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1.
Foodborne Pathog Dis ; 19(1): 56-61, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34665017

RESUMO

In recent years, methicillin-resistant Staphylococcus aureus (MRSA) has gained interest in veterinary medicine due to its zoonotic potential. Currently, little information is available on the genotypic and virulence characteristics of MRSA isolates detected in Nigerian abattoirs. To better understand the epidemiology of MRSA associated with the abattoir food chain environment in Nigeria, a total of 18 isolates (humans: n = 5, slaughter animals: n = 5, and environment: n = 8), previously spa typed, were recovered and characterized by Staphylococcus cassette chromosome mec (SCCmec) typing, and phenotypic and genotypic antimicrobial susceptibility testing. In addition, 10 of the 18 MRSA strains with a new spa type (t16571) were subjected to multilocus sequence typing. The similarity of strains was analyzed based on the results of the DNA microarray analysis. The 18 MRSA strains harbored two distinct SCCmec types (IVa and V) and belonged to four clonal clusters (CC1, CC7, CC88, and CC152). All MRSA of the new spa type t16571 (n = 10) harbored the SCCmec type IVa. Seven of the MRSA t16571 strains belonged to ST88, while three other strains were assigned to ST3614. The 18 MRSA isolates were categorized into six virulence profiles, and the detection rate for the Panton-Valentine Leukocidin gene was high (33.3%). The antimicrobial susceptibility profiles of the 18 MRSA varied widely between strains, but phenotypic resistance corresponded to relevant resistance genes harbored. The detection of highly similar MRSA strains in slaughter animals, abattoir workers, and the environment underlines the need to use adequate measures at Nigerian abattoirs to prevent further spread and transmission of MRSA to humans or food.


Assuntos
Staphylococcus aureus Resistente à Meticilina , Infecções Estafilocócicas , Matadouros , Animais , Antibacterianos/farmacologia , Células Clonais , Humanos , Staphylococcus aureus Resistente à Meticilina/genética , Testes de Sensibilidade Microbiana , Nigéria/epidemiologia , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/veterinária
2.
J Dairy Sci ; 101(4): 3380-3386, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29395146

RESUMO

Methicillin-resistant Staphylococcus aureus (MRSA) have been described repeatedly in dairy herds. In this study, we compared the prevalence and antimicrobial resistance of MRSA in bulk tank milk from conventional and organic dairy herds in Germany. Samples were collected from 372 conventional and 303 organic dairy herds throughout Germany. Bulk tank milk (25 mL) was tested for MRSA using an established double selective enrichment method. The MRSA isolates were typed using spa typing and tested for resistance to 19 antimicrobials using the broth microdilution method. Methicillin-resistant Staph. aureus was detected more frequently in bulk tank milk from conventional (9.7%) than from organic (1.7%) dairy herds. Herd size and region were associated with differences in prevalence. Most isolates (38/41) were from spa types associated with the livestock-associated clonal complex CC398. Isolates from conventional herds tended to be more resistant to antimicrobials; however, because of the limited number of isolates from organic herds, no statistical tests were performed. In conclusion, prevalence of MRSA in dairy herds in Germany seems to be increasing and is more prevalent in regions with high livestock density. Organic herds are also affected although at a lower level. Therefore, MRSA should be specifically included in biosecurity protocols for dairy herds, and effective control measures need to be investigated.


Assuntos
Bovinos/microbiologia , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Animais , Indústria de Laticínios , Feminino , Alemanha/epidemiologia , Leite/microbiologia , Agricultura Orgânica , Prevalência , Infecções Estafilocócicas/veterinária
3.
Clin Infect Dis ; 63(10): 1349-1352, 2016 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-27655995

RESUMO

We investigated the evolution and epidemiology of a novel livestock-associated methicillin-resistant Staphylococcus aureus strain, which colonizes and infects urban-dwelling Danes even without a Danish animal reservoir. Genetic evidence suggests both poultry and human adaptation, with poultry meat implicated as a probable source.


Assuntos
Doenças Transmitidas por Alimentos/microbiologia , Gado/microbiologia , Staphylococcus aureus Resistente à Meticilina/genética , Infecções Estafilocócicas , Adulto , Idoso , Animais , DNA Bacteriano/genética , Dinamarca , Feminino , Microbiologia de Alimentos , Humanos , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Vison/microbiologia , Polimorfismo de Nucleotídeo Único/genética , Aves Domésticas/microbiologia , Estudos Retrospectivos , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/transmissão , Infecções Estafilocócicas/veterinária
4.
Int J Med Microbiol ; 303(3): 124-7, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23517689

RESUMO

Methicillin-resistant Staphylococcus aureus sequence type 398 (MRSA ST398) has emerged in livestock worldwide. In particular, areas in Europe with high densities of livestock farming are affected. Consequently, the incidence of human colonization and infection with ST398 is rapidly increasing. Distinguishing different ST398 isolates with standard typing tools is problematic. The objective of this study was to examine the discriminatory power of Multiple-Locus Variable number tandem repeat Fingerprinting (MLVF) on a highly diverse ST398 collection. Our data show that MLVF combined with spa-typing is an attractive approach for high-resolution typing of ST398 isolates and unveiling their relatedness.


