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.

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.

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.

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.

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.

Bacteriemias e infección endovascular por Campylobacter spp: nuestra experiencia en un cuarto de siglo de historia / Bacteremia and endarteritis cases secondary to Campylobacter spp. in a metropolitan hospital: Our experience along a quarter of a century

Six cases of bacteremia and one of endarteritis were identified between 1986 and 2010 in a general hospital in Chile. Five of these cases occurred during the second half of this timeframe, Campylobacterfetus predominated (5 out of 7) and the majority of the infections presented during warmer months. The mean age was 32.4 years (range 19 to 63) all had comorbidities, and main clinical manifestations included fever with diarrhea. Four patients developed hypotension and two septic shock. The latter, associated to C. fetus bacteremia, died before microbiological diagnosis. Six out of 7 patients received antimicrobial therapy. During 2004 and 2010, the rates of Campylobacter spp. positive stool cultures in the same hospital increased 4 times, suggesting an emerging profile. Bacteremia and endarteritis by Campylobacter spp. can develop in vulnerable patients and manifest as fever with or without diarrhea. Finding curved or spiral shaped gram negative rods in blood cultures leads to suspect this pathogen. Species identification is of utmost importance due to antimicrobial resistance especially in C. jejuni. Prognosis is unfavorable due to host characteristics, and case-fatality rate is high.

Se presentan seis casos de bacteriemia y uno de infección vascular por Campylobacter spp, observados en 25 años, con el fin de describir sus características clínicas. Cinco de ellos se registraron en la segunda mitad del período, en concomitancia con el incremento de este agente en el porcentaje de coprocultivos, lo que sugiere un perfil emergente. Las infecciones fueron más frecuentes en los meses cálidos, asociadas principalmente a C. fetus (5 de 7) y a co-morbilidad. La edad promedio de los pacientes fue de 32,4 años (rango 19 a 63 años) y todos tenían comorbilidades. Las manifestaciones clínicas más frecuentes fueron diarrea y fiebre (5 de 7 casos) y dos pacientes cursaron con shock séptico (28,6 por ciento). La evolución fue favorable en cinco pacientes pero los dos que presentaron shock asociado a C. fetus fallecieron (28,6 por ciento). Las bacteriemias o infecciones vasculares por Campylobacter spp., aunque infrecuentes, pueden presentarse en pacientes vulnerables y debutar como cuadros febriles, en presencia o ausencia de diarrea. La identificación de la especie involucrada es de suma importancia debido a la escasa actividad terapéutica de cefalosporinas de tercera generación y quinolonas. El pronóstico de estas bacteriemias es grave debido a las características del hospedero y a su elevada letalidad.

Campylobacter y campylobacteriosis: una mirada desde América del Sur / Campylobacter and campylobacteriosis: a view from South America

Las especies termotolerantes de Campylobacter han adquirido gran importancia en la salud pública, por ser considerados agentes de diarrea infecciosa para el ser humano. En esta breve revisión se presenta información sobre aspectos epidemiológicos, clínicos y bacteriológicos de campylobacteriosis en América del Sur. Asimismo, se señalan algunas diferencias con relación a su presentación en países industrializados.

The thermotolerant species of Campylobacter have become very important in public health, especially as agents of infectious diarrhea in human beings. In this brief revision we present part of the available information generated in South America about epidemiological, clinical and bacteriological aspects of campylobacteriosis and we identify some differences between the observed and documented campylobacteriosis in South America compared to those described in industrialized countries.

Susceptibilidad antimicrobiana de Campylobacter jejuni aislado de coprocultivos en Santiago de Chile / Antimicrobial susceptibility of Campylobacter jejuni isolates from stool cultures in Santiago, Chile

Campylobacter jejuni is a common agent of enterocolitis in humans. Campy lobacteriosis has been recognized as a zoonotic disease whose reservoir is the intestinal flora of poultry. The reposition of fluid and electrolytes is the recommended treatment, and antimicrobials are required only in severe and/or in prolonged disease. Given the emergence of resistance to drugs commonly used in the treatment of acute diarrhea, we studied the antimicrobial susceptibility of 73 strains oí Campylobacter jejuni isolated from stool culture. The antimicrobials tested were: erythromycin, azithromycin, ampicillin and ciprofloxacin. Of the 73 strains tested by E-test, 32.4 percent were resistant to ciprofloxacin and 6.4 percent were resistant to ampicillin. Resistance to erythromycin and azithromycin was not detected. The surveillance of antimicrobial resistance of Campylobacter jejuni is important in the evaluation of empirically used antimicrobials in the treatment of bacterial enterocolitis.

