Survival capacity of Arcobacter butzleri at different incubation temperatures in drinking water

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.

Arcobacter butzleri, un agente zoonótico en leche bovina / Arcobacter butzleri, a zoonotic agent in bovine milk / Arcobacter butzleri, um agente zoonótico em leite Bovino

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.

Isolation and molecular characterization of Arcobacter butzleri and Arcobacter cryaerophilus from the pork production chain in Brazil / Isolamento e caracterização molecular de Arcobacter butzleri e Arcobacter cryaerophilus de linhas de abate suíno e do comércio de carne no Brasil

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)

Biotyping of Arcobacter butzleri isolated from poultry products: rescuing a phenotypic method / Biotipagem de Arcobacter butzleri desde produtos aviares: resgatando um método fenotípico

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.

Arcobacter butzleri and A. cryaerophilus in human, animals and food sources, in southern Chile

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.

Arcobacter: un patógeno emergente de origen alimentario / Arcobacter: a foodborne emerging pathogen

En las últimas tres décadas, se han identificado gran número de enfermedades emergentes que afectan al ser humano, la mayoría de las cuales son de origen infeccioso e incluyen enfermedades bacterianas, virales, parasitarias, entre otras. Muchas de éstas son de origen zoonótico, tal es el caso de algunas especies de Arcobacter, actualmente consideradas bacterias emergentes y, también, asociadas a transmisión alimentaria y de creciente importancia en salud pública. El incremento en los datos de prevalencia e incidencia de casos asociados a esta sugiere que la infección en humanos y animales ha sido subestimada, debido a la carencia de conocimientos al respecto y de un protocolo estándar, universalmente aceptado, para el aislamiento primario de este organismo y al uso de correctos métodos y técnicas de identificación. El incremento en el hallazgo de Arcobacter en alimentos derivados de animales y en muestras tomadas durante el proceso de producción de alimentos, hace que aumente la preocupación en materia de salud pública, ya que aún se conoce muy poco del potencial patogénico de las especies Arcobacter y los pocos estudios que se han llevado a cabo, muestran una gran cantidad de especies hospederas y rutas de transmisión. Dado lo anterior, el objetivo de la presente revisión es el actualizar al lector en las características más destacadas de esta bacteria en cuanto a su morfología, distribución, clasificación, transmisión, asociación con aguas, alimentos, mascotas y animales de crianza, como también sobre su aislamiento en laboratorio, factores de virulencia y patrones de sensibilidad a antibióticos.

In the last three decades, several emergent diseases affecting human beings have been identified, most of them from infectious origin including bacterial, viral, parasitic and even difficult to classify as spongiform encephalopathy. Most of these are zoonotic as it is the case of Arcobacter, currently considered as an emerging and food borne pathogen, of growing importance for public health. The increase in the prevalence and incidence of cases associated to this bacteria as well as in the number of actual researches and reports, suggest that the infection in human beings and animals has been underestimated due to a lack in knowledge about this bacteria and of a standardized isolation protocols, as well as the use of correct identification methods and techniques. Increasing trends in the isolation of Arcobacter from animal derivates used as food and from samples taken during production processes, cause an augment in public health awareness, since there is little knowledge about the pathogenic potential of Arcobacter species and the few focused in this bacterial group, show many different transmission routes and host species. Given this, the objective of the present review is to actualize the reader in the most important characteristics of this bacterium, including its morphology, distribution, classification, transmission, association with water, food, pets and animals, as well as the laboratory isolation techniques, virulence factors and their antibiotic susceptibility patterns.

Arcobacter butzleri: first isolation report from chicken carcasses in costa rica

Arcobacter butzleri isolation from chicken carcasses in Costa Rica is reported for the first time. The isolated strains (P and R) were presumptively identified by their phenotypic characteristics. Definitive identification was made using a multiplex PCR assay for the simultaneous detection and identification of Arcobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii. These first isolations indicate the necessity of further investigation about the prevalence, distribution, ecology and interactions with human beings of this and other Arcobacter species.

Susceptibility of Arcobacter butzleri to human blood serum / Susceptibilidade de Arcobacter butzleri ao soro humano

A susceptibilidade de 10 amostras de Arcobacter butzleri ao soro humano foi estudada. A maior atividade bactericida foi encontrada no soro humano normal, com taxas de sobrevivência bacterianainversamente proporcionais à diluição do soro. As maiores taxas de sobrevivência foram obtidas com o soro inativado pelo calor. As taxas de sobrevivência decresceram com a adição de soro fresco ao inativado. O soro com valores reduzidos de gamaglobulinas e valores normais de complemento mostrou ativo efeito bactericida. Os resultados demonstraram que A. butzleri é altamente susceptível ao efeito bactericida do soro humano, sugerindo que pode ser capaz de ativar diretamente o complemento pela via alternativa.

Persistence of Arcobacter butzleri CCUG 30484 on plastic, stainless steel and glass surfaces / Persistência de Arcobacter butzleri CCUG 30404 em superfícies de plástico, aço inox e vidro

The persistence of A. butzleri CCUG 30484 on various surfaces under 32 percent and 64 percent relative humidity suspended in physiological saline or nutrient broth to simulate relatively clean or soiled conditions was studied using various isolation techniques. Our study revealed that A. butzleri CCUG 30484 cells were able to survive for a considerable period of time, even after the droplet of suspending medium has been visibly dried. An extended survival on polypropylene coupons at both humidity levels was observed, particularly at soiled conditions.

Estudou-se a persistência de Arcobacter butzleri CCUG 30404 em várias superfícies de contato com alimentos a 32 por cento e 64 por cento de umidade relativa, suspenso em salina fisiológica e caldo nutriente para simular condições limpas e sujas. Nosso estudo indicou que A. butzleri CCUG 30404 foi capaz de sobreviver por longo tempo, mesmo após a secagem da gota. Observou-se que a sobrevivência for mais prolongada nos cupons de polipropileno, especialmente em condições sujas.

A Case of Bacteremia Caused by Arcobacter butzleri / 대한진단검사의학회지

Arcobacter butzleri, a gram-negative, slightly curved bacterium previously described as an aerotolerant Campylobacter, was isolated from the blood of a 53-year-old male patient with alcoholic cirrhosis. The isolate was identified by various phenotypic tests and 16S rRNA sequencing analysis. The patient was treated with amikacin and recovered uneventfully. To our knowledge, this is the first case of A. butzleri bacteremia reported in Korea.

Isolamento de Aerobacter (Campylobacter) cryaerophila de fetos suínos abortados / Isolation of Aerobacter (Campylobacter) cryaerophila from aborted pig fetuses

Foram examinados 6 fetos suínos abortados no terço final da gestação, provenientes de duas granjas do Rio Grande do Sul, Brasil. Os órgãos internos (rim e fígado) de cada feto foram inoculados em meio semi-sólido de Tween 80/40 LH, visando o isolamento de leptospiras. Os tubos foram incubados a 30°C. Foi isolado de todos os materiais um microorganismo, classificado como Arcobacter cryaerophila. Estes foram os primeiros isolamentos desta bactéria no Brasil.

Samples of liver and kidney from aborted pig fetuses from two different farms were examined. Samples were cultured in semi-solid leptospira isolation media and incubated at 30°C. Arcobacter cryaerophila was isolated from all the specimens. These were the first reports on the isolation of the microorganism from pigs in Brazil.