Detecção de Campylobacter jejuni em produtos de frango utilizando separação imunomagnética / Detection of Campylobacter jejuni in chicken products using immunomagnetic separation

Campylobacter jejuni é o principal causador de gastroenterite bacteriana aguda, e a carne de frango tem se mostrado uma importante fonte de transmissão. Este microrganismo é de difícil isolamento e os métodos convencionais muitas vezes não são eficientes, podendo levar a resultados errôneos. O objetivo deste trabalho foi desenvolver e testar a técnica de separação imunomagnética (IMS) na detecção de C. jejuni em produtos de frango. Micropartículas magnéticas ligadas a anticorpos policlonais anti-C. jejuni foram utilizadas para concentrar C. jejuni antes da semeadura em ágar. O protocolo foi comparado com o método convencional. C. jejuni foi recuperado do alimento experimentalmente contaminado por ambos os métodos, entretanto, quando foi usada a IMS, a presença de microrganismos contaminantes nos meios de cultura foi menor. C. jejuni foi isolado de 7% das amostras de alimento naturalmente contaminadas, usando IMS, e de 3% pelo método convencional. C. coli foi isolado de uma amostra pelo método convencional, mas não foi detectado pelo protocolo com IMS. A técnica de IMS pode ser usada para isolamento de C. jejuni de alimentos, oferecendo a vantagem de detectar em amostras o microrganismo cujo isolamento não é obtido por meio do método convencional.(AU)

Campylobacter jejuni is the main cause of acute bacterial gastroenteritis and chicken meat has shown to be an important source of infection. This microorganism is difficult to isolate and the conventional methods are often inefficient and may lead to erroneous results. This study aimed at developing and testing the technique of immunomagnetic separation (IMS) in the detection of C. jejuni in chicken products. Microparticles magnetically connected anti-C. jejuni polyclonal antibodies were used to concentrate C. jejuni before agar seeding. The protocol was compared with the conventional method. C. jejuni was recovered from experimentally contaminated food for both methods, however, when the IMS was used, the presence of contaminating microorganisms in the means of culture was smaller. C. jejuni was isolated from 7% of samples of food naturally contaminated, using IMS, and 3% by conventional method. C. coli was isolated from a sample by conventional method, but it was not detected by protocol with IMS. The IMS technique can be used for isolation of C. jejuni in food, offering the advantage of detecting the microorganism in samples from which the isolation is not obtained with the use of the conventional method.(AU)

Bacteriemia por Campylobacter jejuni en un paciente con infección por VIH en etapa SIDA / Campylobacter jejuni bacteremia in a patient with HIV infection in AIDS stage

Resumen La bacteriemia es una presentación atípica de la infección por Campylobacter jejuni, y es más frecuente en pacientes con inmunodepresión avanzada debido al VIH u otras enfermedades sistémicas. Debido a la terapia anti-retroviral, en las últimas décadas el número de casos ha disminuido. Presentamos el caso de una mujer en situación de calle, con VIH en etapa C3, que cursó con una bacteriemia por C. jejuni durante su hospitalización por una tuberculosis pulmonar. Realizamos una breve revisión de bacteriemia por C. jejuni en pacientes con VIH.

Bacteremia is an atypical presentation of Campylobacter jejuni infection and it is more frequent in patients with advanced inmunodepression due to HIV or other sistemic diseases. Because of the highly active antiretroviral therapy, in the last decades the number of cases had declined. We report a case of a homeless woman with HIV in C3 stage who was diagnosed with the bacteremia during her hospitalization for pulmonary tuberculosis, and a brief review of C. jejuni bacteremia in HIV patients.

Campylobacter jejuni and Campylobacter coli originated from chicken carcasses modulate their transcriptome to translate virulence genes in human cells / Campylobacter jejuni e Campylobacter coli originadas de carcaças de frango modulam seu transcriptoma para traduzir genes de virulência em células humanas

The aim was to determine the spread of genetically similar profiles of Campylobacter in chicken carcasses and evaluate their ability to produce transcripts for ciaB, dnaJ, p19 and sodB genes, before and after cultivation in Caco-2 cells. The strains used were isolated from 420 samples of chicken carcasses chilled and frozen ready for marketing. The species were identified by PCR-multiplex, the phylogeny was determined by RAPD-PCR and the presence of transcripts was performed by RT-PCR. We identified 74 (17.6%) of Campylobacter strains, being 55 (74.3%) C. jejuni and 19 (25.7%) C. coli. The phylogenetic relationship demonstrated heterogeneity between isolates of the same species, with absence of clones, indicating the high level of diversity of circulating genotypes. The gene transcription showed conflicting results before and after the culture in Caco-2 cell, so that before cultivation isolates showed greater capacity to transcribe genes related to survival and after the interaction with human cells, the strains showed higher potential to transcribe genes associated with virulence. The result of this study contributes to the understanding of how these seemingly fragile microorganisms are the most prevalent bacterial agents in human gastroenteritis.(AU)

