns in the domestic dog: A retrospective study of the Drever breed. Theriogenology , 70(5), 783-794. Golińska E, Sowińska N, Tomusiak-Plebanek A, Szydło M, Witka N, Lenarczyk J, Strus M (2021) The vaginal microflora changes in various stages of the estrous cycle of healthy female dogs and the ones with genital tract infections. BMCVeterinaryResearch, 17(1), 1-8. Groppetti D, Pecile A, Barbero C, Martino PA (2012) Vaginal bacterial flora and cytology in proestrous bitches: Role on fertility. Theriogenology 77(8), 1549-1556 Hollinshead FK, Hanlon DW (2017) Factors affecting the reproductive performance of bitches: A prospective cohort study involving 1203 inseminations with fresh and frozen semen. Theriogenology, 101, 62–72. Hutchins RG, Vaden SL, Jacob ME, Harris TL, Bowles KD, Wood MW, Bailey CS (2014) Vaginal microbiota of spayed dogs with or without recurrent urinary tract infections. Journal of Veterinary Internal Medicine, 28(2), 300–304. John A, Nair DR, Sreejith JR, Deepthi L, Praseeda R, Ajitkumar G (2011) Vaginal Microflora and its Antibiogram in Infertile Bitches. Online Veterinary Journal, 6(1), e79. Johnston SD, Kustritz MVR, Olson PNS (2001) Canine and Feline Theriogenology. Saunders, Philadelphia, USA, 592p Kimura MA (1980) Simple Method for Estimating Evolutionary Rates of Base Substitutions Through Comparative Studies of Nucleotide Sequences. Journal of Molecular Evolution, 16, 111-120. Lartey PA, Nellans HN, Tanaka SK (1994) New developments in macrolides: structures and antibacterial and prokinetic activities. Advances in Pharmacology (San Diego, Calif.) , Elsevier Academic Press, Inc. Pp. 307-343. Lyman CC, Holyoak GR, Meinkoth K, Wieneke X, Chillemi KA, DeSilva U (2019) Canine endometrial and vaginal microbiomes reveal distinct and complex ecosystems. PLoS ONE 14(1), e0210157. doi.org/10.1371/journal.pone.0210157 Ogwuche A, Ekiri AB, Endacott I, Maikai BV, Idoga ES, Alafiatayo R, Cook AJ (2021) Antibiotic use practices of veterinarians and para-veterinarians and the implications for antibiotic stewardship in Nigeria. Journal of the South African Veterinary Association,92, 1-14. Olson PS (1975) Steptococcuscanis : An isolate from the canine uterus. Veterinary Medicine Small Animal Clinician, 70, 933–934. Olson PS, Mather EC (1978) Canine vaginal and uterine bacterial flora. Journal of the American Veterinary Medical Association, 172, 708–711. Rossolini GM, Arena F,Giani T (2017) Mechanisms of Antibacterial Resistance. In: Jonathan Cohen, William Powderly, Steven Opal (Eds.) Infectious Diseases (4 th Edition), Elsevier Ltd. 2, Pp. e1181-1196. Runcan EE, da Silva MAC (2018) Whelping and dystocia: maximizing success of medical management. Topics in Companion Animal Medicine, 33(1), 12-16. Thomas J, Thanigaivel S, Vijayakumar S, Acharya K, Shinge D, Seelan TSJ, Mukherjee A, Chandrasekaran N (2014) Pathogenecity of Pseudomonas aeruginosa in Oreochromis mossambicus and treatment using lime oil nanoemulsion. Colloids and Surfaces B: Biointerfaces, 116, 372-377. Wagner J, Short K, Catto-Smith AG, Cameron DJ, Bishop RF, Kirkwood CD (2008) Identification and characterisation of Pseudomonas 16S ribosomal DNA from ileal biopsies of children with Crohn's disease. PloS one , 3(10), e3578. Zubair M, Ubaid-ur-Rehman S, Sajid SM (2014) Causes of infertility in bitch. Advances in Animaland Veterinary Sciences, 2(10), 565-573. Additional Declarations No competing interests reported. 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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-1580205","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":100382353,"identity":"fb4fde1e-ae60-46f0-a3d9-f5108abfaad1","order_by":0,"name":"Paul Habila Mamman","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzklEQVRIiWNgGAWjYFACHgjFDyISCkjRItkA0mJAihaDA2CSCA38YmcPPviYYydvfH514ocHBgzy/GIH8GuRnJ2XbDhzW7LhthtvN0sAHWY4c3YCfi0Gt3PMpHm3MTNuu3F2A0hLgsFtAlrsb+eY//67rd5+84yzm38QpcVAOscMaMXhxA38vduIs0Xidl6yZO+248kzbvBus0gwkCDsF/7ZuQc//NxWbdvff3bzzR8VNvL80gS0INkHVilBrHKwfQdIUT0KRsEoGAUjCQAACHNF8GZNAsgAAAAASUVORK5CYII=","orcid":"","institution":"Ahmadu Bello University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Paul","middleName":"Habila","lastName":"Mamman","suffix":""},{"id":100382354,"identity":"61793891-b449-42eb-923e-ab19a86bbc70","order_by":1,"name":"Bala Ningi Umar","email":"","orcid":"","institution":"Ahmadu Bello University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Bala","middleName":"Ningi","lastName":"Umar","suffix":""},{"id":100382355,"identity":"67251a1d-af5e-4143-855c-b5806fa92a61","order_by":2,"name":"Abubakar Nafi'u Kakudi","email":"","orcid":"","institution":"Ahmadu Bello University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Abubakar","middleName":"Nafi'u","lastName":"Kakudi","suffix":""},{"id":100382356,"identity":"1f544837-1012-4327-aa64-b07c3265892b","order_by":3,"name":"Akeem Olayiwola Ahmed","email":"","orcid":"","institution":"University of Ilorin","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Akeem","middleName":"Olayiwola","lastName":"Ahmed","suffix":""},{"id":100382357,"identity":"a28949e7-43ad-4a27-aed5-be044a878ed6","order_by":4,"name":"Kennedy Chah","email":"","orcid":"","institution":"University of Nigeria, Nsukka","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Kennedy","middleName":"","lastName":"Chah","suffix":""},{"id":100382358,"identity":"be040900-19e0-4b79-b524-bada12510488","order_by":5,"name":"Haruna Makanjuola Kazeem","email":"","orcid":"","institution":"Ahmadu Bello University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Haruna","middleName":"Makanjuola","lastName":"Kazeem","suffix":""}],"badges":[],"createdAt":"2022-04-21 10:14:25","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-1580205/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-1580205/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":20747623,"identity":"b02edf90-b5d8-4193-99c8-e4f7fc2bad72","added_by":"auto","created_at":"2022-04-25 21:25:29","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":10800,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree with boostrap values showing the relationships between some selected bacteria in the gene bank (each having its accession number in parenthesis) and the five aerobic bacterial isolates(each having a red square box) from the vagina of infertile bitches in Zaria, Nigeria.\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-1580205/v1/b2f7a79af279764742032cd0.png"},{"id":27537951,"identity":"6e1080b6-346a-43b6-93bd-9a1b087668f8","added_by":"auto","created_at":"2022-10-10 08:14:24","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":439384,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1580205/v1/99d903a9-92a4-4360-8313-f405be50270e.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Phenotypic and Molecular Characterization of Aerobic Bacteria from the Vagina of Apparently Healthy and Infertile Bitches in Zaria, Nigeria","fulltext":[{"header":"Introduction","content":"\u003cp\u003eReproduction in dogs is a complex biological process that is governed by various external and internal parameters having different influencing factors. Similarly, reproductive disorders in bitches are usually multifactorial in origin, involving multiple predisposing factors (de Paula Antunes et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Hollinshead and Hanlon, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Failure to detect and correct these anomalies associated factors which predispose bitches to infertility may lead to irreversible syndromes of the reproductive tract (Gavrilovic et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Runcan and da Silva, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eInfertility is one of the common reproductive disorders affecting dogs; which comprise a long list of anatomical, hormonal, nutritional, genetic, management and infectious causes (Fontbonne, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Zubair et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Hollinshead and Hanlon, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Anon, 2021). Generally, these are broadly categorized as infectious and non-infectious causes. Bacteria are the leading cause of infectious canine infertility (Zubair et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Hollinshead and Hanlon, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).Infertility of infectious origin poses a serious threat to the breeding and management of bitches; inflammatory diseases associated with the reproductive system of bitches are significant causes of such reproductive failure. Consequently, male dog contract infection from the infected female and rapidly spread it to other females through coitus. These infections are therefore of vital interest for economic and public health reasons and may create serious problems for the commercial litter producers; hence the need for an in-depth knowledge about the bacterial flora colonizing the reproductive tract of bitch.