Laboratory Analysis of the Resistance Spectrum and Antibacterial Susceptibility of Vibrio cholerae Strains Isolated in Kazakhstan from 1970 to 2024

Laboratory Analysis of the Resistance Spectrum and Antibacterial Susceptibility of Vibrio cholerae Strains Isolated in Kazakhstan from 1970 to 2024 | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Laboratory Analysis of the Resistance Spectrum and Antibacterial Susceptibility of Vibrio cholerae Strains Isolated in Kazakhstan from 1970 to 2024 Zyat Abdel Z., Zauresh Zhumadilova B., Raikhan Mussagalieva S., and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6902069/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract C holera is an acute diarrheal disease caused by the bacterium Vibrio cholerae , transmitted via the fecal–oral route, primarily through contaminated water or food. Despite progress in vaccination and antibiotic use, cholera continues to pose a significant public health threat, especially in regions with inadequate sanitation and unsafe drinking water. A major challenge in the management of cholera is the emergence of antibiotic-resistant V. cholerae strains, which compromise the effectiveness of standard therapeutic regimens. These resistant strains necessitate the development of novel treatment and prevention strategies. In 2018, studies on phenotypic resistance markers in Vibrio cholerae isolates collected in Kazakhstan revealed that 38.4% of the examined strains exhibited resistance, with monoresistant strains predominating (23.1%). Additionally, 9.6% of the isolates carried two or more resistance markers. Among the Vibrio cholerae O1 serogroup isolates, 38.5% were resistant, while in the non-O1 serogroup, resistance was observed in 40.0% of cases. Objectives: In this study, a comprehensive analysis was conducted on the antibiotic resistance of Vibrio cholerae strains isolated in Kazakhstan from 1970 to 2024, focusing on their susceptibility to various classes of antimicrobial agents. The molecular and biochemical mechanisms underlying the development of resistance were also investigated, and potential impacts on the epidemiological situation and biosafety were assessed. Methods : In this study, a total of 26 Vibrio cholerae strains isolated in Kazakhstan between 1970 and 2024 from clinical cases and environmental sources were used to screen for antimicrobial susceptibility and resistance profiles. Susceptibility testing was performed using the Kirby–Bauer disk diffusion method and E-test. To detect resistance genes, phenotypic assays and real-time Polymerase Chain Reaction were applied. One reference strain, 59 antibacterial agents across major drug classes, and a BacResista GLA Real-Time PCR Detection Kit were employed. Results : Phenotypic susceptibility testing of V. cholerae (n = 26) conducted in vitro demonstrated high sensitivity to cefotaxime, tetracycline, doxycycline, ciprofloxacin, and kanamycin. A similarly high level of susceptibility was observed for gentamicin, chloramphenicol, ampicillin, and rifampicin (96.2% of isolates). Real-time PCR results revealed no presence of resistance genes to glycopeptide or beta-lactam antibiotics in the tested V. cholerae strains. However, van A/B genes (Ct = 9.166, FAM channel) and the tem gene (Ct = 34.60, CY5 channel) were detected in the control strains Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 9027 (Ct = 8.954 and 24.85, respectively). Conclusions : The absence of resistance to major classes of antimicrobial agents among all 26 V. cholerae isolates indicates the continued high clinical efficacy of these antibiotics in the treatment of cholera. These findings are of critical importance in the context of potential epidemic outbreaks, as they provide clinicians with a reliable basis for selecting empirical therapy. Infectious Diseases cholera strains antibiotics resistance sensitivity statistics Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 1. Introduction Cholera is a rapidly progressing and potentially fatal disease that requires prompt intervention for effective control and treatment. Key strategies for managing cholera include rehydration therapy, antibiotic administration, and improvements in sanitation, all of which significantly reduce mortality and disease transmission. Cholera caused by toxigenic Vibrio cholerae strains remains one of the most dangerous infectious diseases, particularly in low- and middle-income countries where sanitation and hygiene conditions are inadequate. The disease also poses a serious threat in areas affected by natural disasters and humanitarian crises, where access to clean water and food supplies is compromised. V. cholerae is a Gram-negative, comma-shaped bacterium that causes acute watery diarrhea, leading to rapid dehydration and hypovolemia. Without timely treatment, cholera can lead to death within hours or days, with case fatality rates exceeding 50%. However, early rehydration and antibiotic therapy can reduce mortality to below 1% [ 1 – 4 ]. Antibiotic resistance is a global public health concern that affects not only well-known pathogens but also less-studied organisms such as Vibrio cholerae . Cholera has traditionally been treated with antibiotics such as tetracycline, doxycycline, and fluoroquinolones. However, the increasing use of these agents has led to the emergence of V. cholerae strains resistant to these drugs, posing significant challenges for disease management and increasing the risk of transmission [ 5 ]. According to the World Health Organization (WHO), Vibrio cholerae has demonstrated increasing resistance to conventional antibiotics, such as ampicillin and tetracycline, in several regions of Africa and Southeast Asia in recent decades [ 6 ]. Studies have shown that antibiotic-resistant strains can persist in the environment and be transmitted from person to person, thereby facilitating the spread of infection. A critical aspect of resistance research is understanding the mechanisms that enable V. cholerae to maintain viability under antimicrobial pressure. These mechanisms include both genetic adaptation and the horizontal transfer of resistance genes among bacteria [ 7 ]. For instance, resistance to tetracycline may be associated with genes encoding active efflux pumps or ribosomal protection proteins that prevent antibiotic binding [ 8 ]. Additionally, epidemiological and social factors—such as high population density, inadequate sanitation, and the widespread, unregulated use of antibiotics—play a major role in the dissemination of resistance [ 9 ]. Thus, understanding the causes of Vibrio cholerae resistance and elucidating its underlying mechanisms are essential for developing effective strategies for cholera treatment and prevention [ 10 ]. In 2018, studies were conducted to investigate phenotypic markers of antibiotic resistance among Vibrio cholerae isolates collected in Kazakhstan. Of the isolates analyzed, 38.4% exhibited resistance, with monoresistant strains predominating (23.1%), while 9.6% harbored two or more resistance markers [ 11 ]. Phenotypic resistance was most frequently observed against kanamycin (17.3%). Among Vibrio cholerae isolates of the O1 serogroup, 38.5% were resistant, compared to 40.0% of non-O1 serogroup isolates. A notably high proportion of resistant strains—45.8%—was recorded in the Kyzylorda region. Furthermore, resistant strains isolated from water sources in Almaty city, as well as in the Almaty and Zhambyl regions, shared a common phenotype characterized by resistance to erythromycin [ 11 ]. In this study, we conducted a comprehensive analysis of the antibiotic resistance of V. cholerae strains isolated in Kazakhstan between 1970 and 2024, examining their susceptibility to various classes of antimicrobial agents. Additionally, we investigated the molecular and biochemical mechanisms contributing to resistance, and we assessed their potential impact on the epidemiological situation and public biosafety. 2. Materials and Methods 2.1. Bacterial Strains and Isolates This study included 26 collection strains of Vibrio cholerae from different serogroups (O1 and non-O1), isolated from clinical samples and surface water in Almaty city and the Almaty, Turkistan, Mangystau, West Kazakhstan, and Karaganda regions of the Republic of Kazakhstan. These strains were collected between 1970 and 2024. Among them, 25 strains belonged to the V. cholerae O1 serogroup, biovar El Tor, and 1 strain was classified as V. cholerae non-O1. Specifically, 5 strains were isolated in Almaty, 1 from the Almaty region, 6 from the Turkistan region, 2 from the Mangystau region, 8 from the West Kazakhstan region, and 4 from the Karaganda region (Table 1 ). Table 1 List of Vibrio cholerae strains examined by serogroup, serovar, pathogenicity gene profile, Kazakhstan region, time, and source of isolation. Strain Serogroup Serovariant Pathogenicity Gene Profile Region, Year, and Place of Strain Isolation V. cholerae KZ-07-23 O1 Ogawa ctxAB + , tcpA + − Almaty, 2009, human V. cholerae KZ-07-22 O1 Ogawa ctxAB + , tcpA + − Almaty, 2024, human V. cholerae KZ-07-05 non O1 Ogawa ctxAB + , tcpA + − Almaty, 1998, water V. cholerae KZ-09-15 O1 Ogawa ctx + , ctx − Turkistan, 2003, water V. cholerae KZ-07-08 O1 Inaba ctx + , ctx − Turkistan, 2005, water V. cholerae KZ-07-26 O1 Inaba ctx + , ctx − Turkistan, 2005, water V. cholerae KZ-07-24 O1 Inaba ctx + , ctx − Turkistan, 2005, water V. cholerae KZ-05-27 O1 Ogawa ctx + , ctx − Turkistan, 2010, water V. cholerae KZ-01-12 O1 Ogawa ctxA − , tcpA − Turkistan, 2023, water V. cholerae KZ-17-13 O1 Ogawa ctxA − , tcpA − Mangystau, 1997, human V. cholerae KZ-19-21 O1 Ogawa ctxA − , tcpA − Mangystau, 1997, human V. cholerae KZ-07-03 O1 Inaba ctxA − , tcp − West Kazakhstan, 2016, water V. cholerae KZ-07-10 O1 Ogawa ctxA − , tcp − West Kazakhstan, 2016, water V. cholerae KZ-13-09 O1 Ogawa ctxA − , tcp − West Kazakhstan, 2016, water V. cholerae KZ-09-17 O1 Ogawa ctxA − , tcp − West Kazakhstan, 2016, water V. cholerae KZ-07-02 O1 Hikojima ctxA − , tcp − West Kazakhstan, 2016, water V. cholerae KZ-07-01 O1 Ogawa ctxA − West Kazakhstan, 2023, water V. cholerae KZ-12-04 O1 Ogawa ctxA − West Kazakhstan, 2023, water V. cholerae KZ-09-07 O1 Inaba ctxA − West Kazakhstan, 2023, water V. cholerae KZ-11-06 O1 Ogawa ctxA − , tcpA − Karaganda, 1997, human V. cholerae KZ-07-18 O1 Ogawa ctxA − , tcpA − Karaganda, 1997, human V. cholerae KZ-13-16 O1 Ogawa ctxA − , tcpA − Karaganda, 1997, human V. cholerae KZ-12-14 O1 Ogawa ctxA − , tcpA − Karaganda, 2000, human V. cholerae KZ-07-19 O1 Ogawa ctxA − , tcpA − Almaty, 1970, human V. cholerae KZ-02-20 O1 Ogawa ctxA − , tcpA − Almaty, 1970, human V. cholerae KZ-07-11 O1 Ogawa ctxA − , tcpA − Almaty, 1971, human 2.2. Antimicrobial Susceptibility Testing The susceptibility/resistance of Vibrio cholerae strains to antibacterial agents—including doxycycline (30 µg), ciprofloxacin (10 µg), tetracycline (30 µg), cefotaxime (30 µg), kanamycin (30 µg), nalidixic acid (30 µg), trimethoprim (5 µg), furazolidone (50 µg), gentamicin (10 µg), chloramphenicol (10 µg), ampicillin (25 µg), streptomycin (300 µg), and rifampicin (15 µg)—was assessed in accordance with established methodological guidelines [ 12 ]. A total of 59 antibiotics in disk form and 50 in E-test strips, representing 25 main groups, were used. Susceptibility testing was performed using the standard disk diffusion method (Kirby–Bauer test) and the E-test. Resistance genes were detected using an extended-spectrum β-lactamase (ESBL) phenotypic method [ 32 – 36 ] and real-time PCR for screening resistance to glycopeptides and beta-lactams [37]. Reference control strains included V. cholerae KA-37, Escherichia coli ATCC 25922, Salmonella typhimurium ATCC 14025, and Pseudomonas aeruginosa ATCC 9027. The strains were cultured on Mueller–Hinton agar (pH 7.3 ± 0.2) and Hottinger agar (pH 7.2 ± 0.1) at 37°C. For susceptibility testing, bacterial suspensions were prepared from 24-hour agar cultures in 0.85% isotonic sodium chloride solution, standardized using the McFarland standard (R092-1NO LOT0000633797; exp. 02/2026, HiMedia Laboratories Pvt. Ltd., Maharashtra, India), corresponding to 1.5 × 10⁸ CFU/mL (standards R092A and R092B). After 10–15 minutes, antibiotic disks were applied, and cultures were incubated at 37°C. Preliminary measurements were taken after 12 hours and finalized after 18 hours. Control assays included the reference strains and plates with sterile disks. The diameters of the inhibition zones around the disks were measured to the nearest millimeter using a precision measuring template from HiMedia Laboratories Pvt. Ltd., India. 