Occurrence of Aliarcobacter spp. in fresh and pre-cut vegetables of common use in San José, Costa Rica.

Aliarcobacter is a Gram-negative rod that can cause disease in both animals and humans. Several studies have evidenced its presence in a wide variety of foods. Given that the number of foodborne illness outbreaks linked to the consumption of vegetables has increased worldwide and that there is a lack of information about the occurrence of Aliarcobacter spp. in these, the aim of this study was to evaluate its presence and the occurrence of virulence factors in both fresh and ready-to-eat vegetable samples. 180 vegetable samples from Costa Rica were analyzed for the presence of Aliarcobacter spp., including 90 pre-cut vegetable packages and 90 fresh vegetables. Two (2.2%) of the isolates from pre-cut vegetables and 19 (21.1%) of the isolates obtained from fresh vegetables were confirmed as Aliarcobacter spp. One of the isolates from the pre-cut vegetable samples was identified as A. butzleri. For the fresh vegetables, 11 isolates were identified as A. skirrowii, one as A. butzleri, and the 7 remaining isolates could not be identified at a species level. There is an 87.5% positivity for hecA and 93.8% for pldA, virulence genes in strains isolated from fresh produce, contrasting with an absolute absence from pre-cut vegetable-isolated strains. These results evidence the presence of Aliarcobacter on fresh and pre-cut vegetables from Costa Rica and the potential hazard it might represent for public health.

The ability of Aliarcobacter butzleri strains isolated from foods of animal origin in Costa Rica to form biofilm.

Aliarcobacter butzleri is a zoonotic emerging food and waterborne pathogen widely distributed in nature. It is present in food processing environments and can easily be spread through the food industry because of its ability to form biofilm. The aim of this work was to determine the ability of strains isolated in Costa Rica from different food matrixes of animal origin to form biofilm. Thirty-eight A. butzleri strains previously isolated and identified from animal origin products were analyzed using the method described by Stepmovic et al. (2000), in three culture broths, brain heart infusion broth, Boer broth and Houf broth. Results showed that 67% of poultry origin strains, 62.5% of meat origin strains and just 8% of milk origin strains showed ability to form biofilm. The findings of this study confirm the adherence ability of A. butzleri to form biofilm, a characteristic that can promote dispersion and cross contamination along food industry processing lines.

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.

First isolation report of Arcobacter cryaerophilus from a human diarrhea sample in Costa Rica.

Arcobacter cryaerophilus is an emerging enteropathogen and potential zoonotic agent transmitted by food and water. In Costa Rica, this bacterium has not been associated with cases of human gastroenteritis, even though it has been isolated from farm animals, especially poultry. This paper reports the first isolation of A. cryaerophilus from a human case of bloody watery diarrhea and the virulence genes associated with this isolate.

Genotipificación de Rotavirus en niños menores de cinco años en la región sur del Ecuador / Genotyping of Rotavirus in children under five years of age in the Southern region of Ecuador / Genotipagem de Rotavírus em crianças menores de cinco anos na região sul do Equador

Fueron estudiadas, para la búsqueda de rotavirus, 341 muestras fecales obtenidas de sendos niños menores de cinco años con diarrea aguda, atendidos en dos centros de salud de la ciudad de Loja - Ecuador. De ellas, 56 (16%) fueron positivas para rotavirus, siendo genotipificadas 33 muestras para la determinación de genotipos G y P. Los genotipos más frecuentes fueron G4 (42%), P[6] (36%) y la combinación mixta G4+9P[6] (21%). Este es el primer estudio de carácter molecular en rotavirus realizado en el sur de Ecuador y los datos obtenidos ratifican la variabilidad de cepas de rotavirus circulantes en este país.