Occurrence of Campylobacter jejuni in Shiraz, Southwest Iran

To investigate the occurrence of Campylobacter jejuni among other bacterial agents of acute diarrhea. Over a period of 3 months, 114 specimens of acute diarrhea were collected from patients aged 2-58 years referred to Nemazi Hospital, Shiraz, Iran. The samples were tested for bacterial pathogens causing acute diarrhea. Of the 114 samples, 40 proved to be positive for bacterial enteric pathogens: C. jejuni 11 [9.6%], Salmonella spp.18 [15.8%] and Shigella spp. 11 [9.6%]. Maximum prevalence of C. jejuni was observed in patients aged 11-15 years. Significantly more females than males were infected with all diarrheal agents [p <0.05]. This study confirmed the presence of C. jejuni as a significant cause of gastroenteritis in Shiraz, Southwest Iran

Aislamiento, caracterización bioquímica y sensibilidad de cepas de Campylobacter sp. en diarreas agudas en el Paraguay / Isolation, biochemical characterization and sensitivity of stocks of Campylobacter sp. in acute diarrhoeas in Paraguay

Estudio de aislamiento y caracterización de Campylobacter como agente etiológico de diarreas agudas de 331 muestras de heces diarreicas obtenidas de pacientes ambulatorios e internados que fueron analizadas en el Laboratorio Central de Salud Pública en el periodo diciembre 2006 al mes de agosto de 2007

Bacteriemia por Campylobacter fetus subsp fetus en un paciente inmunosuprimido / Bacteraemia due to Campylobacter fetus in an immune suppressed patient

We report a case of bacteraemia by Campylobacter fetus subsp. fetus in a 77 year-old woman with immunosuppression secondary to steroid use. Diagnosis was suspected by finding Gram negative curved rods in blood cultures taken after 4 days of a febrile illness without local findings. Diarrhea was not present. There was no consumption of undercooked meat or non-pasteurized milk and no contact with pets. The patient was treated with sulbactam-cefoperazone due to the coexistence of urinary tract infection by multiresistant E. coli. The outcome was favorable and albeit susceptibility was not assessed, quinolone resistance was presumed because illness appeared during ciprofloxacin prophylaxis for urinary tract infection. In contrast to C. jejuni infections, C. fetus infections are associated to debilitated or immunosuppressed patients, bacteraemia is predominant, diarrhea is rarely observed and disease is not self-limited.

Comunicamos un caso de bacteriemia por Campylobacter fetus subsp. fetus en una paciente inmunosuprimida de 77 años, usuaria de corticoesteroides. La sospecha fue planteada por el hallazgo de bacilos gramnegativos curvos en los hemocultivos tomados al cuarto día de evolución de un cuadro febril sin diarrea. La paciente fue tratada con sulbactam/ cefoperazona debido a la coexistencia de una infección urinaria por Escherichia coli multiresistente. No hubo antecedentes de consumo de alimentos crudos o parcialmente cocidos en las semanas recientes y tampoco había contacto con mascotas. La evolución fue favorable. Aunque no se estudió la susceptibilidad del aislado, se presumió resistencia a quinolonas ya que el cuadro apareció durante el uso profiláctico de ciprofloxacina para infección urinaria. En contraste con C. jejuni, las infecciones por C. fetus se asocian a pacientes debilitados o inmunosuprimidos, se manifiestan predominantemente como bacteriemias sin diarrea y no tiene un perfil autolimitado.

A 16S rDNA-based nested PCR protocol to detect Campylobacter gracilis in oral infections

The aim of this study was to describe a 16S rDNA-based nested polymerase chain reaction (nPCR) assay to investigate the occurrence of Campylobacter gracilis in oral infections. Samples were collected from ten infected root canals, ten cases of acute periradicular abscesses and eight cases of adult marginal periodontitis. DNA extracted from the samples was initially amplified using universal 16S rDNA primers. A second round of amplification used the first PCR products to detect C. gracilis using oligonucleotide primers designed from species-specific 16S rDNA signature sequences. The nPCR assay used in this study showed a detection limit of 10 C. gracilis cells and no cross-reactivity was observed with nontarget bacteria. C. gracilis was detected in the three types of oral infections investigated - 4/10 infected root canals; 2/10 acute periradicular abscesses; and 1/8 subgingival specimens from adult periodontitis. The method proposed in this study showed both high sensitivity and high specificity to directly detect C. gracilis in samples from root canal infections, abscesses, and subgingival plaque. Our findings confirmed that C. gracilis may be a member of the microbiota associated with distinct oral infections, and its specific role in such diseases requires further clarification

Isolation of nalidixic acid resistant Campylobacters from cases of paediatric diarrhoea in Chennai.