Assuntos
Portador Sadio/veterinária , Gado/microbiologia , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/genética , Epidemiologia Molecular/métodos , Tipagem Molecular/métodos , Infecções Estafilocócicas/veterinária , Animais , Portador Sadio/epidemiologia , Impressões Digitais de DNA/métodos , Europa (Continente)/epidemiologia , Genótipo , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Repetições Minissatélites , Infecções Estafilocócicas/epidemiologia , Proteína Estafilocócica A/genética
5.
Arch Virol ; 158(11): 2341-50, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23760627

RESUMO

Some years ago, MRSA clonal complex (CC) 398 emerged, which spread extensively in livestock animals. People in contact with food production animals are at high risk of colonization. A reduction of MRSA CC398 in livestock might be achieved by application of virulent phages. However, there have not yet been any reports published on phages lysing MRSA CC398 strains. In this study, three virulent phages (PSa1, PSa2 and PSa3) with lytic activity against MRSA CC398 strains were isolated from German pig farms. Morphologically, the phages are members of the family Podoviridae, and they exhibited an identical host range. They lysed 52 (60 %) out of 86 tested MRSA CC398 strains representing 18 different spa types. While the PSa1 and PSa3 genomes have a similar size of approximately 17.5 kb, the PSa2 genome is somewhat larger (ca. 18.5 kb). Southern hybridization revealed strong DNA homologies between the phages, which was confirmed by sequence analysis of cloned restriction fragments and PCR products. Moreover, the whole PSa3 genomic sequence (17,602 bp) showed a close relationship to 44AHJD-like phages, which are not known to contain virulence-associated genes. To assess whether these phages might be candidates for applications, in vitro experiments were carried out in which the number of MRSA CC398 cells could be reduced by up to four log10 units. The phages were stable at a wide range of temperatures and pH values.


Assuntos
Staphylococcus aureus Resistente à Meticilina/virologia , Podoviridae/isolamento & purificação , Fagos de Staphylococcus/isolamento & purificação , Suínos/virologia , Criação de Animais Domésticos , Animais , Técnicas de Tipagem Bacteriana , Bacteriólise , Biologia Computacional/métodos , DNA Viral/análise , DNA Viral/genética , DNA Viral/isolamento & purificação , Poeira/análise , Fezes/virologia , Humanos , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/genética , Testes de Sensibilidade Microbiana , Podoviridae/classificação , Podoviridae/genética , Podoviridae/fisiologia , Análise de Sequência de DNA , Fagos de Staphylococcus/classificação , Fagos de Staphylococcus/genética , Fagos de Staphylococcus/fisiologia
6.
Foodborne Pathog Dis ; 10(3): 263-9, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23268760

RESUMO

The Shiga toxin-producing Escherichia coli O104:H4 outbreak in Germany in 2011 required the development of appropriate tools in real-time for tracing suspicious foods along the supply chain, namely salad ingredients, sprouts, and seeds. Food commodities consumed at locations identified as most probable site of infection (outbreak clusters) were traced back in order to identify connections between different disease clusters via the supply chain of the foods. A newly developed relational database with integrated consistency and plausibility checks was used to collate these data for further analysis. Connections between suppliers, distributors, and producers were visualized in network graphs and geographic projections. Finally, this trace-back and trace-forward analysis led to the identification of sprouts produced by a horticultural farm in Lower Saxony as vehicle for the pathogen, and a specific lot of fenugreek seeds imported from Egypt as the most likely source of contamination. Network graphs have proven to be a powerful tool for summarizing and communicating complex trade relationships to various stake holders. The present article gives a detailed description of the newly developed tracing tools and recommendations for necessary requirements and improvements for future foodborne outbreak investigations.


