Post-outbreak monitoring of Clostridium botulinum types C and D using Polymerase chain reaction in a dairy farm in Kanchanaburi province, Thailand
Article Sidebar
Main Article Content
Abstract
Objective: This study aimed to investigate the presence of C. botulinum on dairy farms with a history of outbreaks to support animal health monitoring and improve preventive strategies.
Materials and Methods: Fecal samples were randomly collected from 400 dairy cattle, including lactating cows, dry cows, pregnant heifers, heifer weaners, and calves, along with 10 feedstuff samples. Approximately 1-2 g of each sample was inoculated into cooked meat medium and incubated anaerobically at 35 ºC for 5 days. DNA was extracted from the enrichment culture and analyzed for C. botulinum types C and D using Polymerase chain reaction. Positive PCR products were confirmed by Sanger DNA sequencing.
Results: C. botulinum types C and D were detected in fecal samples at a rate of 2.75% (11/400). These were found in the feces of lactating cows (1.80%, 3/167), dry cows (6.98%, 6/86), pregnant heifers (2.00%, 1/50), and calves (2.70%,1/37). No fecal samples from heifer weaners (0/60) and feedstuffs (0/10) were positive.
Conclusion: The persistence of C. botulinum one year after an outbreak suggests possible gastrointestinal colonization and environmental contamination. Given the farm’s open system and presence of vectors such as flies, birds, rodents, cats, and dogs, biosecurity measures such as disinfection stations, sanitation equipment, and designated footwear at entrances are recommended to reduce pathogen spread.
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
References
Abdel-Moein KA, Hamza DA, 2016. Occurrence of human pathogenic Clostridium botulinum among healthy dairy animals: an emerging public health hazard. Pathog Glob Health 110(1), 25-29.
Alemu B, Ayele M, 2018. Review on botulism in cattle. Appl J Hygiene 7(2), 15-17.
Bacteriological Analytical Manual (BAM). 2001. Chapter 17: Clostridium botulinum. [Cited 2024 Aug 2]. Available from: https://www.fda.gov/food/laboratory-methods-food/bam-chapter-17-clostridium-botulinum
Carter GR, John R, Cole Jr, 1990. Diagnostic procedures in veterinary bacteriology and mycology, 5th edition. Academic press, Inc.
Center for Food Security and Public Health (CFSPH). 2018. Botulism [internet]. [Cited 2024 Aug 18]. Available from: https://www.cfsph.iastate.edu/Factsheets/pdfs/botulism.pdf
Dahlenborg M, Borch E, Radstrom P, 2003. Prevalence of Clostridium botulinum types B, E and F in faecal samples from Swedish cattle. J Food Protect 82(2), 105-110.
Driehuis F, Wilkinson JM, Jiang Y, Ogunade I, Adesogan TA, 2018. Silage review: Animal and human health risks from silage. J Dairy Sci 101: 4093-4110.
Espelund M, Klaveness D, 2014. Botulism outbreaks in natural environments-an update. Front Microbiol 5, 287.
Franciosa G, Fenicia L, Caldiani C, Aureli P, 1996. PCR for detection of Clostridium botulinum type C in avian and environmental samples. J Clin Microbiol 34(4), 882-885.
Goldsztejn M, Grenda T, Koziel N, Sapala M, Mazur M, Sieradzki Z, Krol B, Kwiatek K, 2020. Potential determinants of Clostridium spp. Occurrence in Polish silage. J Vet Res 64, 549-555.
Heffron A, Poxton IR, 2007. A PCR approach to determine the distribution of toxin genes in closely related Clostridium species: Clostridium botulinum type C and D neurotoxins and C2 toxin, and Clostridium novyi a toxin. J Med Microbiol 56, 196-201.
Meurens F, Carlin F, Federighi M, Filippitzi ME, Fournier M, Fravalo P, Ganiere JP, Grisot L, Guillier L, Hilaire D, Kooh P, Le Bouquin-Leneveu S, Le Marechal C, Mazuet C, Morvan H, Petit K, Vaillancourt JP, Woudstra C, 2023. Clostridium botulinum type C, D, C/D, and D/C: An update. Front Microbiol 13, 1099184.
Narongwanichgarn W, Ngernbaht S, Tunak C, Paethaisong T, Naksuwan R, Namkratok R, 2023. Diagnosis of botulism in a dairy farm in Kanchanaburi province in 2023[internet]. [Cited 2024 Dec 20]. Available from: https://expert.dld.go.th/webnew/index.php/th/news-menu
/e-book-menu/697-publish2527-12-03
Ngamjarus C, Chongsuvivatwong V, 2016. n4Studies: Sample size calculation for an epidemiological study on a smart device. Siriraj Med J 68, 160-170.
Notermans S, Dufrenne J, Oosterom J, 1981. Persistence of Clostridium botulinum type B on a cattle farm after an outbreak of botulism. Appl Environ Microbiol 41(1), 179-183.
Park HY, Lee K, Jung SC, Cho YS, 2022. Prevalent toxin types of Clostridium botulinum in South Korean cattle farms. Vet Anim Sci 15, 100239.
Quinn PJ, Carter ME, Markey BK, Carter GR, 1994. Clinical veterinary microbiology, Wolfe Publishing.
Rasetti-Escargueil C, Lemichez E, Popof MR, 2019. Public health risk associated with botulism as foodborne zoonoses. Toxins 12(1), 17.
Relun A, Dorso L, Douart A, Chartier C, Guatteo R, Mazuet C, Popoff MR, Assie S, 2017. A large outbreak of bovine botulism possibly linked to a massive contamination of grass silage by type D/C Clostridium botulinum spores on a farm with dairy and poultry operations. Epidemiol Infect, 145, 3477-3485.
Souillard R, Grosjean D, Le Gratiet T, Poezevara T, Rouxel S, Balaine L, Mace S, Martin L, Anniballi F, Chemaly M, Le Bouquin S, Le Marechal C, 2021. Asymptomatic carriage of C. botulinum type D/C in broiler flocks as the source of contamination of a massive botulism outbreak on a dairy cattle farm. Front Microbiol 12, 679377.
Souillard R, Le Marechal C, Hollebecque F, Rouxel S, Barbe A, Houard E, Leon D, Poezevara T, Fach P, Woudstra C, Mahe F, Chemaly M, Le Bouquin S, 2015. Occurrence of C. botulinum in healthy cattle and their environment following poultry botulism outbreaks in mixed farms. Vet Microbiol 180(1-2), 142-145.
Sunagawa H, Ohyama T, Watanabe T, Inoue K, 1992. The complete amino acid sequence of the Clostridium botulinum type D neurotoxin, deduced by nucleotide sequence analysis of the encoding phage d-16Ø genome. J Vet Med Sci 54(5), 905-913.
Wen C, Wei S, Zong X, Wang Y, Jin M, 2021. Microbiota-gut-brain axis and nutritional strategy under heat stress. Anim Nutr 7(4), 1329-1336.
World Organisation for Animal Health (WOAH). 2022. Botulism [internet]. [Cited 2024 Oct 27]. Available from: https://www.woah.org/ app/uploads/2022/02/botulism-1.pdf
Zhang P, 2022. Influence of foods and nutrition on the gut microbiome and implications for intestinal health. Int J Mol Sci 23, 9588.
