Occurrence of Multi-Resistant Staphylococcus aureus in Artisan Goat Coalho Cheese in Northeastern Brazil


  • Breno Bezerra Aragão Federal Rural University of Pernambuco – UFRPE, Department of Veterinary Medicine - R. Manuel de Medeiros, s/n - Dois Irmãos, CEP. 52171-900, Recife-PE, Brazil. https://orcid.org/0000-0002-9550-3692
  • Sabrina Cândido Trajano Federal Rural University of Pernambuco – UFRPE, Department of Veterinary Medicine - R. Manuel de Medeiros, s/n - Dois Irmãos, CEP. 52171-900, Recife-PE, Brazil. https://orcid.org/0000-0003-1200-1206
  • Renato Amorim da Silva Federal Rural University of Pernambuco – UFRPE, Department of Veterinary Medicine - R. Manuel de Medeiros, s/n - Dois Irmãos, CEP. 52171-900, Recife-PE, Brazil. https://orcid.org/0000-0002-8707-9190
  • Bruno Pajeú e Silva Federal Rural University of Pernambuco – UFRPE, Department of Veterinary Medicine - R. Manuel de Medeiros, s/n - Dois Irmãos, CEP. 52171-900, Recife-PE, Brazil. https://orcid.org/0000-0002-0040-393X
  • Rodolfo de Moraes Peixoto Federal Institute of Education, Science and Technology of the Sertão Pernambucano –IF SERTÃO-PE - R. Maria Luzia de Araújo Gomes Cabral, 791 - João de Deus, CEP. 56316-686, Petrolina-PE, Brazil. https://orcid.org/0000-0002-5757-5935
  • Rinaldo Aparecido Mota Federal Rural University of Pernambuco – UFRPE, Department of Veterinary Medicine - R. Manuel de Medeiros, s/n - Dois Irmãos, CEP. 52171-900, Recife-PE, Brazil. https://orcid.org/0000-0002-2844-5509




Background: The artisanal goat coalho cheese is one of the products obtained that stand out in 11 the dairy goat farming of the Northeast of Brazil. Despite its importance, goat cheese is often 12 made under inadequate hygienic-sanitary conditions and usually uses raw goat's milk, 13 increasing the risk of product contamination. Among the pathogens carried by goat coalho 14 cheese, Staphylococcus aureus stands out, being responsible for cases of food poisoning and 15 persistent infections that are difficult to treat. This study aimed to evaluate the contamination, 16 genotypic and phenotypic resistance of Staphylococcus aureus isolated from artisanal coalho 17 cheese made with goat milk produced in the Northeast region of Brazil.

Materials, Methods & Results: This study analyzed only artisanal coalho cheeses made with  raw goat's milk and purchased directly from farms. Twelve samples of artisanal coalho cheeses  made with raw goat's milk were collected (1 sample per property) in 8 municipalities in the  state of Pernambuco, Northeast region of Brazil. For microbiological analysis of enumeration  of Colony Forming Units (CFU/g) of Staphylococcus spp. the methodology recommended by the International Organization for Standardization (2019) and recognized by the Ministry of Agriculture, Livestock and Supply was used. After enumeration, 5 colonies were selected per enumerated plate, a total of 180 Staphylococcus spp. was obtained. These were subjected to thermal extraction of genetic material to search for the nuc gene by Polymerase Chain Reaction, the isolates carrying the nuc gene were subjected to genotypic and phenotypic evaluation of  antimicrobial resistance. After the phenotypic analysis, the Multiple Antimicrobial Resistance Index was evaluated. In all samples, Staphylococcus spp. and were considered unfit for consumption, with the lowest count being 9.4x103 CFU/g and the highest 6.4x106 CFU /g. Of the 180 isolates, 28.34% (51/180) were positive for the detection of the nuc gene. All resistance  genes except mecA, mecC, and norB were detected. Of the 51 S. aureus isolates, 31.37% (16/51) were considered multi-resistant and presented a Multiple Antimicrobial Resistance Index above 0.2.

Discussion: After microbiological analysis it was found that all samples of coalho cheese were out of standards and unfit for human consumption in accordance with Ordinance n° 146/1996 of the Ministry of Agriculture, Livestock and Supply of Brazil. Furthermore, the contamination of goat coalho cheeses is a risk to public health. During sample collection were found inadequate hygiene conditions in the environment used for cheese production. The presence of Staphylococcus aureus can be attributed to hygienic-sanitary failures in cheese production. From a health point of view, it is even more alarming when it comes to S. aureus carrying resistance genes. Although the 51 S. aureus isolates did not carry the mecA, mecC, norB genes and did not show phenotypic resistance to cefoxitin and oxacillin, all other genes were detected, indicating the circulation of S. aureus carrying the tet(L) genes, tet(M), tet-38, msrA, norA, and norC, which so far had not been reported in the production chain of goat coalho cheese in Brazil. Furthermore, the evaluation of the Multiple Antimicrobial Resistance Index identified the occurrence of multiple resistance to antimicrobials in 31.37% (16/51) of S. aureus at high risk to human health. The results obtained are quite worrying and serve as a warning to the scientific community and the Food Safety and Hygiene Inspection Services.