Assuntos
Surtos de Doenças , Infecções por Enterobacteriaceae/epidemiologia , Doenças Transmitidas por Alimentos/epidemiologia , Escherichia coli Shiga Toxigênica/patogenicidade , Análise por Conglomerados , Egito , Infecções por Enterobacteriaceae/microbiologia , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/microbiologia , Alemanha/epidemiologia , Humanos , Extratos Vegetais , Escherichia coli Shiga Toxigênica/isolamento & purificação , Trigonella/microbiologia
8.
Appl Environ Microbiol ; 78(2): 541-8, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22081568

RESUMO

In this study we investigated the kinetics of colonization, the host susceptibility and transmissibility of methicillin-resistant Staphylococcus aureus (MRSA) after nasal treatment of pigs with three different MRSA strains of distinctive clonal lineages (sequence type 398 [ST398], ST8, and ST9), and origin in weaning piglets. The colonization dose of 5.0 × 10(8) CFU/animal was determined in preliminary animal studies. A total of 57 piglets were randomly divided into four test groups and one control group. Each of three test groups was inoculated intranasally with either MRSA ST8, MRSA ST9, or MRSA ST398. The fourth group was a mixture of animals inoculated with MRSA ST398 and noninoculated "sentinel" animals. Clinical signs, the nasal, conjunctival, and skin colonization of MRSA, fecal excretion, and organ distribution of MRSA, as well as different environmental samples were examined. After nasal inoculation with MRSA piglets of all four test groups showed no clinical signs of an MRSA infection. MRSA was present on the nasal mucosa, skin, and conjunctiva in all four test groups, including sentinel animals. Likewise, fecal excretion and internal colonization of MRSA ST8, ST9, and ST398 could be shown in each group. However, fecal excretion and the colonization rate of the nasal mucosa with MRSA ST9 were significantly lower in the first days after infection than in test groups infected with ST8 and ST398. The results of this study suggest differences in colonization potential of the different MRSA types in pigs. Furthermore, colonization of lymph nodes (e.g., the ileocecal lymph node) with MRSA of the clonal lineage ST398 was demonstrated.


Assuntos
Portador Sadio/veterinária , Predisposição Genética para Doença , Staphylococcus aureus Resistente à Meticilina/patogenicidade , Infecções Estafilocócicas/veterinária , Doenças dos Suínos/microbiologia , Animais , Portador Sadio/microbiologia , Túnica Conjuntiva/microbiologia , Fezes/microbiologia , Genótipo , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/genética , Tipagem Molecular , Mucosa Nasal/microbiologia , Pele/microbiologia , Infecções Estafilocócicas/microbiologia , Suínos
9.
Appl Environ Microbiol ; 78(16): 5666-71, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22685139

RESUMO

During 1 year, samples were taken on 4 days, one sample in each season, from pigs, the floor, and the air inside pig barns and from the ambient air and soil at different distances outside six commercial livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA)-positive pig barns in the north and east of Germany. LA-MRSA was isolated from animals, floor, and air samples in the barn, showing a range of airborne LA-MRSA between 6 and 3,619 CFU/m(3) (median, 151 CFU/m(3)). Downwind of the barns, LA-MRSA was detected in low concentrations (11 to 14 CFU/m(3)) at distances of 50 and 150 m; all upwind air samples were negative. In contrast, LA-MRSA was found on soil surfaces at distances of 50, 150, and 300 m downwind from all barns, but no statistical differences could be observed between the proportions of positive soil surface samples at the three different distances. Upwind of the barns, positive soil surface samples were found only sporadically. Significantly more positive LA-MRSA samples were found in summer than in the other seasons both in air and soil samples upwind and downwind of the pig barns. spa typing was used to confirm the identity of LA-MRSA types found inside and outside the barns. The results show that there is regular airborne LA-MRSA transmission and deposition, which are strongly influenced by wind direction and season, of up to at least 300 m around positive pig barns. The described boot sampling method seems suitable to characterize the contamination of the vicinity of LA-MRSA-positive pig barns by the airborne route.


Assuntos
Microbiologia do Ar , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Microbiologia do Solo , Suínos/microbiologia , Animais , Carga Bacteriana , Genótipo , Alemanha , Abrigo para Animais , Estudos Longitudinais , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/genética , Tipagem Molecular , Estações do Ano , Vento
10.
Int J Food Microbiol ; 375: 109737, 2022 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-35635992