\u003c/p\u003e \u003cp\u003eGlobally, efforts have been made over the last 30 years to address the genital flora of normal fertile dogs as well as those with reproductive tract diseases (Olson and Mather, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e1978\u003c/span\u003e; Allen and Dagnall, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e1982\u003c/span\u003e; Groppetti et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Golińska et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), however, results might differ due to the animals\u0026rsquo; immune status, nutrition, age, breed, and geographical location. It has been documented that the species of bacteria isolated from cranial and caudal vagina of bitchesare similar between normal individuals and those with reproductive anomalies (Doig et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1981\u003c/span\u003e; Hutchins et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Likewise, the type of bacteria isolated do not seem to vary with the stages of estrus cycle, though an increased number of organisms appear to be present during pro-estrus and estrus (Allen and Dagnall, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e1982\u003c/span\u003e; Golińska et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eNotwithstanding, in spite of the vast advances in the knowledge of infectious agents affecting fertility in bitches, the relationship between the presence of bacteria in the reproductive tract and infertility in the bitch remains poorly understood (Golińska et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). However, a heavy growth of a uniform bacterium or potentially fatal organisms like \u003cem\u003eBrucella canis\u003c/em\u003e needs to be investigated as similarly reported by Freshman (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e1991\u003c/span\u003e). Likewise, culturing the vaginal swabs during an overt disease process is valuable in choosing an appropriate therapy. The culture results can be combined with a thorough history, physical examination and vaginal cytology, to adequately assess clinical significance.\u003c/p\u003e \u003cp\u003eIn addition, some clinicians advocate the employment of systemic antibiotic therapy in bitches when a heavy growth of bacterium is identified at the anterior vagina (Johnston et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). Moreover, in Nigeria, dog breeders often treat bitches with antibiotics at the time of mating to guard against genital infections, and this indiscriminate use may result in development of partial or complete antimicrobial resistance. Hence, antibiogram studies on bacterial isolates assume great importance, and such studies are essential prerequisites for rational use of antimicrobial agents. This study therefore, was aimed at determining the presence and antimicrobial profile of vaginal aerobic bacterial flora from both apparently healthy and infertile bitches in Zaria, Nigeria.\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eExperimental Dogs\u003c/h2\u003e \u003cp\u003eThe study was carried out on both clinical and non-clinical cases (presented for vaccination) of bitches presented to the Small Animal Clinic of the Veterinary Teaching Hospital, Ahmadu Bello University (VTH-ABU), Zaria; as well as bitches at their homes within Zaria metropolis. Medical history of each bitch was taken. Bitches were termed healthy or infertile based on the history of previous mating and outcomes. They were considered infertile when presented with a history of having been mated three or more times during previous estrous cycles and had not conceived to a healthy male dog that had sired pups before. Based on these inclusion criteria, available 70 healthy, and 30 infertile bitches were evaluated.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eSample Collection\u003c/h2\u003e \u003cp\u003eA total of 100 samples were obtained from the anterior vagina of the bitches using sterile disposable swabs. Each bitch was restrained in a standing position on the examination table and the tail deflected laterally to expose the peri-vulvar area. The peri-vulvar area was initially cleaned with 5% povidone iodine antiseptic solution (WOSAN\u0026reg;). An eight-inch sterile swab was then carefully introduced, directing the swab initially dorso-cranially to avoid the clitoral fossa and urethral orifice and then longitudinally as previously described (Olson, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e1975\u003c/span\u003e). The tip of the swab was dipped into the anterior vagina then softly rolled around the vaginal wall. Thereafter, the swab was withdrawn gently into its labeled sterile container. The samples collected were packed in a cool box containing ice pack and transported to the Department of Veterinary Microbiology laboratory, ABU Zaria for analyses.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eBacterial Isolation and Phenotypic Identification\u003c/h2\u003e \u003cp\u003eSamples collected were inoculated on prepared plates of blood agar base (Oxoid, Hampshire, UK) enriched with 10% sheep blood for primary isolation. The inoculated plates were incubated aerobically at 37 \u003csup\u003e0\u003c/sup\u003eC for 24 hours, and the isolated colonies were Gram stained for preliminary identification. Gram negative organisms from the primary culture were sub-cultured on MacConkey agar (Oxoid, Hampshire, UK) for differentiation based on their lactose fermentation properties (Cottral, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e1978\u003c/span\u003e). The resulting lactose and non-lactose fermenters were sub-cultured on eosin methylene blue agar (Oxoid, Hampshire, UK) and xylose lysine deoxycholate agar (Oxoid, Hampshire, UK) for probable identification of \u003cem\u003eEscherichia coli\u003c/em\u003e and \u003cem\u003eSalmonella\u003c/em\u003e species respectively. The isolates were then subjected to biochemical characterization, including sugar fermentation, oxidase, triple sugar iron, urease, citrate, methyl red, indole, Voges\u0026ndash;Proskauer, phenylalanine deaminase, catalase and motility tests as previously described (Cowan and Steel, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2002\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eGenetic Identification of Some Isolates\u003c/h2\u003e \u003cp\u003eFive of the bacterial isolates (1ZN2019\u0026ndash;5ZN2019) not clearly identified phenotypically were characterized by molecular means. DNA extraction was carried out in accordance with manufacturer\u0026rsquo;s instruction using ZR fungal/bacterial DNA miniPrep\u0026trade; as previously described (Ahmed et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2017\u003c/span\u003e); and conventional polymerase chain reactions (PCR) assay targeting the 16S rRNA gene was done as par standard protocol described by Wagner et al. (\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2008\u003c/span\u003e) using Ribose 1: 5\u0026rsquo;-GGACTACAGGGTATCTAAT-3\u0026rsquo;, and Ribose 2: 5\u0026rsquo;-AGAGTTTGATCCTGG-3\u0026rsquo; primers. Partial genome sequencing of the amplified product was done at Inqaba laboratory (South Africa). The obtained 16S rDNA sequences were compared with sequences available in GenBank database of National Center for Biotechnology Information (NCBI) using the algorithm BLASTn program (Wagner et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Thomas et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Sequences obtained were edited using BioEdit version 10.2. Alignment of the sequences was done using Clustal W; while the phylogenetic analyses was done using MEGA version 6.0. The resulting phylogenetic tree was an Unrooted Neighbour-Joining Tree built with Kimura 2 Parameter with Gama distribution (K 2-G) (Kimura, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e1980\u003c/span\u003e). Boostrap replicate of 1000 was used to test the robustness of the tree. Seven hundred and eight (708) nucleotide regions were used for construction of the phylogenetic tree.