Diversity and emergence of multi-resistant Staphylococcus spp. isolated from subclinical mastitis in cows in of the state of Piauí, Brazil.

This research aimed to identify the diversity of bacterial species of the genus Staphylococcus spp. in subclinical mastitis in dairy herds in the state of Piauí, Northeastern Brazil, and to evaluate the phenotypic and genotypic resistance profile. Samples were obtained from a total of 17 dairy farms, amounting to 321 positive samples in the California Mastitis Test. Staphylococcus spp. were identified by matrix-assisted laser desorption ionization time-of-flight mass spectroscopy. Subsequently, an antibiogram was performed, and a polymerase chain reaction was carried out to screen for resistance genes in the isolates. Among all the isolates, 59.45% (110/185) belonged to the Staphylococcus genus. Moreover, the following Staphylococcus spp. were identified Staphylococcus aureus, 68.1% (75/110); Staphylococcus chromogenes, 12.7% (14/110); Staphylococcus epidermidis, 5.4% (6/110); Staphylococcus sciuri, 4.5% (5/110); Staphylococcus warneri, 2.7% (3/110); Staphylococcus haemolyticus, 1.8% (2/110); Staphylococcus hominis, 1.8% (2/110); Staphylococcus arlettae, 0.9% (1/110); Staphylococcus capitis, 0.9% (1/110); and Staphylococcus gallinarum, 0.9% (1/110). The antibiogram showed a high frequency of resistance to penicillin and ampicillin, 70.0% (77/110) and 61.8% (68/110), respectively, and a low frequency of resistance to gentamicin and vancomycin, 10.9% (12/110) and 11.8% (13/110), respectively. In the genotypic tests for the different species of Staphylococcus spp., the occurrence of the blaZ gene was observed in 60.9% (67/110) of the isolates, followed by tetL and tetM, both with 20.0% (22/110) each, and the mecA and vanB genes were detected in 0.9% (1/110) of the samples. The identification of all Staphylococcus species isolated from subclinical mastitis cases and the phenotypic and genotypic resistance characterization in these isolates is of great importance for dairy farming in the state of Piauí, as well as for public health.

Bovine mastitis in northeastern Brazil: Occurrence of emergent bacteria and their phenotypic and genotypic profile of antimicrobial resistance.

The aim of this study was to identify emergent pathogens associated with bovine mastitis in northeastern Brazil and to characterize them for phenotypic and genotypic resistance to antimicrobials. A total of 321 milk samples from cows with subclinical mastitis were collected, and the isolates obtained in culture were identified using matrix-associated laser desorption-ionization - time of flight mass spectrometry. Phenotypic and genotypic antimicrobial resistance tests were performed. We identified 72 bacteria considered emergent in the study region: Enterococcus faecalis (26.3%; 19/72), Streptococcus agalactiae (22.2%; 16/72), Enterococcus faecium (20.0%; 15/72), Escherichia coli (6.9%; 5/72), 6.9% (5/72) Lactococcus garvieae (6.9%; 5/72), Acinetobacter baumannii (5.5%; 4/72), Bacillus subtilis (1.3%; 1/72), Kocuria marina (1.3%; 1/72), Macrococcus caseolyticus (1.3%; 1/72), Microbacterium resistens (1.3%; 1/72), Micrococcus luteus (1.3%; 1/72), Streptococcus dysgalactiae (1.3%; 1/72), Streptococcus hyovaginalis (1.3%; 1/72) and Streptococcus pluranimalium (1.3%; 1/72). The antibiogram revealed the following resistance profiles: ampicillin (77.7%; 56/72), cefoxitin (69.4%; 50/72), erythromycin (61.1%; 44/72), oxacillin (63.8%; 46/72), penicillin (79.1%; 57/72), tetracycline (63.8%; 46/72), gentamicin (25.0%; 18/72), and vancomycin (20.8%; 15/72). Of the isolates, 83.4% (60/72) showed multiple resistance to antimicrobials. The tetM gene was identified in 43.0% (31/72) of the isolates, followed by tetL (31.9%; 23/72), and blaZ (26.3%; 19/72). 83.4% (60/72) of the isolates presented a multiple antimicrobial resistance index higher than 0,2. Emergent bacteria with zoonotic and multiresistant potential occur in cows with mastitis in northeastern Brazil. It is necessary to monitor the occurrence of these and other bacteria in livestock environments and develop control strategies to prevent their spread.