2.3. Molecular Genetic Screening DNA extraction was performed using the commercial kit “RealBest UniMag” (Series C-8883, expiration date: July 22, 2025), produced by Vector-BEST, Russia, as well as the “RIBO-prep” kit (Cat. No. K2-9-Et-100, Russia), designed for automated DNA/RNA extraction systems. To detect antibiotic resistance determinants in bacterial lysates, a BacResista GLA Real-Time PCR Detection Kit (DNA-Technology LLC, Moscow, Russia) was used. This kit targets genes encoding resistance to glycopeptide and beta-lactam antibiotics, including vanA/B (vancomycin and teicoplanin); mecA (methicillin and oxacillin); tem , ctx-M-1 , and shv (penicillins and cephalosporins); and oxa-40-like , oxa-48-like , oxa-23-like , oxa-51-like , imp , kpc , ges , ndm , and vim (carbapenems). 3. Results The results of the laboratory screening for the antimicrobial susceptibility and resistance profiling of Vibrio cholerae strains (n = 26) isolated in the Republic of Kazakhstan between 1970 and 2024 from clinical and environmental sources are presented in Table 2 . Based on their cultural, morphological, biochemical, and serological characteristics, all strains were identified as typical representatives of the family Vibrionaceae, genus Vibrio , species cholerae , belonging to both O1 and non-O1 serogroups. Table 2 Ranges and diameters (in mm) of growth inhibition zones for 26 Vibrio cholerae strains isolated in Kazakhstan between 1970 and 2024 from clinical and environmental sources. Strain Isolation Area Number of V. cholerae Isolates Range of Minimum Inhibitory Concentration Values of Antibacterial Drugs by Main Group β-lactams Macroleades Tetracyclines Aminoglycosides Amphenicols Glycopeptides Lincosamides Fluoroquinolones Antibiotics of Different Groups Almaty city 5 16–40 20–27 24–29 19–28 25–28 23–28 21–29 30–40 0–33 Almaty region 1 19–39 23–24 26–30 18–29 28–29 25–26 25–26 30–35 16–28 Turkestan region 6 16–40 21–28 21–30 18–28 26–30 23–28 24–27 30–40 15–32 Mangistau region 2 17–40 25–26 24–30 17–26 25–26 24–26 25–28 30–39 16–31 West Kazakhstan region 8 16–40 20–26 21–30 17–30 25–30 20–25 21–29 29–40 16–33 Karaganda region 4 16–40 23–28 23–0 17–29 25–29 24–25 22–27 29–40 11–33 Total 26 16.6–39.8 22.1–26.5 23.1–24.8 17.6–28.3 25.8–28.6 21.1–26.1 23–27.5 29.6–39 5.3–31.6 Of the 26 V. cholerae O1 and non-O1 strains examined, 100% (26/26) were susceptible to the following antimicrobial agents: doxycycline (30 µg), ciprofloxacin (10 µg), tetracycline (30 µg), cefotaxime (30 µg), and kanamycin (30 µg). With respect to nalidixic acid (30 µg), 65.4% (17/26) of the strains demonstrated high susceptibility, 30.8% (8/26) exhibited intermediate susceptibility, and 3.8% (1/26) showed resistance. For trimethoprim (5 µg), 73.1% (19/26) of the isolates were highly susceptible, while 26.9% (7/26) showed intermediate susceptibility. For furazolidone (50 µg), 73.1% (19/26) of the V. cholerae strains were highly susceptible, while 26.9% (7/26) exhibited intermediate susceptibility. With respect to gentamicin (10 µg), 96.2% (25/26) of the isolates were highly susceptible, and 3.8% (1/26) showed intermediate susceptibility. Similarly, for chloramphenicol (10 µg) and ampicillin (25 µg), 96.2% (25/26) of the strains demonstrated high susceptibility, with 3.8% (1/26) showing intermediate susceptibility to each antibiotic. Regarding streptomycin (300 µg), 88.5% (23/26) of the V. cholerae strains were highly susceptible, 7.7% (2/26) showed intermediate susceptibility, and 3.8% (1/26) exhibited low susceptibility. For rifampicin (15 µg), 96.2% (25/26) of the strains demonstrated high susceptibility, while 3.8% (1/26) showed intermediate susceptibility. The results of the study, including the strain numbers and the names of the antibacterial agents tested on Mueller–Hinton and Hottinger agars, are presented in summary charts in Figs. 1 and 2 , along with selected individual results in Figs. 3 and 4 . The most active antibiotics belonged to the β-lactam group, including cefotaxime, cefixime, chloramphenicol, and cefamandole, followed by cefazolin, tetracycline, and ampicillin. Agents traditionally used for cholera treatment—such as ciprofloxacin, doxycycline, and azithromycin—demonstrated comparatively lower activity. Furazolidone exhibited the weakest activity against all 26 strains, with an average inhibition zone diameter of 11.1 mm (range: 10–18 mm). An in vitro phenotypic susceptibility analysis of V. cholerae (n = 26) demonstrated 100% sensitivity to β-lactams, tetracyclines, aminoglycosides, amphenicols, glycopeptides, lincosamides, and quinolones. Additionally, 96.5% of the strains were susceptible to antibiotics from other classes. The results of the susceptibility and resistance testing were confirmed using the standard E-test method, with strips indicating the minimum inhibitory concentration (Fig. 4 ). The results of the antimicrobial resistance gene detection in 26 V. cholerae strains using the BacResista GLA Real-Time PCR Detection Kit with real-time PCR are presented in Table 3 and Fig. 5 . Table 3 Results of antimicrobial resistance gene detection in 26 Vibrio cholerae strains using the BacResista GLA Real-Time PCR Detection Kit and real-time PCR. Name of Genetic Determinants and Verification Controls and Indicators by Species V. cholerae KA-37 Escherichia coli ATCC 25922 Salmonella typhimurium ATCC 14 , 025 Pseudomonas aeruginosa ATCC 9027 V. cholerae KZ00-00 *** Control « – » Control « + » FAM TBM * 7.507 8.029 15.33 7.648 ‾x = 7.17 – 23.27 imp – – – – – – 23.63 ctx-M-1 – – – – – – 22.20 van A/B – 9.166 – 8.954 – – 24.40 oxa-48-like – – – – – – 24.07 vim – – – – – – 24.31 oxa-23-like – – – – – – 23.43 shv – – – – – – 23.61 HEX IC ** 25.73 25.99 25.87 25.99 ‾x = 25.78 26.21 26.70 IC 26.91 26.79 25.94 26.79 ‾x = 24.43 25.58 25.81 IC 24.57 25.24 25.13 25.24 ‾x = 24.56 25.13 25.27 IC 24.37 24.39 24.93 24.39 ‾x = 24.60 24.94 25.10 IC 24.44 24.39 24.67 24.39 ‾x = 24.45 24.98 25.12 IC 24.62 – 24.96 – ‾x = 24.68 25.15 25.03 IC 24.50 24.37 24.56 24.37 ‾x = 24.63 24.67 24.65 CY5 oxa-51-like – – – – – – 24.69 Tem – 34.60 – 24.85 – – 20.20 mec A – – – – – – 24.40 oxa-40-like – – – – – – 24.79 Kpc – – – – – – 24.12 Ndm – – – – – – 23.53 Ges – – – – – – 23.24 * TBM—total bacterial mass; ** IC—internal control; *** KZ-00-00—average value based on results from 26 V. cholerae strains. Currently, research on the genetic determinants of bacterial resistance to glycopeptide and beta-lactam antibiotics is receiving significant attention, as these classes of antimicrobial agents are widely used for the treatment of complicated and/or severe infections. Investigating the most common types of beta-lactamases produced by various pathogenic bacteria can aid in interpreting their antibiotic susceptibility profiles, informing therapeutic decision-making, and enhancing infection control practices at the local level. A molecular genetic analysis aiming to detect antibiotic resistance determinants in the genomes of 26 Vibrio cholerae isolates, including clinical strains collected in Kazakhstan from 1970 to 2024, revealed no presence of resistance genes to glycopeptides ( vanA/B —vancomycin and teicoplanin) or beta-lactam antibiotics ( mecA —methicillin and oxacillin; tem , ctx-M-1 , and shv —penicillins and cephalosporins; oxa-40-like , oxa-48-like , oxa-23-like , oxa-51-like , imp , kpc , ges , ndm , and vim —carbapenems). The control strains used in this study included Escherichia coli ATCC 25922, Salmonella typhimurium ATCC 14025, and Pseudomonas aeruginosa ATCC 9027, in which the vanA/B gene (Ct = 9.166, FAM channel) and the tem gene (Ct = 34.60, CY5 channel) were detected in E. coli ATCC 25,922 and in P. aeruginosa ATCC 9027 (Ct = 8.954 and 24.85, respectively). Thus, the results confirm the absence of genetic markers of resistance to 59 antimicrobial agents in all examined V. cholerae isolates. No resistance was detected to the following major classes of antibiotics: extended-spectrum beta-lactams (penicillins, cephalosporins, and carbapenems), monobactams, macrolides, tetracyclines, aminoglycosides, amphenicols, glycopeptides, lincosamides, fluoroquinolones, and other antibiotic groups. These findings highlight the importance of the regular surveillance of V. cholerae susceptibility to antimicrobial agents, which enables timely adjustments to therapeutic strategies and effective responses to changes in the epidemiological situation. 4. Discussion Vibrio species undergo genetic changes to enhance their adaptation and resistance to antibiotics. A study on the susceptibility of V. cholerae O1 El Tor strains to polymyxin B (PB) in Odisha, India, between 1995 and 2019 found that 89.4% of 1,200 strains were resistant, while 10.6% were susceptible. Susceptibility began to appear from 2005 onward, with the exception of 2015. An E-test analysis revealed that strains with the ctxB7 genotype had a lower minimum inhibitory concentration (MIC ≤ 4 µg/mL), whereas ctxB1 genotypes exhibited higher MIC values (24 and 32 µg/mL) [ 13 ]. The study conducted in Odisha, India, between 1995 and 2019, examined changes in antibiotic susceptibility, as well as the presence of virulence and resistance genes, in Vibrio cholerae O1 strains. Antimicrobial susceptibility was assessed using the disk diffusion method, and resistance and virulence genes were identified using PCR. All strains were susceptible to gentamicin, chloramphenicol, norfloxacin, and ciprofloxacin but exhibited resistance to one or more other antibiotics. Resistance genes (SulII, dfrA1, and strB) and SXT elements were detected in 90% of the isolates. The Haitian variant of tcpA emerged in 1999 and gradually increased in prevalence. Multiplex PCR confirmed the presence of virulence genes (toxR, ompU, ace, rtxC, ctxA, tcpA, rfbO1, and ompW) in all strains [ 14 , 15 ]. Antibiotic resistance among epidemic Vibrio cholerae strains is increasing. A study analyzing V. cholerae O1 El Tor strains isolated in China from 1961 to 2010 showed that resistance levels were generally low, except for nalidixic acid (45.9%), tetracycline (11%), and trimethoprim/sulfamethoxazole (38.5%). All strains from the 1960s were susceptible, whereas resistance levels rose significantly during the 1990s and beyond. Class I integrons were more frequently found among strains from 1993–1998, and the SXT element was commonly detected in isolates collected after 1993. These findings highlight the growing resistance of epidemic V. cholerae strains in China [ 16 ]. In Odisha, cholera has remained a significant threat, causing outbreaks with high morbidity and mortality. A study characterized Vibrio cholerae O1 strains from the 2018 and 2019 outbreaks in Bargarh and Rayagada. Multidrug-resistant strains were isolated and analyzed using PCR and pulsed-field gel electrophoresis (PFGE). All strains carried both virulence and resistance genes, including dfrA1 (100%), strB (76.9%), and intSXT (61.5%). The outbreaks were driven by multidrug-resistant strains, underscoring the need for continuous monitoring to prevent future epidemics [ 17 ]. In 2022, the number of cholera cases in the WHO European Region more than doubled. In one study, a total of 49 Vibrio cholerae O1 isolates collected in Europe were analyzed to confirm their affiliation with the seventh pandemic El Tor lineage (7PET) and to assess their virulence and antibiotic resistance. All isolates were found to belong to the Pre - AFR15 sublineage, which is likely associated with the global surge in cholera cases. Antibiotic resistance analysis revealed no abnormal resistance profiles. Genomic sequencing was recommended for accurate strain identification and the monitoring of their evolutionary dynamics [ 18 ]. Vibrio cholerae O139, first identified in 1992, has been recognized as a causative agent of cholera epidemics. Since 1993, O139 strains have been isolated in China, leading to sporadic cases and foodborne outbreaks. In a study conducted in China, the antibiotic resistance of strains collected between 1993 and 2009 was analyzed. Initially, the strains showed resistance to only a few antibiotics; however, after 1998, multidrug resistance increased significantly. In contrast, V. cholerae O1 strains exhibited lower levels of resistance. Toxigenic O139 strains showed a high prevalence of resistance elements, indicating complex resistance patterns. The high level of multidrug resistance poses a growing public health threat, highlighting the need for continuous surveillance and control [ 19 , 20 ]. In Africa, Vibrio cholerae O1 strains with multidrug resistance have been reported for many years, and during recent epidemics in Kenya, new resistance phenotypes have emerged. One study aimed to analyze the epidemiological features and antimicrobial resistance of strains isolated in Kenya during 2007–2010 and 2015–2016. A total of 228 strains were examined, including 226 clinical and 2 environmental isolates. All strains belonged to the El Tor biotype and were found to be susceptible to ceftriaxone, gentamicin, and ciprofloxacin, while showing resistance to trimethoprim–sulfamethoxazole, streptomycin, and nalidixic acid. The SXT integrative element was detected in 95.5% of the strains, whereas class 1, 2, and 3 integrons were not detected. Overall, 64.5% of the isolates exhibited multidrug resistance. During the 2015–2016 outbreaks, strains with chromosomal mutations were identified that remained susceptible to most antibiotics except nalidixic acid. The study revealed two distinct resistance patterns and emphasized the importance of continuous surveillance to prevent the emergence of new resistant strains [ 21 – 23 ]. Vibrio cholerae is a common inhabitant of aquatic ecosystems, and the state of West Bengal in India—particularly the Ganges Delta region—is known for having a high cholera burden. In a study conducted in Midnapore, V. cholerae isolates were obtained from wastewater samples. All isolates belonged to non-O1/non-O139 serogroups. The majority (74.7%) exhibited moderate biofilm-forming ability, while 6.3% demonstrated strong biofilm formation. PCR screening revealed that most isolates carried genes associated with biofilm formation (such as cdgH , cdgM , cdgK , etc.). Two strains (3.17%) contained the cholera toxin genes ctxA and ctxB . Approximately 24.8% of the isolates carried the ompU gene, while other virulence factors were detected less frequently. All isolates tested positive for toxT , toxR , and hapR , which regulate virulence expression. Resistance profiling indicated both increased antibiotic susceptibility and the presence of multidrug resistance (MDR). The detection of such strains in the environment is a concern. The high prevalence of diguanylate cyclases (DGCs) suggests their potential as alternative therapeutic targets against MDR strains [ 24 , 25 ]. A molecular genetic analysis conducted on 26 Vibrio cholerae O1 isolates, including clinical strains collected in Kazakhstan between 1970 and 2024, revealed no resistance genes to glycopeptide and beta-lactam antibiotics ( vanA/B, mecA, tem, ctx-M-1, shv, oxa-40-like, oxa-48-like, oxa-23-like, oxa-51-like, imp, kpc, ges, ndm , and vim ). These findings indicate the absence of genetically mediated resistance to major classes of antimicrobial agents in the analyzed isolates [ 24 , 29 , 30 ]. Such results underscore the importance of the regular epidemiological monitoring of pathogen susceptibility to antibiotics, enabling timely responses to potential shifts in resistance patterns [ 25 , 26 , 31 ]. The absence of resistance to major antibiotic classes among all examined V. cholerae isolates indicates the continued high clinical efficacy of these drugs. These findings are particularly significant in the context of potential cholera outbreaks, as they provide clinicians with a reliable foundation for selecting empirical therapy [ 27 , 28 ]. Furthermore, phenotypic susceptibility testing showed that 100% of the strains were sensitive to doxycycline, ciprofloxacin, tetracycline, cefotaxime, and kanamycin. A high susceptibility was also observed in 96.2% of the isolates to chloramphenicol, ampicillin, gentamicin, and rifampicin; 88.5% to streptomycin; and 73.1% to trimethoprim and furazolidone. At the same time, intermediate susceptibility and isolated cases of resistance were observed for nalidixic acid, streptomycin, and furazolidone [ 29 ]. 5. Conclusions An analysis of antibiotic susceptibility in 26 Vibrio cholerae O1 strains isolated from various regions of Kazakhstan over nearly five decades demonstrated the sustained efficacy of the most commonly used antimicrobial agents. Isolated cases of resistance—particularly to nalidixic acid, streptomycin, and trimethoprim—highlight the importance of continued systematic surveillance of antimicrobial resistance in V. cholerae . The findings of this study may inform treatment planning, preventive measures, and local microbiological surveillance efforts in cholera-endemic areas. All Vibrio cholerae O1 strains isolated in Kazakhstan between 1970 and 2024 demonstrated a high susceptibility to several critically important antimicrobial agents, including doxycycline, ciprofloxacin, tetracycline, cefotaxime, and kanamycin. High levels of susceptibility were also observed for gentamicin, chloramphenicol, ampicillin, and rifampicin (96.2% of strains). Moderate resistance was observed against nalidixic acid, trimethoprim, furazolidone, chloramphenicol, ampicillin, streptomycin, and rifampicin. All strains, despite variations in serovar and pathogenicity profiles ( ctxAB⁺ and tcpA⁺/⁻ ), remain susceptible to the primary drugs used in cholera treatment, indicating their continued potential clinical effectiveness at the present stage. No resistance genes to glycopeptide or beta-lactam antibiotics were detected via real-time PCR in the tested strains. However, the vanA/B (Ct = 9.166, FAM channel) and tem (Ct = 34.60, CY5 channel) genes were identified in Escherichia coli ATCC 25922 and in Pseudomonas aeruginosa ATCC 9027, with Ct values of 8.954 and 24.85, respectively. Over the study period (1970–2024), no significant increase in the proportion of polyresistant strains was observed. However, the presence or emergence of even a single resistant isolate suggests the possibility of gene transfer under natural conditions, indicating the ongoing circulation of Vibrio cholerae O1 between humans and environmental sources. This highlights the need for continuous epidemiological and microbiological surveillance. Declarations Author Contributions: Conceptualization, Z.A., Z.Z. and R.M.; methodology, Z.A., R.M. and B.T.; software, D.O.; validation, Z.A., R.M., B.B., B.T., Z.B., D.O., N.S., Z.D., and B.A.; formal analysis, B.T., B.B., D.O., N.S. and B.A.; investigation, Z.A., Z.Z., and R.M.; resources, Z.D., B.B., B.T., D.O. and N.S.; data curation, Z.A., R.M., D.O. and N.S.; writing—original draft preparation, Z.A.; writing—review and editing, Z.Z., D.O. and R.M.; visualization, Z.A. and D.O.; supervision, Z.Z., R.M. and A.A.; project administration, D.O.; funding acquisition, Z.A. All authors have read and agreed to the published version of the manuscript Funding: This research was conducted within the framework of the project funded by the Ministry of Science and Higher Education of the Republic of Kazakhstan, titled “Investigation of Antibiotic Resistance Genes in Plague and Cholera Pathogens and the Development of a PCR Test System” (Project IRN: AP19679355). The project was supported by the Committee of Science of the Ministry of Science and Higher Education of the Republic of Kazakhstan. Institutional Review Board Statement: Not applicable. Informed Consent Statement: Not applicable. Data Availability Statement: The original contributions presented in this study are included in the article. Further inquiries can be directed to the corresponding authors Conflicts of Interest: The authors declare no conflicts of interest. References Ojeda Rodriguez JA, Hashmi MF, Kahwaji CI Vibrio cholerae Infection. In StatPearls ; StatPearls Publishing: Treasure Island, FL, USA. Available online: https://www.ncbi.nlm.nih.gov/books/NBK526099/ (cited 2025 Jun 5) Igere BE, Onohuean H, Iwu DC, Igbinosa EO (2023) Polymyxin sensitivity/resistance cosmopolitan status, epidemiology and prevalence among O1/O139 and non-O1/non-O139 Vibrio cholerae : A meta-analysis. 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Decod Infect Transm 3:100042. https://doi.org/10.1016/j.dcit.2025.100042 Kolobe MT (2025) Cholera Outbreak in Chienge District, Luapula Province, Zambia—31st May–9th June 2023: Re-emerging threat. Int J Infect Dis 152:107433. https://doi.org/10.1016/j.ijid.2024.107433 Boué Y, Niang M, Lapostolle A, Chamouine A, Cattin TB, Favre M, Rouard C, Mortier C, Piarroux R, Carvelli J (2025) Cholera outbreak in Mayotte (France): A retrospective description of 16 patients treated for hypovolemia in the ICU. Infect Dis Now 55:105020. https://doi.org/10.1016/j.idnow.2024.105020 Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing. Sanitary and epidemiological rules SP 3.1.7.2826-10. Cholera prevention. Moscow (2010) Available online: https://docs.cntd.ru/document/1200080421 (accessed on) World Health Organization (2014) Antimicrobial Resistance: Global Report on Surveillance. WHO, Geneva, Switzerland Okeke IN, Laxminarayan R, Bhutta ZA, Duse AG, Jenkins P, O'Brien TF, Pablos-Mendez A, Klugman KP (2005) Antimicrobial resistance in developing countries. Part I: Recent trends and current status. Lancet Infect Dis 5:481–493 Boucher HW, Talbot GH, Bradley JS, Edwards JE, Gilbert D, Rice LB, Scheld M, Spellberg B, Bartlett J (2009) Bad bugs, no drugs: no ESKAPE! An update from the Infectious Diseases Society of America. Clin Infect Dis 48:1–12 Hasan NA, Choi SY, Eppinger M, Clark PW, Chen A, Alam M, Haley BJ, Taviani E, Hine E, Su Q, Tallon LJ (2012) Genomic diversity of 2010 Haitian cholera outbreak strains. Proc. Natl. Acad. Sci. USA 109 , E2010–E2017 Faruque SM, Nair GB (2014) Epidemiology, genetics, and ecology of toxigenic Vibrio cholerae . Microbiol Spectr 2:VE–0004 CLSI (2023) Performance Standards for Antimicrobial Susceptibility Testing , 33rd ed., CLSI Supplement M100; Clinical and Laboratory Standards Institute: Wayne, PA, USA Davies J, Davies D (2010) Origins and evolution of antibiotic resistance. Microbiol Mol Biol Rev 74:417–433 Igbinosa EO, Okoh AI (2010) Antibiotic susceptibility profile o f Vibrio cholerae O1 strains isolated from wastewater final effluents in South Africa. Environ Monit Assess 168:321–329 Federal Center for Hygiene and Epidemiology of Rospotrebnadzor (2010) Guidelines for determining the susceptibility of causative agents of dangerous bacterial infections (plague, anthrax, etc.) to antibacterial drugs: Methodical recommendations 4.2.2495-09. Russia, Moscow, p 59. (In Russian) Abdirasilova AA, Abdel ZZ (2023) Methodological Recommendations for Real-Time Polymerase Chain Reaction (RT-PCR) for the Detection of Plague Microbe DNA. Almaty, Kazakhstan, p 37. (In Russian)KazBookExport Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6902069","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":471730180,"identity":"3419dc87-1a03-4ee0-a355-61f4c9a99744","order_by":0,"name":"Zyat Abdel Z.","email":"","orcid":"","institution":"Masgut Aikimbayev’s National Scientific Center Especially Dangerous Infections","correspondingAuthor":false,"prefix":"","firstName":"Zyat","middleName":"Abdel","lastName":"Z.","suffix":""},{"id":471730181,"identity":"72612956-934d-456a-87c8-47490b9f8fba","order_by":1,"name":"Zauresh Zhumadilova 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06:19:37","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-6902069/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6902069/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":84921234,"identity":"2d51c08d-12a7-46c3-80d3-354714693010","added_by":"auto","created_at":"2025-06-18 19:47:22","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":2912430,"visible":true,"origin":"","legend":"\u003cp\u003eRange of inhibition zone diameters for 26 \u003cem\u003eVibrio cholerae\u003c/em\u003e strains on Hottinger agar, in millimeters.