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eAntimicrobial Susceptibility Test\u003c/h2\u003e \u003cp\u003eThe pure bacterial isolates were subjected to antimicrobial sensitivity test using the Kirby-Bauer disk diffusion method (Bauer et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e1966\u003c/span\u003e), then interpreted based on Clinical Laboratory Standard Institute Guidelines (CLSI, 2016). The drug impregnated disks (Oxoid, Hampshire, UK) included: tetracycline (30\u0026micro;g), gentamicin (10\u0026micro;g), streptomycin (10\u0026micro;g), amoxycilin/clavulanic acid (30\u0026micro;g), ampicillin (10\u0026micro;g), penicillin (10 units), oxacillin (1\u0026micro;g), cefoxitin (30\u0026micro;g), vancomycin (30\u0026micro;g), erythromycin (15\u0026micro;g), and ciprofloxacin (5\u0026micro;g).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eData Analyses\u003c/h2\u003e \u003cp\u003eDescriptive statistics was applied to calculate frequencies, and percentages of bacterial occurrence rates using IBM SPSS software version 26 (NY, USA). Statistical analysis was performed to determine significant differences in the occurrence rate of each microbial species within the tested groups using Fisher\u0026rsquo;s exact test. P-values less than 0.05 were considered statistically significant.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eA total of 169 bacterial isolates were identified from the entire samples analyzed, out of which 103 were obtained from apparently healthy bitches, while 66from infertile ones. \u003cem\u003eStaphylococcus saprophyticus\u003c/em\u003e had the highest occurrence rate of 27.2%, followed by \u003cem\u003eStreptococcus\u003c/em\u003e species (14.8%), and \u003cem\u003eCorynebacterium\u003c/em\u003e species (13.6%). While the lowest occurrence rates were the molecularly identified \u003cem\u003eSchaalia canis\u003c/em\u003e (1.3%) and \u003cem\u003eGemella palaticanis\u003c/em\u003e (1.8%) both of which were isolated from infertile bitches only (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Broadly, 15 different categories of bacteria were isolated with an overall occurrence rate of 11.3%. Samples from the healthy bitches yielded 9.8% isolation rate while 14.7% was recorded from the infertile bitches. Therefore, infertile bitches had significantly higher (P\u0026thinsp;=\u0026thinsp;0.007) bacterial occurrence rate than the healthy bitches. Among the different bacteria isolated, only \u003cem\u003eG. palaticanis\u003c/em\u003e and \u003cem\u003eStreptococcus\u003c/em\u003e species (other than \u003cem\u003eStrept\u003c/em\u003e. \u003cem\u003ehyovaginalis\u003c/em\u003e) differ significantly (P ˂ 0.05) between the infertile and healthy bitches (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Similarly, majority of the isolates were Gram positive with only \u003cem\u003eEscherichia coli\u003c/em\u003e and \u003cem\u003eProteus vulgaris\u003c/em\u003e beingGram negative (Tables\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and \u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eBased on the sequence results of the16S rRNA gene, isolates 1ZN2019 with 762bp sequence size was identified as \u003cem\u003eEnterococcus dispar\u003c/em\u003e (Accession No: MN585805); 2ZN2019 with 739bp sequence size as \u003cem\u003eSchaalia canis\u003c/em\u003e (Accession No: MN585806); and 3ZN2019 with 761bp sequence size as \u003cem\u003eStreptococcus hyovaginalis\u003c/em\u003e (Accession No: MN585807). Isolates 4ZN2019 which has 768bp sequence, and 5ZN2019 with 757bp sequence sizes were respectively identified as \u003cem\u003eEnterococcus faecium\u003c/em\u003e (Accession No: MN585808) and \u003cem\u003eGemella palaticanis\u003c/em\u003e (Accession No: MN585809). The obtained bacterial sequences were deposited in the GeneBank (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://submit.ncbi.nlm.nih.gov/subs/?search=SUB6437530\u003c/span\u003e\u003cspan address=\"https://submit.ncbi.nlm.nih.gov/subs/?search=SUB6437530\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). The dendogram of the phylogenetic tree drawn from the five identified bacterial isolates and other related organisms earlier deposited from different parts of the world is presented as Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003eThe bacteria isolated in the present study exhibited varying degree of sensitivity to different antimicrobials. Among all the 11 antimicrobials tested, gentamicin proved to be the most effective as 89.25% of all the isolates tested were sensitive to it. This is followed by a cephalosporin, cefoxitin which showed 70.00% sensitivity, while the highest rate of resistance was recorded among the penicillins (penicillin, oxacillin, amoxicillin/clavulanic acid, and ampicillin) which had about 73.00% average resistance to all the isolates tested (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The detailed antimicrobial resistance pattern of the isolates is shown in Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e. Several categories of the isolates particularly \u003cem\u003eE. coli, Streptococcus\u003c/em\u003e species (other than \u003cem\u003eStrep\u003c/em\u003e.\u003cem\u003ehyovaginalis\u003c/em\u003e), and \u003cem\u003eEnterococcus\u003c/em\u003e species (other than\u003cem\u003eE. faecium\u003c/em\u003e) exhibited multidrug resistance (MDR), resisting at leastthree different classes of antimicrobials.Similarly, \u003cem\u003eMicrococcus\u003c/em\u003e species showed an extended spectrum of drug resistance (XDR) as they were resistant to at least one agent in nearly all the classes of antimicrobial tested.\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\u003eOccurrence of aerobic bacteria recovered from the vagina of apparently healthy and infertile bitches in Zaria, Nigeria\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\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=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eS/No\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eName of isolate\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003eHealthy Bitches (n\u0026thinsp;=\u0026thinsp;70)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003eInfertile Bitches (n\u0026thinsp;=\u0026thinsp;30)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eP-values\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFrequency\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePercentage (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eFrequency\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003ePercentage (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eStaphylococcus saprophyticus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e45.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e46.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e1.000\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eStaphylococcus aureus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e14.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e20.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.554\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eStaphylococcus schleiferi\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e8.