Prevalence and Characterization of Pathogenic Bacteria in Bulk Tank Raw Milk, Thailand

Chanida  KUPRADIT; Sasidhorn  INNOK; Jirayus  WORARATPHOKA; Mariena  KETUDAT-CAIRNS

doi:10.48048/wjst.2020.4177

Authors

Chanida KUPRADIT Department of Applied Biology, Faculty of Sciences and Liberal Arts, Rajamangala University of Technology Isan, Nakhon Ratchasima 30000, Thailand
Sasidhorn INNOK Department of Applied Biology, Faculty of Sciences and Liberal Arts, Rajamangala University of Technology Isan, Nakhon Ratchasima 30000, Thailand
Jirayus WORARATPHOKA Department of Applied Biology, Faculty of Sciences and Liberal Arts, Rajamangala University of Technology Isan, Nakhon Ratchasima 30000, Thailand
Mariena KETUDAT-CAIRNS School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand

DOI:

https://doi.org/10.48048/wjst.2020.4177

Keywords:

Foodborne pathogens, Raw milk, Raw milk collection center, Virulence genes

Abstract

Although there are a number of raw milk collection centers in Nakhon Ratchasima, there is a lack of information with regard to the process of isolation and characterization of foodborne pathogens in raw milk. Therefore, the purpose of this research was to investigate the prevalence and characterization of foodborne pathogens, including Bacillus cereus, Escherichia coli, Listeria monocytogenes, Salmonella spp. and Staphylococcus aureus from 33 raw milk samples from 9 different raw milk collection centers located in 8 districts in Nakhon Ratchasima, Thailand. This study was conducted from January to March 2016. Results revealed that the contaminations of L. monocytogenes and Salmonella spp. were not detected in any of the raw milk samples tested. The prevalence of B. cereus, E. coli, and S. aureus in raw milk samples was found to be 9 % (10 - 2.0 ×10⁴ CFU/ml), 42.4 and 54 % (85 - 2.7 ×10⁴ CFU/ml), respectively. The distribution of virulence genes was tested in B. cereus and S. aureus using gene specific primers by polymerase chain reaction. Out of the 29 analyzed coagulase-positive S. aureus isolates, 27 isolates (93 %) were positive for eap gene amplification and 14 isolates (48 %) showed amplicon of eap gene and all 5 enterotoxin genes, including seG, seGV, seI, seIV, and seM genes. All 8 B. cereus isolates tested showed positive PCR result with enterotoxin FM (entFM) gene but they showed negative with hemolysin gene (hblA and hblD genes) amplifications. It was inferred from these findings that bulk tank milk is a potential source of S. aureus and B. cereus in milk.

Downloads

Download data is not yet available.

Metrics

Metrics Loading ...

Author Biographies

Chanida KUPRADIT, Department of Applied Biology, Faculty of Sciences and Liberal Arts, Rajamangala University of Technology Isan, Nakhon Ratchasima 30000, Thailand

Department of Applied Biology

Sasidhorn INNOK, Department of Applied Biology, Faculty of Sciences and Liberal Arts, Rajamangala University of Technology Isan, Nakhon Ratchasima 30000, Thailand

Department of Applied Biology

Jirayus WORARATPHOKA, Department of Applied Biology, Faculty of Sciences and Liberal Arts, Rajamangala University of Technology Isan, Nakhon Ratchasima 30000, Thailand

Department of Applied Biology

Mariena KETUDAT-CAIRNS, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand

School of Biotechnology

References

JS Van Kessel, JS Karns, L Gorski, BJ McCluskey and ML Perdue. Prevalence of Salmonellae, Listeria monocytogenes and fecal coliforms in bulk tank milk on US Dairies. J. Dairy Sci. 2004; 87, 2822-30.

R Dewanti-Hariyadi and D Gitapratiwi. Prevalence of foodborne diseases in South East and Central Asia. In: Y Motarjemi, G Moy, E Cameron and D Todd (Eds.). Encyclopedia of Food Safety. 1st ed. Elsevier Science & Technology, San Diego, 2014, p. 287-94.

