RESUMO
Considering that plasmid conjugation is a major driver for the dissemination of antimicrobial resistance in bacteria, this study aimed to investigate the effects of residual concentrations of antimicrobial growth promoters (AGPs) in poultry litter on the frequencies of IncFII-FIB plasmid conjugation among Escherichia coli organisms. A 2 × 5 factorial trial was performed in vitro, using two types of litter materials (sugarcane bagasse and wood shavings) and five treatments of litter: non-treated (CON), herbal alkaloid sanguinarine (SANG), AGPs monensin (MON), lincomycin (LCM) and virginiamycin (VIR). E. coli H2332 and E. coli J62 were used as donor and recipient strains, respectively. The presence of residues of monensin, lincomycin and virginiamycin increased the frequency of plasmid conjugation among E. coli in both types of litter materials. On the contrary, sanguinarine significantly reduced the frequency of conjugation among E. coli in sugarcane bagasse litter. The conjugation frequencies were significantly higher in wood shavings compared with sugarcane bagasse only in the presence of AGPs. Considering that the presence of AGPs in the litter can increase the conjugation of IncFII-FIB plasmids carrying antimicrobial resistance genes, the real impact of this phenomenon on the dissemination of antimicrobial resistant bacteria in the poultry production chain must be investigated.
Assuntos
Anti-Infecciosos , Infecções por Escherichia coli , Saccharum , Animais , Antibacterianos/farmacologia , Anti-Infecciosos/farmacologia , Celulose/farmacologia , Conjugação Genética , Escherichia coli/genética , Infecções por Escherichia coli/microbiologia , Lincomicina/farmacologia , Monensin , Plasmídeos/genética , Aves Domésticas/microbiologia , Virginiamicina/farmacologiaRESUMO
Methicillin resistance mediated by the mecA gene in Staphylococcus aureus, also known as "true MRSA", is typically associated with high oxacillin MIC values (≥8 mg/L). Because non-mecA-mediated oxacillin resistant S. aureus phenotypes can also cause hard-to-treat diseases in humans, their misidentification as methicillin-susceptible S. aureus strains (MSSA) can compromise the efficiency of the antimicrobial therapy. These strains have been refereed as Borderline Oxacillin-Resistant S. aureus (BORSA) but their characterization and role in clinical microbiology have been neglected. Considering the increasing importance of livestock-associated methicillin-resistant S. aureus ST398 (LA-MRSA) as an emerging zoonotic pathogen worldwide, this study aimed to report the genomic context of oxacillin resistance in porcine S. aureus ST398 strains. S. aureus isolates were recovered from asymptomatic pigs from three herds. Oxacillin MIC values ranged from 4 to 32 mg/L. MALDI-TOF-confirmed isolates were screened for mecA and mecC by PCR and genotyped by means of PFGE and Rep-PCR. Seven isolates were whole genome sequenced. None of the isolates harbored the mecA gene or its variants. Although all seven sequenced isolates belonged to one sequence type (ST398), two different spa types (t571 and t1471) were identified. All isolates harbored conserved blaZ gene operon and no mutations on genes encoding for penicillin-binding-proteins were detected. Genes conferring resistance against other drugs such as aminoglycosides, chloramphenicol, macrolide, lincosamide and streptogramin (MLS), tetracycline and trimethoprim were also detected. Isolates also harbored virulence genes encoding for adhesins (icaA; icaB; icaC; icaD; icaR), toxins (hlgA; hlgB; hlgC; luk-PV) and protease (aur). Pigs can serve as reservoirs of non-mecA-mediated oxacillin-resistant ST398 strains potentially pathogenic to humans. Considering that mecA has been the main target to screen methicillin-resistant staphylococci, the occurrence of BORSA phenotypes is probably underestimated in livestock.