Keywords: goat cheese, contamination, resistance, one health


Download data is not yet available.


Brakstad O.G., Aasbakk K. & Maeland J.A. 1992. Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. Journal of Clinical Microbiology. 30:1654-1660.

BRASIL. Ministério da Agricultura, Pecuária e Abastecimento. Portaria nº 146 de 07 de março de 1996. Oficializa os Regulamentos Técnicos de Identidade e Qualidade dos Produtos Lácteos. Brasília: Ministério da Agricultura, Pecuária e Abastecimento, 1996. Available in: < https://www.dourados.ms.gov.br/wp-content/uploads/2016/05/RTIQ-Leite-Completo-PORTARIA-146_96-ok.pdf >. [Accessed online in June 2021].

Chambers H.F. & Deleo, F.R. 2009. Waves of resistance: Staphylococcus aureus in the antibiotic era. Nature Reviews Microbiology. 7:629-641.

- Clinical and Laboratory Standards Institute (CLSI). 2017. Performance Standards for Antimicrobial Susceptibility Testing, 27th ed. Clinical and Laboratory Standards Institute, 950 West Valley Road, Suite 2500, Wayne, Pennsylvania 19087 USA (CLSI supplement M100). (accessed 26 June 2021). Available in: <https://file.qums.ac.ir/repository/mmrc/clsi%202017.pdf>. [Accessed online in June 2021].

Fan H.H., Kleven S.H. & Jackwood, M.W. 1995. Application of polymerase chain reaction with arbitrary primers to strain identification of Mycoplasma gallisepticum. Avian Diseases. 39:729-735.

Field A. 2009. Descobrindo a estatística usando o SPSS. 2.ed. Porto Alegre: Artmed, 688 p.

Giedraitiene A., Vitkauskiene A., Naginiene R. & Pavilonis A. 2011. Antibiotic resistance mechanisms of clinically important bacteria. Medicina. 47:137-146.

ISO - International Organization for Standardization. 2019. Microbiology of food and animal feeding stuffs — Horizontal method for the enumeration of coagulase-positive staphylococci (Staphylococcus aureus and other species) - Part 1: Technique using Baird-Parker agar médium. Available in: <https://www.normas.com.br/visualizar/abnt-nbr-nm/11661/abnt-nbriso6888-1-microbiologia-de-alimentos-para-consumo-humano-e-animal-metodo-horizontal-para-enumeracao-de-estafilococos-coagulase-positiva-Staphylococcus-aureus-e-outras-especies-parte-1-tecnica-usando-agar-baird-parker>. [Accessed online in June 2021].

Johler S., Giannini P., Jermini M., Hummerjohann J., Baumgartner A. & Stephan R. 2015. Further Evidence for Staphylococcal Food Poisoning Outbreaks Caused by egc-Encoded Enterotoxins. Toxins. 7:997-1004.

Krumperman P.H. 1983. Multiple antibiotic resistance indexing of Escherichia coli to identify high-risk sources of fecal contamination of foods. Applied and Environmental Microbiology. 46:165-170.

Laxminarayan R., Duse A., Wattal C., Zaidi A.K.M., Wertheim H.F.L., Sumpradit N., Vlieghe E., Hara G.L., Gould I.M., Goossens H., Greko C.; So A.D., Bigdeli M., Tomson G., Woodhouse W., Ombaka E., Peralta A.Q., Qamar F.N., Mir F., Kariuki S., Bhutta Z.A., Coates A., Bergstrom R., Wright G.D., Brown E.D. & Cars O. 2013. Antibiotic resistance-the need for global solutions. Lancet Infectious Diseases. 13:1057-1098.

Liu M., Wu X., Li J., Liu L., Zhang R., Shao D. & Du X. 2017. The specific anti-biofilm effect of gallic acid on Staphylococcus aureus by regulating the expression of the ica operon. Food Control. 73:613-618.

Magiorakos A.-P., Srinivasan A., Carey R.B., Carmeli Y., Falagas M.E., Giske C.G., Harbarth S., Hindler J.F., Kahlmeter G., Olsson-Liljequist B., Paterson D.L., Rice L.B., Stelling J., Struelens M.J., Vatopoulos A., Weber J.T. & Monnet D.L. 2012. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clinical Microbiology and Infection. 18:268-281.