RESUMO

Staphylococcus aureus is an important opportunistic pathogen of both humans and animals. It can cause several diseases, including mastitis, as well as food poisoning by production of heat-stable enterotoxins in food. The aim of this study was to determine the prevalence of S. aureus and the diversity of strains circulating in the Zambian dairy value chain, which have not been studied in detail before. Three provinces were covered by the study (Lusaka, Southern, and Western) and almost 2000 samples along the dairy value chain, covering both the informal and formal market sectors, were taken at two time points (dry and wet season), with a special focus on raw milk. Nearly 300 presumptive S. aureus isolates were confirmed by MALDI-TOF MS and real-time PCR. Raw milk from traditional and smallholder farms was widely contaminated with S. aureus; prevalence was 33-46% depending on the study province. Raw milk from milk collection centres, informal traders, traditional market sellers, and processors were also frequently contaminated with S. aureus. In addition, S. aureus was detected in several milk bucket swabs and nasal and hand swabs of milkers. From industrially processed (heat-treated) milk and dairy products, no S. aureus was isolated. Methicillin-resistant S. aureus (MRSA) were not detected, but around 10% of the S. aureus isolates carried lukS-PV, a marker gene for the virulence factor Pantone-Valentine leucocidin (PVL), which has been associated with severe diseases in human. Molecular typing identified a total of 44 spa types including 13 novel types: t18396, t18397, t18398, t18399, t18400, t18402, t18416, t20459, t20460, t20461, t20462, t20463, and t20464. Furthermore, 12 novel multi-locus sequence types were identified: ST7012, ST7100, ST7101, ST7177, ST7291, ST7304, ST7305, ST7344, ST7596, ST7597, ST7598, and ST7599, of which ST7012, ST7177, and ST7596 fall into the bovine-associated clonal linage CC97. The spa types t084, t267, t355, and the novel type t20464 were common in all three study provinces. The predominant spa type varied depending on the province. Whole genome sequencing (WGS) and core genome multi-locus sequence typing (cgMLST) indicates transmission of strains along the Zambian dairy chain with possible persistence in the chain over time. cgMLST also revealed a very close relatedness between some isolates from milkers and from raw milk or milk buckets. The high prevalence and wide spa type diversity of S. aureus, as well as possible direct or indirect transmission of (potentially highly virulent) S. aureus to humans along the Zambian dairy value chain, are of public health concern, particularly as milk and milk products are often consumed raw by the Zambian population.


Assuntos
Staphylococcus aureus Resistente à Meticilina , Infecções Estafilocócicas , Animais , Antibacterianos , Bovinos , Feminino , Staphylococcus aureus Resistente à Meticilina/genética , Testes de Sensibilidade Microbiana , Leite , Tipagem de Sequências Multilocus , Prevalência , Saúde Pública , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/veterinária , Staphylococcus aureus/genética , Zâmbia/epidemiologia
11.
BMC Vet Res ; 7: 69, 2011 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-22074403

RESUMO

BACKGROUND: The purpose of this study was to investigate the prevalence of MRSA in herds of fattening pigs in different regions of Germany, and to determine factors associated with the occurrence of this pathogen. For this purpose pooled dust samples were collected, and a questionnaire covered information regarding herd characteristics and management practices. Samples were pre-enriched in high-salt medium followed by selective enrichment containing cefoxitin/aztreonam, and culturing. Presumptive colonies were confirmed by multiplex-PCR targeting nuc-, mecA- and 16S rRNA-genes. Isolates were spa- and SCCmec-, and in selected cases, multilocus sequence-typed. Susceptibilities to 13 antimicrobials were determined by broth microdilution. Statistical analysis was carried out using backward stepwise logistic regression to calculate odds ratios with the MRSA test result as the outcome and herd characteristics as categorical covariates. RESULTS: Overall, 152 of 290 (52%) fattening pig farms tested positive for MRSA. The prevalence in the east, north- and south-west of Germany ranged from 39 to 59%.t011 (66%) and t034 (23%) were the most commonly identified spa-types, and 85% of isolates carried SCCmec Type V. Identified spa-types were all associated with clonal complex CC398. Susceptibility testing revealed that all isolates were resistant to tetracycline. High resistance rates were also found for sulfamethoxazole/trimethoprim (40%), and quinupristin/dalfopristin (32%). In addition, 83% of strains displayed multidrug resistant (> 3 substance classes) phenotypes.Logistic regression revealed herd size (large farms OR: 5.4; CI: 2.7-11.2; p < 0.05), and production type (wean-to-finish OR: 4.0; CI: 1.6-10.4; p < 0.05) as risk factors associated with a positive MRSA finding in fattening pig operations. CONCLUSIONS: MRSA CC398 is widely distributed among herds of fattening pigs in Germany. Farm management plays a crucial role in the dissemination of MRSA with herd size, and production type representing potential major indicators.


