Comprehensive Analysis of Escherichia coli in Bovine Urinary Tract Infections: Serotypes, Antibiotic Resistance, and Virulence Gene Profiles

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Aim of the present study was to characterize Escherichia coli ( E. coli ) strains isolated from urinary tract infected (UTI) bovines and to understand the distribution of antimicrobial resistance (AMR) in northern part of India. A cross-sectional study was carried out with sampling 17 randomly selected husbandry setups in 6 districts representing two agroclimatic zones. Out of 254 suspected animals from UTI, 105 animals found to be affected with UTI based on clinical examination, routine urine, culture testing and ultrasonography. Among the 105 affected animals, E. coli was isolated from 47 animals turning out to be 44.76% positivity. A total of 13 serotypes of E. coli were identified with maximum occurrence of O156 (23.40%) serotype. Among 13 serotypes isolated, five serotypes viz. O5, O128, O83, O11, and O159 are zoonotic important and are of public health concern. Antimicrobial resistance gene profiling depicted 100% occurrence of tet (A) and gyr A phenotype in E. coli isolates. Remarkably, meropenem demonstrated the highest efficacy against the isolates, with 89.6% sensitivity. Unfortunately, 85.10% of the tested isolates displayed multidrug resistance, and all isolates carried the Fim H (100%) gene and two isolates carried hly A gene. Our findings emphasize genetic diversity and wide dispersion of zoonotically significant multidrug-resistant, virulent uropathogenic E. coli strains among dairy animals. Health sciences/Molecular medicine Health sciences/Nephrology Health sciences/Urology Antimicrobial resistance Bovine E. coli Fim H Serotype Urinary tract infections Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 INTRODUCTION In ruminants, Urinary tract infection (UTI) typically occurs as cystitis, ureteritis and pyelonephritis which most often result from ascending infection with Corynebacterium renale or Escherichia coli (Smith et al., 2021 ) causing economic losses via reduction in reproduction and production of the farm (Yeruham et al. 2006 ). Uropathogenic E. coli (UPEC) pathotypes are responsible for causing extra-intestinal E. coli infections, primarily urinary tract infections with specific virulence attributes encoded on genetic elements. Uropathogenic E. coli (UPEC) strains have shown certain virulent properties, including iron uptake systems, adhesins, specific O: K: H serotypes and synthesis of cytotoxins that contribute to colonization and invasion of the bacterium (Janke et al., 2001 ). Also, successful colonization of UPEC in the urinary tract depends on the expression of fimbria adhesion proteins, which facilitate attachment of the bacterium to the urothelium, and on the presence of specific bacterial genes that encode virulence factors (Ristow and Welch, 2016 ). The role of E. coli and various serotypes involved in the pathogenesis of bovine UTI is poorly understood and documented. Traditionally it was believed that urinary tract of healthy individual is sterile but recent studies of the bovine urinary microbiota pointed out the presence of E. coli as resident members of this niche (Giannattasio-Ferraz et al., 2022 ), suggesting dysbiosis as the mechanism of urinary tract infection rather than external sources (Garretto et al., 2020 ). Antimicrobial resistance (AMR) has been recognized as one of the most significant threats to the health of people and milk-producing animals (Gemeda et al ., 2023). Investigating antimicrobial resistance (AMR) in E. coli is crucial because the environmental transmission of AMR is likely more significant in E. coli than in other microbiota members and it can survive in the environment for extended periods (Barrera et al., 2019 ). Moreover, much of the AMR in E. coli is carried on mobile genetic elements that can be transferred between bacteria, facilitating the rapid spread and persistence of resistance genes across different bacterial species (horizontal gene transfer) (Johnson et al., 2012 ). As a result, AMR in E. coli is considered a major public health threat. This study aimed to report the occurrence of E. coli in UTI affected bovines as well as to evaluate the presence of resistance and virulence genes of E. coli isolated from dairy animals suffering from UTI. MATERIAL AND METHODS Study area: Various animal husbandry setups were selected randomly across six Haryana districts spanning two agro-climatic zones (North-eastern and South- western zone) as illustrated in Fig. 1 , along with UTI affected bovine presented at Veterinary Teaching Hospital, Hisar. Present study was conducted with ethical approval from the IAEC, reference number VCC/IAEC/2022/1624 + 51. All methods were performed in accordance with relevant guidelines and regulations (Appendix 1). Animal Selection: Samples were randomly collected from suspected UTI cases across various animal husbandry setups, based on case history and a slightly modified version of the Acute Cystitis Symptom Score (ACSS) Index (Alidjanov et al., 2018 ) (Table 1 ). Each parameter was scored from 0 to 2 and sum of scores yielded a total cumulative score. Patient with cumulative score 6 and above were selected for sample collection. Table 1 Criteria for assessing severity of bovine cystitis (USQOLAT) Domain Symptoms Nil (0)/No Moderate (1)/Yes Severe (2) Typical Frequent voiding of small urine volume Painful/difficult urination/stranguria Hematuria 2-3times/day during urination Every time Suprapubic discomfort/flank pain Diurnal temperature variation Tail swishing Differential Fever Vaginal/urethral discharge Prolapse Ist degree 2nd degree Isolation and identification E. coli : Two hundred and fifty-four urine samples were collected aseptically using urinary catheter and were inoculated on MacConkey agar at 37° C for 24 h. After overnight incubation at 37° C typical pink colonies on MacConkey agar were Gram-stained and then single pure colony was transferred to eosin-methylene-blue (EMB) agar for incubation at 37° C overnight. Colonies with typical metallic sheen on EMB agar, were considered as E. coli -positive and transferred to nutrient agar for additional confirmatory biochemical tests (IMViC tests), molecular tests and serotyping. DNA Extraction and E. coli Confirmation : Presumptive E. coli isolates were molecularly confirmed by PCR amplification of universal stress protein (usp A gene). The PCR was performed using primer pair as depicted in Table 2 . The DNA extracted from ATCC 25922 was used as positive control for E. coli . PCR were carried out in 12.5µl total volume consisted of 7µl of master mix, 0.5 µl of 10 µM solution of each primer (forward and reverse primer), 1µl of DNA sample and remaining nuclease free water to make 12.5 µl. PCR amplifications started with initial denaturation at95 ºC for 2min, 30 cycles of denaturation at 94 ºC for 2 min, annealing at 58 ºC for 1min, extension at 72 ºC for 1min, and final extension at 72 ºC for 5min. Table 2 Primer pairs used for the characterization of E. coli (Osek, 2001) S. No. Description Gene Primers (5’-3’) Product size (bp) 1 Universal stress protein Usp A F-CCGATACGCTGCCAATCAGT R-ACGCAGACCGTAAGGGCCAGAT 884 Serotyping: After confirmation of isolates as E. coli , isolates were sent to the National Escherichia and Salmonella Centre, Kasauli (H.P) for final serotyping. Two individually similar colonies from each of the purified cultures of identified isolates were inoculated in semisolid media and incubated at 37ºC overnight. One tube of each culture was sent to the National Escherichia and Salmonella Centre, Kasauli (H.P) for serotyping, other tube was kept at 4ºC till further use. Kauffman- White scheme was consulted and organisms were assigned to the E . coli serotype. Detection of virulent and antimicrobial resistance genes: Virulent genes including Pap GII, hlyA, fimH, Usp and antimicrobial-resistant genes, including blaSHV, aac (3)-IV, tetA, tetB, gyrA, and floR, were amplified via 30 cycles of PCR amplification using the specific primers as depicted in Table 3 . The temperature profile was as follows: DNA denaturation at 94°C for 1min, followed by annealing at the specific temperature for each primer set for 30 s, extension at 72°C for 60 s, and a final extension at 72°C for 10 min. Electrophoresis was carried out at 85 V/cm of gel in 1X TAE running buffer in mini gel electrophoresis assembly, using power supply for 45 minutes to one hour. The amplified product was detected with the help of good view dye in agarose gel electrophoresis. The DNA bands were observed under the UV transilluminator. Table 3 Annealing temperature and sequence of Oligonucleotide primers used for amplification of antibiotic resistance and virulent gene of E. coli detection S.No. Gene Description Primers (5’-3’) Product size (bp) Annealing temperature Reference 1 Adhesins (papGII) Class II P pili F: GGGATGAGCGGGCCTTTGAT R: CGGGCCCCCAAGTAACTCG 190 57 ̊ C Nam et al. ( 2013 ) 2 Toxins (hlyA) α-Hemolysin F: AACAAGGATAAGCACTGTTCTGGCT R: ACCATATAAGCGGTCATTCCCATCA 1177 54 ̊C Self- designed 3 Adhesins (fimH) Type 1 fimbria F: ATGAAACGAGTTATTACCCTGTT R: TATTGATAAACAAAAGTCACGCC 900 58 ̊C Mohammed et al. ( 2020 ) 4 Miscellaneous (usp) Uropathogen-specific protein F: ACATTCACGGCAAGCCTCAG R: AGCGAGTTCCTGGTGAAAGC 440 55 ̊C Nam et al. ( 2013 ) 5. blaSHV Betalactam (Ampicillin) F: TCGCCTGTGTATTATCTCCC R: CGCAGATAAATCACCACAATG 768 55 ̊C Rahman et al. ( 2022 ) 6. aac (3)- IV Aminoglycosides (Gentamicin) F: CTTCAGGATGGCAAGTTGGT R: TCATCTCGTTCTCCGCTCAT 286 52 ̊C Rahman et al. ( 2022 ) 7. FloR Chloramphenicol F: TATCTCCCTGTCGTTCCAG R: AGAACTCGCCGATCAATG 399 60̊ C Mohammed et al. ( 2020 ) 8. tet (A) Tetracyclines F: GCTACATCCTGCTTGCCTTC R: CATAGATCGCCGTGAAGAGG 210 62 ̊C Mustapha et al . (2018) 9. GyrA Quinolones F: ACGTACTAGGCAATGACTGG R: AGAAGTCGCCGTCGATAGAAC 189 51 ̊C Self- designed Antibiotic Susceptibility Testing: The antimicrobial susceptibility test on the isolates was performed according to the National Committee for Clinical Laboratory Standards (CLSI, 2024) using the Kibry-Bauer disk diffusion test method on Muller-Hinton agar. The diameter of the zones of inhibition was measured and compared with zone size interpretation guidelines described by the Clinical Laboratory Standard Institute (CLSI, 2024) for the family Enterobacteriaceae and determined as sensitive, intermediate, or resistant. The isolated E. coli were tested for sensitivity to the most used antimicrobials. Statistical Analysis All demographic data and results were recorded on Excel spreadsheets. The data were analysed