O objetivo foi determinar a disseminação de perfis geneticamente semelhantes de Campylobacter em carcaças de frango e avaliar sua capacidade de produzir transcritos para os genes ciaB, dnaJ, p19 e sodB, antes e após o cultivo em células Caco-2. As cepas utilizadas foram isoladas de 420 amostras de carcaças de frango resfriadas e congeladas prontas para comercialização. As espécies foram identificadas por PCR-multiplex, a filogenia foi determinada por RAPD-PCR e a presença de transcritos foi realizada por RT-PCR. Identificamos 74 (17,6%) das cepas de Campylobacter, sendo 55 (74,3%) C. jejuni e 19 (25,7%) C. coli. A relação filogenética demonstrou heterogeneidade entre isolados da mesma espécie, com ausência de clones, indicando o alto nível de diversidade dos genótipos circulantes. A transcrição gênica mostrou resultados conflitantes antes e após a cultura em células Caco-2, de modo que, antes do cultivo, os isolados apresentaram maior capacidade de transcrever genes relacionados à sobrevivência e após a interação com células humanas, as linhagens apresentaram maior potencial para transcrever genes associados à virulência. O resultado deste estudo contribui para a compreensão de como esses microrganismos aparentemente frágeis são os agentes bacterianos mais prevalentes na gastroenterite humana.(AU)

Prevalencia y caracterización del perfil de susceptibilidad antimicrobiana de Campylobacter jejuni y Campylobacter coli en niños con diarrea de la ciudad de Loja, Ecuador / Prevalence and antimicrobial behavior of Campylobacter jejuni and Campylobacter coli in children with diarrhea in Loja city, Ecuador

Resumen Introducción: Campylobacter es un importante agente de diarrea en el ser humano. En Ecuador, la información sobre Campylobacter es escasa y no existen antecedentes de susceptibilidad antimicrobiana. Objetivo: Describir la prevalencia de Campylobacter en niños con diarrea y su comportamiento in vitro frente a cinco antimicrobianos. Método: Se estudiaron 253 niños entre siete meses y 9 años de edad, que consultaron por diarrea en dos hospitales de la ciudad de Loja. Se realizó cultivo de muestras fecales e identificación por pruebas fenotípicas y por RPC múltiple. La susceptibilidad antimicrobiana fue determinada por el método de difusión en disco. Resultados: Campylobacter fue diagnosticado en 16 (6,3%) de las muestras, aislándose C. jejuni en 13 (5,1%) y C. coli en 3 (1,2%). Todas las cepas fueron susceptibles a gentamicina y ampicilina/ ácido clavulánico, con baja resistencia a ampicilina y eritromicina y alta resistencia a ciprofloxacina.

Introduction: Campylobacter is an important agent of diarrhea in humans. In Ecuador, the information on Campylobacter is scarce and there are not antecedents about antimicrobial susceptibility. Objective: To describe Campylobacter prevalence in children with diarrhea and their behavior against five antimicrobials in vitro. Method: We studied 253 children with diarrhea aging 7 months to 9 years who consulted for diarrhea in two hospitals in the city of Loja. Fecal samples were cultured and identification by tests by phenotypic tests and multiplex PCR. Susceptibility to 5 antibiotics was determined by the disc-diffusion method. Results: Campylobacter was found in 16 (6.3%) children, being C. jejuni the most frequent one (5.1%), followed by C. coli (1.2%). All strains were susceptible to gentamicin and ampicillin/clavulanic acid, being found low resistance to ampicillin and erythromycin and high resistance to ciprofloxacin.

Caracterización de cepas de Campylobacter jejuni obtenidas desde carne de pollo y heces de aves de corral de la zona central de Chile / Characterization of Campylobacter jejuni samples coming form poultry meat and feces

Background Campylobacter jejuni is one of the main causal agents of food borne diseases. Infections with this pathogen are mainly caused by chicken meat consumption. Aim To characterize antibiotic resistance and virulence factors in C. jejuni strains obtained from chicken meat and poultry feces in Central Chile. Material and Methods The presence of C. jejuni in 30 meat and 40 feces samples from poultry was studied. From these samples, we obtained 40 strains which were characterized at the molecular level for the presence of 16 genes involved in virulence using PCR. In parallel, antibiotic resistance for ciprofloxacin, nalidixic acid, tetracycline, erythromycin, azithromycin, chloramphenicol y ampicillin was analyzed. Results Twenty and 63% of feces and chicken meat samples were positive for C. jejuni, respectively. Moreover, a high percentage of strains showed antibiotic resistance, where 27% of strains were resistant to all tested antibiotics, except for azithromycin. Finally, 10% of the strains coming from feces contained 14 out of 16 virulence genes evaluated. Only 23% of the strains did not contain any of these genes. Conclusions A high percentage of feces and chicken meat samples are contaminated with C. jejuni. Moreover, these strains show a high genetic and phenotypic diversity represented by their antibiotic resistance profiles and the presence of virulence factors.