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.174\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eStreptococcus hyovaginalis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.300\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOther \u003cem\u003eStreptococcus\u003c/em\u003e species\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e17.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e40.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.021٭\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCorynebacterium\u003c/em\u003e species\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e20.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e30.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.306\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eEnterococcus faecium\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.300\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eEnterococcus dispar\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.300\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOther \u003cem\u003eEnterococcus\u003c/em\u003e species\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e11.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.272\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eMicrococcus\u003c/em\u003e species\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e20.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.082\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eProteus vulgaris\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e8.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e16.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.298\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eEscherichia coli\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e10.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e1.000\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eBaccilus\u003c/em\u003especies\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e6.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.635\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eGemellapalaticanis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e10.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.025٭\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eSchaaliacanis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e6.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.088\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e103\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e66\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003eP-values with an asterisk٭are statistically significant (P˂0.05).\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\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\u003eMorphological and Biochemical characteristics of some aerobic Gram-positive bacteria isolated from the vagina of apparently healthy and infertile bitches in Zaria, Nigeria\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\u003eS/No\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eColonial morphology\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMicroscopic features\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eBiochemical tests\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eOrganism identified\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSmall, raised, β-hemolytic colonies on blood agar\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGram positive cocci in cluster\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCatalase, coagulase, glucose, trehalose, maltose, mannitol, and sucrose positive; while negative for xylose, and arabinose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eStaphylococcus aureus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSmall, raised, β-hemolytic colonies on blood agar\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGram positive cocci in cluster\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCatalase, glucose, trehalose, mannitol, and sucrose positive; while negative for coagulase, maltose, xylose, and arabinose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eS. saprophyticus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSmall, raised, whitish β-hemolytic colonies\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGram positive cocci in cluster\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCatalase, glucose, and sucrose positive; while negative for coagulase, trehalose, mannitol, maltose, xylose, and arabinose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eS. schleiferi\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGlossy-dew drop-like, α, and β-hemolytic colonies\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGram positive cocci in short and long chains\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePositive for glucose; and negative for catalase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eStreptococcus\u003c/em\u003e sppother than \u003cem\u003eSt. hyovaginalis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGlossy-dew drop-like, γ-hemolytic colonies\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGram positive cocci in short chains and branches\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eGlucose, lactose, dextrose, raffinose, arabinose, and urease positive; while negative for catalase, coagulase, trehalose, mannitol, maltose, xylose, and sucrose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eEnterococcus\u003c/em\u003e species other than \u003cem\u003eE\u003c/em\u003e. \u003cem\u003efaecium\u003c/em\u003e and \u003cem\u003eE. dispar\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSmall, raised, β-hemolytic colonies with little yellowish pigmentation on blood agar\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGram positive cocci with tetrad appearance\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePositive for catalase, and glucose; but negative for coagulase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eMicrococcus\u003c/em\u003e species\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLarge, flat, β-hemolytic colonies with serrated edges\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGram positive rods in short chains\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCatalase, glucose, lactose, and urease positive; while negative for sucrose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eBaccilus\u003c/em\u003especies\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSmall, raised, moist, greyish, γ-hemolytic colonies\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGram positive, pleomorphic rods with Chinese letters appearance\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePositive for catalase, glucose, and lactose; but negative for coagulase, sucrose, indole, and urease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eCorynebacterium\u003c/em\u003e species\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePin-point β-hemolytic colonies on blood agar\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGram positive cocci mostly in pairs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e٭٭٭\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eGemellapalaticanis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTiny granular-like, β-hemolytic colonies\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGram positive coccobaccilli in chains\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e٭٭٭\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eSchaaliacanis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003eNote: Organisms with٭٭٭ were identified via molecular characterization\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \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\u003eBiochemical characteristics of some aerobic Gram-negative bacteria isolated from the vagina of apparently healthy and infertile bitches in Zaria, Nigeria\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"16\"\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 \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c14\" colnum=\"14\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c15\" colnum=\"15\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c16\" colnum=\"16\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eS/No.