with the statistical software, IBM SPSS 25. RESULTS Characteristics of the collected sample: A total of 254 animals were screened for bovine UTI, 134 from tertiary setup of hospital (University referral hospital) and remaining 120 from different animal husbandry setups viz. Gaushala, organized dairy farm, unorganized dairy farm, and field hospitals; 30 animals each as shown in Fig. 2 . Out of 254, 105 animals were found positive for UTI based on culture testing, urinalysis, haemato- biochemical testing, renal biomarkers, and renal ultrasonography. Out of 120 animals suspected of having a urinary tract infection based on clinical severity scores, 60 were confirmed to be infected, emphasizing the sensitivity of the scoring system as a reliable tool for identifying UTIs. Occurrence of Escherichia coli : A total of 157 organisms were isolated from 105 UTI affected animals with 54 animals having single infections and 51 animals comprising mixed infections. Cultural and biochemical testing identified Escherichia coli as the predominant bacterium, accounting for 30% (47/105) of animals as shown in Fig. 3 . All 47 suspected E. coli isolates were subjected to PCR targeting the E. coli -specific Usp A gene for confirmation. PCR results for these 47 presumptive E. coli isolates showed an amplification band of 884 bp, specific to the Usp A gene, confirming them as E. coli illustrated in Fig. 4 . Serotyping results of E. coli isolates : Confirmed 47 Escherichia coli samples were characterized based on Serotyping at CRI Kasauli. Isolates were serotyped into different groups as shown in Table 4 . Out of total 13 serotypes, most frequently encountered serogroups of Escherichia coli was O156 (23.40%) followed by O84 and O101 (10.6%) each, O169 (6.38%), O128, O83, O120 and O18 (4.26%) each, O5, O134, O11, O88 and O159 (2.13%) each. Among the 13 serotypes identified, five serotypes O5, O128, O83, O11, and O159 were of zoonotic significance. Table 4 Relative frequency of serotypes of E. coli isolates (n = 47) based on serotyping from NSEC, Kasauli S. No. Serotypes Number Percentage 1 O169 3 6.38% 2 O84 5 10.6% 3 O5 (EHEC)* 1 2.13% 4 O134 1 2.13% 5 O128 (EPEC + ETEC + EHEC) * 2 4.26% 6 O156 11 23.40% 7 O101 5 10.63% 8 UT 8 17.02% 9 O83 (UPEC)* 2 4.26% 10 O120 2 4.26% 11 O11 (ETEC)* 1 2.13% 12 O18 2 4.26% 13 O88 1 2.13% 14 O159 (ETEC + EIEC) * 1 2.13% 15 Non- viable 2 4.26% * EHEC- Enterohemorrhagic E. coli ; EPEC- Enteropathogenic E. coli ; ETEC- Enterotoxigenic E. coli ; UPEC- Uropathogenic E. coli ; EIEC- Enteroinvasive E. coli Characterization of Uro-pathogenic Escherichia coli (UPEC) using PCR : For UPEC characterization, all the confirmed Escherichia coli isolates (n = 47) were subjected to PCR using a set of four virulence genes (VGs). PCR components and conditions were standardized using DNA extracted from colonies of confirmed Escherichia coli isolates in the current study. Out of 47 Escherichia coli isolates, only 2 isolates had hlyA gene while fimH present in 100% isolates (Fig. 5 ). Invitro antibiotic sensitivity based antimicrobial pattern of Escherichia coli isolates : Antimicrobial sensitivity pattern of E. coli isolates is represented in Fig. 6 . Overall maximum sensitivity of E. coli isolates was found towards meropenem (89.36%), followed by gentamicin (44.68%), amikacin (40.42%), ceftriaxone- sulbactam (34.04%), kanamycin, Co- trimoxazole/sulpha-trimethoprim and azithromycin each (31.91%), chloramphenicol (29.78%), cefoperazone-tazobactam (27.65%), piperacillin-tazobactam and neomycin each (25.33%), amoxicillin-sulbactam (23.40%), amoxicillin- clavulanic acid (17.02%), ceftizoxime (10.63%), cefpodoxime (8.51%), moxifloxacin (6.38%), ciprofloxacin, ampicillin and norfloxacin (4.25%) each, ofloxacin and ceftriaxone each (2.12%) and least towards tetracycline, enrofloxacin, penicillin- G, amoxicillin and oxytetracycline (0%). Molecular characterization of E. coli isolates based on antimicrobial resistance : All 47 PCR confirmed E. coli isolates were subjected to PCR for identification of antibiotic resistant genes. Out of 47 E. coli isolates, all had tet A and Gyr A (100%) gene followed by blaSHV gene (80.85%), floR gene (70.21%) and aac (3)-IV gene in 27.65% isolates as illustrated in Table 5 , Fig. 7 , 8 . Table 5 Occurrence of different antimicrobial resistance gene in E. coli isolates Antibiotic group Gene Product (bp) Number of positive E. coli isolates (n = 47) Percent positivity Tetracycline Tet A 210 47 100% Quinolone Gyr A 189 47 100% Beta- lactams Bla SHV 768 38 80.85% Chloramphenicol floR 399 33 70.21% Aminoglycosides aac (3)- IV 286 13 27.65% DISCUSSION In the present study, among 157 bacterial isolates from 105 UTI affected animals, E. coli was the most frequently found isolate, detected in 30% of urine samples, followed by Staphylococcus spp. in 27%, Streptococcus spp. in 24%, and either Klebsiella or Micrococcus in 7% of urine samples each, with Corynebacterium spp. present in 5% of affected animals' urine samples. Yerhuam et al. (2006) also reported E. coli to be the most frequent cause of UTI in cows which is in accordance with the observations of the present study. E. coli is the resident flora of gut and uro-genital tract. Frequent fecal contamination of uro-genital tract often leads to ascending bacterial infection in urinary system along with its ability of adherence to epithelial cells of urinary bladder with the means of pilli (Karimi et al. 2006 ; Solomon et al. 2020 ). Phenotypic characterization of E. coli isolates : Serotyping at Central Research Institute, Kasauli, India (National Reference Centre for serotyping of Salmonella and Escherichia coli ) revealed presence of different serotypes of E. coli in bovine. Out of total 13 serotypes, most frequently encountered serogroups of Escherichia coli was O156 (23.40%). And O5, O128, O83, O159 and O11 had zoonotic significance (Stenutz et al., 2006 ). O5 and O128 serotypes were associated with enterohaemorrhagic E. coli , while O128 exhibited enteropathogenic characteristics. Additionally, O128 and O11 were linked to enterotoxigenic E. coli , O159 was identified as entero-invasive E. coli , and O83 was attributed to uropathogenic E. coli . (Stenutz et al. 2006 ). Similarly, Jacokbsen et al . (2012) also observed considerable commonality observed between human and animal E. coli isolates from UTIs. Antimicrobial sensitivity pattern of E. coli isolates : Overall maximum Invitro antimicrobial sensitivity pattern of E. coli isolates was found towards meropenem (89.36%), followed by gentamicin (44.68%), amikacin (40.42%), ceftriaxone- sulbactam (34.04%), kanamycin, Co- trimoxazole/sulpha-trimethoprim and azithromycin each (31.91%), chloramphenicol (29.78%), cefoperazone-tazobactam (27.65%), piperacillin-tazobactam and neomycin each (25.33%), amoxicillin-sulbactam (23.40%), amoxicillin- clavulanic acid (17.02%), ceftizoxime (10.63%), cefpodoxime (8.51%), moxifloxacin (6.38%), ciprofloxacin, ampicillin and norfloxacin (4.25%) each, ofloxacin and ceftriaxone each (2.12%) and least towards tetracycline, enrofloxacin, penicillin- G, amoxicillin and oxytetracycline (0%). While for Corynebacterium spp. isolates, maximum sensitivity to Meropenem (100%) followed by Chloramphenicol (87.5%), kanamycin, amikacin (75%) each enrofloxacin, norfloxacin, amoxicillin- sulbactam, gentamicin, neomycin, ceftriaxone or tetracycline (62.5%) each, ofloxacin, amoxicillin- clavulanic acid, Ceftizoxime, Ceftriaxone-sulbactam or Azithromycin (50%) each, moxifloxacin, amoxicillin, piperacillin- tazobactam, cefoperazone- tazobactam, Cefpodoxime or oxytetracycline (37.5%) each, Penicillin- G (25%) each and least sensitivity towards ampicillin (12.5%). In contrast to the current study, Punia ( 2021 ) reported maximum sensitivity of bacterial isolates towards amoxicillin- sulbactam and amoxicillin- clavulanic acid (86.36%). Also, in contrast to our study, Kushawaha et al . (2012) reported cephalosporin and fluoroquinolone group of antibiotics to be most sensitive. While working on determining prevalence of multi-drug resistant E. coli isolated from 200 bovine urine samples, Armanullah et al. ( 2018 ) reported 35.5 percent prevalence of E. coli and observed maximum sensitivity of isolates towards amikacin and chloramphenicol. Such high level of resistance for antibiotics commonly used for treating bovine UTI can be attributed to the natural selection of E. coli isolates competent of evading these antibiotics through different mechanisms. The current study highlights the significance of establishing an antibiogram for urinary tract infections in bovines. The indiscriminate use, irregular dosing, or under-dosing of antibiotics may lead to resistant mutants. Therefore, it is recommended to conduct antimicrobial sensitivity testing before initiating treatment to prevent treatment failure. The present study also proposes a rational approach to antibiotic use and emphasizes investigating multiple bacterial causes to address complicated cases effectively. Virulent gene profile of E. coli isolates : Distribution of virulence gene associated with bovine urinary tract infections has not been previously determined and characterized. Uropathogenic potential of E. coli was assessed using PCR with virulence-specific primers. Among the 47 E. coli isolates tested, all were positive for the fimH gene, while only 2 isolates were positive for the hlyA gene, and Pap G-II and Usp genes were absent in all isolates. In line with our research, Kawamura-Sato et al. ( 2010 ); Abdullah and Mustafa ( 2019 ) and Mohammed et al. ( 2020 ) similarly identified the FimH gene in all 38 E. coli isolates collected from canine affected by urinary tract infections. Babacan and Izgur (2021) also reported fimH gene as predominant (100%) in UTI affected canines while in contrast to our study, he also reported papC in 71.1%, sfaDE in 82.2%, and hly A in 73.33% isolates. Ninety-nine percent of E. coli strains carry genes encoding type 1 fimbriae which, during urinary tract infections, induce heightened inflammation by damaging cells in the urinary tract (Vigil et al. 2011 ). Type 1 fimbriae encoded by FimH function as highly versatile virulence factors of UPEC, aiding in bacterial attachment to diverse cell types throughout the urinary tract, demonstrating a strong affinity for proximal tubules, distal tubules and collecting ducts (Avalos Vizcarra et al. 2016 ). HlyA is a pore-forming toxin and causes inducible nitric-oxide-synthase (iNOS)-mediated cell membrane injury and apoptosis with role in the increased production of IL-6 and IL-8 by inducing Ca2 + oscillations in renal epithelial cells (Ristow & Welch, 2016 ). In the present study, none of these E. coli isolates harboured the gene that encodes for Pap G-II and Usp. Based on the findings of our study, we can infer that fimH is the predominant virulence gene responsible for causing UTI in our research. Like our study, Khalifeh and Obaidat, ( 2022 ) also reported FimH as predominant gene in causing urinary tract infections. Antimicrobial resistance profile of E. coli isolates : Multiple drug resistance has been reported in E. coli and Salmonella spp. of different origins throughout the world (Bodewes et al. 2014 ). Out of total 47 of E. coli isolates, 35 isolates were found to be extreme drug resistance (XDR), four isolates to be multi- drug resistant (MDR) and one isolate showed pan drug resistance (PDR) and remaining seven isolates to be sensitive. Similar to our study, Put et al. ( 2015 ) found E. coli causing pyelonephritis affected cattle to be multidrug resistant. All 47 PCR-confirmed E. coli isolates underwent PCR analysis to detect antibiotic-resistant genes. Among these isolates, 100% harbored tet A and Gyr A genes, followed by the blaSHV gene in 80.85% of cases, the floR gene in 70.21% of cases, and the aac (3)-IV gene in 27.65% of isolates. In the present study, all isolates carried tet (A) resistance gene suggesting that these genes are important for the development of tetracycline resistance. Tet(A) encoding efflux mechanisms, has been reported to be the most common tetracycline resistance determinant in E. coli isolates from humans and animals in many countries (Shin et al. 2015 ). Among the 47 E. coli isolates, 33 were determined to be resistant to Chloramphenicol. The role of active efflux pumps, such as floR, was noted to be significant in both intrinsic and acquired resistance to Chloramphenicol (Chang et al. 2015 ). Moreover, the upregulation of efflux pumps impacting Chloramphenicol has become more prevalent in uropathogenic E. coli (Blickwede and Schwarz, 2004 ). The presence of GyrA genes was observed in all isolates. Additionally, the high-level resistance identified in this study suggests that certain isolates may harbor multiple mutations at various points within their E. coli genomes. This finding aligns with a previous study that revealed enrofloxacin-resistant uropathogenic E. coli isolates possessing two-point mutations, one in ParC and the other in GyrA (Chang et al. 2015 ). Moreover, several researchers have reported that overexpression of the AcrAB-TolC system can lead to multidrug resistance, including resistance to fluoroquinolones (Poole, 2000 ). Presence of bLa- SHV in 80.85% E. coli isolates showing resistance against β -lactam antibiotics by hydrolysis because they contain ESBLs (Extended-spectrum β-lactamases) (Hussain et al. 2021 ). Mechanism underlying aminoglycoside resistance in E. coli isolates (27.65%) of our study involves the production of AMEs (aminoglycosides modifying enzymes) by bacteria, which modify and deactivate aminoglycoside drugs, thus imparting resistance to them. AG N-acetyltransferases (aac-3(IV) each specific to a position on the aminoglycoside (AG), carry out these modifications, as indicated by their nomenclature. This modification reduces the affinity between the modified aminoglycoside antibiotics and bacterial ribosomes, ultimately leading to drug resistance (Zhang et al. 2023 ). So, based on our study, we concluded that there is alarming evidence of antimicrobial resistance and high percentage of E. coli isolates from bovine UTI is resistant to two or more antibiotic groups like findings observed by Mustapha et al. ( 2020 ); Ismail and Abutarbush, ( 2020 ). Declarations Acknowledged: Authors are thankful to HOD, VCC, LUVAS, Hisar. Data Availability Statement: All datasets generated and/or analysed during the current study are presented in the article, the accompanying Source Data or Supplementary Information files, or are available from the corresponding author upon reasonable request. Authors contribution statement: Tarun Kumar, Annu Yadav and Ankit Kumar conceptualized, conceived, and designed the study. Annu Yadav, Pawan Bagri and Dinesh did primer designing, performed molecular work. Pawan Bagri, Vinay Ganesh Rao Joshi and Babu Lal Jangir did laboratory analysis. Tarun Kumar and Neelesh Sindhu finalized the manuscript draft. Ethics Statement: Approval for animals under this experiment was granted by the Institutional Animal Ethics Committee (IAEC) of the institute under the reference IAEC/LUVAS/28/10.Top of Form Conflict of Interest statement: The authors declare no conflict of interest . Funding source: No funding agency. References Abdullah, A. R. & Mustafa, J. Y. 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Bacterial isolation and antibiotic sensitivity test from urine of buffalo calves ( Bubalus bubalis ) affected with urethral obstruction. Buff Bull 31 (2). (2012). Mohammed, Y. J., Mustafa, J. Y. & Abdullah, A. R. Isolation and molecular study of some bacterial of some bacterial urinary tract infections of sheep in Basrah province. Iraq J. Agri Sci. 51 (3), 885–893 (2020). Mustapha, M., Goel, P., Kumar, V. & Maan, S. Detection and phylogenetic characterization of virulence genes of E. coli associated with canine urinary tract infections in India. Iran J. Vet. Med 14 (1). (2020). Nam, E. H., Ko, S., Chae, J. S. & Hwang, C. Y. Characterization and zoonotic potential of uro-pathogenic Escherichia coli isolated from dogs. J. Microbiol. Biotech. 23 (3), 422–429 (2013). Poole, K. Efflux-mediated resistance to fluoroquinolones in gram-negative bacteria. Antimicro Agent Chemo . 44 (9), 2233–2241 (2000). Punia, S. Diagnosis and treatment of bacterial urinary tract infections in an Organized buffalo herd. M.V.Sc. Thesis submitted to Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana. (2021). Put, E. et al. Surgical correction of pyelonephritis caused by multidrug-resistant Escherichia coli in a dairy cow. Vla Dierg Tijd 84 (2). (2015). Rahman, M. M. et al. Association of Antibiotic Resistance Traits in Uropathogenic Escherichia coli (UPEC) Isolates. Canad J. Infect. Dis. Med. Microbiol (2022). Ristow, L. C. & Welch, R. A. Hemolysin of uropathogenic Escherichia coli : A cloak or a dagger? Biochim. Biophys. Act. Biomemb . 1858 (3), 538–545 (2016). Shin, S. W., Shin, M. K., Jung, M., Belaynehe, K. M. & Yoo, H. S. Prevalence of antimicrobial resistance and transfer of tetracycline resistance genes in Escherichia coli isolates from beef cattle. App Env Microbiol. 81 (16), 5560–5566 (2015). Smith, B. P., Van Metre, D. C. & Pusterla, N. Diseases of the renal system. Large Animal Internal Medicine. Sixth edition; 906–908. (2021). Solomon, D., Shpigel, N. Y., Salamon, H. & Goshen, T. Epidemiology and risk factors of pyelonephritis in Israeli dairy cattle. Isr. J. Vet. Med. 75 (1), 6–11 (2020). Stenutz, R., Weintraub, A. & Widmalm, G. The structures of Escherichia coli O-polysaccharide antigens. FEMS Microbiol. Rev. 30 , 382–403 (2006). Vigil, P. D., Alteri, C. J. & Mobley, H. L. Identification of in vivo-induced antigens including an RTX family exoprotein required for uropathogenic Escherichia coli virulence. Inf. Imm . 79 (6), 2335–2344 (2011). Yeruham, I., Elad, D., Avidar, Y. & Goshen, T. A herd level analysis of urinary tract infection in dairy cattle. Vet. J. 171 , 172–176 (2006). Zhang, Y. et al. The prevalence and distribution of aminoglycoside resistance genes. Biosaf. Health . 5 (01), 14–20 (2023). Additional Declarations No competing interests reported. Supplementary Files APPENDIX.docx Cite Share Download PDF Status: Published Journal Publication published 27 Nov, 2025 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 14 Jul, 2025 Reviewers agreed at journal 12 Jul, 2025 Reviews received at journal 10 Jul, 2025 Reviews received at journal 10 Jul, 2025 Reviewers agreed at journal 09 Jul, 2025 Reviewers agreed at journal 09 Jul, 2025 Reviews received at journal 08 Jul, 2025 Reviewers agreed at journal 07 Jul, 2025 Reviewers invited by journal 07 Jul, 2025 Editor assigned by journal 03 Jul, 2025 Editor invited by journal 25 Jun, 2025 Submission checks completed at journal 18 Jun, 2025 First submitted to journal 18 Jun, 2025 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. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6831233","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":483052168,"identity":"444c156d-6645-418d-baf3-ebe5443b81b1","order_by":0,"name":"Annu Yadav","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/UlEQVRIiWNgGAWjYHACAwjFzP/xwYcKEIO5gUgt7AzGhjPOgLQwEquFn8FMmrMNxCKgxeD44Y2feWru5BkcZkiQZpxXG83fDtTyo2Ibbi1n0oqleY49KwZqOWBcuO147ozDjA2MPWdu49ZyIMdAcgbb4cQNQJXJM7cdy20AMpgZ2/BoOf/G+OeMfyAtzAyHeeccy51PUMuNHDOJj20gLWyMzbwNNbkbCGmRvPGszOJj3+HEmYd5mBlnHDuQuxGo5SA+v/CdT958I+Hb4cS+82fYf3yoqcudd/7wwQc/KnBrUTiAyj8MJg9gqEMC8g2o/Dp8ikfBKBgFo2CEAgAyLGYLeQC2PgAAAABJRU5ErkJggg==","orcid":"","institution":"Lala Lajpat Rai University of Veterinary and Animal Sciences","correspondingAuthor":true,"prefix":"","firstName":"Annu","middleName":"","lastName":"Yadav","suffix":""},{"id":483052169,"identity":"f339ffe5-f29b-4324-972b-7251b569ccb5","order_by":1,"name":"Tarun Kumar","email":"","orcid":"","institution":"Lala Lajpat Rai University of Veterinary and Animal Sciences","correspondingAuthor":false,"prefix":"","firstName":"Tarun","middleName":"","lastName":"Kumar","suffix":""},{"id":483052170,"identity":"601ac6f0-e4ee-488f-babd-b6f061a2d4fb","order_by":2,"name":"DINESH Dinesh","email":"","orcid":"","institution":"Lala Lajpat Rai University of Veterinary and Animal Sciences","correspondingAuthor":false,"prefix":"","firstName":"DINESH","middleName":"","lastName":"Dinesh","suffix":""},{"id":483052171,"identity":"d3cd3c9e-29c0-4440-a16c-3f5ef67fe6ba","order_by":3,"name":"Pawan Bagri","email":"","orcid":"","institution":"Lala Lajpat Rai University of Veterinary and Animal Sciences","correspondingAuthor":false,"prefix":"","firstName":"Pawan","middleName":"","lastName":"Bagri","suffix":""},{"id":483052172,"identity":"5dc37c15-a45f-44ff-a5a7-e4b78c883fe6","order_by":4,"name":"Ankit Kumar","email":"","orcid":"","institution":"Lala Lajpat Rai University of Veterinary and Animal Sciences","correspondingAuthor":false,"prefix":"","firstName":"Ankit","middleName":"","lastName":"Kumar","suffix":""},{"id":483052173,"identity":"4adf4a96-2dcb-42f6-9915-1a714d2bf4f6","order_by":5,"name":"Neelesh Sindhu","email":"","orcid":"","institution":"Lala Lajpat Rai University of Veterinary and Animal Sciences","correspondingAuthor":false,"prefix":"","firstName":"Neelesh","middleName":"","lastName":"Sindhu","suffix":""},{"id":483052174,"identity":"6d5c2c91-c88e-4c99-a8a6-6159248997bb","order_by":6,"name":"Vinay