Presencia de serotipos de Campylobacter jejuni O:19 en la cadena avícola Argentina / Campylobacter jejuni O:19 serotype in Argentine poultry meat supply chain

Las especies termotolerantes de Campylobacter han tomado gran relevancia en los últimos años debido a que son los principales agentes zoonóticos causantes de enfermedades transmitidas por alimentos. Adicionalmente, Campylobacter jejuni serotipo O:19 ha sido relacionado con el desarrollo del síndrome posdiarreico de Guillain-Barré. El objetivo de este trabajo fue determinar la proporción de cepas de C. jejuni que se corresponden con el serotipo O:19 dentro de las aisladas en diferentes etapas de 3 cadenas de producción de carne aviar en la provincia de Santa Fe. Se observó que el 18% del total de cepas de C. jejuni recuperadas (10/55) pertenecían al serotipo O:19; estas se aislaron en 4 de las 5 etapas de producción de carne aviar elegidas a los fines de este estudio. Este hallazgo da cuenta de un riesgo importante para la salud pública de los consumidores y debería ser considerado en los estudios epidemiológicos de Campylobacter, para implementar en forma urgente medidas de control sobre este microorganismo.

Thermotolerant species of Campylobacter have been focus of attention in the last years because they are the major agent causing zoonotic foodborne diseases. In addition, Campylobacter jejuni O:19 serotype was associated with Guillain Barré syndrome. The aim of this study was to determine the proportion of C. jejuni O:19 serotype isolated at different stages of three poultry meat supply chain in Santa Fe, Argentina. The analysis showed that 18% of isolated C. jejuni belong to serotype O:19. It was also determined that the presence of these strains is given in almost all production stages. These results reflect a significant risk to public health of consumers. Epidemiological studies of Campylobacter should be considered to establish a risk manager policy.

Aeromonas hydrophila and Campylobacter jejuni isolated in fresh tuna (Thunnus spp) sold in São Paulo, Brazil / Detecção de Aeromonas hydrophila e Campylobacter jejuni em atum (Thunnus spp) fresco comercializado em São Paulo, Brasil

Aeromonas hydrophila and Campylobacter jejuni are bacteria of emerging importance in public health. However, little has been published about fish contaminated by these pathogens. The present study aimed to verify the presence of Aeromonas hydrophila and Campylobacter jejuni in fresh tuna samples (Thunnus spp.) caught off the coast of Santa Catarina State and distributed in the wholesale market of São Paulo/SP. A total of 85 tuna fillet samples were collected and examined by PCR and bacteriological analyses. Aeromonas spp. was detected in 11/85 (13%) samples, with 10/11 (90.9 %) being confirmed as Aeromonas hydrophila by PCR. Campylobacter spp. was found in 10/85 (11.7%) samples, 10/10 (100%) identified as Campylobacter jejuni by PCR and conventional biochemical analyses. Both pathogens were found in 2/85 (2.3%) samples. This is the first report on the contamination of fresh tuna by Campylobacter jejuni and Aeromonas hydrophila in Brazil. In addition to show that tuna can be a vehicle for transmission of pathogens when consumed raw, it emphasizes the importance of further studies to support the control these pathogens in fish...

Aeromonas hydrophila e Campylobacter jejuni são bactérias de importância emergente em saúde pública, porém com escassos trabalhos publicados na área de pescado. O presente estudo investigou a presença de Aeromonas hydrophila e Campylobacter jejuni em amostras de atum (Thunnus spp.) fresco, capturados no litoral de Santa Catarina e distribuídos no comércio atacadista de São Paulo, SP. Foram colhidas 85 amostras de filé de atum e processadas por análises bacteriológicas e PCR. Do total, 11/85 (13%) amostras foram positivas para Aeromonas spp., sendo 10/11 (90,9%) confirmadas como Aeromonas hydrophila pela PCR. Para Campylobacter spp., 10/85 (11,7%) foram positivas, sendo 10/10 (100%) identificadas como Campylobacter jejuni pelas provas bioquímicas tradicionais e PCR ressaltando-se que duas (2/85 - 2,3%) amostras de atum albergavam ambos os patógenos. Trata-se do primeiro relato no Brasil de contaminação de atum fresco por Campylobacter jejuni e Aeromonas hydrophila, indicando que este alimento ingerido in natura pode ser um veículo de transmissão de agentes patogênicos, ressaltando-se a importância de estudos adicionais que deem suporte ao controle desses microrganismos em pescado consumido cru...