Martineau F., Lansac N., Me C., Roy P.H., Ouellette M. & Bergeron M.G. 2000. Correlation between the Resistance Genotype Determined by Multiplex PCR Assays and the Antibiotic Susceptibility Patterns of Staphylococcus aureus and Staphylococcus epidermidis. Antimicrobial Agents and Chemotherapy. 44:231-238.

Mavrogianni V.S., Menzies P.I., Fragkou I.A. & Fthenakis, G.C. 2011. Principles of Mastitis Treatment in Sheep and Goats. Veterinary Clinics of North America: Food Animal Practice. 27:115-120.

Nakagawa S., Taneike I., Mimura D., Iwakura N., Nakayama T., Emura T., Kitatsuji M., Fujimoto A. & Yamamoto T. 2005. Gene sequences and specific detection for Panton-Valentineleukocidin. Biochemical and Biophysical Research Communications. 328:995-1002.

Ng L.K.I., Martin M.A. & Mulvey, M. 2001. Multiplex PCR for the detection of tetracycline resistant genes. Molecular and Cellular Probes. 15:209-215.

Paterson G.K., Larsen A.R., Robb A., Edwards G.E., Pennycott T.W., Foster G., Mot D., Hermans K., Baert K., Peacock S.J., Parkhill J., Zadoks R.N. & Holmes M.A. 2012. The newly described mecA homologue, mecALGA251, is present in methicillin-resistant Staphylococcus aureus isolates from a diverse range of host species. Journal of Antimicrobial Chemotherapy. 67:2809-2813.

Pereira C.S., Santos L.M.M., Machado L.S., Melo D.A., Coelho S.M.O., Pereira V.L.A., Souza M.M.S. & Nascimento E.R. 2021. Proteomics characterization of Staphylococcus spp. From goat mastitis and phenogeno-typical assessment of resistance to beta-lactamics. Pesquisa Veterinária Brasileira. 41:1-6.

Prabhu K., Rao S. & Rao V. 2011. Inducible clindamycin resistance in Staphylococcus aureus isolated from clinical samples. Journal of Laboratory Physicians. 3:25-27.

Queiroga R.C.R.E., Santos B.M., Gomes A.M.P., Monteiro M.J., Teixeira S.M., Souza E.L., Pereira C.J.D. & Pintado M.M.E. 2013. Nutritional, textural and sensory properties of Coalho cheese made of goats’, cows’ milk and their mixture. LWT - Food Science and Technology. 50:538-544.

Sawant A.A., Gillespie B.E. & Oliver, S.P. 2009. Antimicrobial susceptibility of coagulase-negative Staphylococcus species isolated from bovine milk. Veterinary Microbiology. 134:73-81.

Souza V., Martins P.Y.F., Pinto D.S., Fernandes D.R. & Lima A.R. 2017. Sensibilidade antimicrobiana de Staphylococcus aureus isolados no leite de cabras com mastite subclínica. Comunicado Técnico On-line. 167:1-7. Available in: < https://www.embrapa.br/busca-de-publicacoes/-/publicacao/1087468/sensibilidad e-antimicrobiana-de-staphylococcus-aureus-isolados-no-leite-de-cabras-com-mas tite-subclinica>. [Accessed online in June 2021].

Truong-Bolduc Q.C., Dunman P.M., Strahilevitz J., Projan S.J. & Hooper, D.C., 2005. MgrA Is a Multiple Regulator of Two New Efflux Pumps in Staphylococcus aureus. Journal of Bacteriology. 187:2395-2405.

Truong-Bolduc Q.C., Strahilevitz J. & Hooper D.C. 2006. NorC, a new efflux pump regulated by MgrA of Staphylococcus aureus. Antimicrobial Agents and Chemotherapy. 50:1104-1107.

Truong-Bolduc Q.C., Zhang X. & Hooper D.C. 2003. Characterization of NorR Protein, a Multifunctional Regulator of norA Expression in Staphylococcus aureus. Journal of Bacteriology. 185:3127-3138.

Zelazny A.M., Ferraro M.J., Glennen A., Hindler J.F., Mann L.M., Munro S., Murray P.R., Reller L.B., Tenover F.C. & Jorgensen J.H. 2005. Selection of strains for quality assessment of the disk induction method for detection of inducible clindamycin resistance in Staphylococci: a CLSI collaborative study. Journal of Clinical Microbiology. 43:2613-2615.



How to Cite

Aragão, B. B., Trajano, S. C., Silva, R. A. da, Silva, B. P. e, Peixoto, R. de M., & Mota, R. A. (2021). Occurrence of Multi-Resistant Staphylococcus aureus in Artisan Goat Coalho Cheese in Northeastern Brazil. Acta Scientiae Veterinariae, 49. https://doi.org/10.22456/1679-9216.117447




Most read articles by the same author(s)

1 2 > >>