\u003c/p\u003e","description":"","filename":"Figura1.png","url":"https://assets-eu.researchsquare.com/files/rs-6902069/v1/14e1854619238b40418caa94.png"},{"id":84920598,"identity":"a30c3678-b26e-47d3-986a-847b95e8dce4","added_by":"auto","created_at":"2025-06-18 19:39:22","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":3591298,"visible":true,"origin":"","legend":"\u003cp\u003eRange of inhibition zone diameters for 26 \u003cem\u003eVibrio cholerae\u003c/em\u003e strains on Mueller–Hinton agar, in millimeters.\u003c/p\u003e","description":"","filename":"Figura2.png","url":"https://assets-eu.researchsquare.com/files/rs-6902069/v1/f71c6af7ccefe2091423188b.png"},{"id":84920600,"identity":"b3d35ab6-b8f2-4bd6-a5cc-0cf00a211074","added_by":"auto","created_at":"2025-06-18 19:39:22","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":322201,"visible":true,"origin":"","legend":"\u003cp\u003eInhibition zone diameters determined using the disk diffusion method and the detection of ESBL resistance genes using the phenotypic method: (a–d)—susceptibility to antibiotics; (e, f)—absence of resistance genes; Image shows a Petri dish with a visual diameter of 35 cm.\u003c/p\u003e","description":"","filename":"Figura3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6902069/v1/be4485a8300919ecddef9cdc.jpg"},{"id":84920604,"identity":"8c4ada2b-8e9e-495e-80c2-323f836e6c58","added_by":"auto","created_at":"2025-06-18 19:39:22","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":326187,"visible":true,"origin":"","legend":"\u003cp\u003eResults of susceptibility testing using the E-test method: panels (a–f) show the susceptibility of two different Vibrio cholerae strains to gentamicin, cefotaxime, and ofloxacin. Panels (a), (b), and (c) correspond to Strain 1, while panels (d), (e), and (f) correspond to Strain 2.\u003c/p\u003e","description":"","filename":"Figura4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6902069/v1/fafebe07c7b4950991ff7a17.jpg"},{"id":84920607,"identity":"5e579eda-1eff-4b7c-83f3-cc9f9f0a1cfa","added_by":"auto","created_at":"2025-06-18 19:39:22","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":425878,"visible":true,"origin":"","legend":"\u003cp\u003eAmplification curves by fluorophore detection channels: (\u003cstrong\u003ea\u003c/strong\u003e) FAM; (\u003cstrong\u003eb\u003c/strong\u003e) HEX; (\u003cstrong\u003ec\u003c/strong\u003e) CY5. Red and green dots represent PCR amplification curves of different samples. Red dots indicate positive amplification signals, while green dots indicate negative or no amplification.\u003c/p\u003e","description":"","filename":"Figura5.png","url":"https://assets-eu.researchsquare.com/files/rs-6902069/v1/cb3a772d5770c6e8bfb95c5d.png"},{"id":84922010,"identity":"d26402b5-be66-412d-b7e9-84549f661986","added_by":"auto","created_at":"2025-06-18 20:03:26","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":8318300,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6902069/v1/f9521c31-1b46-4afd-a2fd-3bf0273ba48a.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003eLaboratory Analysis of the Resistance Spectrum and Antibacterial Susceptibility of \u003cem\u003eVibrio cholerae\u003c/em\u003e Strains Isolated in Kazakhstan from 1970 to 2024\u003c/p\u003e","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eCholera is a rapidly progressing and potentially fatal disease that requires prompt intervention for effective control and treatment. Key strategies for managing cholera include rehydration therapy, antibiotic administration, and improvements in sanitation, all of which significantly reduce mortality and disease transmission. Cholera caused by toxigenic \u003cem\u003eVibrio cholerae\u003c/em\u003e strains remains one of the most dangerous infectious diseases, particularly in low- and middle-income countries where sanitation and hygiene conditions are inadequate. The disease also poses a serious threat in areas affected by natural disasters and humanitarian crises, where access to clean water and food supplies is compromised. \u003cem\u003eV. cholerae\u003c/em\u003e is a Gram-negative, comma-shaped bacterium that causes acute watery diarrhea, leading to rapid dehydration and hypovolemia.\u003c/p\u003e \u003cp\u003eWithout timely treatment, cholera can lead to death within hours or days, with case fatality rates exceeding 50%. However, early rehydration and antibiotic therapy can reduce mortality to below 1% [\u003cspan additionalcitationids=\"CR2 CR3\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Antibiotic resistance is a global public health concern that affects not only well-known pathogens but also less-studied organisms such as \u003cem\u003eVibrio cholerae\u003c/em\u003e. Cholera has traditionally been treated with antibiotics such as tetracycline, doxycycline, and fluoroquinolones. However, the increasing use of these agents has led to the emergence of \u003cem\u003eV. cholerae\u003c/em\u003e strains resistant to these drugs, posing significant challenges for disease management and increasing the risk of transmission [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAccording to the World Health Organization (WHO), \u003cem\u003eVibrio cholerae\u003c/em\u003e has demonstrated increasing resistance to conventional antibiotics, such as ampicillin and tetracycline, in several regions of Africa and Southeast Asia in recent decades [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Studies have shown that antibiotic-resistant strains can persist in the environment and be transmitted from person to person, thereby facilitating the spread of infection. A critical aspect of resistance research is understanding the mechanisms that enable \u003cem\u003eV. cholerae\u003c/em\u003e to maintain viability under antimicrobial pressure. These mechanisms include both genetic adaptation and the horizontal transfer of resistance genes among bacteria [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. For instance, resistance to tetracycline may be associated with genes encoding active efflux pumps or ribosomal protection proteins that prevent antibiotic binding [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Additionally, epidemiological and social factors\u0026mdash;such as high population density, inadequate sanitation, and the widespread, unregulated use of antibiotics\u0026mdash;play a major role in the dissemination of resistance [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThus, understanding the causes of \u003cem\u003eVibrio cholerae\u003c/em\u003e resistance and elucidating its underlying mechanisms are essential for developing effective strategies for cholera treatment and prevention [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn 2018, studies were conducted to investigate phenotypic markers of antibiotic resistance among \u003cem\u003eVibrio cholerae\u003c/em\u003e isolates collected in Kazakhstan. Of the isolates analyzed, 38.4% exhibited resistance, with monoresistant strains predominating (23.1%), while 9.6% harbored two or more resistance markers [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Phenotypic resistance was most frequently observed against kanamycin (17.3%). Among \u003cem\u003eVibrio cholerae\u003c/em\u003e isolates of the O1 serogroup, 38.5% were resistant, compared to 40.0% of non-O1 serogroup isolates. A notably high proportion of resistant strains\u0026mdash;45.8%\u0026mdash;was recorded in the Kyzylorda region. Furthermore, resistant strains isolated from water sources in Almaty city, as well as in the Almaty and Zhambyl regions, shared a common phenotype characterized by resistance to erythromycin [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn this study, we conducted a comprehensive analysis of the antibiotic resistance of \u003cem\u003eV. cholerae\u003c/em\u003e strains isolated in Kazakhstan between 1970 and 2024, examining their susceptibility to various classes of antimicrobial agents. Additionally, we investigated the molecular and biochemical mechanisms contributing to resistance, and we assessed their potential impact on the epidemiological situation and public biosafety.\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Bacterial Strains and Isolates\u003c/h2\u003e \u003cp\u003eThis study included 26 collection strains of \u003cem\u003eVibrio cholerae\u003c/em\u003e from different serogroups (O1 and non-O1), isolated from clinical samples and surface water in Almaty city and the Almaty, Turkistan, Mangystau, West Kazakhstan, and Karaganda regions of the Republic of Kazakhstan. These strains were collected between 1970 and 2024. Among them, 25 strains belonged to the \u003cem\u003eV. cholerae\u003c/em\u003e O1 serogroup, biovar El Tor, and 1 strain was classified as \u003cem\u003eV. cholerae\u003c/em\u003e non-O1. Specifically, 5 strains were isolated in Almaty, 1 from the Almaty region, 6 from the Turkistan region, 2 from the Mangystau region, 8 from the West Kazakhstan region, and 4 from the Karaganda region (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eList of \u003cem\u003eVibrio cholerae\u003c/em\u003e strains examined by serogroup, serovar, pathogenicity gene profile, Kazakhstan region, time, and source of isolation.\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\u003eStrain\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSerogroup\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSerovariant\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePathogenicity Gene Profile\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eRegion, Year, and Place of Strain Isolation\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-07-23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxAB\u003csup\u003e+\u003c/sup\u003e, tcpA\u003csup\u003e+ \u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAlmaty, 2009, human\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-07-22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxAB\u003csup\u003e+\u003c/sup\u003e, tcpA\u003csup\u003e+ \u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAlmaty, 2024, human\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-07-05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003enon\u003c/em\u003e O1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxAB\u003csup\u003e+\u003c/sup\u003e, tcpA\u003csup\u003e+ \u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAlmaty, 1998, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-09-15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectx\u003csup\u003e+\u003c/sup\u003e, ctx\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTurkistan, 2003, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-07-08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eInaba\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectx\u003csup\u003e+\u003c/sup\u003e, ctx\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTurkistan, 2005, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-07-26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eInaba\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectx\u003csup\u003e+\u003c/sup\u003e, ctx\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTurkistan, 