Assuntos
Staphylococcus aureus Resistente à Meticilina/fisiologia , Infecções Estafilocócicas/veterinária , Doenças dos Suínos/epidemiologia , Criação de Animais Domésticos/normas , Animais , Anti-Infecciosos/farmacologia , Poeira/análise , Alemanha/epidemiologia , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Testes de Sensibilidade Microbiana , Tipagem Molecular , Prevalência , Fatores de Risco , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologia , Inquéritos e Questionários , Suínos , Doenças dos Suínos/microbiologia
12.
Vet Microbiol ; 216: 52-59, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29519525

RESUMO

Staphylococcus aureus is a zoonotic pathogen of significant public health concern. Information on the prevalence and risk factors facilitating bacterial colonization and spread under abattoir settings in Nigeria are scarce. This cross-sectional study was designed to determine prevalence of S. aureus as well as risk factors on knowledge and practices facilitating pathogen carriage among workers and slaughter animals in two municipal abattoirs of Ilorin and Ibadan, Nigeria. Swab samples (n = 1671) from nostrils of cattle, goats, pigs and abattoir workers, and from meat tables and abattoir walls were collected for detection of S. aureus. A questionnaire was administered to 275 workers to elucidate risk factors of pathogen carriage applying a logistic regression model. S. aureus prevalence was 6.5%. In total, MSSA and MRSA were detected at a frequency of 5.4% and 1.1%. Molecular analysis of the isolates revealed 19 different spa types, including a novel spa type (t16751). Gender, marital status, occupation and abattoir location were factors influencing worker's practices in relation to pathogen carriage and spread in the abattoir setting. This present study detected not only low MSSA and MRSA prevalence, in both abattoirs but also low risk perception and hygiene practices employed by abattoir workers. Good practices among workers at Nigerian abattoirs are needed to mitigate S. aureus carriage. Further studies expounding the antibiotic resistance and relationships of MSSA and MRSA strains detected in this study are needed to complement understanding of the spread of S. aureus in the abattoir food chain.


Assuntos
Carne/microbiologia , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Saúde Pública , Infecções Estafilocócicas/veterinária , Staphylococcus aureus/isolamento & purificação , Matadouros , Adulto , Animais , Bovinos , Estudos Transversais , Farmacorresistência Bacteriana , Feminino , Humanos , Higiene , Masculino , Staphylococcus aureus Resistente à Meticilina/genética , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Cavidade Nasal/microbiologia , Nigéria , Prevalência , Fatores de Risco , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/transmissão , Staphylococcus aureus/genética , Suínos , Adulto Jovem , Zoonoses/epidemiologia , Zoonoses/microbiologia , Zoonoses/transmissão
13.
Vet Microbiol ; 200: 25-32, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27318609

RESUMO

Little is known about the characteristics of MRSA occurring along the broiler and turkey production chains. The aim of this study was to characterise and compare MRSA of turkey and broiler origin sampled on different production levels using a DNA microarray and antimicrobial susceptibility testing. Main differences could be observed in the prevalence of the resistance genes erm(C), aacA-aphD and tet(K) and the number of non-wild type strains with minimum inhibitory concentration values (MICs) above the epidemiological cut-off values (ECOFFs) for gentamicin and kanamycin. Overall, MRSA with non-wild type phenotype for the macrolide-lincosamide-streptogramin group, tetracycline, and trimethoprim were found in more than 70% of poultry isolates. Non-wild type isolates carrying the qacC gene conferring resistance to quaternary ammonium compound disinfectants were found at all production stages in similar frequencies. Regarding the presence of enterotoxin genes in poultry-derived MRSA the enterotoxin gene cluster (egc) was only found in Non-CC (clonal complex) 398 strains. Three CC398 strains harboured the genes sed (from turkey at slaughter and broiler meat) and sea-N315 (from a chicken carcass). One Non-CC398 strain isolated from turkey meat was found positive for the seb gene and also enterotoxin production. Similarity analysis based on selected resistance and virulence markers revealed a high clonality among Non-CC398 isolates. Isolates of the same clonal complex clustered together according to their common virulence and resistance profiles. Strains of CC9 were grouped in two subclusters due to different resistance genes. Our results underline, that there are other poultry associated clones of MRSA (mainly CC9 and CC5) besides the predominant CC398 which are similar in both poultry species.


