RESUMO

Due to the lack of knowledge about Campylobacterales in the Chilean poultry industry, the objective of this research was to know the prevalence, resistance, and genotypes of Campylobacter, Arcobacter and Helicobacter in 382 samples of chicken meat purchased in Valdivia, Chile. The samples were analyzed using three isolation protocols. Resistance to four antibiotics was evaluated by phenotypic methods. Genomic analyses were performed on selected resistant strains to detect resistance determinants and their genotypes. A total of 59.2% of the samples were positive. Arcobacter butzleri (37.4%) was the most prevalent species, followed by Campylobacter jejuni (19.6%), C. coli (11.3%), A. cryaerophilus (3.7%) and A. skirrowii (1.3%). Helicobacter pullorum (14%) was detected by PCR in a subset of samples. Campylobacter jejuni was resistant to ciprofloxacin (37.3%) and tetracycline (20%), while C. coli and A. butzleri were resistant to ciprofloxacin (55.8% and 2.8%), erythromycin (16.3% and 0.7%) and tetracycline (4.7% and 2.8%), respectively. Molecular determinants were consistent with phenotypic resistance. The genotypes of C. jejuni (CC-21, CC-48, CC-49, CC-257, CC-353, CC-443, CC-446 and CC-658) and C. coli (CC-828) coincided with genotypes of Chilean clinical strains. These findings suggest that besides C. jejuni and C. coli, chicken meat could play a role in the transmission of other pathogenic and antibiotic-resistant Campylobacterales.

RESUMO

Arcobacter represents a zoonotic emerging pathogen with increasing importance for public health and drinking water has been cited as a major risk factor for its dissemination. The aim of this work was to evaluate the survival capacity of Arcobacter in different water matrixes stored at different temperatures. Three different water matrixes were used, including potable water with a chlorine concentration of 0,5 mg/mL, non-chlorinated water and non-chlorinated water added with an 11% of organic matter. Each matrix was inoculated in a 1/10 proportion with 103 and 105 Arcobacter pools, divided into 4 different subsamples, in order to be incubated at 0°C, 5°C, 12 °C and 25°C by up to 15 days. The presence of Arcobacter in each matrix was determined on days 1, 3, 5, 7, 9, 11, 13 and 15. Results obtained show that this bacterium can survive in all the water matrixes evaluated, regardless of the presence or not of residual disinfecting agent. Also, the amount of CFU/mL inoculated in water correlates with the number of bacteria that can survive on it, and that incubation temperature has a significant effect over the bacterial survival.

Assuntos

RESUMO

Conspicuous egg-shaped, white, and smooth structures were observed at a hydrothermal vent site in the Guaymas Basin, Gulf of California. The gelatinous structures decomposed within hours after sampling. Scanning electron microscopy (SEM) and light microscopy showed that the structure consisted of filaments of less than 0.1 µm thickness, similar to those observed for "Candidatus Arcobacter sulfidicus." SEM-energy-dispersive X-ray spectroscopy (EDS) showed that the filaments were sulfur rich. According to 16S rRNA gene amplicon and fluorescence in situ hybridization (FISH) analyses, Arcobacter, a sulfide oxidizer that is known to produce filamentous elemental sulfur, was among the dominant species in the structure and was likely responsible for its formation. Arcobacter normally produces woolly snowflake like structures in opposed gradients of sulfide and oxygen. In the laboratory, we observed sulfide consumption in the anoxic zone of the structure, suggesting an anaerobic conversion. The sulfide oxidation and decomposition of the structure in the laboratory may be explained by dissolution of the sulfur filaments by reaction with sulfide under formation of polysulfides. IMPORTANCE At the deep-sea Guaymas Basin hydrothermal vent system, sulfide-rich hydrothermal fluids mix with oxygenated seawater, thereby providing a habitat for microbial sulfur oxidation. Microbial sulfur oxidation in the deep sea involves a variety of organisms and processes and can result in the excretion of elemental sulfur. Here, we report on conspicuous white and smooth gelatinous structures found on hot vents. These strange egg-shaped structures were often observed on previous occasions in the Guaymas Basin, but their composition and formation process were unknown. Our data suggest that the notable and highly ephemeral structure was likely formed by the well-known sulfide-oxidizing Arcobacter. While normally Arcobacter produces loose flocs or woolly layers, here smooth gel-like structures were found.