ISOLATION AND MOLECULAR IDENTIFICATION OF POTENTIALLY PATHOGENIC Escherichia coli AND Campylobacter jejuni IN FERAL PIGEONS FROM AN URBAN AREA IN THE CITY OF LIMA, PERU / Isolamento e detecção de Escherichia coli e Campylobacter jejuni potencialmente patogênicos em pombos selvagens de uma área urbana na cidade de Lima, Perú

SUMMARY Feral pigeons (Columbia livia) live in close contact with humans and other animals. They can transmit potentially pathogenic and zoonotic agents. The objective of this study was to isolate and detect strains of diarrheagenic Escherichia coli and Campylobacter jejuniof urban feral pigeons from an area of Lima, Peru. Fresh dropping samples from urban parks were collected for microbiological isolation of E. coli strains in selective agar, and Campylobacterby filtration method. Molecular identification of diarrheagenic pathotypes of E.coliand Campylobacter jejuni was performed by PCR. Twenty-two parks were sampled and 16 colonies of Campylobacter spp. were isolated. The 100% of isolates were identified as Campylobacter jejuni. Furthermore, 102 colonies of E. coli were isolated and the 5.88% resulted as Enteropathogenic (EPEC) type and 0.98% as Shiga toxin-producing E. coli (STEC). The urban feral pigeons of Lima in Peru can act as a reservoir or carriers of zoonotic potentially pathogenic enteric agents.

RESUMO Os pombos selvagens (Columbia livia) vivem em estreito contato com os seres humanos e outros animais. Podem transmitir agentes potencialmente patogênicos e zoonóticos. Os objetivos deste estudo foram isolar e detectar cepas de Escherichia coli diarreiogênica e Campylobacter jejuni de pombos selvagens urbanos de uma área de Lima, Peru. Amostras de fezes frescas foram coletadas em parques urbanos para o isolamento microbiológico para cepas de E. coli em ágar seletivo e Campylobacterpor método de filtração. Identificação molecular de patótipos diarreiogênicos de E. coli e Campylobacter jejuni foi realizado por PCR. Vinte e dois parques foram amostrados e 16 colônias de Campylobacter spp. foram isolados. O 100% dos isolados foram identificados como Campylobacter jejuni. Além disso, 102 colônias de E. coli foram isoladas e 5,88% resultaram como tipo enteropatogênico (EPEC) e 0,98% como produtora de toxina Shiga (STEC). Os pombos selvagens urbanos de Lima no Peru podem atuar como reservatório ou ser portador de agentes zoonóticos entéricos potencialmente patogênicos.

Detecção dos genes codificantes da toxina CDT, e pesquisa de fatores que influenciam na produção de hemolisinas em amostras de Campylobacter jejuni de origem avícola / Detection of the genes encoding the toxin CDT, and research factors which influence the production of hemolysin in Campylobacter jejuni from poultry products

Membros termofílicos do gênero Campylobacter são reconhecidos como importantes enteropatógenos para o ser humano e animais. A grande diversidade ecológica destes micro-organismos em diferentes habitats tais como água, animais e alimentos predispõem ao aparecimento de novos fatores de virulência. Este trabalho teve por objetivo detectar os genes codificantes da Toxina Distensiva Citoletal (CDT) por meio da técnica de PCR, pesquisar a atividade de hemolisinas e a influência de soluções quelantes e de íons nesta atividade. Foram utilizadas 45 amostras de Campylobacter jejuni de origem avícola para pesquisa de atividade hemolítica, cultivadas em Caldo Triptona de Soja (TSB). Após o crescimento bacteriano, as amostras foram semeadas em Ágar tríptico de soja (TSA) contendo 5% de sangue de ovino. Para verificar a influência de agentes quelantes e solução de íons na atividade hemolítica, as amostras de C. jejuni foram cultivadas em TSB contendo separadamente os quelantes EDTA, ácido acético, soluções de íons CaCl2, MgCl2 e FeCl3, em atmosfera de microaerofilia. Quanto à atividade de hemolisina de C. jejuni em placas de TSA - sangue ovino foi possível observar que houve hemólise em 40% das amostras analisadas apenas com caldo TSB. Somente o ácido acético apresentou ação quelante sobre a atividade de hemolisinas em amostras de C. jejuni semeadas em placas de TSA - sangue ovino. Para detecção dos genes cdtA, cdtB e cdtC através da técnica da Reação em Cadeia da Polimerase (PCR) foram utilizadas 119 amostras de C. jejuni de origem avícola. Foi possível observar que 37,8% possuíam o perfil de genes cdtABC. Os resultados demonstraram em amostras avícolas a presença de cepas de C. jejuni com potencial virulento, devido à presença dos genes da toxina CDT e potencial hemolítico, que apresentou ação reduzida in vitro com ácido acético...