Assuntos
Galinhas/microbiologia , Farmacorresistência Bacteriana , Staphylococcus aureus Resistente à Meticilina , Doenças das Aves Domésticas/microbiologia , Infecções Estafilocócicas/veterinária , Perus/microbiologia , Animais , Antibacterianos/farmacologia , Enterotoxinas/genética , Fazendas , Alemanha , Lincosamidas/farmacologia , Macrolídeos/farmacologia , Meticilina/farmacologia , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/patogenicidade , Testes de Sensibilidade Microbiana , Fenótipo , Infecções Estafilocócicas/microbiologia , Estreptograminas/farmacologia , Virulência/genética
14.
Sci Rep ; 7(1): 2004, 2017 05 17.
Artigo em Inglês | MEDLINE | ID: mdl-28515479

RESUMO

Staphylococcus aureus MRSA strains belonging to the clonal complex 398 (CC398) are highly prevalent in livestock and companion animals but may also cause serious infections in humans. CC398 strains in livestock usually do not possess well-known virulence factors that can be frequently found in other MRSA sequence types (ST). Since many staphylococcal virulence genes are residing on the genomes of temperate phages, the question arises why livestock-associated (LA-) CC398 strains are only rarely infected by those phages. We isolated and characterized four temperate phages (P240, P282, P630 and P1105) containing genes of the immune evasion cluster (IEC) and/or for the Panton-Valentine leucocidin (PVL). Sequence analysis of the phage genomes showed that they are closely related to known phages and that the DNA region encoding lysis proteins, virulence factors and the integrase exhibits numerous DNA repeats which may facilitate genomic rearrangements. All phages lysed and lysogenized LA-CC398 strains. Integration of IEC phage P282 was detected at ten sites of the hosts' chromosome. The prophages were stably inherited in LA-CC398 and enterotoxin A, staphylokinase and PVL toxin were produced. The data demonstrate that lysogenic conversion of LA-CC398 strains by virulence-associated phages may occur and that new pathotypes may emerge by this mechanism.


Assuntos
Doenças dos Animais/microbiologia , Gado/microbiologia , Staphylococcus aureus Resistente à Meticilina/genética , Infecções Estafilocócicas/veterinária , Fatores de Virulência/genética , Animais , Bacteriólise , Bacteriófagos/fisiologia , Bacteriófagos/ultraestrutura , Sequência de Bases , Cromossomos Bacterianos , Ordem dos Genes , Genes Virais , Genoma Bacteriano , Genômica/métodos , Staphylococcus aureus Resistente à Meticilina/patogenicidade , Staphylococcus aureus Resistente à Meticilina/virologia , Virulência
15.
Vet Microbiol ; 200: 46-51, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26923248

RESUMO

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) isolates of clonal complex 398 (CC398) are frequently found in Europe, and recent studies highlighted the importance of mobile genetic element (MGE) exchange for host adaptation of this lineage. Of note, one of the MGEs commonly found in human S. aureus isolates, the immune evasion cluster (IEC) harboring bacteriophage Saint3, is very rarely found in LA-MRSA CC398 isolates obtained from farm animals, but more frequently found in LA-MRSA CC398 that were retransmitted to humans. Here, we analyzed with a set of S. aureus CC398 isolates harboring/lacking φSaint3 how this MGE affects (i) phagocytosis of CC398 isolates by polymorphonuclear neutrophils (PMNs), and (ii) hemolysis of human and livestock-derived erythrocytes. Isolates lacking φSaint3 were more efficiently phagocytosed by human PMNs in whole blood phagocytosis assays than isolates harboring this bacteriophage, irrespective of their origin. Notably, a similar effect was observed when equine blood was utilized, but not detected with porcine blood. Integration of φSaint3 into LA-MRSA CC398 strains lacking this MGE confirmed these findings, as φSaint3-harboring recipients were again less efficiently ingested by PMNs in equine and human blood than their parental strains. Integration of φSaint3 strongly reduced the hemolytic potential of the culture supernatants against human-derived erythrocytes, and to a smaller extent also against porcine-derived erythrocytes, while φSaint3 integration only slightly affected the hemolytic capacities against equine-derived red blood cells. The significant protective effect of φSaint3 against phagocytosis by equine PMNs suggests that the host specificity of the IEC components might be broader than currently assumed.


Assuntos
Toxinas Bacterianas/genética , Bacteriófagos/fisiologia , Proteínas Hemolisinas/genética , Evasão da Resposta Imune , Staphylococcus aureus Resistente à Meticilina/virologia , Esfingomielina Fosfodiesterase/genética , Infecções Estafilocócicas/microbiologia , Animais , Eritrócitos/microbiologia , Hemólise , Cavalos , Especificidade de Hospedeiro , Humanos , Gado , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/imunologia , Neutrófilos , Fagocitose , Infecções Estafilocócicas/imunologia , Suínos
16.
Vet Microbiol ; 185: 7-14, 2016 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-26931385