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"14\" nameend=\"c15\" namest=\"c2\"\u003e \u003cp\u003eBiochemical Tests Conducted\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c16\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eName of Bacterium\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCAT\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOF\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTSI\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eNIT\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eIND\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eURE\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eCIT\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eMR\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eVP\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003eMOT\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e \u003cp\u003eOXD\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c13\"\u003e \u003cp\u003eGLU\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c14\"\u003e \u003cp\u003eSUC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c15\"\u003e \u003cp\u003eLAC\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eF\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eK/A.H\u003csub\u003e2\u003c/sub\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e-/+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e\u003cem\u003eProteus vulgaris\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eF\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eA/A. G\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e\u003cem\u003eEscherichia coli\u003c/em\u003e\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\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eKey: CAT\u0026thinsp;=\u0026thinsp;catalase, OF\u0026thinsp;=\u0026thinsp;oxidative/fermentative, TSI\u0026thinsp;=\u0026thinsp;triple sugar iron, NIT\u0026thinsp;=\u0026thinsp;nitrate reduction, IND\u0026thinsp;=\u0026thinsp;indole, URE\u0026thinsp;=\u0026thinsp;urease, CIT\u0026thinsp;=\u0026thinsp;citrate, MR\u0026thinsp;=\u0026thinsp;methyl red, VP\u0026thinsp;=\u0026thinsp;Voges Proskauer, MOT\u0026thinsp;=\u0026thinsp;motility, OXD\u0026thinsp;=\u0026thinsp;oxidase, GLU\u0026thinsp;=\u0026thinsp;glucose, SUC\u0026thinsp;=\u0026thinsp;sucrose, LAC\u0026thinsp;=\u0026thinsp;lactose, F\u0026thinsp;=\u0026thinsp;fermenter, A\u0026thinsp;=\u0026thinsp;acid, K\u0026thinsp;=\u0026thinsp;alkaline, G\u0026thinsp;=\u0026thinsp;gas, H\u003csub\u003e2\u003c/sub\u003eS\u0026thinsp;=\u0026thinsp;hydrogen sulphide, -=negative, and +\u0026thinsp;=\u0026thinsp;positive\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003cp\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\u003ePercentage antimicrobial susceptibility profile of aerobic bacteria isolated from the vagina of apparently healthy and infertile bitches in Zaria, Nigeria\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"25\"\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 \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c14\" colnum=\"14\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c15\" colnum=\"15\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c16\" colnum=\"16\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c17\" colnum=\"17\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c18\" colnum=\"18\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c19\" colnum=\"19\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c20\" colnum=\"20\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c21\" colnum=\"21\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c22\" colnum=\"22\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c23\" colnum=\"23\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c24\" colnum=\"24\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c25\" colnum=\"25\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAntibiotic (\u0026micro;g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e \u003cp\u003e\u003cem\u003eStaphylococcus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c7\" namest=\"c5\"\u003e \u003cp\u003e\u003cem\u003eStreptococcus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c10\" namest=\"c8\"\u003e \u003cp\u003e\u003cem\u003eEnterococcus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c13\" namest=\"c11\"\u003e \u003cp\u003e\u003cem\u003eMicrococcus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c16\" namest=\"c14\"\u003e \u003cp\u003e\u003cem\u003eCorynebacterium\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c19\" namest=\"c17\"\u003e \u003cp\u003e\u003cem\u003eSchaalia\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c22\" namest=\"c20\"\u003e \u003cp\u003e\u003cem\u003eGemella\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c25\" namest=\"c23\"\u003e \u003cp\u003e\u003cem\u003eEscherichia\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eS\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eI\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003eR\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003eS\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003eI\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003eR\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003eS\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003eI\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003eR\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u003cb\u003eS\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cb\u003eI\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003eR\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e\u003cb\u003eS\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e\u003cb\u003eI\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e\u003cb\u003eR\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e\u003cb\u003eS\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e\u003cb\u003eI\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003e\u003cb\u003eR\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003e\u003cb\u003eS\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e\u003cb\u003eI\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e\u003cb\u003eR\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003e\u003cb\u003eS\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003e\u003cb\u003eI\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003e\u003cb\u003eR\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTE (30)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCN (10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e94\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\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAMC (30)\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\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFOX (30)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOX (1)\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\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVA (30)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003e70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAMP (10)\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\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e80\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\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eP (10)\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\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eE (15)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eS (10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCIP (5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\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\u003eNote: Staphylococcus = (\u003cem\u003eS. aureus\u003c/em\u003e \u0026amp; \u003cem\u003eS. saprophyticus\u003c/em\u003e), Streptococcus = (\u003cem\u003eStrep. Hyovaginalis\u003c/em\u003e \u0026amp; other species), Enterococcus = (\u003cem\u003eE. faecium, E. dispar\u003c/em\u003e, \u0026amp; other species), Micrococcus = (\u003cem\u003eMicrococcus\u003c/em\u003e