Assuntos

RESUMO

The first case of acute watery diarrhea disease due to Aliarcobacter butzleri (formerly Arcobacter butzleri) in Ecuador is reported. An infant presented with moderate protein-calorie malnutrition, dehydration and anemia. A curved Gram-negative organism was isolated from stools, having been preliminarily identified by phenotypic characteristics. Definitive identification was achieved by multiplex PCR. Aliarcobacter butzleri was the only pathogenic microorganism isolated. No other entero-pathogens, enterovirus or parasites were found. Our findings strongly suggest that in this specific case, A. butzleri was the etiological agent. Further investigations are needed to develop standardized diagnostic protocols and to establish the prevalence and significance of Aliarcobacter infections in humans.

Assuntos

RESUMO

This research aims to investigate the presence and pathogenic potential of Arcobacter in poultry meat samples purchased in the retail market of Valdivia (South of Chile) as well as in faecal samples from backyard chickens from rural areas around this city. The isolates obtained were identified by molecular methods. Furthermore, putative virulence genes were assessed by PCR and the antimicrobial resistance was tested by phenotypic methods. Arcobacter was present in 41·6% of the samples, with the highest value in retail poultry meat (55·7%) followed by backyard production (28·0%). Arcobacter butzleri was the most prevalent species (75·6%) followed by Arcobacter skirrowii (14·8%) and Arcobacter cryaerophilus (9·6%). An 8·5% of A. butzleri strains from meat were resistant to both ciprofloxacin and tetracycline and 6·1% were resistant to erythromycin, while none was resistant to gentamycin, unlike strains from domestic chickens, which showed no resistance. Furthermore, A. butzleri strains from chicken meat presented a higher prevalence of virulence genes than strains from domestic chickens. In fact, in this last group, some genes (hecA, hecB and irgA) were completely absent. Therefore, this study provides insight on the epidemiology of Arcobacter in Chilean poultry and suggests that under traditional breeding conditions strains are, apparently, less pathogenic and drug resistant.

Assuntos

RESUMO

Para la búsqueda de especies de Arcobacter fueron estudiadas 50 muestras de leche bovina obtenidas de los centros de acopio de la empresa láctea Ecolac, de las provincias de Loja y Zamora Chinchipe, Ecuador, y se aisló un total de ocho cepas de Arcobacter butzleri (16%). Esta frecuencia de aislamiento concuerda y está dentro de los rangos descriptos en la literatura. Todas las cepas aisladas fueron sensibles a gentamicina. Se encontró alta frecuencia de resistencia a tetraciclina (6/8 cepas) y a ciprofloxacina (4/8 cepas). Se verificó la ocurrencia de multirresistencia en tres de las ocho cepas aisladas.

A total of 50 samples of bovine milk obtained from bulk tanks milk of the collection centers belonging to the company ECOLAC, of the provinces of Loja and Zamora Chinchipe, Ecuador, were studied for Arcobacter species diagnosis, being isolated 8 strains of Arcobacter butzleri (16%). This frequency of isolation agrees and falls within the ranges described in the literature. All the isolated strains were susceptible to gentamicin. High resistance levels to tetracycline and ciprofloxacin were found with 6/8 and 4/8 resistant strains respectively. Multi-resistance was found in three of the eight isolated strains.

Foram estudadas, para a pesquisa de espécies de Arcobacter, 50 amostras de leite bovino, obtidas dos centros de coleta da empresa de laticínios ECOLAC, das províncias de Loja e Zamora Chinchipe, Equador, sendo isoladas em total 8 cepas de Arcobacter butzleri (16%). Esta frequência de isolamento concorda e está dentro dos níveis descritos na literatura. Todas as cepas isoladas foram sensíveis à gentamicina. Foi encontrada alta frequência de resistência à tetraciclina (6/8 cepas) e à ciprofloxacina (4/8 cepas), sendo verificada a ocorrência de multirresistência em três das oito cepas isoladas.