Ganesh","email":"","orcid":"","institution":"Lala Lajpat Rai University of Veterinary and Animal Sciences","correspondingAuthor":false,"prefix":"","firstName":"Vinay","middleName":"","lastName":"Ganesh","suffix":""},{"id":483052175,"identity":"ea7519a4-5949-4975-be9c-42ff3d0bb709","order_by":7,"name":"Babu Lal Jangir","email":"","orcid":"","institution":"Lala Lajpat Rai University of Veterinary and Animal Sciences","correspondingAuthor":false,"prefix":"","firstName":"Babu","middleName":"Lal","lastName":"Jangir","suffix":""}],"badges":[],"createdAt":"2025-06-05 17:23:23","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6831233/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6831233/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41598-025-26316-2","type":"published","date":"2025-11-27T15:58:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":86449825,"identity":"5a105027-f9b2-44cc-bfb2-610dd31ef610","added_by":"auto","created_at":"2025-07-10 18:50:55","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":191974,"visible":true,"origin":"","legend":"\u003cp\u003eHaryana political map depicting sample collection sites from various animal husbandry setups\u003c/p\u003e","description":"","filename":"FIGURE1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6831233/v1/ce6308b7fafc6905a09e1581.jpg"},{"id":86450323,"identity":"80cae743-2130-45a7-95a9-8d9cf1bf1811","added_by":"auto","created_at":"2025-07-10 18:58:55","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":149092,"visible":true,"origin":"","legend":"\u003cp\u003eDetail of animals screened at different animal husbandry setups from different agro-climatic zones of Haryana\u003c/p\u003e","description":"","filename":"Figure2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6831233/v1/95ef64eaf07f4da5a5a3d70c.jpg"},{"id":86450325,"identity":"3e74c730-1c45-4ee4-a48f-0b72258f4875","added_by":"auto","created_at":"2025-07-10 18:58:55","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":216733,"visible":true,"origin":"","legend":"\u003cp\u003eBiochemical identification of \u003cem\u003eEscherichia coli \u003c/em\u003eisolates using ready to use test kit\u003c/p\u003e","description":"","filename":"Figure3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6831233/v1/7d70d47b2c97152949da6f12.jpg"},{"id":86449829,"identity":"9510cc5d-711e-4989-b4b0-a2d4baa0a7c5","added_by":"auto","created_at":"2025-07-10 18:50:55","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":62161,"visible":true,"origin":"","legend":"\u003cp\u003eSimplex PCR- based identification of \u003cem\u003eEscherichia coli\u003c/em\u003e isolates; Lane L- 1kbp DNA Ladder- ng/5µl (HI media Gene Ruler), PC positive control- Usp A gene with \u003cem\u003eE. Coli\u003c/em\u003e (ATCC 25922), NC negative control, Lane 1-31- show results for Usp A gene with clinical isolates)\u003c/p\u003e","description":"","filename":"FIGURE4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6831233/v1/3e3502d2fb9b12bfa56a0070.jpg"},{"id":86450751,"identity":"992119d7-be29-489c-a0ec-27486410af5b","added_by":"auto","created_at":"2025-07-10 19:14:55","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":45277,"visible":true,"origin":"","legend":"\u003cp\u003eDetermination of FimH gene indicated by simplex PCR; {Ladder (Thermo Scientific, SM0243, 1 kbp DNA Ladder) NTC- negative template control, 1-15 \u003cem\u003eE. coli\u003c/em\u003e isolates)\u003c/p\u003e","description":"","filename":"FIGURE5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6831233/v1/d59520c7ffba888227926ad6.jpg"},{"id":86450328,"identity":"9c588d46-c3c9-4756-9d24-9f946ccc3d74","added_by":"auto","created_at":"2025-07-10 18:58:55","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":135999,"visible":true,"origin":"","legend":"\u003cp\u003eHeat map showing antibiotic sensitivity pattern of \u003cem\u003eE. coli \u003c/em\u003eisolates\u003c/p\u003e","description":"","filename":"FIGURE6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6831233/v1/87141a7589edc244b70ac0f8.jpg"},{"id":86449833,"identity":"950379fe-bfe9-4223-b559-ae80ffb434b3","added_by":"auto","created_at":"2025-07-10 18:50:55","extension":"jpg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":78928,"visible":true,"origin":"","legend":"\u003cp\u003eDetermination of gyr A gene (189bp) representing quinolone resistance by simplex PCR {Ladder- L (Thermo Scientific, SM0243, 100 bp DNA Ladder}, 1-34 \u003cem\u003eE. coli \u003c/em\u003eisolates\u003c/p\u003e","description":"","filename":"FIGURE7.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6831233/v1/e3298d6bf1bdf5bda707d6c8.jpg"},{"id":86450752,"identity":"476a25ba-13fa-4f97-96da-ad9257fdb788","added_by":"auto","created_at":"2025-07-10 19:14:55","extension":"jpg","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":60501,"visible":true,"origin":"","legend":"\u003cp\u003eDetermination of bla- SHV gene (768bp) representing beta-lactams resistance by simplex PCR {Ladder (Thermo Scientific, SM0243, 100 bp DNA Ladder) NTC- negative template control; 33-47 \u003cem\u003eE. coli\u003c/em\u003e isolates)\u003c/p\u003e","description":"","filename":"FIGURE8.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6831233/v1/45f0ab3759ae0811e5143ae5.jpg"},{"id":97178524,"identity":"9ec20a9b-af01-4b5b-9de2-ac06bcecace6","added_by":"auto","created_at":"2025-12-01 16:10:38","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2226379,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6831233/v1/d2af6c8f-6377-432b-80ac-c7bda5d7ded1.pdf"},{"id":86450322,"identity":"e8916675-2f9f-4d80-8c04-c7dacd9ef96c","added_by":"auto","created_at":"2025-07-10 18:58:55","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1179598,"visible":true,"origin":"","legend":"","description":"","filename":"APPENDIX.docx","url":"https://assets-eu.researchsquare.com/files/rs-6831233/v1/0f414ff40712a60b479087f1.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Comprehensive Analysis of Escherichia coli in Bovine Urinary Tract Infections: Serotypes, Antibiotic Resistance, and Virulence Gene Profiles","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eIn ruminants, Urinary tract infection (UTI) typically occurs as cystitis, ureteritis and pyelonephritis which most often result from ascending infection with \u003cem\u003eCorynebacterium renale\u003c/em\u003e or \u003cem\u003eEscherichia coli\u003c/em\u003e (Smith et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) causing economic losses via reduction in reproduction and production of the farm (Yeruham et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Uropathogenic \u003cem\u003eE. coli\u003c/em\u003e (UPEC) pathotypes are responsible for causing extra-intestinal \u003cem\u003eE. coli\u003c/em\u003e infections, primarily urinary tract infections with specific virulence attributes encoded on genetic elements. Uropathogenic \u003cem\u003eE. coli\u003c/em\u003e (UPEC) strains have shown certain virulent properties, including iron uptake systems, adhesins, specific O: K: H serotypes and synthesis of cytotoxins that contribute to colonization and invasion of the bacterium (Janke et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). Also, successful colonization of UPEC in the urinary tract depends on the expression of fimbria adhesion proteins, which facilitate attachment of the bacterium to the urothelium, and on the presence of specific bacterial genes that encode virulence factors (Ristow and Welch, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The role of \u003cem\u003eE. coli\u003c/em\u003e and various serotypes involved in the pathogenesis of bovine UTI is poorly understood and documented. Traditionally it was believed that urinary tract of healthy individual is sterile but recent studies of the bovine urinary microbiota pointed out the presence of \u003cem\u003eE. coli\u003c/em\u003e as resident members of this niche (Giannattasio-Ferraz et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), suggesting dysbiosis as the mechanism of urinary tract infection rather than external sources (Garretto et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAntimicrobial resistance (AMR) has been recognized as one of the most significant threats to the health of people and milk-producing animals (Gemeda \u003cem\u003eet al\u003c/em\u003e., 2023). Investigating antimicrobial resistance (AMR) in \u003cem\u003eE. coli\u003c/em\u003e is crucial because the environmental transmission of AMR is likely more significant in \u003cem\u003eE. coli\u003c/em\u003e than in other microbiota members and it can survive in the environment for extended periods (Barrera et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Moreover, much of the AMR in \u003cem\u003eE. coli\u003c/em\u003e is carried on mobile genetic elements that can be transferred between bacteria, facilitating the rapid spread and persistence of resistance genes across different bacterial species (horizontal gene transfer) (Johnson et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). As a result, AMR in \u003cem\u003eE. coli\u003c/em\u003e is considered a major public health threat.\u003c/p\u003e\u003cp\u003eThis study aimed to report the occurrence of \u003cem\u003eE. coli\u003c/em\u003e in UTI affected bovines as well as to evaluate the presence of resistance and virulence genes of \u003cem\u003eE. coli\u003c/em\u003e isolated from dairy animals suffering from UTI.\u003c/p\u003e"},{"header":"MATERIAL AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eStudy area:\u003c/h2\u003e\u003cp\u003eVarious animal husbandry setups were selected randomly across six Haryana districts spanning two agro-climatic zones (North-eastern and South- western zone) as illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, along with UTI affected bovine presented at Veterinary Teaching Hospital, Hisar. Present study was conducted with ethical approval from the IAEC, reference number VCC/IAEC/2022/1624\u0026thinsp;+\u0026thinsp;51. All methods were performed in accordance with relevant guidelines and regulations (Appendix 1).\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eAnimal Selection:\u003c/h3\u003e\n\u003cp\u003eSamples were randomly collected from suspected UTI cases across various animal husbandry setups, based on case history and a slightly modified version of the Acute Cystitis Symptom Score (ACSS) Index (Alidjanov et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Each parameter was scored from 0 to 2 and sum of scores yielded a total cumulative score. Patient with cumulative score 6 and above were selected for sample collection.