species), Corynebacterium (\u003cem\u003eCorynebacterium\u003c/em\u003e species), Schaalia = (\u003cem\u003eS\u003c/em\u003e. \u003cem\u003ecanis\u003c/em\u003e), Gemella = (\u003cem\u003eG\u003c/em\u003e. \u003cem\u003epalaticanis\u003c/em\u003e), Escherichia = (\u003cem\u003eE\u003c/em\u003e. \u003cem\u003ecoli\u003c/em\u003e), TE\u0026thinsp;=\u0026thinsp;Tetracycline, CN\u0026thinsp;=\u0026thinsp;Gentamicin, AMC\u0026thinsp;=\u0026thinsp;Amoxicillin/Clavulanicacid, FOX\u0026thinsp;=\u0026thinsp;Cefoxitin, OX\u0026thinsp;=\u0026thinsp;Oxacillin, VA\u0026thinsp;=\u0026thinsp;Vancomycin, AMP\u0026thinsp;=\u0026thinsp;Ampicillin, P\u0026thinsp;=\u0026thinsp;Penicillin, E\u0026thinsp;=\u0026thinsp;Erythromycin, S\u0026thinsp;=\u0026thinsp;Streptomycin, CIP\u0026thinsp;=\u0026thinsp;Ciprofloxacin. Using the CLSI, 2016: R\u0026thinsp;=\u0026thinsp;Resistant, I\u0026thinsp;=\u0026thinsp;Intermediate, and S\u0026thinsp;=\u0026thinsp;Sensitive\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\u003eAntimicrobial resistance patterns of some aerobic bacteria isolated from the vagina of apparently healthy and infertile bitches in Zaria, Nigeria\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=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eS/No\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eName of Isolate\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eResistance Pattern\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eClass of Antimicrobials Resisted\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eStaphylococcus saprophyticus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTE-AMC-OX-AMP-P\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eStaphylococcus aureus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAMC-AMP-P-OX\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eStreptococcus hyovaginalis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAMC-OX-AMP-P\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOther \u003cem\u003eStreptococcus\u003c/em\u003e species\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTE-AMC-OX-AMP-P-E\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCorynebacterium\u003c/em\u003e species\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAMC-OX-AMP-P\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eEnterococcus faecium\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAMC-OX-AMP-P\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eEnterococcus dispar\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTE-AMC-OX-VA-AMP-P-S\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOther \u003cem\u003eEnterococcus\u003c/em\u003e species\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTE-AMC-OX-VA-AMP-P\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eMicrococcus\u003c/em\u003e species\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTE-CN-AMC-FOX-OX-AMP-P-E-S-CIP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eEscherichia coli\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAMC-OX-VA-AMP-P-E\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eGemellapalaticanis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTE-AMP-P\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eSchaaliacanis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAMC-OX-VA-AMP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eTE\u0026thinsp;=\u0026thinsp;Tetracycline, AMC\u0026thinsp;=\u0026thinsp;Amoxicillin/Clavulanicacid, OX\u0026thinsp;=\u0026thinsp;Oxacillin, AMP\u0026thinsp;=\u0026thinsp;Ampicillin, P\u0026thinsp;=\u0026thinsp;Penicillin, E\u0026thinsp;=\u0026thinsp;Erythromycin, VA\u0026thinsp;=\u0026thinsp;Vancomycin,S\u0026thinsp;=\u0026thinsp;Streptomycin, CN\u0026thinsp;=\u0026thinsp;Gentamicin,FOX\u0026thinsp;=\u0026thinsp;Cefoxitin, and CIP\u0026thinsp;=\u0026thinsp;Ciprofloxacin\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThis study assessed the presence and antibiogram of aerobic bacterial populations in the vagina of apparently healthy and infertile bitches. The findings showed some level of variation in number and type of bacteria between the two different groups. The incidence of common pathogens, particularly, \u003cem\u003eStaphylococcus saprophyticus, S. aureus, E. coli, Streptococcus\u003c/em\u003es pecies, \u003cem\u003eBacillus\u003c/em\u003e species, \u003cem\u003eEnterococcus\u003c/em\u003e, and \u003cem\u003eCorynebacterium\u003c/em\u003e species was similar between the healthy and infertile bitches. This outcome corelates well with the results of other parallel studies (Findik et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Groppetti et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Antonov, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Golińskaet al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) which documented \u003cem\u003eStaphylococcus, Streptococcus, Enterococcus, Corynebacterium, Proteus\u003c/em\u003e and \u003cem\u003eEscherichia\u003c/em\u003e to be the most common genera colonizing the vagina of dogs. However, the present study established significant difference in the overall bacterial occurrence rate between healthy and infertile bitches; This contradicts the studies of Hutchins et al. (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) and Golińska et al. (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) who reported that the total number of bacteria in the vagina of healthy and non-healthy dogs at different stages of estrus cycle was almost the same when conducting similar studies. The exact reason for this variation is not clear, but one possible explanation is the relatively large sample size (100) of the current study as against the 44 and 39 respectively utilized by the two previous studies. This is to say that, a probable significant difference in the prevalence of specific bacteria would have been identified if much higher sample sizes were employed in those preceding research.\u003c/p\u003e \u003cp\u003eThe restricted vaginal presence of \u003cem\u003eStaphylococcus schleiferi\u003c/em\u003e within healthy bitches could have a protective competitive role against potentially more dangerous pathogens like \u003cem\u003eGemella palaticanis\u003c/em\u003e and \u003cem\u003eSchaalia canis\u003c/em\u003e which were never isolated from the healthy bitches in the present study. This statement is in line with the findings of Feldman and Nelson (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2003\u003c/span\u003e) who documented that normal vaginal microbiota has been shown to protect genito-urinary tract against pathogenic bacteria through nutrient and epithelial cell receptors competition. Conversely, the relative higher incidence of \u003cem\u003eProteus vulgaris\u003c/em\u003e observed in infertile bitches agrees with the findings of Golińska et al. (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) who reported significantly higher prevalence of Gram-negative rods (other than \u003cem\u003eE. coli\u003c/em\u003e) in the vagina of dogs with genital tract infections. Likewise, the significantly higher occurrence rate of \u003cem\u003eStreptococcus\u003c/em\u003e species in infertile bitches might be responsible for the infertility in those dogs. This is because β-haemolytic \u003cem\u003eStreptococci\u003c/em\u003e, as well as \u003cem\u003eProteus, E. coli\u003c/em\u003e and \u003cem\u003eEnterococcus\u003c/em\u003e species were previously associated with infertility in bitches (John et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Similarly, the significantly higher occurrence of \u003cem\u003eG\u003c/em\u003e. \u003cem\u003epalaticanis\u003c/em\u003e in the anterior vagina of infertile bitches could be associated with the development of genital tract infection that might eventually result in infertility, however, a focused case control study would be required to ascertain this hypothesis.