Occurrence of emerging multiresistant pathogens in the production chain of artisanal goat coalho cheese in Brazil.

Sanitary-hygienic failures in cheese making can pose health risks to consumers. This study aimed to identify multiresistant pathogens in different production stages of artisanal goat coalho cheese in Brazil and characterize their phenotypic and genotypic resistance. Eleven properties in the state of Pernambuco, Brazil, participated in the study. Samples were obtained from different stages of production and the humans involved. The samples obtained were submitted to microbiological culture, then all the isolated microorganisms were submitted to the Matrix Associated Laser Desorption-Ionization - Time of Flight technique for the microbiological identification of the species. Subsequently, Staphylococcus spp., Enterococcus spp. and Macrococcus caseolyticus were subjected to polymerase chain reaction to search for resistance genes and disc diffusion technique to evaluate the resistance profile. A total of 111 isolates were obtained and 31 species were identified, with the frequency of Staphylococcus spp. (62.20%; 69/111), Enterococcus spp. (11.60%; 13/111), Macrococcus caseolyticus (10%; 11/111), Bacillus spp. (3.60%; 4/111), Enterobacter spp. (3.60%; 4/111), Aureobasidium pullulans (1.80%; 2/111), Corynebacterium camporealensis (1.80%; 2/111), Issatchenkia occidentalis (1.80%; 2/111), Kocuria kristinae (1.80%; 2/111), Aerococcus viridans (0.90%; 1/111) and Filifactor villosus (0.90%; 1/111). Phenotypic and genotypic resistance was also detected with the occurrence of 15.90% (7/44) of the mecA gene, 4% (1/25) vanA, and 4% (1/25) vanB in Staphylococcus spp. and 20% (2/10) vanB in and Enterococcus spp. Emerging multiresistant pathogens are present in the production chain of artisanal goat cheese and humans, who exert an important role in disseminating these bacteria with imminent risks to human health.

Multiresistant zoonotic pathogens isolated from goat milk in Northeastern Brazil.

Dairy goats play a significant role in socio-economic, cultural, and nutritional development in many countries. This study aimed to identify multiresistant zoonotic pathogens causing mastitis in goats, in addition to characterizing them for the presence of resistance genes and phenotypic resistance. A total of 714 milk samples from 357 lactating goats in 12 farms in the Northeast region of Brazil were analyzed. The isolates were submitted to Matrix Associated Laser Desorption-Ionization - Time of Flight to identify bacterial species, Polymerase Chain Reaction (PCR) to search for resistance genes, and an antibiogram to evaluate the phenotypic profile of antimicrobial resistance. A total of 214 pathogens were identified and bacterial prevalence was 83.29 % (178/214) Staphylococcus spp.; 6.50 % (14/214) Micrococcus luteus; 3.73 % (8/214) Corynebacterium spp.; 2.80 % (6/214) Bacillus spp.; 1.38 % (3/214) Escherichia coli; 0.92 % (2/214) Enterobacter cloacae; 0.46 % (1/214) Aerococcus viridans; 0.46 % (1/214) Morganella morganii; and 0.46 % (1/214) Turicella otitidis. As for gene frequency, 64.60 % (115/178) of the isolates carried the blaZ gene; 37.07 % (66/178) norA; 22.47 % (40/178) tet(L); 16.85 % (30/178) tet(M); 14.04 % (25/178) norB; 8.42 % (15/178) vanA; 7.30 % (13/178) msrA; 6.41 % (5/178) tet-38; 4.49 % (8/178) norC; 2.25 % (4/178) mecA; and 0.56 % (1/178) vanB. Emerging multiresistant zoonotic pathogens are present in the goat milk production chain, especially the coagulase-negative Staphylococcus species that pose a risk to human and animal health.