\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\u003eCriteria for assessing severity of bovine cystitis (USQOLAT)\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\u003eDomain\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSymptoms\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNil (0)/No\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eModerate (1)/Yes\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eSevere (2)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTypical\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eFrequent voiding of small urine volume\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\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\u003ePainful/difficult urination/stranguria\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\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\u003eHematuria\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e2-3times/day during urination\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eEvery\u003c/p\u003e\u003cp\u003etime\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\u003eSuprapubic discomfort/flank pain\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\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\u003eDiurnal temperature variation\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\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\u003eTail swishing\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eDifferential\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eFever\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\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\u003eVaginal/urethral discharge\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\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\u003eProlapse\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eIst degree\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2nd degree\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\u003cb\u003eIsolation and identification\u003c/b\u003e \u003cb\u003eE. coli\u003c/b\u003e:\u003c/p\u003e\u003cp\u003eTwo hundred and fifty-four urine samples were collected aseptically using urinary catheter and were inoculated on MacConkey agar at 37\u0026deg; C for 24 h. After overnight incubation at 37\u0026deg; C typical pink colonies on MacConkey agar were Gram-stained and then single pure colony was transferred to eosin-methylene-blue (EMB) agar for incubation at 37\u0026deg; C overnight. Colonies with typical metallic sheen on EMB agar, were considered as \u003cem\u003eE. coli\u003c/em\u003e-positive and transferred to nutrient agar for additional confirmatory biochemical tests (IMViC tests), molecular tests and serotyping.\u003c/p\u003e\u003cp\u003e\u003cb\u003eDNA Extraction and\u003c/b\u003e \u003cb\u003eE. coli\u003c/b\u003e \u003cb\u003eConfirmation\u003c/b\u003e:\u003c/p\u003e\u003cp\u003ePresumptive \u003cem\u003eE. coli\u003c/em\u003e isolates were molecularly confirmed by PCR amplification of universal stress protein (usp A gene). The PCR was performed using primer pair as depicted in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The DNA extracted from ATCC 25922 was used as positive control for \u003cem\u003eE. coli\u003c/em\u003e. PCR were carried out in 12.5\u0026micro;l total volume consisted of 7\u0026micro;l of master mix, 0.5 \u0026micro;l of 10 \u0026micro;M solution of each primer (forward and reverse primer), 1\u0026micro;l of DNA sample and remaining nuclease free water to make 12.5 \u0026micro;l. PCR amplifications started with initial denaturation at95 \u0026ordm;C for 2min, 30 cycles of denaturation at 94 \u0026ordm;C for 2 min, annealing at 58 \u0026ordm;C for 1min, extension at 72 \u0026ordm;C for 1min, and final extension at 72 \u0026ordm;C for 5min.\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\u003ePrimer pairs used for the characterization of \u003cem\u003eE. coli\u003c/em\u003e (Osek, 2001)\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\u003eDescription\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eGene\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003ePrimers (5\u0026rsquo;-3\u0026rsquo;)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eProduct size (bp)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eUniversal stress protein\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eUsp A\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eF-CCGATACGCTGCCAATCAGT\u003c/p\u003e\u003cp\u003eR-ACGCAGACCGTAAGGGCCAGAT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e884\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\n\u003ch3\u003eSerotyping:\u003c/h3\u003e\n\u003cp\u003eAfter confirmation of isolates as \u003cem\u003eE. coli\u003c/em\u003e, isolates were sent to the National \u003cem\u003eEscherichia\u003c/em\u003e and \u003cem\u003eSalmonella\u003c/em\u003e Centre, Kasauli (H.P) for final serotyping. Two individually similar colonies from each of the purified cultures of identified isolates were inoculated in semisolid media and incubated at 37\u0026ordm;C overnight. One tube of each culture was sent to the National \u003cem\u003eEscherichia\u003c/em\u003e and \u003cem\u003eSalmonella\u003c/em\u003e Centre, Kasauli (H.P) for serotyping, other tube was kept at 4\u0026ordm;C till further use. Kauffman- White scheme was consulted and organisms were assigned to the \u003cem\u003eE\u003c/em\u003e. \u003cem\u003ecoli\u003c/em\u003e serotype.\u003c/p\u003e\n\u003ch3\u003eDetection of virulent and antimicrobial resistance genes:\u003c/h3\u003e\n\u003cp\u003eVirulent genes including Pap GII, hlyA, fimH, Usp and antimicrobial-resistant genes, including blaSHV, aac (3)-IV, tetA, tetB, gyrA, and floR, were amplified via 30 cycles of PCR amplification using the specific primers as depicted in Table \u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. The temperature profile was as follows: DNA denaturation at 94\u0026deg;C for 1min, followed by annealing at the specific temperature for each primer set for 30 s, extension at 72\u0026deg;C for 60 s, and a final extension at 72\u0026deg;C for 10 min. Electrophoresis was carried out at 85 V/cm of gel in 1X TAE running buffer in mini gel electrophoresis assembly, using power supply for 45 minutes to one hour. The amplified product was detected with the help of good view dye in agarose gel electrophoresis. The DNA bands were observed under the UV transilluminator.\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\u003eAnnealing temperature and sequence of Oligonucleotide primers used for amplification of antibiotic resistance and virulent gene of \u003cem\u003eE. coli\u003c/em\u003e detection\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=\"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=\"char\" char=\".\" 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\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\u003eGene\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eDescription\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003ePrimers (5\u0026rsquo;-3\u0026rsquo;)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eProduct size (bp)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eAnnealing temperature\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eReference\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\u003eAdhesins (papGII)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eClass II P pili\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eF: GGGATGAGCGGGCCTTTGAT\u003c/p\u003e\u003cp\u003eR: CGGGCCCCCAAGTAACTCG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e190\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e57 ̊ C\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eNam et al. (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2013\u003c/span\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\u003eToxins (hlyA)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eα-Hemolysin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eF: AACAAGGATAAGCACTGTTCTGGCT\u003c/p\u003e\u003cp\u003eR: ACCATATAAGCGGTCATTCCCATCA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1177\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e54 ̊C\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eSelf- designed\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\u003eAdhesins (fimH)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eType\u003c/p\u003e\u003cp\u003e1 fimbria\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eF: ATGAAACGAGTTATTACCCTGTT\u003c/p\u003e\u003cp\u003eR: TATTGATAAACAAAAGTCACGCC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e900\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e58 ̊C\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eMohammed et al. (\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2020\u003c/span\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\u003eMiscellaneous (usp)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eUropathogen-specific protein\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eF: ACATTCACGGCAAGCCTCAG\u003c/p\u003e\u003cp\u003eR: AGCGAGTTCCTGGTGAAAGC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e440\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e55 ̊C\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eNam et al. (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2013\u003c/span\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\u003eblaSHV\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eBetalactam\u003c/p\u003e\u003cp\u003e(Ampicillin)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eF: TCGCCTGTGTATTATCTCCC\u003c/p\u003e\u003cp\u003eR: CGCAGATAAATCACCACAATG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e768\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e55 ̊C\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eRahman et al. (\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2022\u003c/span\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\u003eaac (3)- IV\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAminoglycosides (Gentamicin)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eF: CTTCAGGATGGCAAGTTGGT\u003c/p\u003e\u003cp\u003eR: TCATCTCGTTCTCCGCTCAT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e286\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e52 ̊C\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eRahman et al. (\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2022\u003c/span\u003e)\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\u003eFloR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eChloramphenicol\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eF: TATCTCCCTGTCGTTCCAG\u003c/p\u003e\u003cp\u003eR: AGAACTCGCCGATCAATG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e399\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e60̊ C\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eMohammed et al. (\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2020\u003c/span\u003e)\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\u003etet (A)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eTetracyclines\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eF: GCTACATCCTGCTTGCCTTC\u003c/p\u003e\u003cp\u003eR: CATAGATCGCCGTGAAGAGG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e210\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e62 ̊C\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eMustapha \u003cem\u003eet al\u003c/em\u003e. (2018)\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\u003eGyrA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eQuinolones\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eF: ACGTACTAGGCAATGACTGG\u003c/p\u003e\u003cp\u003eR: AGAAGTCGCCGTCGATAGAAC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e189\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e51 ̊C\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eSelf- designed\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\n\u003ch3\u003eAntibiotic Susceptibility Testing:\u003c/h3\u003e\n\u003cp\u003eThe antimicrobial susceptibility test on the isolates was performed according to the National Committee for Clinical Laboratory Standards (CLSI, 2024) using the Kibry-Bauer disk diffusion test method on Muller-Hinton agar. The diameter of the zones of inhibition was measured and compared with zone size interpretation guidelines described by the Clinical Laboratory Standard Institute (CLSI, 2024) for the family Enterobacteriaceae and determined as sensitive, intermediate, or resistant. The isolated \u003cem\u003eE. coli\u003c/em\u003e were tested for sensitivity to the most used antimicrobials.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eStatistical Analysis\u003c/strong\u003e\u003cp\u003eAll demographic data and results were recorded on Excel spreadsheets. The data were analysed with the statistical software, IBM SPSS 25.\u003c/p\u003e\u003c/p\u003e"},{"header":"RESULTS","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\u003ch2\u003eCharacteristics of the collected sample:\u003c/h2\u003e\u003cp\u003eA total of 254 animals were screened for bovine UTI, 134 from tertiary setup of hospital (University referral hospital) and remaining 120 from different animal husbandry setups viz. Gaushala, organized dairy farm, unorganized dairy farm, and field hospitals; 30 animals each as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. Out of 254, 105 animals were found positive for UTI based on culture testing, urinalysis, haemato- biochemical testing, renal biomarkers, and renal ultrasonography. Out of 120 animals suspected of having a urinary tract infection based on clinical severity scores, 60 were confirmed to be infected, emphasizing the sensitivity of the scoring system as a reliable tool for identifying UTIs.\u003c/p\u003e\u003cp\u003e\u003cb\u003eOccurrence of\u003c/b\u003e \u003cb\u003eEscherichia coli\u003c/b\u003e:\u003c/p\u003e\u003cp\u003eA total of 157 organisms were isolated from 105 UTI affected animals with 54 animals having single infections and 51 animals comprising mixed infections. Cultural and biochemical testing identified \u003cem\u003eEscherichia coli\u003c/em\u003e as the predominant bacterium, accounting for 30% (47/105) of animals as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. All 47 suspected \u003cem\u003eE. coli\u003c/em\u003e isolates were subjected to PCR targeting the \u003cem\u003eE. coli\u003c/em\u003e-specific Usp A gene for confirmation. PCR results for these 47 presumptive \u003cem\u003eE. coli\u003c/em\u003e isolates showed an amplification band of 884 bp, specific to the Usp A gene, confirming them as \u003cem\u003eE. coli\u003c/em\u003e illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e.\u003c/p\u003e\u003cp\u003e\u003cb\u003eSerotyping results of\u003c/b\u003e \u003cb\u003eE. coli\u003c/b\u003e \u003cb\u003eisolates\u003c/b\u003e:\u003c/p\u003e\u003cp\u003eConfirmed 47 \u003cem\u003eEscherichia coli\u003c/em\u003e samples were characterized based on Serotyping at CRI Kasauli. Isolates were serotyped into different groups as shown in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. Out of total 13 serotypes, most frequently encountered serogroups of \u003cem\u003eEscherichia coli\u003c/em\u003e was O156 (23.40%) followed by O84 and O101 (10.6%) each, O169 (6.38%), O128, O83, O120 and O18 (4.26%) each, O5, O134, O11, O88 and O159 (2.13%) each. Among the 13 serotypes identified, five serotypes O5, O128, O83, O11, and O159 were of zoonotic significance.\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\u003eRelative frequency of serotypes of \u003cem\u003eE. coli\u003c/em\u003e isolates (n\u0026thinsp;=\u0026thinsp;47) based on serotyping from NSEC, Kasauli\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=\"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\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\u003eSerotypes\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNumber\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\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\u003e\u003cb\u003e1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eO169\u003c/b\u003e\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\u003e6.38%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eO84\u003c/b\u003e\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\u003e10.6%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eO5 (EHEC)*\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e2.13%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e4\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eO134\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e2.13%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e5\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eO128 (EPEC\u0026thinsp;+\u0026thinsp;ETEC\u0026thinsp;+\u0026thinsp;EHEC) *\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e4.26%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e6\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eO156\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e23.40%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e7\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eO101\u003c/b\u003e\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\u003e10.63%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e8\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eUT\u003c/b\u003e\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\u003e17.02%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e9\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eO83 (UPEC)*\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e4.26%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e10\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eO120\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e4.26%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e11\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eO11 (ETEC)*\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e2.13%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e12\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eO18\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e4.26%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e13\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eO88\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e2.13%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e14\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eO159 (ETEC\u0026thinsp;+\u0026thinsp;EIEC) *\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e2.13%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e15\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eNon- viable\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e4.26%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"4\"\u003e\u003cb\u003e* EHEC- Enterohemorrhagic\u003c/b\u003e \u003cb\u003eE. coli\u003c/b\u003e; \u003cb\u003eEPEC- Enteropathogenic\u003c/b\u003e \u003cb\u003eE. coli\u003c/b\u003e; \u003cb\u003eETEC- Enterotoxigenic\u003c/b\u003e \u003cb\u003eE. coli\u003c/b\u003e; \u003cb\u003eUPEC- Uropathogenic\u003c/b\u003e \u003cb\u003eE. coli\u003c/b\u003e; \u003cb\u003eEIEC- Enteroinvasive\u003c/b\u003e \u003cb\u003eE. coli\u003c/b\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eCharacterization of Uro-pathogenic\u003c/b\u003e \u003cb\u003eEscherichia coli\u003c/b\u003e \u003cb\u003e(UPEC) using PCR\u003c/b\u003e:\u003c/p\u003e\u003cp\u003eFor UPEC characterization, all the confirmed \u003cem\u003eEscherichia coli\u003c/em\u003e isolates (n\u0026thinsp;=\u0026thinsp;47) were subjected to PCR using a set of four virulence genes (VGs). PCR components and conditions were standardized using DNA extracted from colonies of confirmed \u003cem\u003eEscherichia coli\u003c/em\u003e isolates in the current study. Out of 47 \u003cem\u003eEscherichia coli\u003c/em\u003e isolates, only 2 isolates had hlyA gene while fimH present in 100% isolates (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cb\u003eInvitro\u003c/b\u003e \u003cb\u003eantibiotic sensitivity based antimicrobial pattern of\u003c/b\u003e \u003cb\u003eEscherichia coli\u003c/b\u003e \u003cb\u003eisolates\u003c/b\u003e:\u003c/p\u003e\u003cp\u003eAntimicrobial sensitivity pattern of \u003cem\u003eE. coli\u003c/em\u003e isolates is represented in Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e. Overall maximum sensitivity of \u003cem\u003eE. coli\u003c/em\u003e isolates was found towards meropenem (89.36%), followed by gentamicin (44.68%), amikacin (40.42%), ceftriaxone- sulbactam (34.04%), kanamycin, Co- trimoxazole/sulpha-trimethoprim and azithromycin each (31.91%), chloramphenicol (29.78%), cefoperazone-tazobactam (27.65%), piperacillin-tazobactam and neomycin each (25.33%), amoxicillin-sulbactam (23.40%), amoxicillin- clavulanic acid (17.02%), ceftizoxime (10.63%), cefpodoxime (8.51%), moxifloxacin (6.38%), ciprofloxacin, ampicillin and norfloxacin (4.25%) each, ofloxacin and ceftriaxone each (2.12%) and least towards tetracycline, enrofloxacin, penicillin- G, amoxicillin and oxytetracycline (0%).