Ecological and laboratory assessment of the resistance spectrum and antibacterial susceptibility of Vibrio cholerae strains isolated in the Caspian region of Kazakhstan

Document Type : Research Paper

Authors

1 Masgut Aikimbayev’s National Scientific Center Especially Dangerous Infections of the National Holding “QazBioPharm” of the Ministry of Health of the Republic of Kazakhstan, 14 Zhakhanger St., Almaty 050054, Republic of Kazakhstan

2 Scientific Center for Anti-Infective Drugs of the National Holding “QazBioPharm”, Ministry of Health of the Republic of Kazakhstan, Astana 010000, Kazakhstan; LLP Research and Production Association Kazpharmacom, Spasskaya str., 64A., Alamty 050028, Kazakhstan

10.22124/cjes.2025.9161

Abstract

Given Kazakhstan’s geographical proximity to the Caspian Sea and the detection of V. cholerae in river systems connected to the Caspian basin, this study also contributes to a broader understanding of cholera ecology and antimicrobial resistance within the region. For the first time, this study identified and thoroughly examined the role of genetic and biochemical factors contributing to the development of resistance in Vibrio cholerae strains under the conditions of the Republic of Kazakhstan. A comprehensive analysis was conducted on the antibiotic resistance of V. cholerae strains isolated in Kazakhstan from 1997 to 2023, with a focus on their potential for persistence, adaptation, and spread within the broader Caspian ecological network. Molecular and biochemical mechanisms underlying the development of resistance were investigated, and their potential impact on the epidemiological situation and biological safety was assessed.  Phenotypic susceptibility testing of V. cholerae (n = 13) 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, vanA/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). In conclusion, the absence of resistance to major classes of antimicrobial agents among all 13 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 and environmental persistence of V. cholerae in Caspian-associated aquatic systems, as they provide clinicians and public health officials with a reliable basis for empirical therapy selection and regional preparedness planning.