Assuntos
Staphylococcus aureus Resistente à Meticilina , Infecções Estafilocócicas , Doenças dos Suínos , Animais , Staphylococcus aureus Resistente à Meticilina/genética , Testes de Sensibilidade Microbiana/veterinária , Oxacilina/farmacologia , Infecções Estafilocócicas/veterinária , Staphylococcus aureus , SuínosRESUMO
In light of the scarcity of information about the occurrence and epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) and coagulase-negative staphylococci (MRCNS) in small ruminants in general, and particularly dairy goats, we launched this limited-scope study. The findings reported here show the detection of MRSA and MRCNS in goat milk and teat skin samples from dairy goat herds in the state of Ohio. A total of 120 milk samples and 120 teat-swab samples were collected from 5 farms. After conventional isolation and phenotypic characterization of the staphylococci colonies, bacterial isolates were tested by PCR assay targeting the genes nuc to identify Staphylococcus aureus and mecA to detect MRSA and MRCNS. The clonal complexes of MRSA isolates was also determined by multiloccus sequence typing. Fifteen (6.2%) positive S. aureus samples were found in this study: 9 from milk and 6 from teat skin samples. Four (2%) MRSA isolates were detected and, using multiloccus sequence typing genotyping, these were designated to clonal complexes CC133 (n = 2; milk samples) and CC5 (n = 2; teat skin). Three (1.25%) coagulase-negative staphylococci isolates from the teat skin also harbored the mecA gene. Although, the MRSA isolated from milk samples is not a typical human-associated lineage, the CC5 clone isolated from teat skin is a common and widespread clonal complex associated with humans, suggesting that this extramammary niche could be a relevant reservoir of methicillin-resistant staphylococci. Furthermore, the fact that 75% of MRSA were recovered from 1 farm showing poor hygiene practices strengthens the hypothesis that good hygiene practices could be useful to prevent persistence and spread of MRSA at a farm level.
Assuntos
Doenças das Cabras/epidemiologia , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Infecções Estafilocócicas/veterinária , Staphylococcus aureus/isolamento & purificação , Animais , Antibacterianos/farmacologia , Coagulase/metabolismo , Doenças das Cabras/tratamento farmacológico , Cabras , Meticilina/farmacologia , Testes de Sensibilidade Microbiana , Ohio , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/epidemiologiaRESUMO
The aim of this study was to investigate the phenotypic and genotypic diversity and anti-microbial resistance among staphylococci of dairy herds that originated from Paraiba State, north-eastern Brazil, a region where such studies are rare. Milk samples (n = 552) were collected from 15 dairy farms. Isolates were evaluated for anti-microbial susceptibility by Kirby-Bauer disc diffusion method. Confirmation of methicillin-resistant Staphylococcus aureus (MRSA) was performed using multiplex PCR targeting mecA and nuc genes in addition to phenotypic assay based on PBP-2a latex agglutination. Clonal relatedness of isolates was determined by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) genotyping. Staphylococci were detected in 269 (49%) of the samples. Among these, 65 (24%) were S. aureus. The remaining 204 isolates were either coagulase-negative staphylococci (n = 188; 70%) or coagulase positive other than S. aureus (n = 16; 6%). Staphylococci were cultured in seven (35%) of the 20 hand swab samples, from which five isolates were S. aureus. The isolates were most commonly resistant against penicillin (43%), ampicillin (38%) and oxacillin (27%). The gene mecA was detected in 21 S. aureus from milk and in one isolate from a milker's hand. None of the isolates were resistant to vancomycin. PFGE findings showed high clonal diversity among the isolates. Based on MLST, we identified a total of 11 different sequence types (STs 1, 5, 6, 83, 97, 126, 1583, 1622, 1623, 1624 and 1625) with four novel STs (ST1622-ST1625). The findings show that MRSA is prevalent in milk from semi-extensive dairy cows in north-eastern Brazil, and further investigation on its extent in various types of milk production systems and the farm-to-table continuum is warranted.
Assuntos
Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Leite/microbiologia , Intoxicação Alimentar Estafilocócica/veterinária , Animais , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Biodiversidade , Brasil/epidemiologia , Bovinos , Indústria de Laticínios , Farmacorresistência Bacteriana Múltipla , Feminino , Contaminação de Alimentos , Microbiologia de Alimentos , Variação Genética , Genótipo , Humanos , Staphylococcus aureus Resistente à Meticilina/classificação , Testes de Sensibilidade Microbiana , Nuclease do Micrococo/genética , Tipagem de Sequências Multilocus , Proteínas de Ligação às Penicilinas/genética , Fenótipo , Reação em Cadeia da Polimerase , Intoxicação Alimentar Estafilocócica/epidemiologia , Staphylococcus aureus/efeitos dos fármacosRESUMO
The purpose of this study was to investigate the occurrence, antimicrobial resistance patterns, phenotypic and genotypic relatedness of Salmonella enterica recovered from captive wildlife host species and in the environment in Ohio, USA. A total of 319 samples including faecal (n = 225), feed (n = 38) and environmental (n = 56) were collected from 32 different wild and exotic animal species in captivity and their environment in Ohio. Salmonellae were isolated using conventional culture methods and tested for antimicrobial susceptibility with the Kirby-Bauer disc diffusion method. Salmonella isolates were serotyped, and genotyping was performed using the pulsed-field gel electrophoresis (PFGE). Salmonella was detected in 56 of 225 (24.9%) faecal samples; six of 56 (10.7%) environmental samples and six of 38 (15.8%) feed samples. Salmonella was more commonly isolated in faecal samples from giraffes (78.2%; 36/46), cranes (75%; 3/4) and raccoons (75%; 3/4). Salmonella enterica serotypes of known public health significance including S. Typhimurium (64.3%), S. Newport (32.1%) and S. Heidelberg (5.3%) were identified. While the majority of the Salmonella isolates were pan-susceptible (88.2%; 60 of 68), multidrug-resistant strains including penta-resistant type, AmStTeKmGm (8.8%; six of 68) were detected. Genotypic diversity was found among S. Typhimurium isolates. The identification of clonally related Salmonella isolates from environment and faeces suggests that indirect transmission of Salmonella among hosts via environmental contamination is an important concern to workers, visitors and other wildlife. Results of this study show the diversity of Salmonella serovars and public health implications of human exposure from wildlife reservoirs.