\u003c/p\u003e\u003cp\u003e\u003cb\u003eMolecular characterization of\u003c/b\u003e \u003cb\u003eE. coli\u003c/b\u003e \u003cb\u003eisolates based on antimicrobial resistance\u003c/b\u003e:\u003c/p\u003e\u003cp\u003eAll 47 PCR confirmed \u003cem\u003eE. coli\u003c/em\u003e isolates were subjected to PCR for identification of antibiotic resistant genes. Out of 47 \u003cem\u003eE. coli\u003c/em\u003e isolates, all had tet A and Gyr A (100%) gene followed by blaSHV gene (80.85%), floR gene (70.21%) and aac (3)-IV gene in 27.65% isolates as illustrated in Table \u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e, \u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003e.\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\u003eOccurrence of different antimicrobial resistance gene in \u003cem\u003eE. coli\u003c/em\u003e isolates\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=\"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=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAntibiotic group\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGene\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eProduct (bp)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNumber of positive\u003c/p\u003e\u003cp\u003e\u003cem\u003eE. coli\u003c/em\u003e isolates (n\u0026thinsp;=\u0026thinsp;47)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePercent positivity\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTetracycline\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eTet A\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e210\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e47\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eQuinolone\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGyr A\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e189\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e47\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eBeta- lactams\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBla SHV\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e768\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e38\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e80.85%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eChloramphenicol\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003efloR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e399\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e70.21%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eAminoglycosides\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eaac (3)- IV\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e286\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e27.65%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eIn the present study, among 157 bacterial isolates from 105 UTI affected animals, \u003cem\u003eE. coli\u003c/em\u003e was the most frequently found isolate, detected in 30% of urine samples, followed by \u003cem\u003eStaphylococcus\u003c/em\u003e spp. in 27%, \u003cem\u003eStreptococcus\u003c/em\u003e spp. in 24%, and either \u003cem\u003eKlebsiella\u003c/em\u003e or \u003cem\u003eMicrococcus\u003c/em\u003e in 7% of urine samples each, with \u003cem\u003eCorynebacterium\u003c/em\u003e spp. present in 5% of affected animals' urine samples. Yerhuam et al. (2006) also reported \u003cem\u003eE. coli\u003c/em\u003e to be the most frequent cause of UTI in cows which is in accordance with the observations of the present study. \u003cem\u003eE. coli\u003c/em\u003e is the resident flora of gut and uro-genital tract. Frequent fecal contamination of uro-genital tract often leads to ascending bacterial infection in urinary system along with its ability of adherence to epithelial cells of urinary bladder with the means of pilli (Karimi et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Solomon et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cb\u003ePhenotypic characterization of\u003c/b\u003e \u003cb\u003eE. coli\u003c/b\u003e \u003cb\u003eisolates\u003c/b\u003e:\u003c/p\u003e\u003cp\u003eSerotyping at Central Research Institute, Kasauli, India (National Reference Centre for serotyping of \u003cem\u003eSalmonella\u003c/em\u003e and \u003cem\u003eEscherichia coli\u003c/em\u003e) revealed presence of different serotypes of \u003cem\u003eE. coli\u003c/em\u003e in bovine. Out of total 13 serotypes, most frequently encountered serogroups of \u003cem\u003eEscherichia coli\u003c/em\u003e was O156 (23.40%). And O5, O128, O83, O159 and O11 had zoonotic significance (Stenutz et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). O5 and O128 serotypes were associated with enterohaemorrhagic \u003cem\u003eE. coli\u003c/em\u003e, while O128 exhibited enteropathogenic characteristics. Additionally, O128 and O11 were linked to enterotoxigenic \u003cem\u003eE. coli\u003c/em\u003e, O159 was identified as entero-invasive \u003cem\u003eE. coli\u003c/em\u003e, and O83 was attributed to uropathogenic \u003cem\u003eE. coli\u003c/em\u003e. (Stenutz et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Similarly, Jacokbsen \u003cem\u003eet al\u003c/em\u003e. (2012) also observed considerable commonality observed between human and animal \u003cem\u003eE. coli\u003c/em\u003e isolates from UTIs.\u003c/p\u003e\u003cp\u003e\u003cb\u003eAntimicrobial sensitivity pattern of\u003c/b\u003e \u003cb\u003eE. coli\u003c/b\u003e \u003cb\u003eisolates\u003c/b\u003e:\u003c/p\u003e\u003cp\u003eOverall maximum \u003cem\u003eInvitro\u003c/em\u003e antimicrobial sensitivity pattern of \u003cem\u003eE. coli\u003c/em\u003e isolates was found towards meropenem (89.36%), followed by gentamicin (44.68%), amikacin (40.42%), ceftriaxone- sulbactam (34.04%), kanamycin, Co- trimoxazole/sulpha-trimethoprim and azithromycin each (31.91%), chloramphenicol (29.78%), cefoperazone-tazobactam (27.65%), piperacillin-tazobactam and neomycin each (25.33%), amoxicillin-sulbactam (23.40%), amoxicillin- clavulanic acid (17.02%), ceftizoxime (10.63%), cefpodoxime (8.51%), moxifloxacin (6.38%), ciprofloxacin, ampicillin and norfloxacin (4.25%) each, ofloxacin and ceftriaxone each (2.12%) and least towards tetracycline, enrofloxacin, penicillin- G, amoxicillin and oxytetracycline (0%). While for \u003cem\u003eCorynebacterium\u003c/em\u003e spp. isolates, maximum sensitivity to Meropenem (100%) followed by Chloramphenicol (87.5%), kanamycin, amikacin (75%) each enrofloxacin, norfloxacin, amoxicillin- sulbactam, gentamicin, neomycin, ceftriaxone or tetracycline (62.5%) each, ofloxacin, amoxicillin- clavulanic acid, Ceftizoxime, Ceftriaxone-sulbactam or Azithromycin (50%) each, moxifloxacin, amoxicillin, piperacillin- tazobactam, cefoperazone- tazobactam, Cefpodoxime or oxytetracycline (37.5%) each, Penicillin- G (25%) each and least sensitivity towards ampicillin (12.5%). In contrast to the current study, Punia (\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) reported maximum sensitivity of bacterial isolates towards amoxicillin- sulbactam and amoxicillin- clavulanic acid (86.36%). Also, in contrast to our study, Kushawaha \u003cem\u003eet al\u003c/em\u003e. (2012) reported cephalosporin and fluoroquinolone group of antibiotics to be most sensitive. While working on determining prevalence of multi-drug resistant \u003cem\u003eE. coli\u003c/em\u003e isolated from 200 bovine urine samples, Armanullah et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) reported 35.5 percent prevalence of \u003cem\u003eE. coli\u003c/em\u003e and observed maximum sensitivity of isolates towards amikacin and chloramphenicol. Such high level of resistance for antibiotics commonly used for treating bovine UTI can be attributed to the natural selection of \u003cem\u003eE. coli\u003c/em\u003e isolates competent of evading these antibiotics through different mechanisms. The current study highlights the significance of establishing an antibiogram for urinary tract infections in bovines. The indiscriminate use, irregular dosing, or under-dosing of antibiotics may lead to resistant mutants. Therefore, it is recommended to conduct antimicrobial sensitivity testing before initiating treatment to prevent treatment failure. The present study also proposes a rational approach to antibiotic use and emphasizes investigating multiple bacterial causes to address complicated cases effectively.\u003c/p\u003e\u003cp\u003e\u003cb\u003eVirulent gene profile of\u003c/b\u003e \u003cb\u003eE. coli\u003c/b\u003e \u003cb\u003eisolates\u003c/b\u003e:\u003c/p\u003e\u003cp\u003eDistribution of virulence gene associated with bovine urinary tract infections has not been previously determined and characterized. Uropathogenic potential of \u003cem\u003eE. coli\u003c/em\u003e was assessed using PCR with virulence-specific primers. Among the 47 \u003cem\u003eE. coli\u003c/em\u003e isolates tested, all were positive for the fimH gene, while only 2 isolates were positive for the hlyA gene, and Pap G-II and Usp genes were absent in all isolates. In line with our research, Kawamura-Sato et al. (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2010\u003c/span\u003e); Abdullah and Mustafa (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) and Mohammed et al. (\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) similarly identified the FimH gene in all 38 \u003cem\u003eE. coli\u003c/em\u003e isolates collected from canine affected by urinary tract infections. Babacan and Izgur (2021) also reported fimH gene as predominant (100%) in UTI affected canines while in contrast to our study, he also reported papC in 71.1%, sfaDE in 82.2%, and hly A in 73.33% isolates.\u003c/p\u003e\u003cp\u003eNinety-nine percent of \u003cem\u003eE. coli\u003c/em\u003e strains carry genes encoding type 1 fimbriae which, during urinary tract infections, induce heightened inflammation by damaging cells in the urinary tract (Vigil et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Type 1 fimbriae encoded by FimH function as highly versatile virulence factors of UPEC, aiding in bacterial attachment to diverse cell types throughout the urinary tract, demonstrating a strong affinity for proximal tubules, distal tubules and collecting ducts (Avalos Vizcarra et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). HlyA is a pore-forming toxin and causes inducible nitric-oxide-synthase (iNOS)-mediated cell membrane injury and apoptosis with role in the increased production of IL-6 and IL-8 by inducing Ca2\u0026thinsp;+\u0026thinsp;oscillations in renal epithelial cells (Ristow \u0026amp; Welch, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn the present study, none of these \u003cem\u003eE. coli\u003c/em\u003e isolates harboured the gene that encodes for Pap G-II and Usp. Based on the findings of our study, we can infer that fimH is the predominant virulence gene responsible for causing UTI in our research. Like our study, Khalifeh and Obaidat, (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) also reported FimH as predominant gene in causing urinary tract infections.