RESUMO

Methicillin-susceptible Staphylococcus (S.) aureus (MSSA) and methicillin-resistant S. aureus (MRSA) are colonizers of skin and mucosa. In humans, MSSA and MRSA compete for colonization space in the anterior nares of pig farmers; however, it was also shown that MSSA/MRSA co-colonization is common and one clone can be found rather than differing types of MSSA and MRSA. We investigated the colonization and clonality of both, MSSA and MRSA in pigs over a longer time. Eighteen sows were nasally sampled three times every ten weeks. Additionally, environmental samples were taken. Samples were investigated for MSSA and MRSA, respectively. The spa type was defined from up to five MRSA and MSSA isolates found per sample and sampling time; selected isolates were further investigated by microarray. Three sows (16.7%) were completely negative for MSSA and MRSA. Twelve pigs (66.7%) were irregularly positive for both, MSSA and MRSA over the time, whereas seven out of them (38.9%) were simultaneously colonized. CC398 (t034, t011) MRSA and CC9 (t337, t1430, and t13816) MSSA associated spa types were exclusively found. In 44.4% (n=8) of sows up to two different types of MSSA were present at the same time and sample. Strains of the same clonal lineage showed a high genetic identity despite their origin. Highly identic clones were present in sows and their environment. As conclusion, MSSA/MRSA may not exclude each other in the anterior nares of pigs. Pigs may also carry different clones at the same time.


Assuntos
Biodiversidade , Coinfecção/veterinária , Staphylococcus aureus Resistente à Meticilina/fisiologia , Infecções Estafilocócicas/veterinária , Staphylococcus aureus/fisiologia , Doenças dos Suínos/microbiologia , Animais , Técnicas de Tipagem Bacteriana , Células Clonais , Coinfecção/microbiologia , Feminino , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Cavidade Nasal/microbiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/classificação , Staphylococcus aureus/genética , Staphylococcus aureus/isolamento & purificação , Suínos
17.
Berl Munch Tierarztl Wochenschr ; 129(7-8): 290-5, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27529990

RESUMO

In the autumn 2013, the National Reference Laboratory for coagulase positive staphylococci (CPS) including Staphylococcus (S.) aureus (NRL-Staph) at the Federal Institute for Risk Assessment has organized its first interlaboratory profiency testing (ILPT) trial for the detection of staphylococcal enterotoxins (SE) types SEA to SEE in food. The purpose of the ILPT was to assess the analytical competence of the official laboratories of the Federal German "Länder"authorities. Moreover, it was the intention to gain an overview of the standard methods implemented in the participating laboratories for the purpose of SE detection in food. Five cream cheese samples at three different contamination levels (blank, low, and high) were sent to each participant. In total, 15 laboratories participated to the ILPT: 14 laboratories from 11 Federal German "Länder", and the European Reference Laboratory for CPS including S. aureus (EU-RL for CPS). Data sets from 14 participating laboratories were included in the analysis. Overall, a specificity of 100% (14/14 true negative results), a sensitivity of 55% (31/56 true positive results), and an accuracy of 64% (45/60 true results) was achieved. The majority of participants (9/15) used other analytical methods for the detection of SE in food than the suggested European Screening Method (ESM) v5. To conclude on the ILPT in general it is to state that the majority of participating laboratories failed to correctly identify SE-low-contaminated samples. Further efforts are necessary to improve the analytical capacity and sensitivity as regards the detection of SE in food in Germany.


Assuntos
Queijo/análise , Enterotoxinas/análise , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Staphylococcus aureus , Queijo/microbiologia , Alemanha
18.
Vet Microbiol ; 191: 88-95, 2016 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-27374912

RESUMO

Livestock associated methicillin-resistant Staphylococcus aureus (LA-MRSA) has emerged in animal production worldwide. Most LA-MRSA in Europe belong to the clonal complex (CC) 398. The reason for the LA-MRSA emergence is not fully understood. Besides antimicrobial agents used for therapy, other substances with antimicrobial activity applied in animal feed, including metal-containing compounds might contribute to their selection. Some of these genes have been found in various novel SCCmec cassettes. The aim of this study was to assess the occurrence of metal-resistance genes among a LA-S. aureus collection [n=554, including 542 MRSA and 12 methicillin-susceptible S. aureus (MSSA)] isolated from livestock and food thereof. Most LA-MRSA isolates (76%) carried at least one metal-resistance gene. Among the LA-MRSA CC398 isolates (n=456), 4.8%, 0.2%, 24.3% and 71.5% were positive for arsA (arsenic compounds), cadD (cadmium), copB (copper) and czrC (zinc/cadmium) resistance genes, respectively. In contrast, among the LA-MRSA non-CC398 isolates (n=86), 1.2%, 18.6% and 16.3% were positive for the cadD, copB and czrC genes, respectively, and none were positive for arsA. Of the LA-MRSA CC398 isolates, 72% carried one metal-resistance gene, and the remaining harboured two or more in different combinations. Differences between LA-MRSA CC398 and non-CC398 were statistically significant for arsA and czrC. The czrC gene was almost exclusively found (98%) in the presence of SCCmec V in both CC398 and non-CC398 LA-MRSA isolates from different sources. Regarding the LA-MSSA isolates (n=12), some (n=4) were also positive for metal-resistance genes. This study shows that genes potentially conferring metal-resistance are frequently present in LA-MRSA.