2005, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-07-24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eInaba\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectx\u003csup\u003e+\u003c/sup\u003e, ctx\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTurkistan, 2005, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-05-27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectx\u003csup\u003e+\u003c/sup\u003e, ctx\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTurkistan, 2010, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-01-12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcpA\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTurkistan, 2023, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-17-13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcpA\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMangystau, 1997, human\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-19-21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcpA\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMangystau, 1997, human\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-07-03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eInaba\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcp\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eWest Kazakhstan, 2016, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-07-10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcp\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eWest Kazakhstan, 2016, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-13-09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcp\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eWest Kazakhstan, 2016, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-09-17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcp\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eWest Kazakhstan, 2016, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-07-02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eHikojima\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcp\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eWest Kazakhstan, 2016, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-07-01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eWest Kazakhstan, 2023, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-12-04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eWest Kazakhstan, 2023, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-09-07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eInaba\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eWest Kazakhstan, 2023, water\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-11-06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcpA\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eKaraganda, 1997, human\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-07-18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcpA\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eKaraganda, 1997, human\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-13-16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcpA\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eKaraganda, 1997, human\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-12-14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcpA\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eKaraganda, 2000, human\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-07-19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcpA\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAlmaty, 1970, human\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-02-20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcpA\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAlmaty, 1970, human\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ-07-11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eO1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOgawa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ectxA\u003csup\u003e\u0026minus;\u003c/sup\u003e, tcpA\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAlmaty, 1971, human\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 \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Antimicrobial Susceptibility Testing\u003c/h2\u003e \u003cp\u003eThe susceptibility/resistance of \u003cem\u003eVibrio cholerae\u003c/em\u003e strains to antibacterial agents\u0026mdash;including doxycycline (30 \u0026micro;g), ciprofloxacin (10 \u0026micro;g), tetracycline (30 \u0026micro;g), cefotaxime (30 \u0026micro;g), kanamycin (30 \u0026micro;g), nalidixic acid (30 \u0026micro;g), trimethoprim (5 \u0026micro;g), furazolidone (50 \u0026micro;g), gentamicin (10 \u0026micro;g), chloramphenicol (10 \u0026micro;g), ampicillin (25 \u0026micro;g), streptomycin (300 \u0026micro;g), and rifampicin (15 \u0026micro;g)\u0026mdash;was assessed in accordance with established methodological guidelines [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. A total of 59 antibiotics in disk form and 50 in E-test strips, representing 25 main groups, were used. Susceptibility testing was performed using the standard disk diffusion method (Kirby\u0026ndash;Bauer test) and the E-test. Resistance genes were detected using an extended-spectrum β-lactamase (ESBL) phenotypic method [\u003cspan additionalcitationids=\"CR33 CR34 CR35\" citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e] and real-time PCR for screening resistance to glycopeptides and beta-lactams [37]. Reference control strains included \u003cem\u003eV. cholerae\u003c/em\u003e KA-37, \u003cem\u003eEscherichia coli\u003c/em\u003e ATCC 25922, \u003cem\u003eSalmonella typhimurium\u003c/em\u003e ATCC 14025, and \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e ATCC 9027. The strains were cultured on Mueller\u0026ndash;Hinton agar (pH 7.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2) and Hottinger agar (pH 7.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1) at 37\u0026deg;C.\u003c/p\u003e \u003cp\u003eFor susceptibility testing, bacterial suspensions were prepared from 24-hour agar cultures in 0.85% isotonic sodium chloride solution, standardized using the McFarland standard (R092-1NO LOT0000633797; exp. 02/2026, HiMedia Laboratories Pvt. Ltd., Maharashtra, India), corresponding to 1.5 \u0026times; 10⁸ CFU/mL (standards R092A and R092B). After 10\u0026ndash;15 minutes, antibiotic disks were applied, and cultures were incubated at 37\u0026deg;C. Preliminary measurements were taken after 12 hours and finalized after 18 hours. Control assays included the reference strains and plates with sterile disks. The diameters of the inhibition zones around the disks were measured to the nearest millimeter using a precision measuring template from HiMedia Laboratories Pvt. Ltd., India.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3. Molecular Genetic Screening\u003c/h2\u003e \u003cp\u003eDNA extraction was performed using the commercial kit \u0026ldquo;RealBest UniMag\u0026rdquo; (Series C-8883, expiration date: July 22, 2025), produced by Vector-BEST, Russia, as well as the \u0026ldquo;RIBO-prep\u0026rdquo; kit (Cat. No. K2-9-Et-100, Russia), designed for automated DNA/RNA extraction systems.\u003c/p\u003e \u003cp\u003eTo detect antibiotic resistance determinants in bacterial lysates, a BacResista GLA Real-Time PCR Detection Kit (DNA-Technology LLC, Moscow, Russia) was used. This kit targets genes encoding resistance to glycopeptide and beta-lactam antibiotics, including \u003cb\u003evanA/B\u003c/b\u003e (vancomycin and teicoplanin); \u003cb\u003emecA\u003c/b\u003e (methicillin and oxacillin); \u003cb\u003etem\u003c/b\u003e, \u003cb\u003ectx-M-1\u003c/b\u003e, and \u003cb\u003eshv\u003c/b\u003e (penicillins and cephalosporins); and \u003cb\u003eoxa-40-like\u003c/b\u003e, \u003cb\u003eoxa-48-like\u003c/b\u003e, \u003cb\u003eoxa-23-like\u003c/b\u003e, \u003cb\u003eoxa-51-like\u003c/b\u003e, \u003cb\u003eimp\u003c/b\u003e, \u003cb\u003ekpc\u003c/b\u003e, \u003cb\u003eges\u003c/b\u003e, \u003cb\u003endm\u003c/b\u003e, and \u003cb\u003evim\u003c/b\u003e (carbapenems).\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cp\u003eThe results of the laboratory screening for the antimicrobial susceptibility and resistance profiling of \u003cem\u003eVibrio cholerae\u003c/em\u003e strains (n\u0026thinsp;=\u0026thinsp;26) isolated in the Republic of Kazakhstan between 1970 and 2024 from clinical and environmental sources are presented in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003cp\u003eBased on their cultural, morphological, biochemical, and serological characteristics, all strains were identified as typical representatives of the family Vibrionaceae, genus \u003cem\u003eVibrio\u003c/em\u003e, species \u003cem\u003echolerae\u003c/em\u003e, belonging to both O1 and non-O1 serogroups.\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\u003eRanges and diameters (in mm) of growth inhibition zones for 26 \u003cem\u003eVibrio cholerae\u003c/em\u003e strains isolated in Kazakhstan between 1970 and 2024 from clinical and environmental sources.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"11\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eStrain Isolation Area\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eNumber of \u003cem\u003eV. cholerae\u003c/em\u003e Isolates\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"9\" nameend=\"c11\" namest=\"c3\"\u003e \u003cp\u003eRange of Minimum Inhibitory Concentration Values of Antibacterial Drugs by Main Group\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eβ-lactams\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMacroleades\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTetracyclines\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAminoglycosides\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eAmphenicols\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eGlycopeptides\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eLincosamides\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eFluoroquinolones\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003eAntibiotics of Different Groups\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAlmaty city\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20\u0026ndash;27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e24\u0026ndash;29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e19\u0026ndash;28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e25\u0026ndash;28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e23\u0026ndash;28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e21\u0026ndash;29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e30\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e0\u0026ndash;33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAlmaty region\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e19\u0026ndash;39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e23\u0026ndash;24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e26\u0026ndash;30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e18\u0026ndash;29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e28\u0026ndash;29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e25\u0026ndash;26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e25\u0026ndash;26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e30\u0026ndash;35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e16\u0026ndash;28\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTurkestan region\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e21\u0026ndash;28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e21\u0026ndash;30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e18\u0026ndash;28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e26\u0026ndash;30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e23\u0026ndash;28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e24\u0026ndash;27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e30\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e15\u0026ndash;32\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMangistau region\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e25\u0026ndash;26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e24\u0026ndash;30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e17\u0026ndash;26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e25\u0026ndash;26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e24\u0026ndash;26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e25\u0026ndash;28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e30\u0026ndash;39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e16\u0026ndash;31\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWest Kazakhstan region\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20\u0026ndash;26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e21\u0026ndash;30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e17\u0026ndash;30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e25\u0026ndash;30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e20\u0026ndash;25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e21\u0026ndash;29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e29\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e16\u0026ndash;33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eKaraganda region\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e23\u0026ndash;28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e23\u0026ndash;0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e17\u0026ndash;29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e25\u0026ndash;29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e24\u0026ndash;25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e22\u0026ndash;27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e29\u0026ndash;40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e11\u0026ndash;33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTotal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16.6\u0026ndash;39.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e22.1\u0026ndash;26.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e23.1\u0026ndash;24.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e17.6\u0026ndash;28.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e25.8\u0026ndash;28.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e21.1\u0026ndash;26.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e23\u0026ndash;27.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e29.6\u0026ndash;39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e5.3\u0026ndash;31.6\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\u003eOf the 26 \u003cem\u003eV. cholerae\u003c/em\u003e O1 and non-O1 strains examined, 100% (26/26) were susceptible to the following antimicrobial agents: doxycycline (30 \u0026micro;g), ciprofloxacin (10 \u0026micro;g), tetracycline (30 \u0026micro;g), cefotaxime (30 \u0026micro;g), and kanamycin (30 \u0026micro;g). With respect to nalidixic acid (30 \u0026micro;g), 65.4% (17/26) of the strains demonstrated high susceptibility, 30.8% (8/26) exhibited intermediate susceptibility, and 3.8% (1/26) showed resistance. For trimethoprim (5 \u0026micro;g), 73.1% (19/26) of the isolates were highly susceptible, while 26.9% (7/26) showed intermediate susceptibility.\u003c/p\u003e \u003cp\u003eFor furazolidone (50 \u0026micro;g), 73.1% (19/26) of the \u003cem\u003eV. cholerae\u003c/em\u003e strains were highly susceptible, while 26.9% (7/26) exhibited intermediate susceptibility. With respect to gentamicin (10 \u0026micro;g), 96.2% (25/26) of the isolates were highly susceptible, and 3.8% (1/26) showed intermediate susceptibility. Similarly, for chloramphenicol (10 \u0026micro;g) and ampicillin (25 \u0026micro;g), 96.2% (25/26) of the strains demonstrated high susceptibility, with 3.8% (1/26) showing intermediate susceptibility to each antibiotic.\u003c/p\u003e \u003cp\u003eRegarding streptomycin (300 \u0026micro;g), 88.5% (23/26) of the \u003cem\u003eV. cholerae\u003c/em\u003e strains were highly susceptible, 7.7% (2/26) showed intermediate susceptibility, and 3.8% (1/26) exhibited low susceptibility. For rifampicin (15 \u0026micro;g), 96.2% (25/26) of the strains demonstrated high susceptibility, while 3.8% (1/26) showed intermediate susceptibility.\u003c/p\u003e \u003cp\u003eThe results of the study, including the strain numbers and the names of the antibacterial agents tested on Mueller\u0026ndash;Hinton and Hottinger agars, are presented in summary charts in Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, along with selected individual results in Figs.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e and \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe most active antibiotics belonged to the β-lactam group, including cefotaxime, cefixime, chloramphenicol, and cefamandole, followed by cefazolin, tetracycline, and ampicillin. Agents traditionally used for cholera treatment\u0026mdash;such as ciprofloxacin, doxycycline, and azithromycin\u0026mdash;demonstrated comparatively lower activity. Furazolidone exhibited the weakest activity against all 26 strains, with an average inhibition zone diameter of 11.1 mm (range: 10\u0026ndash;18 mm).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eAn in vitro phenotypic susceptibility analysis of \u003cem\u003eV. cholerae\u003c/em\u003e (n\u0026thinsp;=\u0026thinsp;26) demonstrated 100% sensitivity to β-lactams, tetracyclines, aminoglycosides, amphenicols, glycopeptides, lincosamides, and quinolones. Additionally, 96.5% of the strains were susceptible to antibiotics from other classes.\u003c/p\u003e \u003cp\u003eThe results of the susceptibility and resistance testing were confirmed using the standard E-test method, with strips indicating the minimum inhibitory concentration (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe results of the antimicrobial resistance gene detection in 26 \u003cem\u003eV. cholerae\u003c/em\u003e strains using the \u003cem\u003eBacResista GLA Real-Time PCR Detection Kit\u003c/em\u003e with real-time PCR are presented in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eResults of antimicrobial resistance gene detection in 26 \u003cem\u003eVibrio cholerae\u003c/em\u003e strains using the \u003cem\u003eBacResista GLA Real-Time PCR Detection Kit\u003c/em\u003e and real-time PCR.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" morerows=\"1\" nameend=\"c2\" namest=\"c1\" rowspan=\"2\"\u003e \u003cp\u003eName of Genetic Determinants and Verification\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"7\" nameend=\"c9\" namest=\"c3\"\u003e \u003cp\u003eControls and Indicators by Species\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae KA-37\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003eEscherichia coli\u003c/em\u003e ATCC 25922\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eSalmonella typhimurium ATCC 14\u003c/em\u003e,\u003cem\u003e025\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003ePseudomonas aeruginosa ATCC 9027\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cem\u003eV. cholerae\u003c/em\u003e KZ00-00 ***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eControl \u0026laquo; \u0026ndash; \u0026raquo;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eControl \u0026laquo; + \u0026raquo;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"7\" rowspan=\"8\"\u003e \u003cp\u003eFAM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTBM *\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7.507\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8.029\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e15.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7.648\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026oline;x\u0026thinsp;=\u0026thinsp;7.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e23.27\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eimp\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e23.63\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ectx-M-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e22.20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003evan A/B\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.166\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.954\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e24.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eoxa-48-like\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e24.07\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003evim\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e24.31\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eoxa-23-like\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e23.43\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eshv\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e23.61\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"6\" rowspan=\"7\"\u003e \u003cp\u003eHEX\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIC **\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e25.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e25.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e25.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e25.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026oline;x\u0026thinsp;=\u0026thinsp;25.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e26.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e26.70\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e26.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e26.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e25.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e26.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026oline;x\u0026thinsp;=\u0026thinsp;24.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e25.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e25.81\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e24.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e25.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e25.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e25.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026oline;x\u0026thinsp;=\u0026thinsp;24.56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e25.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e25.27\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e24.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e24.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e24.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e24.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026oline;x\u0026thinsp;=\u0026thinsp;24.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e24.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e25.10\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e24.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e24.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e24.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e24.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026oline;x\u0026thinsp;=\u0026thinsp;24.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e24.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e25.12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e24.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e24.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026oline;x\u0026thinsp;=\u0026thinsp;24.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e25.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e25.03\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e24.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e24.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e24.56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e24.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026oline;x\u0026thinsp;=\u0026thinsp;24.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e24.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e24.65\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"6\" rowspan=\"7\"\u003e \u003cp\u003eCY5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eoxa-51-like\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e24.69\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTem\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e34.