\u003c/p\u003e\u003cp\u003e\u003cb\u003eAntimicrobial resistance profile of\u003c/b\u003e \u003cb\u003eE. coli\u003c/b\u003e \u003cb\u003eisolates\u003c/b\u003e:\u003c/p\u003e\u003cp\u003eMultiple drug resistance has been reported in \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eSalmonella\u003c/em\u003e spp. of different origins throughout the world (Bodewes et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Out of total 47 of \u003cem\u003eE. coli\u003c/em\u003e isolates, 35 isolates were found to be extreme drug resistance (XDR), four isolates to be multi- drug resistant (MDR) and one isolate showed pan drug resistance (PDR) and remaining seven isolates to be sensitive. Similar to our study, Put et al. (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) found \u003cem\u003eE. coli\u003c/em\u003e causing pyelonephritis affected cattle to be multidrug resistant.\u003c/p\u003e\u003cp\u003eAll 47 PCR-confirmed \u003cem\u003eE. coli\u003c/em\u003e isolates underwent PCR analysis to detect antibiotic-resistant genes. Among these isolates, 100% harbored tet A and Gyr A genes, followed by the blaSHV gene in 80.85% of cases, the floR gene in 70.21% of cases, and the aac (3)-IV gene in 27.65% of isolates. In the present study, all isolates carried \u003cem\u003etet\u003c/em\u003e (A) resistance gene suggesting that these genes are important for the development of tetracycline resistance. Tet(A) encoding efflux mechanisms, has been reported to be the most common tetracycline resistance determinant in \u003cem\u003eE. coli\u003c/em\u003e isolates from humans and animals in many countries (Shin et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Among the 47 \u003cem\u003eE. coli\u003c/em\u003e isolates, 33 were determined to be resistant to Chloramphenicol. The role of active efflux pumps, such as floR, was noted to be significant in both intrinsic and acquired resistance to Chloramphenicol (Chang et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Moreover, the upregulation of efflux pumps impacting Chloramphenicol has become more prevalent in uropathogenic \u003cem\u003eE. coli\u003c/em\u003e (Blickwede and Schwarz, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). The presence of GyrA genes was observed in all isolates. Additionally, the high-level resistance identified in this study suggests that certain isolates may harbor multiple mutations at various points within their \u003cem\u003eE. coli\u003c/em\u003e genomes. This finding aligns with a previous study that revealed enrofloxacin-resistant uropathogenic \u003cem\u003eE. coli\u003c/em\u003e isolates possessing two-point mutations, one in ParC and the other in GyrA (Chang et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Moreover, several researchers have reported that overexpression of the AcrAB-TolC system can lead to multidrug resistance, including resistance to fluoroquinolones (Poole, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). Presence of bLa- SHV in 80.85% \u003cem\u003eE. coli\u003c/em\u003e isolates showing resistance against \u003cem\u003eβ\u003c/em\u003e-lactam antibiotics by hydrolysis because they contain ESBLs (Extended-spectrum β-lactamases) (Hussain et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eMechanism underlying aminoglycoside resistance in \u003cem\u003eE. coli\u003c/em\u003e isolates (27.65%) of our study involves the production of AMEs (aminoglycosides modifying enzymes) by bacteria, which modify and deactivate aminoglycoside drugs, thus imparting resistance to them. AG N-acetyltransferases (aac-3(IV) each specific to a position on the aminoglycoside (AG), carry out these modifications, as indicated by their nomenclature. This modification reduces the affinity between the modified aminoglycoside antibiotics and bacterial ribosomes, ultimately leading to drug resistance (Zhang et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSo, based on our study, we concluded that there is alarming evidence of antimicrobial resistance and high percentage of \u003cem\u003eE. coli\u003c/em\u003e isolates from bovine UTI is resistant to two or more antibiotic groups like findings observed by Mustapha et al. (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2020\u003c/span\u003e); Ismail and Abutarbush, (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledged:\u0026nbsp;\u003c/strong\u003eAuthors are thankful to HOD, VCC, LUVAS, Hisar.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement:\u0026nbsp;\u003c/strong\u003eAll datasets generated and/or analysed during the current study are presented in the article, the accompanying Source Data or Supplementary Information files, or are available from the corresponding author upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors contribution statement:\u0026nbsp;\u003c/strong\u003eTarun Kumar, Annu Yadav and Ankit Kumar conceptualized, conceived, and designed the study. Annu Yadav, Pawan Bagri and Dinesh did primer designing, performed molecular work. Pawan Bagri, Vinay Ganesh Rao Joshi and Babu Lal Jangir did laboratory analysis. Tarun Kumar and Neelesh Sindhu finalized the manuscript draft.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics Statement:\u003c/strong\u003e Approval for animals under this experiment was granted by the Institutional Animal Ethics Committee (IAEC) of the institute under the reference IAEC/LUVAS/28/10.Top of Form\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest statement:\u0026nbsp;\u003c/strong\u003eThe authors declare no conflict of interest\u003cstrong\u003e. \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding source:\u0026nbsp;\u003c/strong\u003eNo funding agency.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbdullah, A. R. \u0026amp; Mustafa, J. Y. Isolation and molecular detection study of bacterial causes pyelonephritis of cattle in Basrah province. \u003cem\u003eBiochem. Cell. 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The prevalence and distribution of aminoglycoside resistance genes. \u003cem\u003eBiosaf. Health\u003c/em\u003e. \u003cb\u003e5\u003c/b\u003e (01), 14\u0026ndash;20 (2023).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Antimicrobial resistance, Bovine, E. coli, Fim H, Serotype, Urinary tract infections","lastPublishedDoi":"10.21203/rs.3.rs-6831233/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6831233/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eWidespread emergence of multidrug-resistant and highly virulent uro- pathogenic \u003cem\u003eEscherichia coli\u003c/em\u003e (UPEC) strains pose a significant crisis for both human and animal populations worldwide. Aim of the present study was to characterize \u003cem\u003eEscherichia coli\u003c/em\u003e (\u003cem\u003eE. coli\u003c/em\u003e) strains isolated from urinary tract infected (UTI) bovines and to understand the distribution of antimicrobial resistance (AMR) in northern part of India. A cross-sectional study was carried out with sampling 17 randomly selected husbandry setups in 6 districts representing two agroclimatic zones. Out of 254 suspected animals from UTI, 105 animals found to be affected with UTI based on clinical examination, routine urine, culture testing and ultrasonography. Among the 105 affected animals, \u003cem\u003eE. coli\u003c/em\u003e was isolated from 47 animals turning out to be 44.76% positivity. A total of 13 serotypes of \u003cem\u003eE. coli\u003c/em\u003e were identified with maximum occurrence of O156 (23.40%) serotype. Among 13 serotypes isolated, five serotypes viz. O5, O128, O83, O11, and O159 are zoonotic important and are of public health concern. Antimicrobial resistance gene profiling depicted 100% occurrence of tet (A) and gyr A phenotype in \u003cem\u003eE. coli\u003c/em\u003e isolates. Remarkably, meropenem demonstrated the highest efficacy against the isolates, with 89.6% sensitivity. Unfortunately, 85.10% of the tested isolates displayed multidrug resistance, and all isolates carried the Fim H (100%) gene and two isolates carried hly A gene. Our findings emphasize genetic diversity and wide dispersion of zoonotically significant multidrug-resistant, virulent uropathogenic \u003cem\u003eE. coli\u003c/em\u003e strains among dairy animals.\u003c/p\u003e","manuscriptTitle":"Comprehensive Analysis of Escherichia coli in Bovine Urinary Tract Infections: Serotypes, Antibiotic Resistance, and Virulence Gene Profiles","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-10 18:50:50","doi":"10.21203/rs.3.rs-6831233/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-07-14T04:18:17+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"138184407286902123186511104354202020820","date":"2025-07-12T11:53:46+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-07-10T15:18:28+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-07-10T05:40:13+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"44560882112211008155144328237937929226","date":"2025-07-09T15:25:06+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"6837841440062527823630552447998874934","date":"2025-07-09T12:46:13+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-07-08T08:10:05+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"71144918623008655415347128985386677326","date":"2025-07-07T15:36:04+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-07-07T08:25:22+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-07-03T18:43:54+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-06-25T17:28:06+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-06-18T08:11:56+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-06-18T08:02:04+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"ec84f636-37d2-4bd3-83a1-b61085d0f104","owner":[],"postedDate":"July 10th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":51291770,"name":"Health sciences/Molecular medicine"},{"id":51291771,"name":"Health sciences/Nephrology"},{"id":51291772,"name":"Health sciences/Urology"}],"tags":[],"updatedAt":"2025-12-01T16:02:52+00:00","versionOfRecord":{"articleIdentity":"rs-6831233","link":"https://doi.org/10.1038/s41598-025-26316-2","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2025-11-27 15:58:00","publishedOnDateReadable":"November 27th, 2025"},"versionCreatedAt":"2025-07-10 18:50:50","video":"","vorDoi":"10.1038/s41598-025-26316-2","vorDoiUrl":"https://doi.org/10.1038/s41598-025-26316-2","workflowStages":[]},"version":"v1","identity":"rs-6831233","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6831233","identity":"rs-6831233","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}