Occurrence of multi-resistant Staphylococcus aureus in artisan goat coalho cheese in northeastern Brazil

The artisanal goat coalho cheese is one of the products obtained that stand out in the dairy goat farming of the Northeast of Brazil. Despite its importance, goat cheese is often made under inadequate hygienic-sanitary conditions and usually uses raw goat's milk, increasing the risk of product contamination. Among the pathogens carried by goat coalho cheese, Staphylococcus aureus stands out, being responsible for cases of food poisoning and persistent infections that are difficult to treat. This study aimed to evaluate the contamination, genotypic and phenotypic resistance of Staphylococcus aureus isolated from artisanal coalho cheese made with goat milk produced in the Northeast region of Brazil. 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. 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 no 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.(AU)

Methicillin-resistant Staphylococcus epidermidis in handler of cheese made with goat's milk in Brazil / Staphylococcus epidermidis resistente à meticilina em manipulador de queijo elaborado com leite de cabra no Brasil

ABSTRACT: In the Northeastern Brazil, artisanal cheese production is an important local economic activity for small producers. Methicillin-resistant Staphylococcus is responsible for causing infection in animals and humans. This study described the first detection of methicillin-resistant S. epidermidis isolated in the nasal cavity of a handler of coalho cheese made with goat's milk in Northeastern Brazil. This brief communication highlighted the importance of adopting biosafety measures by cheese handlers, in order to reduce possible contamination and the spread of pathogens in the production chain of this type of artisanal cheese in Brazil.

RESUMO: Na região Nordeste do Brasil, a cadeia de produção de queijo artesanal é uma atividade local importante para pequenos produtores. Staphylococcus resistentes à meticilina são responsáveis por causar infecções em animais e seres humanos. Neste estudo descreve-se a primeira detecção de S. epidermidis resistente à meticilina isolado da cavidade nasal de um manipulador de queijo coalho elaborado com leite de cabra no Nordeste do Brasil. Este relato destaca a importância da adoção de medidas de biossegurança por manipuladores de queijo, a fim de reduzir possíveis contaminações e a disseminação de patógenos na cadeia produtiva deste tipo de queijo artesanal no Brasil.

Genetic traceability of Staphylococcus aureus strains isolated from primiparous dairy cows mastitis, humans and environment in the Northeast region of Brazil / Rastreabilidade genética de cepas de Staphylococcus aureus isoladas de mastite de vacas primíparas, ordenhadores e ambiente de ordenha na região Nordeste do Brasil

ABSTRACT: This research aimed to investigate the genotypic relatedness of 18 Staphylococcus aureus strains isolated from intramammary infections in primiparous cows and extramammary sites on five dairy herds by rep-PCR using RW3A primers, and by PFGE using the endonuclease SmaI. The isolates were also evaluated in vitro for the susceptibility against beta-lactam antimicrobials drugs (penicillin and oxacillin), considering that beta-lactams are frequently used for treating staphylococcal intrammamary infections. The rep-PCR typing was highly discriminatory (D value= 0.9804) and a total of 15 patterns were detected. The PFGE method was also highly discriminatory (D value= 0.9667) and a total of 13 patterns were observed. A total of 15 out of 18 (83%) isolates were resistant to penicillin and one out of 18 (6%) to oxacillin. In conclusion, these findings confirmed the occurrence of a high genetic diversity of S. aureus strains at the herds and the presence of clonally-related strains only at the same herd, emphasizing a variety of genotypic profiles among the isolates.