Thermophilic members of the Campylobacter genus are recognized as important enteropathogenics for humans and animals. The great variety of ecological habitats, such as water, food and milk, may promote new virulence factors. To detect the encoding genes distending cytolethal toxin (CDT) by PCR and study the hemolytic activity with influence of chelation solutions and ions, 45 Campylobacter jejuni samples from poultry production origin were used to perform the hemolytic research. To check the influence of chelation agents and solution of ions in the hemolytic activity, samples of C. jejuni strains were grown in tryptone soy broth TSB containing chelation agents separately EDTA, acetic acid, CaCl2, MgCl2 and FeCl3 ions solutions in microaerophilic atmosphere and then streaked on 5% sheep blood tryptic soy agar (TSA). To perform the detection of cdtA, cdtB and cdtC genes the technique of Polymerase Chain Reaction (PCR) was used in 119 samples of C. jejuni from poultry production origin. We found 40% of samples showing hemolysis after growing with TSB. Only the acetic acid showed reduction in hemolysis. The prevalent gene profile was cdtABC in 37.8 % of the samples. It was observed that the results showed the presence of C. jejuni strains with virulent potential, due to presence of the CDT toxin genes and the hemolytic activity, which showed in vitro reduced when acetic acid was added...

Occurrence of Campylobacter jejuni and C. coli on broiler carcasses after chilling in southern Brazil / Campylobacter jejuni e C. coli em carcaças de frangos após a refrigeração por imersão no sul do Brasil

Campylobacter jejuni and C. coli have been associated with gastrointestinal disorders in human beings, due mainly to the consumption of chicken meat. Despite control measures for reducing contamination by these bacteria, the detection of Campylobacter in carcasses after chilling remains high. A total of 105 carcasses were assessed by the horizontal detection method in five federally inspected slaughterhouses in southern Brazil in 2012 and in the first three months of 2013. Campylobacter was isolated in 37.1% of the carcasses, of which 97.5% contained C. jejuni and 2.5% were infected by C. coli. The rate of positive carcasses across the slaughterhouses ranged from 0 to 71.4%. Determining the occurrence of Campylobacter among flocks is crucial for estimating the microbial load at specific points along the slaughtering process and for minimizing the risk of contamination of end products by Campylobacter.(AU)

Campylobacter jejuni e C. coli têm sido associados a problemas gastroentéricos em seres humanos principalmente devido ao consumo de carne de frango. Embora medidas de controle sejam realizadas para reduzir a contaminação por estas bactérias, a identificação de Campylobacter em carcaças após a refrigeração por imersão é alto. Foram analisadas 105 carcaças pelo método de detecção horizontal em cinco abatedouros sob Inspeção Federal no sul do Brasil em 2012 e nos três primeiros meses de 2013. Campylobacter foi isolada em 37,1% das carcaças analisadas, as quais 97,5% foram identificados como C. jejuni e 2,5% como C. coli. A ocorrência de carcaças positivas entre matadouros variou de zero a 71,4%. O conhecimento sobre a ocorrência de Campylobacter entre os lotes é fundamental para estimar a extensão da carga microbiana em pontos específicos do abate e consequentemente minimizar o risco de contaminação por Campylobacter em produtos finais de frangos.(AU)

Characterization of the virulence, growth temperature and antibiotic resistance of the Campylobacter jejuni IAL 2383 strain isolated from humans

The objective of this study was to characterize the C. jejuni IAL2383 strain isolated from humans in Brazil. Transcripts for the racR, dnaJ and ciaB genes were found and flaA, plda and cadF genes were present in the genome and bacteria was sensitive to most of the important antimicrobials used to treat humans. C. jejuni IAL2383 is a good experimental model to analyze the interactions with cells.

Campylobacter jejuni in commercial eggs

This study evaluated the ability of Campylobacter jejuni to penetrate through the pores of the shells of commercial eggs and colonize the interior of these eggs, which may become a risk factor for human infection. Furthermore, this study assessed the survival and viability of the bacteria in commercial eggs. The eggs were placed in contact with wood shavings infected with C. jejuni to check the passage of the bacteria. In parallel, the bacteria were inoculated directly into the air chamber to assess the viability in the egg yolk. To determine whether the albumen and egg fertility interferes with the entry and survival of bacteria, we used varying concentrations of albumen and SPF and commercial eggs. C. jejuni was recovered in SPF eggs (fertile) after three hours in contact with contaminated wood shavings but not in infertile commercial eggs. The colonies isolated in the SPF eggs were identified by multiplex PCR and the similarity between strains verified by RAPD-PCR. The bacteria grew in different concentrations of albumen in commercial and SPF eggs. We did not find C. jejuni in commercial eggs inoculated directly into the air chamber, but the bacteria were viable during all periods tested in the wood shavings. This study shows that consumption of commercial eggs infected with C. jejuni does not represent a potential risk to human health.