Assuntos

RESUMO

Acanthamoeba castellanii is a free-living amoeba found mainly in humid environments and Arcobacter butzleri is an emerging zoonotic pathogen, both can establish in vitro endosymbiotic relationships in the absence of bacterial replication. We analyzed the localization of A. butzleri within A. castellanii establishing their association with endoplasmic reticulum vesicles and mitochondria. Through confocal microscopy, we observed that during the early stages of endosymbiosis, there is not colocalization between amoebic vacuoles containing A. butzleri and mitochondria or ER vesicles of A. castellanii. Considering that energy production of this bacterium occurs via metabolism of amino acids or the tricarboxylic acid cycle, these results contribute to explain the absence of bacterial replication, since A. butzleri would not have access to the nutrients found in endoplasmic reticulum vesicles and mitochondria. In addition, we observe that A. butzleri induces significantly the actin polymerization of A. castellanii during the early stages of endosymbiosis.

Assuntos

RESUMO

Arcobacter butzleri is an emerging foodborne zoonotic pathogen that has been isolated from environmental water sources. This pathogen establishes in vitro endosymbiotic relationships with Acanthamoeba castellanii, a free-living amoeba found in environmental matrices such as soil and water. The principal aim of this study was to analyse the transcriptional pattern of flagellar (flaA-flaB-flgH-motA) and other putative virulence genes (ciaB-cadF-mviN-pldA) of A. butzleri during its interaction with A. castellanii by quantitative real-time PCR. The transcriptional analysis showed up-regulation of all genes analysed before A. butzleri became established as an endocytobiont of A. castellanii. In contrast, while A. butzleri remains an endocytobiont, a significant and sustained decrease in the transcription of all analysed genes was observed. Our findings suggest that A. butzleri requires a biphasic transcriptional pattern of flagellar and other putative virulence genes to establish an endosymbiotic relationship with A. castellanii.

Assuntos

RESUMO

Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii are Gram-negative pathogenic microorganisms that cause watery diarrhea and septicemia in humans. The aims of this study were to detect the presence of Arcobacter spp. in chicken meat from butcher shops in São Paulo (Brazil) and to verify their virulence genes and genotypic profiles. A total of 300 chicken cuts were analyzed. The results show the presence of Arcobacter spp. in 18.3% of samples, which were identified as A. butzleri (63.6%) and A. cryaerophilus (36.3%). All strains were positive for the virulence genes ciaB and mviN, followed by cj1349 (98%), pldA (94.4%), cadF (72.7%), tlyA (92.7%), hecA (49%), irgA (47.2%), and hecB (34.5%). These strains were subjected to single-enzyme amplified fragment length polymorphism. Nineteen genotypic profiles were obtained for A. butzleri, and 17 for A. cryaerophilus. These results confirm the presence of virulent strains of A. butzleri and A. cryaerophilus in the chicken meat in Brazil. The presence of potentially virulent strains of Arcobacter highlights a possible public health risk, particularly with respect to ingestion of undercooked foods and cross-contamination from uncooked foods during food preparation and contaminated utensils.

Assuntos

RESUMO

Arcobacter is an emerging zoonotic pathogen, and the major transmission routes to humans are the handling or consumption of contaminated raw/undercooked food products of animal origin, water and seafood. The isolation and identification of Arcobacter species are not routine in clinical laboratories; therefore, its true incidence in human infections may be underestimated. The present study aimed to isolate and characterize Arcobacter from carcasses and fecal samples collected at swine slaughterhouses and from meat markets in São Paulo State, Brazil. The isolates were identified using multiplex-PCR to differentiate the species and analyzed by single-enzyme amplified fragment length polymorphism (SE-AFLP). Arcobacter spp. were isolated from 73.0% of swine carcasses, 4% of fecal samples and 10% of pork samples. A. butzleri was the most prevalent species identified, followed by A. cryaerophilus. Interestingly, the carcasses presented higher frequency of A. butzleri isolation, whereas only A. cryaerophilus was isolated from fecal samples. SE-AFLP enabled the characterization of A. butzleri and A. cryaerophilus into 51 and 63 profiles, respectively. The great genetic heterogeneity observed for both species corroborates previous reports. This study confirms the necessity for a standard isolation protocol and the improvement of molecular tools to further elucidate Arcobacter epidemiology.(AU)