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e24.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e20.20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003emec A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e24.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eoxa-40-like\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e24.79\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKpc\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e24.12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNdm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e23.53\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e23.24\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* TBM\u0026mdash;total bacterial mass; ** IC\u0026mdash;internal control; *** KZ-00-00\u0026mdash;average value based on results from 26 \u003cem\u003eV. cholerae\u003c/em\u003e strains.\u003c/p\u003e \u003cp\u003eCurrently, research on the genetic determinants of bacterial resistance to glycopeptide and beta-lactam antibiotics is receiving significant attention, as these classes of antimicrobial agents are widely used for the treatment of complicated and/or severe infections. Investigating the most common types of beta-lactamases produced by various pathogenic bacteria can aid in interpreting their antibiotic susceptibility profiles, informing therapeutic decision-making, and enhancing infection control practices at the local level.\u003c/p\u003e \u003cp\u003eA molecular genetic analysis aiming to detect antibiotic resistance determinants in the genomes of 26 \u003cem\u003eVibrio cholerae\u003c/em\u003e isolates, including clinical strains collected in Kazakhstan from 1970 to 2024, revealed no presence of resistance genes to glycopeptides (\u003cem\u003evanA/B\u003c/em\u003e\u0026mdash;vancomycin and teicoplanin) or beta-lactam antibiotics (\u003cem\u003emecA\u003c/em\u003e\u0026mdash;methicillin and oxacillin; \u003cem\u003etem\u003c/em\u003e, \u003cem\u003ectx-M-1\u003c/em\u003e, and \u003cem\u003eshv\u003c/em\u003e\u0026mdash;penicillins and cephalosporins; \u003cem\u003eoxa-40-like\u003c/em\u003e, \u003cem\u003eoxa-48-like\u003c/em\u003e, \u003cem\u003eoxa-23-like\u003c/em\u003e, \u003cem\u003eoxa-51-like\u003c/em\u003e, \u003cem\u003eimp\u003c/em\u003e, \u003cem\u003ekpc\u003c/em\u003e, \u003cem\u003eges\u003c/em\u003e, \u003cem\u003endm\u003c/em\u003e, and \u003cem\u003evim\u003c/em\u003e\u0026mdash;carbapenems). The control strains used in this study included \u003cem\u003eEscherichia coli\u003c/em\u003e ATCC 25922, \u003cem\u003eSalmonella typhimurium\u003c/em\u003e ATCC 14025, and \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e ATCC 9027, in which the \u003cem\u003evanA/B\u003c/em\u003e gene (Ct\u0026thinsp;=\u0026thinsp;9.166, FAM channel) and the \u003cem\u003etem\u003c/em\u003e gene (Ct\u0026thinsp;=\u0026thinsp;34.60, CY5 channel) were detected in \u003cem\u003eE. coli\u003c/em\u003e ATCC 25,922 and in \u003cem\u003eP. aeruginosa\u003c/em\u003e ATCC 9027 (Ct\u0026thinsp;=\u0026thinsp;8.954 and 24.85, respectively).\u003c/p\u003e \u003cp\u003eThus, the results confirm the absence of genetic markers of resistance to 59 antimicrobial agents in all examined \u003cem\u003eV. cholerae\u003c/em\u003e isolates. No resistance was detected to the following major classes of antibiotics: extended-spectrum beta-lactams (penicillins, cephalosporins, and carbapenems), monobactams, macrolides, tetracyclines, aminoglycosides, amphenicols, glycopeptides, lincosamides, fluoroquinolones, and other antibiotic groups. These findings highlight the importance of the regular surveillance of \u003cem\u003eV. cholerae\u003c/em\u003e susceptibility to antimicrobial agents, which enables timely adjustments to therapeutic strategies and effective responses to changes in the epidemiological situation.\u003c/p\u003e"},{"header":"4. Discussion","content":"\u003cp\u003e \u003cem\u003eVibrio\u003c/em\u003e species undergo genetic changes to enhance their adaptation and resistance to antibiotics. A study on the susceptibility of \u003cem\u003eV. cholerae\u003c/em\u003e O1 El Tor strains to polymyxin B (PB) in Odisha, India, between 1995 and 2019 found that 89.4% of 1,200 strains were resistant, while 10.6% were susceptible. Susceptibility began to appear from 2005 onward, with the exception of 2015. An E-test analysis revealed that strains with the \u003cem\u003ectxB7\u003c/em\u003e genotype had a lower minimum inhibitory concentration (MIC\u0026thinsp;\u0026le;\u0026thinsp;4 \u0026micro;g/mL), whereas \u003cem\u003ectxB1\u003c/em\u003e genotypes exhibited higher MIC values (24 and 32 \u0026micro;g/mL) [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe study conducted in Odisha, India, between 1995 and 2019, examined changes in antibiotic susceptibility, as well as the presence of virulence and resistance genes, in \u003cem\u003eVibrio cholerae\u003c/em\u003e O1 strains. Antimicrobial susceptibility was assessed using the disk diffusion method, and resistance and virulence genes were identified using PCR. All strains were susceptible to gentamicin, chloramphenicol, norfloxacin, and ciprofloxacin but exhibited resistance to one or more other antibiotics. Resistance genes (SulII, dfrA1, and strB) and SXT elements were detected in 90% of the isolates. The Haitian variant of tcpA emerged in 1999 and gradually increased in prevalence. Multiplex PCR confirmed the presence of virulence genes (toxR, ompU, ace, rtxC, ctxA, tcpA, rfbO1, and ompW) in all strains [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAntibiotic resistance among epidemic \u003cem\u003eVibrio cholerae\u003c/em\u003e strains is increasing. A study analyzing \u003cem\u003eV. cholerae\u003c/em\u003e O1 El Tor strains isolated in China from 1961 to 2010 showed that resistance levels were generally low, except for nalidixic acid (45.9%), tetracycline (11%), and trimethoprim/sulfamethoxazole (38.5%). All strains from the 1960s were susceptible, whereas resistance levels rose significantly during the 1990s and beyond. Class I integrons were more frequently found among strains from 1993\u0026ndash;1998, and the SXT element was commonly detected in isolates collected after 1993. These findings highlight the growing resistance of epidemic \u003cem\u003eV. cholerae\u003c/em\u003e strains in China [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn Odisha, cholera has remained a significant threat, causing outbreaks with high morbidity and mortality. A study characterized \u003cem\u003eVibrio cholerae\u003c/em\u003e O1 strains from the 2018 and 2019 outbreaks in Bargarh and Rayagada. Multidrug-resistant strains were isolated and analyzed using PCR and pulsed-field gel electrophoresis (PFGE). All strains carried both virulence and resistance genes, including \u003cem\u003edfrA1\u003c/em\u003e (100%), \u003cem\u003estrB\u003c/em\u003e (76.9%), and \u003cem\u003eintSXT\u003c/em\u003e (61.5%). The outbreaks were driven by multidrug-resistant strains, underscoring the need for continuous monitoring to prevent future epidemics [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn 2022, the number of cholera cases in the WHO European Region more than doubled. In one study, a total of 49 \u003cem\u003eVibrio cholerae\u003c/em\u003e O1 isolates collected in Europe were analyzed to confirm their affiliation with the seventh pandemic El Tor lineage (7PET) and to assess their virulence and antibiotic resistance. All isolates were found to belong to the Pre - AFR15 sublineage, which is likely associated with the global surge in cholera cases. Antibiotic resistance analysis revealed no abnormal resistance profiles. Genomic sequencing was recommended for accurate strain identification and the monitoring of their evolutionary dynamics [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003eVibrio cholerae\u003c/em\u003e O139, first identified in 1992, has been recognized as a causative agent of cholera epidemics. Since 1993, O139 strains have been isolated in China, leading to sporadic cases and foodborne outbreaks. In a study conducted in China, the antibiotic resistance of strains collected between 1993 and 2009 was analyzed. Initially, the strains showed resistance to only a few antibiotics; however, after 1998, multidrug resistance increased significantly. In contrast, \u003cem\u003eV. cholerae\u003c/em\u003e O1 strains exhibited lower levels of resistance. Toxigenic O139 strains showed a high prevalence of resistance elements, indicating complex resistance patterns. The high level of multidrug resistance poses a growing public health threat, highlighting the need for continuous surveillance and control [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn Africa, \u003cem\u003eVibrio cholerae\u003c/em\u003e O1 strains with multidrug resistance have been reported for many years, and during recent epidemics in Kenya, new resistance phenotypes have emerged. One study aimed to analyze the epidemiological features and antimicrobial resistance of strains isolated in Kenya during 2007\u0026ndash;2010 and 2015\u0026ndash;2016. A total of 228 strains were examined, including 226 clinical and 2 environmental isolates. All strains belonged to the El Tor biotype and were found to be susceptible to ceftriaxone, gentamicin, and ciprofloxacin, while showing resistance to trimethoprim\u0026ndash;sulfamethoxazole, streptomycin, and nalidixic acid. The SXT integrative element was detected in 95.5% of the strains, whereas class 1, 2, and 3 integrons were not detected. Overall, 64.5% of the isolates exhibited multidrug resistance. During the 2015\u0026ndash;2016 outbreaks, strains with chromosomal mutations were identified that remained susceptible to most antibiotics except nalidixic acid. The study revealed two distinct resistance patterns and emphasized the importance of continuous surveillance to prevent the emergence of new resistant strains [\u003cspan additionalcitationids=\"CR22\" citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003eVibrio cholerae\u003c/em\u003e is a common inhabitant of aquatic ecosystems, and the state of West Bengal in India\u0026mdash;particularly the Ganges Delta region\u0026mdash;is known for having a high cholera burden. In a study conducted in Midnapore, \u003cem\u003eV. cholerae\u003c/em\u003e isolates were obtained from wastewater samples. All isolates belonged to non-O1/non-O139 serogroups. The majority (74.7%) exhibited moderate biofilm-forming ability, while 6.3% demonstrated strong biofilm formation. PCR screening revealed that most isolates carried genes associated with biofilm formation (such as \u003cem\u003ecdgH\u003c/em\u003e, \u003cem\u003ecdgM\u003c/em\u003e, \u003cem\u003ecdgK\u003c/em\u003e, etc.). Two strains (3.17%) contained the cholera toxin genes \u003cem\u003ectxA\u003c/em\u003e and \u003cem\u003ectxB\u003c/em\u003e. Approximately 24.8% of the isolates carried the \u003cem\u003eompU\u003c/em\u003e gene, while other virulence factors were detected less frequently. All isolates tested positive for \u003cem\u003etoxT\u003c/em\u003e, \u003cem\u003etoxR\u003c/em\u003e, and \u003cem\u003ehapR\u003c/em\u003e, which regulate virulence expression. Resistance profiling indicated both increased antibiotic susceptibility and the presence of multidrug resistance (MDR). The detection of such strains in the environment is a concern. The high prevalence of diguanylate cyclases (DGCs) suggests their potential as alternative therapeutic targets against MDR strains [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eA molecular genetic analysis conducted on 26 \u003cem\u003eVibrio cholerae\u003c/em\u003e O1 isolates, including clinical strains collected in Kazakhstan between 1970 and 2024, revealed no resistance genes to glycopeptide and beta-lactam antibiotics (\u003cem\u003evanA/B, mecA, tem, ctx-M-1, shv, oxa-40-like, oxa-48-like, oxa-23-like, oxa-51-like, imp, kpc, ges, ndm\u003c/em\u003e, and \u003cem\u003evim\u003c/em\u003e). These findings indicate the absence of genetically mediated resistance to major classes of antimicrobial agents in the analyzed isolates [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eSuch results underscore the importance of the regular epidemiological monitoring of pathogen susceptibility to antibiotics, enabling timely responses to potential shifts in resistance patterns [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe absence of resistance to major antibiotic classes among all examined \u003cem\u003eV. cholerae\u003c/em\u003e isolates indicates the continued high clinical efficacy of these drugs. These findings are particularly significant in the context of potential cholera outbreaks, as they provide clinicians with a reliable foundation for selecting empirical therapy [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFurthermore, phenotypic susceptibility testing showed that 100% of the strains were sensitive to doxycycline, ciprofloxacin, tetracycline, cefotaxime, and kanamycin. A high susceptibility was also observed in 96.2% of the isolates to chloramphenicol, ampicillin, gentamicin, and rifampicin; 88.5% to streptomycin; and 73.1% to trimethoprim and furazolidone. At the same time, intermediate susceptibility and isolated cases of resistance were observed for nalidixic acid, streptomycin, and furazolidone [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e].