Prevalence of virulence genes in strains of Campylobacter jejuni isolated from human, bovine and broiler

Campylobacter jejuni isolates of different origins (bovine, broiler meat, human) were screened by polymerase chain reaction for the presence of 4 genes cdtB, cst-II, ggt, and virB11, previously linked to virulence such as adherence, invasion, colonization, molecular mimicry, and cytotoxin production. In addition, the isolates were screened for the presence of the global gene regulator csrA linked to oxidative stress responses, biofilms formation, and cell adhesion. All the C. jejuni isolates were positive for cdtB gene. The csrA gene was detected in 100% and 92% of C. jejuni isolates from human and animal origin and the virB11 gene was detected in 7.3% and 3.6% isolates from chicken and human respectively. All isolates from bovine were negative for the virB11 gene. The isolates showed a wide variation for the presence of the remaining genes. Of the C. jejuni recovered from human 83.6%, and 32.7% were positive for cst-II, and ggt respectively. Out of the isolates from chicken 40% and 5.5% isolates revealed the presence of cst-II, and ggt, respectively. Finally of the C. jejuni isolates from bovine, 97.7% and 22.7% were positive for cst-II, and ggt respectively. We conclude that the genes of this study circulate among humans and animals. These results led us to hypothesize that the isolates associated with enteritis (cdtB positives) are not selected by environmental or host-specific factors. On the other hand, the high frequencies of csrA gene in C. jejuni show that this gene is important for the survival of C. jejuni in animals and humans.

Detection of CDT toxin genes in Campylobacter spp. strains isolated from broiler carcasses and vegetables in São Paulo, Brazil

Campylobacteriosis is a worldwide distributed zoonosis. One of the main virulence factors related to Campylobacter spp. in animals and humans is the cytolethal distending toxin (CDT), encoded by three adjacent genes (cdtA, cdtB, cdtC). The occurrence of Campylobacter spp. in samples of vegetables has not been reported in Brazil yet, and has seldom been described in the international literature. The detection of CDT in these strains has not been reported, either. The objectives of the present study were to determine the occurrence of Campylobacter spp. strains carrying virulence factors in samples of poultry and vegetables (lettuce and spinach) from different points of sale, thus verifying if vegetables are as an important vehicle for potentially virulent Campylobacter spp. strains as poultry. Twenty four strains were identified as Campylobacter jejuni by phenotypic and genotypic methods: 22 from broiler carcasses and two from lettuce samples. Three strains were identified as Campylobacter coli: two from broiler carcasses and one from lettuce. The presence of the cdt genes were detected in 20/24 (83.3%) C. jejuni strains, and 3/3 (100%) C. coli strains. The isolation of Campylobacter spp. strains with the cdt gene cluster in lettuce samples points to a new possible source of contamination, which could have an impact in the vegetable production chain and risk to public health. Results show that potentially virulent C. jejuni and C. coli strains remain viable in samples of broiler carcasses and vegetables at the points of sale.

Análisis molecular de la resistencia a fluoroquinolonas y macrólidos en aislados de Campylobacter jejuni de humanos, bovinos y carne de ave / Molecular analysis of fluoroquinolones and macrolides resistance in Campylobacter jejuni isolates from humans, bovine and chicken meat

Background: Campylobacter sp.- one of the leading causes of bacterial food-borne gastrointestinal illness worldwide- is increasingly resistant to fluoroquinolone and macrolide antimicrobials, which has become a major concern for public health. Objective: To describe the susceptibility patterns of Campylobacter jejuni strains to erythromycin and ciprofloxacin and to explore the origin of its resistance in human isolates. Material and Method: In this study, fifty-five ciprofloxacin and erythromycin susceptibility patterns of C. jejuni strains isolated from humans with diarrheal disease, performed by broth microdilution MIC, were compared with 55 and 44 isolates from chicken meat and bovines respectively, obtained from the Metropolitan Region, Chile. Results: Of the 55 human isolates of C. jejuni, 33 (60%) were resistant to ciprofloxacin and all were sensitive to erythromycin. Of the 55 isolates from chicken meat, 32 (58.2%) were resistant to ciprofloxacin and 1.8% were resistant to erythromycin. Of the 44 isolates of C. jejuni from cattle, 8 (18.2%) were resistant to ciprofloxacin and all were sensitive to erythromycin. Four PFGE patterns matched with certain resistance profiles and grouped isolates from human and animal. Conclusion: The findings showed continued effectiveness of erythromycin for campylobacteriosis and a high percentage of C. jejuni strains ciprofloxacin-resistant. This is interesting because it is considered that the presence of ciprofloxacin resistant strains in broiler meat can be in part the source of resistance to this antimicrobial in humans.