RESUMO: Objetivou-se com este estudo investigar a correlação genética de 18 cepas de Staphylococcus aureus isoladas de infecções intramamárias em vacas primíparas e de locais extramamários em cinco propriedades leiteiras através das técnicas de PCR por sequências palindrômicas extragênicas repetitivas (rep-PCR), usando iniciadores RW3A, e de eletroforese em gel de campo pulsado (PFGE), usando a endonuclease SmaI. Os isolados também foram avaliados in vitro quanto à suscetibilidade aos antimicrobianos beta-lactâmicos (penicilina e oxacilina). A tipagem por rep-PCR foi altamente discriminatória (valor D = 0,9804) e um total de 15 padrões foram detectados. Os isolados de S. aureus foram agrupados em três grupos diferentes (A a C), com 80% de similaridade. A técnica de PFGE também foi altamente discriminatória (valor D = 0,9667) e um total de 13 padrões foi observado. A análise do dendrograma com um coeficiente de similaridade de 80% gerou dois grupos diferentes (A e B). Além disso, cepas clonais isoladas do leite foram identificadas na mesma propriedade pelos dois métodos de tipificação e, apesar da presença de cepas dominantes, nossos resultados sugerem uma alta diversidade genética dentre as cepas de S. aureus analisadas. Um total de 15, dos 18 (83%) isolados, eram resistentes à penicilina e um dos 18 (6%) à oxacilina. Assim, esses achados confirmam a ocorrência de uma alta diversidade genética de cepas de S. aureus nas propriedades e a presença de cepas clonalmente relacionadas apenas na mesma propriedade, enfatizando uma variedade de perfis genotípicos entre os isolados.

Serological evidence of Leptospira sp. in humans from Fernando de Noronha Island, Brazil.

The prevalence of leptospirosis in humans is highly variable, being influenced by climatic factors, the presence of reservoirs, occupational exposure, recreational activity, and socioeconomic conditions. The objective of this study was to estimate the prevalence of Leptospira sp. and identify the predominant human serovars on the island of Fernando de Noronha, Brazil, based on a microscopic agglutination test. The prevalence of anti-Leptospira antibodies was 1.17% (4/341; I.C. 0.46%-2.98%), with the predominance of serovars Icterohaemorrhagiae, Javanica, Mini and Louisiana. This is the first study on the occurrence of antibodies against Leptospira sp. in humans in Fernando de Noronha and highlights the need to implement control and prevention strategies in this island environment.

Soroprevalence of bovine alphaherpesvirus type 1 (BoHV-1) and risk factors associated with dairy properties of the municipality of Senador Guiomard, Acre, Brazil / Soroprevalência de alfaherpesvírus bovino tipo 1 (BoHV-1) e fatores de risco associados em propriedades leiteiras do município de Senador Guiomard, Acre, Brasil

Bovine alphaherpesvirus type 1 (BoHV-1) is the etiological agent responsible for serious infections that compromise the respiratory and genital tracts of affected cattle. In order to estimate the seroprevalence of BoHV-1 and to identify the associated risk factors in dairy farms in the city of Senador Guiomard, Acre, Brazil, the present study was carried out through the analysis of 180 blood serum samples submitted to the screening of anti-BoHV-1 by the virus neutralization test (VN) and by means of the evaluation of the epidemiological questionnaire applied in the eight investigated properties. The prevalence of seropositivity for BoHV-1 was 61.1%, ranging from 43.3 to 86.2% among the analyzed properties. The variable absence of veterinary assistance showed statistically significant association (odds ratio ­ OR = 2.10; p < 0.001) with alphaherpesvirus infection. The results demonstrate that the frequency of BoHV-1 is high and needs to be controlled through prophylactic and health management measures.(AU)

O alfaherpesvírus bovino tipo 1 (BoHV-1) é o agente etiológico responsável por graves infecções que comprometem os tratos respiratório e genital dos bovinos acometidos. Com o objetivo de estimar a soroprevalência de BoHV-1 e identificar os fatores de risco associados em propriedades leiteiras do município de Senador Guiomard, Acre, foi realizado o presente estudo, por meio da análise de 180 amostras de soro sanguíneo submetidas à pesquisa de anticorpos anti-BoHV-1 pelo teste de vírus neutralização (VN) e por meio da avaliação do questionário epidemiológico aplicado nas oito propriedades investigadas. A prevalência de soropositividade para BoHV-1 foi de 61,1% variando de 43,3 a 86,2% entre as propriedades analisadas. A variável ausência de assistência veterinária apresentou associação estatisticamente significativa (odds ratio ­ OR = 2,10; p < 0,001) com a infecção pelo alfaherpesvírus. Os resultados demonstram que a frequência de BoHV-1 é alta e precisa ser controlada através de medidas profiláticas e de manejo sanitário.(AU)