A total of 341 fecal samples obtained from children under five years of age with acute diarrhea disease, attending two nursing homes in Loja city, Ecuador were studied for rotavirus diagnosis. Out of them, 56 (16%) were rotavirus positive and 33 samples were genotyped for the determination of genotypes G and P. The most frequent genotypes were G4 (42%), P [6] (36%) and +9 mixed combination G4 P [6] (21%). This is the first molecular study on rotavirus carried out in Southern Ecuador and the obtained data confirms the variability of the rotavirus circulating strains in Ecuador.

Foram estudadas, para a pesquisa de rotavírus, 341 amostras fecais obtidas de igual número de crianças menores de cinco anos, com diarreia aguda, atendidos em dois centros de saúde da cidade de Loja - Equador. Delas, 56 (16%) foram positivas para rotavírus, sendo genotipadas 33 amostras para a determinação de genótipos G e P. Os genótipos mais frequentes foram G4 (42%), P[6] (36%) e a combinação mista G4+9P[6] (21%). Este é o primeiro estudo de caráter molecular em rotavírus realizado no sul do Equador. Os dados obtidos ratificam a variabilidade de cepas de rotavírus circulantes neste país.

SURVIVAL CAPACITY OF Arcobacter butzleri INOCULATED IN POULTRY MEAT AT TWO DIFFERENT REFRIGERATION TEMPERATURES.

Arcobacter spp. are emerging enteropathogens and potential zoonotic agents that can be transmitted by food and water, being considered a public health risk. The high isolation rate of these bacteria from poultry products suggests that it may be a major source of human infections. One hallmark for differentiating the genus Arcobacter from Campylobacter includes their growing capacity at low temperatures (15-30 °C) under aerobic conditions. However, little is known about the population density variation of these bacteria at different refrigeration temperatures. The aim of this study was to determine the survival behavior of two different Arcobacter butzleri concentrations (10(4) CFU/mL and 10(7) CFU/mL) inoculated on chicken legs and held at two different refrigeration temperatures (4 and 10 °C) throughout storage time. Results have shown that A. butzleri had growing capacity both at 4 and 10 °C. No statistical difference between the survival trends was found for both bacterial concentrations and temperatures tested. This study shows that A. butzleri is a robust species with regard to storage temperature, and represents a potential health risk for poultry meat consumers.

Arcobacter: comparison of isolation methods, diversity, and potential pathogenic factors in commercially retailed chicken breast meat from Costa Rica.

Arcobacter species have been recognized as potential food- and waterborne pathogens. The lack of standardized isolation methods and the relatively scarce knowledge about their prevalence and distribution as emerging pathogens are due to the limitations in their detection and identification. This study aimed to determine the presence and the identification of Arcobacter in chicken breast samples commercially retailed in San José, Costa Rica, as well as to describe the adherence and invasive potential of the strains to human cells (HEp-2). Fifty chicken breast samples were collected from retail markets in the metropolitan area of the country. Six different isolation methodologies were applied for the isolation of Arcobacter. Isolation strategies consisted of combinations of enrichments in de Boer or Houf selective broths and subsequent isolation in blood agar (directly or with a previous passive membrane filtration step) or Arcobacter selective agar. Suspicious colonies were identified with a genus-specific PCR, whereas species-level identification was achieved with a multiplex PCR. The overall isolation frequency of Arcobacter was 56%. From the isolation strategies, the combination of enrichment in Houf selective broth followed by filtration on blood agar showed the best performance, with a sensitivity of 89% and a specificity of 84%. A total of 46 isolates were confirmed as Arcobacter with the genus-specific PCR, from which 27 (59%) corresponded to Arcobacter butzleri, 9 (19%) to Arcobacter cryaerophilus, and 10 (22%) were not identified with this multiplex PCR. Regarding the potential pathogenicity, 75% of the isolates presented adherence to HEp-2 cells, while only 22% were invasive to that cell line. All invasive strains were A. butzleri or nonidentified strains. The results show the presence of potentially pathogenic Arcobacter in poultry and recognize the importance it should receive as a potential foodborne pathogen from public health authorities.