Assuntos
Animais Exóticos , Animais Selvagens/microbiologia , Salmonelose Animal/epidemiologia , Salmonella enterica/isolamento & purificação , Ração Animal/microbiologia , Animais , Animais de Zoológico , Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Eletroforese em Gel de Campo Pulsado , Meio Ambiente , Microbiologia Ambiental , Fezes/microbiologia , Genótipo , Ohio/epidemiologia , Fenótipo , Salmonella enterica/classificação , Salmonella enterica/genéticaRESUMO
The aim of this study was to investigate the adulteration of goat milk produced by smallholders in semiarid northeastern Brazil with bovine milk as an adulterant. The study was requested by the association of smallholder producers in the region to investigate and to inhibit adulteration practices as a need to ensure the quality and safety of goat milk. A duplex PCR assay has been developed and standardized. Further validation was performed in 160 fresh bulk goat milk samples. The detection limit of the duplex PCR was 0.5% bovine milk in goat milk and the results indicated that 41.2% of the goat milk presented to market was positive for bovine milk. Making the test available to the association of producers, together with extension activities, have been applied to reduce adulteration in goat milk sold to small-scale dairy plants and to ensure the species origin for goat milk in the state of Paraíba.
Assuntos
Contaminação de Alimentos/análise , Leite/normas , Reação em Cadeia da Polimerase Multiplex/veterinária , Animais , Brasil , Bovinos , Indústria de Laticínios , Feminino , Cabras , Leite/química , Reprodutibilidade dos TestesRESUMO
The aim of this study was to gain information on quality traits, mainly bacterial and somatic cell counts of bulk milk, produced by small- and medium-scale producers in a semiarid northeastern region of Brazil and to identify and characterize possible risk factors associated with those quality traits. A cross-sectional study was performed on 50 farms. Bulk milk samples were collected for bacterial and somatic cell counts. Additionally, information about farm demographics, general management practices, hygiene, and milking procedures was also obtained. Multivariable analysis using logistic regression was performed with predictors previously identified by univariate analysis using a Fisher's Exact test. Aerobic mesophilic bacteria counts varied from 3.59 log to 6.95 log cfu/mL, with geometric mean of 5.27 log cfu/mL. Mean total coliform count was 3.27 log (1.52 log to 5.89 log) most probable number (MPN)/mL, whereas mean thermotolerant coliforms was 2.38 log (1.48 log to 4.75 log) MPN/mL. A high positive correlation was observed between aerobic mesophilic bacteria and coliform counts. Although most farms met the standard for the current regulations for total bacteria (88%) and somatic cell counts (94%), nearly half of the producers (46%) would have problems in achieving the 2012 threshold limit for total bacteria count if no improvement in milk quality occurs. Mean value for staphylococci was 3.99 log (2.31 log to 6.24 log) cfu/mL, and Staphylococcus aureus was detected in 33 (66%) farms. Premilking teat-end wash procedure (odds ratio=0.191) and postmilking teat dip (odds ratio=0.67) were associated with lower aerobic mesophilic bacteria and Staphylococcus aureus counts in bulk milk, respectively. Considering that the farm characteristics in this study are representative of the semiarid northeastern region, these findings encourage further investigations for supporting intervention measures intended to improve the quality of milk produced by smallholders.