Introducción: El incremento de la resistencia en Campylobacter sp. (una de las principales causas de gastroenteritis bacteriana de origen alimentario) a fluoro-quinolonas y macrólidos, es un problema en salud pública. Objetivo: Conocer los patrones de susceptibilidad in vitro de Campylobacter jejuni a eritromicina y ciprofloxacina y conocer el origen de su resistencia en aislados de humanos. Material y Método: En este estudio, se compararon las susceptibilidades a ciprofloxacina y eritromicina -CIM efectuadas por microdilución en caldo- de 55 aislados de C. jejuni provenientes de humanos con enterocolitis, con 55 aislados de carne de pollo y 44 de bovinos obtenidos en la Región Metropolitana, Chile. Resultados: De 55 aislados de C. jejuni de humanos, 33(60%) presentaron resistencia a ciprofloxacina y todos presentaron susceptibilidad a eritromicina. De 55 aislados procedentes de carne de pollo, 32 (58,2%) presentaron resistencia a ciprofloxacina y un aislado resultó resistente a eritromicina (1,8%). De 44 aislados de bovinos, 8(18,2%) presentaron resistencia a ciprofloxacina y todos resultaron sensibles a eritromicina. Cuatro patrones de electroforesis a campo pulsado coincidieron en sus perfiles de resistencia y agruparon aislados de origen humano y animal. Conclusiones: Los resultados muestran que eritromicina continúa siendo efectiva para el tratamiento de la campilobacteriosis y que existe un alto porcentaje de cepas resistentes a ciprofloxacina. Se considera probable que la presencia de cepas resistentes a ciprofloxacina en la carne de pollo puede ser en parte el origen de la resistencia a este fármaco en humanos.

A case of myopericarditis associated to Campylobacter jejuni infection in the southern hemisphere

Myopericarditis is an infrequent complication of acute diarrheal illness due to Campylobacter jejuni, and it has been mainly reported in developed nations. The first case detected in Chile - an upper-middle income country -, that is coincidental with the increasing importance of acute gastroenteritis associated to this pathogen, is described. Recognition of this agent in stools requires special laboratory techniques not widely available, and it was suspected when a young patient presented with acute diarrhea, fever, and chest pain combined with electrocardiogram (EKG) abnormalities and elevated myocardial enzymes. C. jejuni myopericarditis can easily be suspected but its detection requires dedicated laboratory techniques.

Detection and species identification of Campylobacter in stool samples of children and animals from Vellore, south India.

Campylobacter spp. are an important cause of bacterial gastroenteritis frequently isolated from animal, poultry and environmental samples. In this study, we investigated the zoonotic potential of Campylobacter spp. by comparing prevalence rates and species in 394 children with diarrhoea and 652 animals in Vellore using PCR-based tools. Eighteen children (4.5%) had campylobacteriosis, a majority of whom had co-pathogens (15/18) and most were infected with Campylobacter jejuni (16/18). A few C. coli and mixed infections with both species were also seen. Among the animal samples, 16/25 chicken samples (64%) were positive and all were found to be C. jejuni.

Bactriological studies on some diseases causing broilers mortality with the possibility of vaccine production

Two hundred and seventy samples were randomly collected from Sharkia Government and Peasantry Farms in various localities. These samples were subjected from bacteriological investigation allover the year to explore the actual bacterial causative agents. The obtained data revealed that, 65 isolates were E. coll, 20 isolates were Salmonella spp, 15 isolates were Staph aureus and 15 isolates were Campylobacter jejuni with incidence percentages of 24%, 7.4%, 5.5%, and 5.5% respectively. From the results of this study it can be concluded that the E. coli infection is incorporated in complicated respiratory infection and almost need for use of sensitive antibiotic for control measures considered and an important direction to avoid loss of body weight, prolonged respiratory signs and economic losses. Vaccination against E. coli infection induced protection immune response especially vaccinated at 6 days of age. Sensitivity tests related that E. coli were sensitive for lincospectin, doxycydine, gentamicin, colistin sulphat, chloramphicol and amoxicillin while it resistance to erthromycic. Concerning salmonella were sensitive to chloramphenicol, lincospectin, doxycyclin and less sensitive to erythromycin and colistin sulphate. In case of Staph. aureus were sensitive to amoxicillin, gentamycin and lincospectin and less sensitive to erythromycin and resistance to colistin sulphate. In case of Campylobacter jejuni were sensitive to lincospectin, doxycycline, amoxicillin and gentamycin, while no effect could be observed with erythromycin

Genotipificación y resistencia antibacteriana de cepas de Campylobacter spp aisladas en niños y en aves de corral / Genotyping and antibacterial resistance of Campylobacter spp strains isolated in children and in free range poultry