\u003c/p\u003e \u003cp\u003eBacteria of the Genera \u003cem\u003ePasteurella, Pseudomonas, Klebsiella\u003c/em\u003e, and \u003cem\u003eEnterobacter\u003c/em\u003e which were previously reported to reside within dog\u0026rsquo;s reproductive tract (John et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Antonov, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) were not identified in the present study. This variation could be due to the canine intrinsic and genetic variability, indiscriminate use of antibiotics in the study area, or inherent limitations of culture-based systems compared to molecular identification as similarly documented by Lyman et al. (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Another likely cause of this disparity in the type of vaginal microflora is the possible sample contamination from the vestibule or vulva especially in the case of John et al. (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) who used vaginal discharges rather than guarded aseptic swab collection from the anterior vagina.\u003c/p\u003e \u003cp\u003eVarying grades of sensitivity to different antimicrobials were reported in the isolates from both healthy and infertile bitches with aminoglycoside, gentamicin being the most effective as equally observed by John et al. (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Likewise, several isolates displayed resistance to at least two classes of antibiotics termed multidrug resistant phenotypes. In this study, the β-lactam antibiotics; ampicillin, penicillin, oxacillin, and amoxicillin/clavulanic acid which are known to be effective against many Gram-positive pathogens were surprisingly found to be resistant (with average resistance of 81.25%, 68.75%, 62.50% and 79.38% respectively). Gram-negative bacteria, (such as \u003cem\u003eE. coli\u003c/em\u003e) have been established to develop resistance to multiple antibiotics mostly due to their inherent resistance to macrolide antibiotics (such as erythromycin), efflux system, as well as decreased drug uptake mediated by chromosomal mutations or via acquisition of resistance genes (Lartey et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e1994\u003c/span\u003e; Rossolini et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). However, the non-susceptibility of \u003cem\u003eE. coli\u003c/em\u003e to multiple antimicrobials observed in the current study is alarming. Consequently, the extremely drug-resistant \u003cem\u003eMicrococcus\u003c/em\u003e species most likely resulted from the acquisition of multiple resistance determinants over a long period sequel to the known indiscriminate prescription, acquisition and usage of antimicrobials in the study area (FMAEH, 2017; Ogwucheet al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Therefore, the multidrug resistance nature of \u003cem\u003eStreptococcus\u003c/em\u003e species (other than \u003cem\u003eStrep. hyovaginalis\u003c/em\u003e), \u003cem\u003eEnterococcus\u003c/em\u003e species (other than \u003cem\u003eE. faecium \u0026amp; E. dispar\u003c/em\u003e), as well as the extensive antimicrobial resistance of \u003cem\u003eMicrococcus\u003c/em\u003e species recorded in the current study highlight the need for an improved fight against antimicrobial resistance in the study area.\u003c/p\u003e \u003cp\u003eIn conclusion, this study has established the most common bacteria colonizing the anterior vagina of bitches in Zaria, Nigeria. The occurrence of aerobic bacteria was significantly higher in infertile bitches compared to the healthy ones suggesting the possibility of some identified pathogens being responsible for the infertility. Furthermore, varying degree of sensitivity to antimicrobials, and development of multiple drug resistance were identified among the isolates highlighting the need for rational and judicious selection of drugs when managing infertility in dogs. We recommend dog breeders to screen their bitches and studs before breeding especially for the presence of possible multi-drug resistant bacteria which can pose a threat to pregnancy. This should be preceded by thorough clinical and vaginal-cytological examinations.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors wish to thank Dr. K. H. Ahmad for his help in reading and correcting the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eThe authors declare that no specific funds, grants, or other support were received\u0026nbsp;\u003c/em\u003efrom funding agencies in the public, commercial, or not-for-profit sectors \u003cem\u003eduring the preparation of this manuscript.\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eThe authors have no relevant financial or non-financial interests to disclose.\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eAuthors\u0026rsquo; Contributions\u0026nbsp;\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eAll\u003c/em\u003e the authors have contributed substantially to the concept, design, and execution of the study.\u0026nbsp;\u003cem\u003eMaterial preparation, data collection and analysis were performed by Dr. B.N. Umar, Dr. A.N. Kakudi, and Dr. A.O. Ahmed. The first draft of the manuscript was written by Dr. P.H. Mamman and all the authors reviewed the previous versions of the manuscript. All authors read and approved the final manuscript.\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eThe datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics Approval\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe bitches used in this study were managed in accordance with standard ethics. \u003cem\u003eThe study was performed in line with the principles of the declaration of Helsinki;\u0026nbsp;\u003c/em\u003eand it was approved by the Ahmadu Bello University Committee on Animal Use and Care (ABUCAUC).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformed Consent\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAhmed OA, Mamman PH, Raji MA, Kwanashie, CN, Raufu IA,Aremu A (2017) Distribution of Virulence Genes in Salmonella Serovars isolated from poultry farms in Kwara State, Nigeria. Ceylon Journal of Science, 46, 69\u0026ndash;76. https://doi.org/10.4038/cjs.v46i4.7469.\u003c/li\u003e\n \u003cli\u003eAllen WE,Dagnall\u0026nbsp;GJR (1982) Some observations on the aerobic bacterial flora of the genital tract of the dog and bitch. Journal of Small Animal Practice, 23, 325\u0026ndash;335.\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eAnonymous (2021)High Street Epping Veterinary Clinic\u003c/strong\u003e\u003cstrong\u003e:\u0026nbsp;\u003cem\u003eFactors Leading\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eto Infertility\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003ein Bitches\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003e.\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003eRetrieved on October 28, 2021 from\u0026nbsp;\u003c/strong\u003ehttps://www.highstreeteppingvetclinic.com/factors-leading-to-infertility-in-bitch.pml\u003c/li\u003e\n \u003cli\u003eAntonov A (2019) Normal and Pathological Vaginal Microflora in the Bitch-A Review. Journal of Advances in Microbiology,15, 1-5.DOI: 10.9734/JAMB/2019/v15i130074\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eBauer\u0026nbsp;AW, Kirby WM, Sherris JC,Turck M (1966) Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology, 45, 493\u0026ndash;496.\u003c/li\u003e\n \u003cli\u003eCLSI. M100S27: (2016) Performance Standards for Antimicrobial Susceptibility Testing. Clinical Laboratory Standard Institute2016;M100S27\u003c/li\u003e\n \u003cli\u003eCottral GE (1978) \u003cem\u003eManual of Standardised Methods for Veterinary Microbiology\u003c/em\u003e. National Research Council. Subcommittee on Standardized Methods for Veterinary Microbiology. Comelll University Press, Ithaca, New York, Pp. 731.\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eCowan ST, Steel KJ (2002) Manual for The Identification of Medical Bacteria. 2nd edition. Cambridge University press, United Kindom, Pp. 217.\u003c/li\u003e\n \u003cli\u003ede Paula Antunes JMA, de Carvalho Freire DA, de Medeiros Oliveira IVP, Moura GHF, de Castro Demoner L, Ferreira HIP (2016) Infectious Causes of Abortion, Stillbirth and Neonatal Death in Bitches. In: Abdelhay Essayed K.H (Ed). \u003cem\u003eCanine Medicine: Recent Topics and Advanced Research\u003c/em\u003e. Janeza Trdine 9, 5100, Rijeka, Croatia. pp.55-73.