Arcobacter é um patógeno zoonótico emergente e as principais formas de transmissão para humanos são a manipulação e o consumo de água ou alimentos contaminados crus ou mal cozidos. O isolamento e a identificação das espécies de Arcobacter não fazem parte da rotina dos laboratórios clínicos; dessa forma, a real incidência da infecção em humanos é subestimada. O presente estudo teve o objetivo de isolar e caracterizar Arcobacter de carcaças e amostras de fezes coletadas em dois abatedouros de suínos e de carne suína de dois açougues no Estado de São Paulo, Brasil. As estirpes foram identificadas utilizando multiplex-PCR para diferenciar as espécies e foram analisadas por polimorfismo no comprimento de fragmentos amplificados (SE-AFLP). Arcobacter spp. foi isolado de 73% das carcaças, 4% das amostras de fezes e de 10% das amostras de carne suína avaliadas. A. butzleri foi a espécie mais prevalente, seguida por A. cryaerophilus. As carcaças apresentaram a maior taxa de isolamento de A. butzleri enquanto que apenas A. cryaerophilus foi isolado das amostras de fezes. SE-AFLP possibilitou a caracterização de A. butzleri e A. cryaerophilus em 51 e 63 perfis de bandas, respectivamente. A grande heterogeneidade genética observada para ambas as espécies corrobora estudos previous. Estes resultados confirmam a necessidade de protocolos de isolamento padronizados e o aperfeiçoamento das ferramentas moleculares para aprofundar os conhecimetos sobre epidemiologia das infecções pelo gênero Arcobacter.(AU)

Assuntos

RESUMO

Arcobacter is an emerging zoonotic pathogen, and the major transmission routes to humans are the handling or consumption of contaminated raw/undercooked food products of animal origin, water and seafood. The isolation and identification of Arcobacter species are not routine in clinical laboratories; therefore, its true incidence in human infections may be underestimated. The present study aimed to isolate and characterize Arcobacter from carcasses and fecal samples collected at swine slaughterhouses and from meat markets in São Paulo State, Brazil. The isolates were identified using multiplex-PCR to differentiate the species and analyzed by single-enzyme amplified fragment length polymorphism (SE-AFLP). Arcobacter spp. were isolated from 73.0% of swine carcasses, 4% of fecal samples and 10% of pork samples. A. butzleri was the most prevalent species identified, followed by A. cryaerophilus. Interestingly, the carcasses presented higher frequency of A. butzleri isolation, whereas only A. cryaerophilus was isolated from fecal samples. SE-AFLP enabled the characterization of A. butzleri and A. cryaerophilus into 51 and 63 profiles, respectively. The great genetic heterogeneity observed for both species corroborates previous reports. This study confirms the necessity for a standard isolation protocol and the improvement of molecular tools to further elucidate Arcobacter epidemiology.(AU)

Arcobacter é um patógeno zoonótico emergente e as principais formas de transmissão para humanos são a manipulação e o consumo de água ou alimentos contaminados crus ou mal cozidos. O isolamento e a identificação das espécies de Arcobacter não fazem parte da rotina dos laboratórios clínicos; dessa forma, a real incidência da infecção em humanos é subestimada. O presente estudo teve o objetivo de isolar e caracterizar Arcobacter de carcaças e amostras de fezes coletadas em dois abatedouros de suínos e de carne suína de dois açougues no Estado de São Paulo, Brasil. As estirpes foram identificadas utilizando multiplex-PCR para diferenciar as espécies e foram analisadas por polimorfismo no comprimento de fragmentos amplificados (SE-AFLP). Arcobacter spp. foi isolado de 73% das carcaças, 4% das amostras de fezes e de 10% das amostras de carne suína avaliadas. A. butzleri foi a espécie mais prevalente, seguida por A. cryaerophilus. As carcaças apresentaram a maior taxa de isolamento de A. butzleri enquanto que apenas A. cryaerophilus foi isolado das amostras de fezes. SE-AFLP possibilitou a caracterização de A. butzleri e A. cryaerophilus em 51 e 63 perfis de bandas, respectivamente. A grande heterogeneidade genética observada para ambas as espécies corrobora estudos previous. Estes resultados confirmam a necessidade de protocolos de isolamento padronizados e o aperfeiçoamento das ferramentas moleculares para aprofundar os conhecimetos sobre epidemiologia das infecções pelo gênero Arcobacter.(AU)