Assuntos
Desinfetantes/farmacologia , Farmacorresistência Bacteriana/genética , Gado/microbiologia , Metais Pesados/farmacologia , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/genética , Ração Animal/análise , Animais , Europa (Continente) , Staphylococcus aureus/genética
19.
Int J Food Microbiol ; 186: 68-73, 2014 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-25016468

RESUMO

Staphylococcus aureus is an important food-borne pathogen due to the ability of enterotoxigenic strains to produce staphylococcal enterotoxins (SEs) in food. Methicillin-resistant S. aureus (MRSA) is also an important pathogen for humans, causing severe and hard to treat diseases in hospitals and in the community due to its multiresistance against antimicrobials. In particular, strains harbouring genes encoding for the Panton-Valentine leukocidin (PVL) toxin are of concern from a public health perspective as they are usually capable of causing severe skin and soft tissue infections (sSSTIs) and occasionally necrotizing pneumonia which is associated with high mortality. This is the first report on the detection of MRSA with genes encoding for PVL in wild boar meat. Among the 28 MRSA isolated from wild boar meat in the course of a national monitoring programme in Germany, seven harboured PVL-encoding genes. Six of the isolates were identical according to the results of spa-, MLST-, microarray- and PFGE-typing. They could be assigned to the epidemic MRSA clone USA300. Epidemiological investigations revealed that people handling the food were the most likely common source of contamination with these MRSA. These findings call again for suitable hygienic measures at all processing steps of the food production chain. The results of the study underline that monitoring along the food chain is essential to closely characterise the total burden of MRSA for public health.


Assuntos
Microbiologia de Alimentos , Carne/microbiologia , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Animais , Técnicas de Tipagem Bacteriana , Farmacorresistência Bacteriana/genética , Manipulação de Alimentos/normas , Abastecimento de Alimentos/normas , Alemanha , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/genética , Tipagem de Sequências Multilocus , Sus scrofa
20.
Berl Munch Tierarztl Wochenschr ; 127(3-4): 129-34, 2014.
Artigo em Alemão | MEDLINE | ID: mdl-24693658

RESUMO

The use of antimicrobial agents is responsible for the emergence and spread of antibiotic resistant bacteria. Nevertheless, multiresistant bacteria have been found in animals that have never been exposed to antimicrobial agents. Wild animals that are carriers of methicillin-resistant organisms represent a hazard since they can transmit their bacteria to other animals and to humans. In the hunting season 2009/2010 nasal swabs of 98 red deer and 109 wild boars were examined for the presence of methicillin-sensitive and methicillin-resistant staphylococci. From each wild boar methicillin-susceptible staphylococci (Staphylococcus aureus in 28% and Staphylococcus spp. in 72% of the animals) were isolated. In red deer the detection rate of Staphylococcus (S.) aureus and methicillin-susceptible staphylococci was 49% and 17%, respectively. The occurrence of S. aureus was significantly higher (p < 0.05) in red deer than in wild boars. Methicillin-resistant staphylococci were not found. However, in one third of the red deer, methicillin-resistant bacteria of the genus Enterococcus spp. and Bacillus spp. were isolated. The results of the present study indicate that wildlife, especially red deer are an important reservoir for S. aureus and that the upper respiratory tract of red deer is regularly colonised with methicillin-resistant bacteria such as Bacillus spp. and Enterococcus spp. Primarily, commensal bacteria are harmless to human health, however, red deer may be a reservoir for antibiotic-resistant bacteria.


Assuntos
Animais Selvagens/microbiologia , Infecções Bacterianas/microbiologia , Infecções Bacterianas/veterinária , Reservatórios de Doenças/microbiologia , Farmacorresistência Bacteriana , Animais , Antibacterianos/farmacologia , Bacillus/efeitos dos fármacos , Portador Sadio/microbiologia , Portador Sadio/veterinária , Cervos/microbiologia , Enterococcus/efeitos dos fármacos , Alemanha , Meticilina/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Sus scrofa/microbiologia
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