\u003c/p\u003e"},{"header":"5. Conclusions","content":"\u003cp\u003eAn analysis of antibiotic susceptibility in 26 \u003cem\u003eVibrio cholerae\u003c/em\u003e O1 strains isolated from various regions of Kazakhstan over nearly five decades demonstrated the sustained efficacy of the most commonly used antimicrobial agents. Isolated cases of resistance\u0026mdash;particularly to nalidixic acid, streptomycin, and trimethoprim\u0026mdash;highlight the importance of continued systematic surveillance of antimicrobial resistance in \u003cem\u003eV. cholerae\u003c/em\u003e. The findings of this study may inform treatment planning, preventive measures, and local microbiological surveillance efforts in cholera-endemic areas.\u003c/p\u003e \u003cp\u003eAll \u003cem\u003eVibrio cholerae\u003c/em\u003e O1 strains isolated in Kazakhstan between 1970 and 2024 demonstrated a high susceptibility to several critically important antimicrobial agents, including doxycycline, ciprofloxacin, tetracycline, cefotaxime, and kanamycin. High levels of susceptibility were also observed for gentamicin, chloramphenicol, ampicillin, and rifampicin (96.2% of strains).\u003c/p\u003e \u003cp\u003eModerate resistance was observed against nalidixic acid, trimethoprim, furazolidone, chloramphenicol, ampicillin, streptomycin, and rifampicin.\u003c/p\u003e \u003cp\u003eAll strains, despite variations in serovar and pathogenicity profiles (\u003cem\u003ectxAB⁺\u003c/em\u003e and \u003cem\u003etcpA⁺/⁻\u003c/em\u003e), remain susceptible to the primary drugs used in cholera treatment, indicating their continued potential clinical effectiveness at the present stage.\u003c/p\u003e \u003cp\u003eNo resistance genes to glycopeptide or beta-lactam antibiotics were detected via real-time PCR in the tested strains. However, the \u003cem\u003evanA/B\u003c/em\u003e (Ct\u0026thinsp;=\u0026thinsp;9.166, FAM channel) and \u003cem\u003etem\u003c/em\u003e (Ct\u0026thinsp;=\u0026thinsp;34.60, CY5 channel) genes were identified in \u003cem\u003eEscherichia coli\u003c/em\u003e ATCC 25922 and in \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e ATCC 9027, with Ct values of 8.954 and 24.85, respectively.\u003c/p\u003e \u003cp\u003eOver the study period (1970\u0026ndash;2024), no significant increase in the proportion of polyresistant strains was observed. However, the presence or emergence of even a single resistant isolate suggests the possibility of gene transfer under natural conditions, indicating the ongoing circulation of \u003cem\u003eVibrio cholerae\u003c/em\u003e O1 between humans and environmental sources. This highlights the need for continuous epidemiological and microbiological surveillance.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor Contributions:\u003c/strong\u003e\u0026nbsp; Conceptualization, Z.A., Z.Z. and R.M.; methodology, Z.A., R.M. and B.T.; software, D.O.; validation, Z.A., R.M., B.B., B.T., Z.B., D.O., N.S., Z.D., and B.A.; formal analysis, B.T., B.B., D.O., N.S. and B.A.; investigation, Z.A., Z.Z., and R.M.; resources, Z.D., B.B., B.T., D.O. and N.S.; data curation, Z.A., R.M., D.O. and N.S.; writing—original draft preparation, Z.A.; writing—review and editing, Z.Z., D.O. and R.M.; visualization, Z.A. and D.O.; supervision, Z.Z., R.M. and A.A.; project administration, D.O.; funding acquisition, Z.A. All authors have read and agreed to the published version of the manuscript\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e\u0026nbsp; This research was conducted within the framework of the project funded by the Ministry of Science and Higher Education of the Republic of Kazakhstan, titled “Investigation of Antibiotic Resistance Genes in Plague and Cholera Pathogens and the Development of a PCR Test System” (Project IRN: AP19679355). The project was supported by the Committee of Science of the Ministry of Science and Higher Education of the Republic of Kazakhstan.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInstitutional Review Board Statement:\u003c/strong\u003e Not applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformed Consent Statement:\u003c/strong\u003e Not applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement:\u003c/strong\u003e The original contributions presented in this study are included in the article. Further inquiries can be directed to the corresponding authors\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of Interest:\u003c/strong\u003e The authors declare no conflicts of interest.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eOjeda Rodriguez JA, Hashmi MF, Kahwaji CI \u003cem\u003eVibrio cholerae\u003c/em\u003e Infection. In \u003cem\u003eStatPearls\u003c/em\u003e; StatPearls Publishing: Treasure Island, FL, USA. 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Environ Monit Assess 168:321\u0026ndash;329\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFederal Center for Hygiene and Epidemiology of Rospotrebnadzor (2010) Guidelines for determining the susceptibility of causative agents of dangerous bacterial infections (plague, anthrax, etc.) to antibacterial drugs: Methodical recommendations 4.2.2495-09. Russia, Moscow, p 59. (In Russian)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAbdirasilova AA, Abdel ZZ (2023) Methodological Recommendations for Real-Time Polymerase Chain Reaction (RT-PCR) for the Detection of Plague Microbe DNA. Almaty, Kazakhstan, p 37. (In Russian)KazBookExport\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"the Ministry of Science and Higher Education of the Republic of Kazakhstan, titled “Investigation of Antibiotic Resistance Genes in Plague and Cholera Pathogens and the Development of a PCR Test System” (Project IRN: AP19679355)","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"cholera strains, antibiotics, resistance, sensitivity, statistics","lastPublishedDoi":"10.21203/rs.3.rs-6902069/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6902069/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003eholera is an acute diarrheal disease caused by the bacterium \u003cem\u003eVibrio cholerae\u003c/em\u003e, transmitted via the fecal–oral route, primarily through contaminated water or food. Despite progress in vaccination and antibiotic use, cholera continues to pose a significant public health threat, especially in regions with inadequate sanitation and unsafe drinking water. A major challenge in the management of cholera is the emergence of antibiotic-resistant \u003cem\u003eV. cholerae\u003c/em\u003estrains, which compromise the effectiveness of standard therapeutic regimens. These resistant strains necessitate the development of novel treatment and prevention strategies.\u003c/p\u003e\n\u003cp\u003eIn 2018, studies on phenotypic resistance markers in \u003cem\u003eVibrio cholerae\u003c/em\u003eisolates collected in Kazakhstan revealed that 38.4% of the examined strains exhibited resistance, with monoresistant strains predominating (23.1%). Additionally, 9.6% of the isolates carried two or more resistance markers. Among the \u003cem\u003eVibrio cholerae\u003c/em\u003e O1 serogroup isolates, 38.5% were resistant, while in the non-O1 serogroup, resistance was observed in 40.0% of cases.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eObjectives: \u003c/strong\u003eIn this study, a comprehensive analysis was conducted on the antibiotic resistance of \u003cem\u003eVibrio cholerae\u003c/em\u003e strains isolated in Kazakhstan from 1970 to 2024, focusing on their susceptibility to various classes of antimicrobial agents. The molecular and biochemical mechanisms underlying the development of resistance were also investigated, and potential impacts on the epidemiological situation and biosafety were assessed.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e: In this study, a total of 26 \u003cem\u003eVibrio cholerae\u003c/em\u003e strains isolated in Kazakhstan between 1970 and 2024 from clinical cases and environmental sources were used to screen for antimicrobial susceptibility and resistance profiles. Susceptibility testing was performed using the Kirby–Bauer disk diffusion method and E-test. To detect resistance genes, phenotypic assays and real-time Polymerase Chain Reaction were applied. One reference strain, 59 antibacterial agents across major drug classes, and a BacResista GLA Real-Time PCR Detection Kit were employed.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e: Phenotypic susceptibility testing of \u003cem\u003eV. cholerae\u003c/em\u003e (n = 26) conducted in vitro demonstrated high sensitivity to cefotaxime, tetracycline, doxycycline, ciprofloxacin, and kanamycin. A similarly high level of susceptibility was observed for gentamicin, chloramphenicol, ampicillin, and rifampicin (96.2% of isolates). Real-time PCR results revealed no presence of resistance genes to glycopeptide or beta-lactam antibiotics in the tested \u003cem\u003eV. cholerae\u003c/em\u003estrains. However, van A/B genes (Ct = 9.166, FAM channel) and the \u003cstrong\u003etem\u003c/strong\u003egene (Ct = 34.60, CY5 channel) were detected in the control strains \u003cem\u003eEscherichia coli\u003c/em\u003e ATCC 25922 and \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e ATCC 9027 (Ct = 8.954 and 24.85, respectively).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions\u003c/strong\u003e: The absence of resistance to major classes of antimicrobial agents among all 26 \u003cem\u003eV. cholerae\u003c/em\u003e isolates indicates the continued high clinical efficacy of these antibiotics in the treatment of cholera. These findings are of critical importance in the context of potential epidemic outbreaks, as they provide clinicians with a reliable basis for selecting empirical therapy.\u003c/p\u003e","manuscriptTitle":"Laboratory Analysis of the Resistance Spectrum and Antibacterial Susceptibility of Vibrio cholerae Strains Isolated in Kazakhstan from 1970 to 2024","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-18 19:39:17","doi":"10.21203/rs.3.rs-6902069/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"5d05e770-377c-49d6-a3b8-75ea9c15e544","owner":[],"postedDate":"June 18th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":50090857,"name":"Infectious Diseases"}],"tags":[],"updatedAt":"2025-06-18T19:39:17+00:00","versionOfRecord":[],"versionCreatedAt":"2025-06-18 19:39:17","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6902069","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6902069","identity":"rs-6902069","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}