Assuntos

RESUMO

The presence of Arcobacter spp. in minced meat (including beef) samples has been well documented in different countries, with varying frequencies. Nevertheless, the only Latin American country reporting this bacterium in minced beef samples is Mexico, with a 28.8% frequency in 2003. Previous studies in Costa Rica have demonstrated the presence of Arcobacter species in samples taken from the poultry production chain, but still there are no studies performed in bovine meat. The aim of this study was to determine the frequency of this bacterium in 120 samples of minced beef acquired from the Central Valley region of Costa Rica and to describe the antibiotic sensitivity pattern of the isolates obtained. A total of 75 different Arcobacter strains were isolated from minced beef samples, for a final frequency of 48.3%. After species PCR identification, the strains were classified as A. butzleri (37.3%), A. cibarius (14.7%), A. thereius (12%), and Arcobacter spp. (36%). All samples were sensitive to gentamicin but were resistant to ampicillin, levofloxacin, nalidixic acid, and ciprofloxacin. The results obtained in this study show that the frequency of isolation of Arcobacter in minced beef samples is high and that there is a high resistance rate for antibiotics in common use. This suggests that Arcobacter represents a health risk for Costa Rica and that control measures should be developed to decrease its potential impact.

RESUMO

Arcobacter is a gram-negative rod recognized as a potential food- and waterborne pathogen; nevertheless, little is known about the effects of this pathogen on human and animal health. Although Arcobacter species are commonly found in nature, poultry is suspected to be the main vehicle for the transmission of this pathogen. The aims of this work were to determine the prevalence of Arcobacter spp. in broilers produced in Costa Rica for human consumption and to analyze the pathogenic capacity of the isolates through the detection of virulence genes. One hundred fifty-two samples of cecal content (87 farms), 104 samples of carcass rinse after chiller (six processing plants), and 96 carcass rinses from as many retail stores were analyzed. The suspicious isolates were identified using genus-specific PCR, and species-level identification was achieved with a multiplex PCR. Virulence genes were identified using the protocol described by L. Douidah, L. de Zutter, J. Baré, P. De Vos, P. Vandamme, O. Vandenberg, A.-M. Van den Abeele, and K. Houf (J. Clin. Microbiol. 50:735-741, 2012), which includes nine different virulence genes. The overall isolation frequency of Arcobacter was 6.5% (n = 23). Eight (34.8%) of the isolates came from cecal content, 2 (8.7%) were isolated from samples taken after chiller, and 13 (56.5%) were from retail stores. The species isolated included A. thereius (30.4%), A. butzleri (21.7%), A. skirrowii (4.3%), and A. cibarius (4.3%). The remaining samples were classified as Arcobacter sp. Gene tlyA was the most prevalent virulence gene, present in 9 of 23 samples analyzed; genes hecA and pldA were present in one only strain each. A strain of A. butzleri isolated from a retail store presented the highest number of virulence genes (five), and 11 samples did not present any of the genes analyzed. The results obtained suggest that the presence of virulent Arcobacter isolates in the poultry production chain from Costa Rica could be a risk for individuals who consume the contaminated product.

RESUMO

Arcobacter butzleri is an emergent zoonotic foodborne pathogen associated to enteritis and occasionally to bacteremia in human beings. Biotyping of this bacterium is important in order to establish the circulating strains and its dissemination routes. The purpose of this work was to determine the circulating A. butzleri biotypes in poultry products for human consumption in Southern Chile using the method proposed by Lior and Woodward, in order to explore the possibility of introducing this biotyping scheme as a routine laboratory tool. From the 60 strains studied the prevalent biotypes were 8A, 8B, 7A, 4A and 4B. The most frequently isolated biotype, independently of the sample of origin, was 8A with (44 strains, 73.3%). The less frequently isolated biotype was 4B (two strains 3.3%). The biotyping method used results to be simple, easy to handle and yields stable results. Therefore, it might be rescued to be used as a phenotypic tool for epidemiological marking of A. butzleri.