FY Chye, A Abdullah and MK Ayob. Bacteriological quality and safety of raw milk in Malaysia. Food Microbiol. 2004; 21, 535-41.

M Bartoszewicz, BM Hansen and I Swiecicka. The members of the Bacillus cereus group are commonly present contaminants of fresh and heat-treated milk. Food Microbiol. 2008; 25, 588-96.

RA Ombarak, A Hinenoya, SP Awasthi, A Iguchi, A Shima, ARM Elbagory and S Yamasaki. Prevalence and pathogenic potential of Escherichia coli isolates from raw milk and raw milk cheese in Egypt. Int. J. Food Microbiol. 2016; 221, 69-76.

SP Oliver, BM Jayarao and RA Almeida. Foodborne pathogens in milk and the dairy farm environment: food safety and public health implications. Foodborne Pathog. Dis. 2005; 2, 115-29.

United States Food and Drug Administration. Bacteriological Analytical Manual / Food and Drug Administration. 8th ed. MD: AOAC International, Gaithersburg, 1998.

JG Cappuccino and N Sherman. Microbiology: A Laboratory Manual. 5th ed. Benjamin/Cummings Science, California, 1998, p. 470.

C Kupradit, S Rodtong and M Ketudat-Cairns. Development of a DNA macroarray for simultaneous detection of multiple foodborne pathogenic bacteria in fresh chicken meat. World J. Microb. Biot. 2013; 29, 2281-91.

C Kupradit, S Innok, J Woraratphoka and M Ketudat-Cairns. Novel multiplex PCR assay for rapid detection of five bacterial foodborne pathogens. Suranaree J. Sci. Tech. 2017; 24, 41-50.

M Hussain, CV Eiff, B Sinha, I Joost, M Herrmann, G Peters and K Becker. eap gene as novel target for specific identification of Staphylococcus aureus. J. Clin. Microbiol. 2008; 46, 470-6.

J Chen and MW Griffiths. PCR differentiation of Escherichia coli from other gram-negative bacteria using primers derived from the nucleotide sequences flanking the gene encoding the universal stress protein. Lett. Appl. Microbiol. 1998; 27, 369-71.

JC Brooks, B Martinez, J Stratton, A Bianchini, R Krokstrom and R Hutkins. Survey of raw milk cheeses for microbiological quality and prevalence of foodborne pathogens. Food Microbiol. 2012; 31, 154-8.

FM Kivaria, JPTM Noordhuizen and AM Kapaga. Evaluation of the hygienic quality and associated public health hazards of raw milk marketed by smallholder dairy producers in the Dar es Salaam region, Tanzania. Trop. Anim. Health Pro. 2006; 38, 185-94.

P Padungtod and JB Kaneene. Salmonella in food animals and humans in northern Thailand. Int. J. Food Microbiol. 2006; 108, 346-54.

O Aygun and S Pehlivanlar. Listeria spp. in the raw milk and dairy products in Antakya, Turkey. Food Control. 2006; 17, 676-9.

S Jami, A Jamshidi and S Khanzadi. The presence of Listeria monocytogenes in raw milk samples in Mashhad, Iran. Iran. J. Vet. Res. 2010; 11, 363-7.

DM Bianchi, A Barbaro, S Gallina, N Vitale, L Chiavacci, M Caramelli and L Decastelli. Monitoring of foodborne pathogenic bacteria in vending machine raw milk in Piedmont, Italy. Food Control. 2013; 32, 435-9.

F Haghi, H Zeighami, G Naderi, A Samei, S Roudashti, S Bahari and P Shirmast. Detection of major food-borne pathogens in raw milk samples from dairy bovine and ovine herds in Iran. Small Ruminant Res. 2015; 131, 136-40.

RM Kamal, MA Bayoumi and SFA Abd El Aal. MRSA detection in raw milk, some dairy products and hands of dairy workers in Egypt, a mini-survey. Food Control. 2013; 33, 49-53.