Keywords


Abdirasilova, AA, & Abdel, ZZh 2023, Methodological Recommendations for Real-Time Polymerase Chain Reaction (RT-PCR) for the Detection of Plague Microbe DNA, KazBookExport: Almaty, Kazakhstan, 37 p. [In Russian].
Aghayev IZ & Guseinova ASH 2005, Environmental conditions and cholera vibrios in the Caspian Sea waters of Azerbaijan. Journal of Epidemiology and Infectious Diseases, 3: 37-41.
Ali, M et al. 2012, The global burden of cholera. Bulletin of the World Health Organization, 90(3): 209–218.
Amanov, KhG & Berdyev, AN 1995, Choleragenic vibrios in the water bodies of Turkmenistan. Bulletin of Sanitation and Hygiene, 2: 28–31, [In Russian].
Boucher, HW, Talbot, GH, Bradley, JS, Edwards, JE, Gilbert, D, Rice, LB, Scheld, M, Spellberg, B & Bartlett, J 2009, Bad bugs, no drugs: No escape! An update from the Infectious Diseases Society of America. Clinical Infectious Diseases, 48: 1–12.
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. Infectious Diseases Now, 55: 105020. https://doi.org/10.1016/ j.idnow.2024.105020. ISSN 2666-9919.
CLSI 2023, Performance Standards for Antimicrobial Susceptibility Testing, 33rd ed., CLSI Supplement M100, Clinical and Laboratory Standards Institute: Wayne, PA, USA.
Connell SR, Tracz DM, Nierhaus KH, & Taylor DE 2003, Ribosomal protection proteins and their mechanism of tetracycline resistance. Antimicrob Agents Chemother. 47(12): 3675-3681. DOI: 10.1128/AAC.47.12.3675-3681.2003. PMID: 14638464, PMCID: PMC296194.
Dairova, AN, Smagulova, RA & Asanova NK 2022, Ecological study of Vibrio cholerae in the water bodies of Kazakhstan. Bulletin of Microbiology, 4: 45–50, [In Russian].
Davies, J & Davies, D 2010, Origins and evolution of antibiotic resistance. Microbiology and Molecular Biology Reviews (MMBR), 74: 417-433.
Faruque, SM & Nair, GB 2013, Epidemiology, genetics, and ecology of toxigenic Vibrio cholerae. Microbiology Spectrum, 2, VE-0004-2013.
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. Moscow, Russia, 59 p, [In Russian].
Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing 2010, Sanitary and epidemiological rules SP 3.1.7.2826-10. Cholera prevention. Moscow, Available online: https://docs.cntd.ru/document/1200080421 (accessed on).
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. Proceedings of the National Academy of Sciences of the United States of America, 109: E2010–E2017.
Hochhut, B, Lotfi, Y, Mazel, D, Faruque, SM, Woodgate, R, Waldor, MK 2001, Molecular analysis of antibiotic resistance gene clusters in Vibrio cholerae O139 and O1 SXT constins. Antimicrobial Agents and Chemotherapy, 45: 2991-3000, https://doi.org/10.1128/AAC.45.11.2991-3000.2001. PMID: 11600347; PMCID: PMC90773.
Huq, A & Colwell, RR 2014, A microbiological perspective on climate change and cholera. Environmental Microbiology Reports, 6(6): 718-725.
Igbinosa, EO & Okoh, AI 2010, Antibiotic susceptibility profile of Vibrio cholerae O1 strains isolated from wastewater final effluents in South Africa. Environmental Monitoring and Assessment, 168: 321–329.
Islam, MT et al. 2021, Understanding the ecology of cholera through environmental and genomic surveillance. Trends in Microbiology, 29(3): 235–245.
Ivanova, IA, Filippenko, AV, Omelchenko, ND, Tyurina, AV & Gaevskaya, NE 2024, Current status of alternative methods for the prevention and treatment of cholera. Immunopathology, Allergology, Infectology, 3: 19–28, (In Russian), https://doi.org/10.14427/jipai.2024.3.19.
Kolobe, MT 2025, Cholera outbreak in Chienge District, Luapula Province, Zambia, 31st May - 9th June 2023: Re-emerging threat. International Journal of Infectious Diseases, 152: 107433, https://doi.org/10.1016/j.ijid.2024.107433. ISSN 1201-9712.
Marin, MA, Fonseca, EL, Andrade, BN, Cabral, AC, Vicente, AC 2014, Worldwide occurrence of integrative conjugative element encoding multidrug resistance determinants in epidemic Vibrio cholerae O1. PLoS ONE, 9: e108728, https://doi.org/10.1371/journal.pone.0108728. PMID: 25265418; PMCID: PMC4181655.
Rituparna De 2021, Mobile genetic elements of Vibrio cholerae and the evolution of its antimicrobial resistance. Frontiers in Tropical Diseases,   https://www.frontiersin.org/articles/10.3389/fitd.2021. 691604/full, (accessed 21 March 2025).
Nayak, AK, Nayak, SR, Behera, DR, Pal, BB 2021, Dissemination of Vibrio cholerae O1 isolated from Odisha, India. Environmental Microbiology Reports, 13: 355-363, https://doi.org/10.1111/1758-2229.12940. PMID: 33660421.
Nayak, SR, Nayak, AK, Biswal, BL, Pati, S, Pal, BB 2021, Spread of Haitian variant Vibrio cholerae O1 causing cholera outbreaks in Odisha, India. Japanese Journal of Infectious Diseases, 74: 137-143. https://doi.org/10.7883/yoken.JJID.2020.364. PMID: 32863351.
Nishanov, RA & Abdurakhmanov, ShE 2002, Vibrios in the estuaries of the Amu Darya and the Aral Delta.  Uzbek Medical Journal, 6: 33–36 [In Russian].
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 Infectious Diseases, 5: 481-493.
Rahbar M, Rezaeian M, Emadi G 2010, Seasonal dynamics of Vibrio cholerae in coastal waters of northern Iran. Iranian Journal of Environmental Health Science & Engineering, 7(2): 145–152.
Sagiyev ZA, Utepova IB, Mussagaliyeva RS, Alybayev SD, Kulbayeva MM, Ismailova AO, Aldibekova AA 2018, Antibiotic resistance of Vibrio cholera strains isolated in Kazakhstan. Antibiot Khimioter = Antibiotics and Chemotherapy, 63(3-4): 12-17, [In Russian].
Shakhnovich, EA, Sturtevant, D & Mekalanos, JJ 2007, Molecular mechanisms of virstatin resistance by non-O1/non-O139 strains of Vibrio cholerae. Molecular Microbiology, 66: 1331–1341. https://doi.org/10.1111/j.1365-2958.2007.05984.x. PMID: 17986190.
Sokolov AA, Yakovleva NA 2008, Isolation of Vibrio cholerae in the Volga Delta and on the Caspian Sea Coast.  Journal of Microbiology, Epidemiology and Immunobiology, 5: 42–47.
Wang, X et al. 2020, Environmental persistence and virulence potential of Vibrio cholerae from aquatic habitats. Frontiers in Microbiology, 11: Art. 580212.
Wang, R, Lou, J, Liu, J, Zhang, L, Li, J, Kan, B 2010, Antibiotic resistance of Vibrio cholerae O1 El Tor strains from the seventh pandemic in China, 1961–2010. International Journal of Antimicrobial Agents, 40: 361-364, https://doi.org/10.1016/j.ijantimicag.2012.06.010. PMID: 22867881.
WHO 2021, Antimicrobial resistance in the WHO African region: A systematic literature review. https://www.afro.who.int/sites/default/files/2021-11/Antimicrobial%20Resistance%20in%20the% 20WHO%20African%20region%20a%20systematic%20literature%20review.pdf, (accessed 7 April 2025).
World Health Organization 2014, Antimicrobial Resistance: Global Report on Surveillance. WHO Press: Geneva, Switzerland.