Arcobacter butzleri é um patógeno emergente, zoonótico e de transmissão alimentar, associado a enterite e, ocasionalmente, a bacteremia em seres humanos. A biotipagem desta bactéria é importante para estabelecer os biótipos circulantes e suas rotas de disseminação. Os objetivos deste trabalho foram determinar os biótipos de A. butzleri circulantes em alimentos de origem aviar para consumo humano, no sul do Chile, explorando a possibilidade de introduzir o método de biotipagem proposto por Lior e Woodward como uma ferramenta de rotina no laboratório. Entre as 60 cepas estudadas, os biótipos 8A, 8B, 7A e 4B foram os mais prevalentes. O biótipo mais frequentemente isolado, independentemente da amostra de origem, foi o biótipo 8A (44 cepas, 73,3%). O biótipo 4B apresentou a menor frequência de isolamento (duas cepas, 3,3%). O método de biotipagem utilizado resultou ser simples de executar, fácil de manusear e produz resultados estáveis. Portanto, pode ser resgatado para ser usado como uma ferramenta fenotípica para marcação epidemiológica de A. butzleri.

Assuntos

RESUMO

The survival of three Arcobacter butzleri strains inside Acanthamoeba castellanii was assessed using axenic cultures of A. castellanii that were inoculated with the tested strains and incubated at 26°C under aerobic conditions for 240h. The behavior of bacteria in contact with amoebae was monitored using phase contrast microscopy. The bacterial survival rate within amoebae was assessed through counting colony forming units, using the gentamicin protection assay. All A. butzleri strains were able to survive during 240h within the amoebae, thus suggesting that (i) A. butzleri resists the amoebic digestion processes at least for the analyzed time; (ii) that A. castellanii could serve as an environmental reservoir for this bacterium, probably acting as a transmission vehicle for A. butzleri.

Assuntos

RESUMO

The colourless mat covering organically enriched sediments underlying an intensive salmon farm in Estero Pichicolo, southern Chile, was surveyed by combined 454 PyroTag and conventional Sanger sequencing of 16S/18S ribosomal RNA genes for Bacteria and Eukarya. The mat was dominated by the sulphide-oxidizing bacteria (SOB) Candidatus Isobeggiatoa, Candidatus Parabeggiatoa and Arcobacter. By order of their abundances, sulphate-reducing bacteria (SRB) were represented by diverse deltaproteobacterial Desulfobacteraceae, but also within Desulfobulbaceae, Desulfuromonadaceae and Desulfovibrionaceae. The eukaryotic PyroTags were dominated by polychaetes, copepods and nematodes, however, ciliated protozoans were highly abundant in microscopy observations, and were represented by the genera Condylostoma, Loxophyllum and Peritromus. Finally, the abundant Sulfurimonas/Sulfurovum also suggest the occurrence of zero-valence sulphur oxidation, probably derived from Beggiatoaceae as a result of bacteriovorus infaunal activity or generated as free S(0) by the Arcobacter bacteria. The survey suggests an intense and complex sulphur cycle within the surface of salmon-farm impacted sediments.

Assuntos

RESUMO

The isolation frequency of Arcobacter species in children with diarrhea, fowls, mammals and food of avian and marine origin was established. In all the samples it was possible to isolate Arcobacter species corresponding 201 (39.4%) to A. butzleri and 24 (4.7) to A. cryaerophilus . Both species were simultaneously isolated in 19 (3.7%) being A. butzleri the most frequently isolated species.

Assuntos

RESUMO

The isolation frequency of Arcobacter species in children with diarrhea, fowls, mammals and food of avian and marine origin was established. In all the samples it was possible to isolate Arcobacter species corresponding 201 (39.4%) to A. butzleri and 24 (4.7) to A. cryaerophilus. Both species were simultaneously isolated in 19 (3.7%) being A. butzleri the most frequently isolated species.

Assuntos

RESUMO

The isolation frequency of Arcobacter species in children with diarrhea, fowls, mammals and food of avian and marine origin was established. In all the samples it was possible to isolate Arcobacter species corresponding 201 (39.4%) to A. butzleri and 24 (4.7) to A. cryaerophilus. Both species were simultaneously isolated in 19 (3.7%) being A. butzleri the most frequently isolated species.(AU)

Assuntos