Poultry is a main reservoir and source of human infection in campylobacteriosis. Three hundred and forty one stool samples (291 human, 50 avian) were analyzed. In the human group, 220 samples were collected from children with acute diarrheal disease (183 inpatients, 37 outpatients) and 71 from healthy children. Erythromycin and ciprofloxacin agar dilution MIC tests, Penner serotyping and RAPD-PCR genotyping were performed on 23 strains isolated. C. jejuni was reported only in patients with acute diarrhea (5.4 percent inpatients, 2.2 percent outpatients). Campylobacter prevalence in poultry was 34 percent. Cross-resistance to nalidixic acid and ciprofloxacin was found in 33.3 percent of human samples and 11.8 percent of animal samples. Human samples could not be typed using the Penner method. F serotype was the most expressed in poultry. We obtained a total of 14 genotypes (4 / 5 human and 10/15 avian). In conclusion, the predominant species in poultry and humans was C. jejuni, a significant amount of quinolone-resistant human and avian samples were obtained, and avian genotypes and serotypes were not found in human samples. The latter would mean that another source of infection could exist; therefore other reservoirs must be studied.

Las aves de consumo constituyen uno de los principales reservorios y fuente de infección humana de la campilo-bacteriosis. Se analizaron 341 muestras de deposiciones, 291 humanas y 50 aviares. De las muestras, 220 de niños con síndrome diarreico agudo-SDA (183 hospitalizados y 37 consultantes ambulatorios) y 71 niños sanos. A las 23 cepas obtenidas se les realizó CIM por dilución en agar a eritromicina y ciprofloxacina, serotipificación de Penner y genotipiicación por RAPD-PCR. Se encontró Campylobacterjejuni sólo en pacientes con SDA, de ellos 5,4 por ciento ambulatorios y 2,2 por ciento hospitalizados. En aves, la prevalencia de Campylobacter spp., fue de 34 por ciento. Hubo resistencia cruzada a ácido nalidixico y ciprofloxacina en 33,3 por ciento cepas de origen humano y 11,8 por ciento animal. Las cepas humanas fueron no tipiicables por el método de Penner. Predominó entre las aves el serotipo F. Se obtuvo un total de 14 genotipos (4/5 humanos y 10/15 aviares). En conclusión, la especie predominante en aves de corral y en humanos fue C. jejuni, existiendo una alta prevalencia de cepas de origen humano y aviar resistentes a quinolonas. Los genotipos y serotipos aviares no fueron encontrados en cepas de origen humano, lo que indica que podría existir otra fuente de infección, por lo que se requiere estudiar otros reservorios.

Cepas de Campylobacter jejuni resistentes a quinolonas aisladas de humanos, gallinas y pollos / Quinolone resistant Campylobacter jejuni strains isolated from humans and from poultry

Se compararon 8 aislamientos de Campylobacter jejuni provenientes de humanos con enfermedad diarreica aguda, con 23 aislamientos de cloaca de gallinas y pollos obtenidos de zonas próximas a la ciudad de Rosario, todos resistentes a la ciprofloxacina. Las muestras se sembraron en agar selectivo y se incubaron en microaerofilia a 42 °C. Las colonias se identificaron con el método tradicional. Los aislamientos se conservaron a -70 °C en caldo cerebro corazón con 17% v/v de glicerina. La clonalidad se determinó por RAPD-PCR, utilizando el primer 1254 (Stern NJ). Se interpretaron los aislamientos como clones distintos cuando diferían en una banda de amplificación. Se obtuvieron 5 clones diferentes. Los patrones I, II y V fueron aislados en criaderos industriales de pollos y en humanos (el II también en un establecimiento de gallinas ponedoras de huevos). En un gallinero familiar se obtuvo el patrón I. El patrón III sólo se obtuvo de humanos. El patrón IV se halló en uno de los criaderos pero no en humanos. Se pudo determinar que 93.5% de las cepas se aislaron tanto de animales como de humanos, por lo que se considera posible que la colonización de criaderos con cepas resistentes a los antimicrobianos pudiera ser el origen de la infección de humanos.

Eight quinolone resistant Campylobacter jejuni strains isolated from humans with diarrheal disease were compared with 23 isolates from chicken and from laying hens. Samples were cultured on selective agar in microaerophilia, identified by conventional tests, and conserved in 17% glycerol at -70 °C. Clones were determined by RAPD-PCR employing the 1254 primer (Stern NJ). Five patterns were obtained. Patterns I, II, and V were found in both poultry and human isolates. Pattern I was obtained from poultry in a domestic henhouse. Pattern III was only obtained from humans whereas pattern IV was only obtained from poultry. A 95.3% of clones were found in both, humans and poultry. According to these results colonization by quinolone resistant strains could be the origin of this human infection, acquired by ingestion.