\u003c/li\u003e\n \u003cli\u003eDoig\u0026nbsp;PA, Ruhnke HL,Bosu WTK (1981) The Genital mycoplasma and ureaplasma flora of healthy and diseased dogs. Canadian Journal of Comparative Medicine, 45, 223\u0026ndash;231.\u003c/li\u003e\n \u003cli\u003eFederal Ministries of Agriculture, Environment and Health; (FMAEH 2017) Antimicrobial Use and Resistance in Nigeria: Situation Analysis and Recommendations. Pp. 1-105\u003c/li\u003e\n \u003cli\u003eFeldman EC, Nelson RW (2003) Canine and feline endocrinology and reproduction. 3rd Ed. Saunders, Philadelphia, USA, Pp. 1044-1104.\u003c/li\u003e\n \u003cli\u003eFindik\u0026nbsp;M, Maral N, Keskin O, Kalender H, Erdeger J, Aslan S (2003) The relationship between the stages of sexual cycle, the pregnancy and postpartum periods and vaginal flora in kangal breed bitches. Turkish Journal of Veterinary and Animal Science, 27, 761-765.\u003c/li\u003e\n \u003cli\u003eFontbonne A (2011) Infertility in bitches and queens: recent advances.\u0026nbsp;\u003cem\u003eRevista\u003c/em\u003e\u003cem\u003eBrasileira de Reprodu\u0026ccedil;\u0026atilde;o\u003cem\u003eAnimal\u003c/em\u003e\u003cem\u003e,\u003c/em\u003e\u003c/em\u003e35(2), 202-209.\u003c/li\u003e\n \u003cli\u003eFreshman JL (1991) Clinical approach to infertility in the cycling bitch. Veterinary Clinics of North America: Small Animal Practice, 21(3), 427-435.\u003c/li\u003e\n \u003cli\u003eGavrilovic BB, Andersson K, Forsberg C (2008) Reproductive patterns in the domestic dog: A retrospective study of the Drever breed. \u003cem\u003eTheriogenology\u003c/em\u003e, 70(5), 783-794.\u003c/li\u003e\n \u003cli\u003eGolińska\u0026nbsp;E, Sowińska N, Tomusiak-Plebanek A, Szydło M, Witka N, Lenarczyk J, Strus M (2021) The vaginal microflora changes in various stages of the estrous cycle of healthy female dogs and the ones with genital tract infections. BMCVeterinaryResearch, 17(1), 1-8.\u003c/li\u003e\n \u003cli\u003eGroppetti D, Pecile A, Barbero C, Martino PA (2012) Vaginal bacterial flora and cytology in proestrous bitches: Role on fertility.\u0026nbsp;\u003cem\u003eTheriogenology\u003c/em\u003e 77(8), 1549-1556\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eHollinshead FK, Hanlon DW (2017) Factors affecting the reproductive performance of bitches: A prospective cohort study involving 1203 inseminations with fresh and frozen semen. Theriogenology, 101, 62\u0026ndash;72.\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eHutchins\u0026nbsp;RG, Vaden SL, Jacob ME, Harris TL, Bowles KD, Wood MW, Bailey CS (2014) Vaginal microbiota of spayed dogs with or without recurrent urinary tract infections. Journal of Veterinary Internal Medicine, 28(2), 300\u0026ndash;304.\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eJohn A, Nair DR, Sreejith JR, Deepthi L, Praseeda R, Ajitkumar G (2011) Vaginal Microflora and its Antibiogram in Infertile Bitches. Online Veterinary Journal, 6(1), e79.\u003c/li\u003e\n \u003cli\u003eJohnston SD, Kustritz MVR, Olson PNS (2001) Canine and Feline Theriogenology. Saunders, Philadelphia, USA, 592p\u003c/li\u003e\n \u003cli\u003eKimura MA (1980) Simple Method for Estimating Evolutionary Rates of Base Substitutions Through Comparative Studies of Nucleotide Sequences. Journal of Molecular Evolution, 16, 111-120.\u003c/li\u003e\n \u003cli\u003eLartey PA, Nellans HN, Tanaka SK (1994) New developments in macrolides: structures and antibacterial and prokinetic activities. \u003cem\u003eAdvances in Pharmacology (San Diego, Calif.)\u003c/em\u003e, Elsevier\u0026nbsp;Academic Press, Inc. Pp. 307-343.\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eLyman CC, Holyoak GR, Meinkoth K, Wieneke X, Chillemi KA, DeSilva U (2019) Canine endometrial and vaginal microbiomes reveal distinct and complex ecosystems. PLoS ONE 14(1), e0210157. doi.org/10.1371/journal.pone.0210157\u003c/li\u003e\n \u003cli\u003eOgwuche\u0026nbsp;A, Ekiri AB, Endacott I, Maikai BV, Idoga ES, Alafiatayo R, Cook AJ (2021) Antibiotic use practices of veterinarians and para-veterinarians and the implications for antibiotic stewardship in Nigeria. Journal of the South African Veterinary Association,92, 1-14.\u003c/li\u003e\n \u003cli\u003eOlson\u0026nbsp;PS (1975) \u003cem\u003eSteptococcuscanis\u003c/em\u003e: An isolate from the canine uterus. Veterinary Medicine Small Animal Clinician, 70, 933\u0026ndash;934.\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eOlson PS, Mather EC (1978) Canine vaginal and uterine bacterial flora.\u0026nbsp;Journal of the American Veterinary Medical Association,\u0026nbsp;172, 708\u0026ndash;711.\u003c/li\u003e\n \u003cli\u003eRossolini GM, Arena F,Giani T (2017)\u0026nbsp;Mechanisms of Antibacterial Resistance.\u0026nbsp;In:\u0026nbsp;Jonathan Cohen, William Powderly, Steven Opal (Eds.)\u0026nbsp;\u003cem\u003eInfectious Diseases\u003c/em\u003e (4\u003csup\u003eth\u003c/sup\u003eEdition), Elsevier Ltd.\u0026nbsp;2, Pp. e1181-1196.\u003c/li\u003e\n \u003cli\u003eRuncan EE, da Silva MAC (2018) Whelping and dystocia: maximizing success of medical management. Topics in Companion Animal Medicine, 33(1), 12-16.\u003c/li\u003e\n \u003cli\u003eThomas J, Thanigaivel S, Vijayakumar S, Acharya K, Shinge D, Seelan TSJ, Mukherjee A, Chandrasekaran N (2014) Pathogenecity of Pseudomonas aeruginosa in Oreochromis mossambicus and treatment using lime oil nanoemulsion. Colloids and Surfaces B: Biointerfaces, 116, 372-377.\u003c/li\u003e\n \u003cli\u003eWagner J, Short K, Catto-Smith AG, Cameron DJ, Bishop RF, Kirkwood CD (2008) Identification and characterisation of Pseudomonas 16S ribosomal DNA from ileal biopsies of children with Crohn\u0026apos;s disease. \u003cem\u003ePloS one\u003c/em\u003e, 3(10), e3578.\u003c/li\u003e\n \u003cli\u003eZubair M, Ubaid-ur-Rehman S, Sajid SM (2014) Causes of infertility in bitch. Advances in Animaland Veterinary Sciences, 2(10), 565-573.\u003c/li\u003e\n\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":"Aerobes, Bitches, Gemella palaticanis, Schaalia canis, 16S rRNA sequencing","lastPublishedDoi":"10.21203/rs.3.rs-1580205/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-1580205/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eInfertility is a major constrain to a successful dog breeding in many developing countries like Nigeria. The aim of this study therefore, was to establish the aerobic microflora of the vagina of Nigerian bitches. Bacteria were isolated and characterized including susceptibility to commonly used antimicrobials. A total of 169 bacteria were isolated; of which 103 (60.9%) were from apparently healthy bitches and 66 (39.1%) from bitches with reproductive anomalies. These include \u003cem\u003eStaphylococcus saprophyticus\u003c/em\u003e (27.2%), \u003cem\u003eStreptococcus\u003c/em\u003e species (14.8%), and \u003cem\u003eCorynebacterium\u003c/em\u003e species (13.6%). Also, \u003cem\u003eGemella palaticanis\u003c/em\u003e (1.8%), and \u003cem\u003eSchaalia canis\u003c/em\u003e (1.3%) which are new to the study area were isolated for the first time in Nigeria. Nearly all the infertile bitches (90.1%) had mixed infection which might be responsible for their illness. Gentamicin was the most effective drug. \u003cem\u003eMicrococcus\u003c/em\u003e species and \u003cem\u003eEscherichia coli\u003c/em\u003e exhibited multiple drug resistance. Thus, screening of dogs for pathogenic bacteria before breeding is recommended.\u003c/p\u003e","manuscriptTitle":"Phenotypic and Molecular Characterization of Aerobic Bacteria from the Vagina of Apparently Healthy and Infertile Bitches in Zaria, Nigeria","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-04-25 21:25:27","doi":"10.21203/rs.3.rs-1580205/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":"95b3aebc-13d3-4821-a1e1-f578bc47fb01","owner":[],"postedDate":"April 25th, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2022-10-10T08:14:13+00:00","versionOfRecord":[],"versionCreatedAt":"2022-04-25 21:25:27","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-1580205","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-1580205","identity":"rs-1580205","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}