VLM Rall, FP Vieira, R Rall, RL Vieitis, AJ Fernandes, JMG Candeias, KFG Cardoso and JPJ Araújo. PCR detection of staphylococcal enterotoxin genes in Staphylococcus aureus strains isolated from raw and pasteurized milk. Vet. Microbiol. 2008; 132, 408-13.

H Jamali, M Paydar, B Radmehr, S Ismail and A Dadrasnia. Prevalence and antimicrobial resistance of Staphylococcus aureus isolated from raw milk and dairy products. Food Control. 2015; 54, 383-8.

D Kanungpean, J Rujit, J Chanda, N Boonyaprapa, P Tonchotiwetch, W Mahanil, A Tain and A Intarapuk. The quality testing of raw goat’s milk in Nongchok District, Bangkok. J. Mahanakorn Vet. Med. 2014; 9, 83-8.

T Chitov, R Dispan and W Kasinrerk. Incidence and diarrhegenic potential of Bacillus cereus in pasteurized milk and cereal products in Thailand. J. Food Saf. 2008; 28, 467-81.

MA Abdou, NM Awny and AAEM Abozeid. Prevalence of toxicogenic bacteria in some foods and detection of Bacillus cereus and Staphylococcus aureus enterotoxin genes using multiplex PCR. Ann. Microbiol. 2012; 62, 569-80.

S Kumari and PK Sarkar. Prevalence and characterization of Bacillus cereus group from various marketed dairy products in India. Dairy Sci. Tech. 2014; 94, 483-97.

A Tewari and S Abdullah. Bacillus cereus food poisoning: international and Indian perspective. J. Food Sci. Tech. 2015; 52, 2500-11.

S Kumari and P Sarkar. Bacillus cereus hazard and control in industrial dairy processing environment. Food Control. 2016; 69, 20-9.

MC Te Giffel, A Wagendorp, A Herrewegh and F Driehuis. Bacterial spores in silage and raw milk. Anton. Van. Leeuw. 2002; 81, 625-30.

MTM Montanhini, R Montanhini Neto and LS Bersot. Enterotoxigenic potential of Bacillus cereus strains isolated from dairy products at different incubation temperature. Int. Food Res. J. 2015; 22, 1315-7.

J Kovac, RA Miller, LM Carroll, DJ Kent, J Jian, SM Beno and M Wiedmann. Production of hemolysin BL by Bacillus cereus group isolates of dairy origin is associated with whole-genome phylogenetic clade. BMC Genomics 2016; 17, 581.

K Hunt, J Schelin, P Rådström, F Butler and K Jordan. Classical enterotoxins of coagulase-positive Staphylococcus aureus isolates from raw milk and products for raw milk cheese production in Ireland. Dairy Sci. Tech. 2012; 92, 487-99.

R Nazari, H Godarzi, FR Baghi and M Moeinrad. Enterotoxin gene profiles among Staphylococcus aureus isolated from raw milk. Iran. J. Vet. Res. 2014; 15, 409-12.

B Khaenda, W Namwat, D Kotimanusvanij and C Chomvarin. Distribution of new Staphylococcal enterotoxin genes (seg, seh, sei, sej, and sel) in Staphylococcus aureus isolated from retail ready-to-eat foods in the Northeast Thailand. Srinagarind Med. J. 2016; 31, 237-44.

G Blaiotta, D Ercolini, C Pennacchia, V Fusco, A Casaburi, O Pepe and F Villani. PCR detection of staphylococcal enterotoxin genes in Staphylococcus spp. strains isolated from meat and dairy products. Evidence for new variants of seG and seI in S. aureus AB-8802. J. Appl. Microbiol. 2004; 97, 719-30.

A Tewari, SP Singh and R Singh. Incidence and enterotoxigenic profile of Bacillus cereus in meat and meat products of Uttarakhand, India. J. Food Sci. Tech. 2015; 52, 1796-801.

JL McKillip. Prevalence and expression of enterotoxins in Bacillus cereus and other Bacillus spp.: A literature review. Anton. Van. Leeuw. 2000; 77, 393-9.