A short term investigation on the Campylobacter enteritis among children under 10 years of age was carried out in Chennai. The study revealed an isolation rate of 11 per cent in 100 patients suffering from acute diarrhoea comprising C. jejuni (8%) and C. coli. (3%). Among the two culture methods used, the candle jar method was found to be superior to plastic bag incubation system in recovering campylobacters on charcoal cefeperazone deoxycholate agar. While all the isolates were sensitive to ciprofloxacin, all of them exhibited resistance to nalidixic acid.

Participacion de epitopes flagelares de Campylobacter jejuni en la adhesion celular / Partipation of Campylobacter jejuni flagellar epitopes in cellular adhesion

Utilizando una cepa flagelada (052) y una aflagelada (T-1), se estudió la participación de epítopes flagelares de C. jejuni en la adhesión a células HEp-2 in vitro. La cepa 052 presentó una capacidad de adhesion significativamente mayor que la cepa T-1. Cuando los ensayos de adhesión fueron realizados en presencia de anticuerpos monoclonales dirigidos contra epítopes flagelares, la capacidad de adhesión de la cepa 052 experimentó una inhibición que fluctuó entre 64,3 y 92,9 por ciento. Mediante una prueba de ELISA se comprobó que estos anticuerpos monoclonales eran específicos y estaban dirigidos exclusivamente contra epítopes flagelares de la cepa 052, no reaccionando con la cepa T-1. Estos resultados demuestran que epítopes flagelares de C. jejuni estarían participando en el proceso de adhesión, sugiriendo la intervención del flagelo en la instalación del proceso infeccioso.

Frequência de enteropatógenos intestinais isolados no período de julho de 1991 a julho de 1993, no laboratório Bio-Ciência/Lavoisier / Frequency of intestinal enteropatogens isolated in Bio-Ciência/Lavoisier Clinical Laboratory from july 1991 to july 1993

Este trabalho tem como finalidade rever a etiologia da diarréia, e no qual pesquisamos todos os agentes da infecçäo intestinal, relacionados a seguir: E. coli enteropatogênica clássica (EPEC), E.coli enteroinvasora (EIEC), E.coli enterotoxigênica (ETEC), E.coli enterohemorrágica (EHEC), Shigella, Salmonella, Yersinia enterocolítica, Campylobacter, Aeromonas e Vibriocholerae

Mannose-resistant haemagglutination by Campylobacter jejuni--a preliminary communication.

Ten strains of C. jejuni each isolated respectively from patients with diarrhoea and from chicken intestine (10 strains from each source) were examined for presumptive colonization factor(s) by measuring their cell surface hydrophobicity and haemagglutination. None of the strains expressed cell surface hydrophobicity. However, 14 strains (7 from either source) showed variable haemagglutination pattern with human, sheep and rabbit erythrocytes in the presence of 0.5 per cent D-mannose. Thus, mannose resistant haemagglutinin(s) may be involved in the colonization of intestinal mucosal surfaces by C. jejuni.

Isolation of Campylobacters from the canals of Bangkok metropolitan area.

The 100 ml of canal water samples of 36 canals in Bangkok Metropolitan Area were examined in three periods starting from July-September 1988, November 1988-January 1989 and February-April 1989. Each time the 52 water samples were checked. Of 156 water samples, 116 strains of Campylobacter species were isolated. They were 63.79 per cent (74 strains) of C. cryaerophila and 36.21 per cent (42 strains) of C. cryaerophila-like organisms. The differentiation was determined by urease activity test. C. cryaerophila was isolated from 44.23 per cent (23 strains), 51.19 per cent (27 strains) and 46.15 per cent (24 strains) and also C. cryaerophila-like organism from 28.85 per cent (15 strains), 19.23 per cent (10 strains) and 32.69 per cent (17 strains) of the 52 samples during each period respectively. Since C. cryaerophila and C. cryaerophila-like are aerotolerant Campylobacter, they grow well in aerobic conditions at 25 degrees-36 degrees C. On the contrary, thermophilic Campylobacter such as C. jejuni, C. coli and C. laridis require atmosphere containing 5 per cent O2, 10 per cent CO2, 85 per cent N2 and temperature at 36 degrees-42 degrees C, so the environment in the canals is unfavorable for their growth. The etiological role of C. cryaerophila in pathogenesis in humans is still unknown, and requires furthers study. This study shows that canals can be an important source of these two Campylobacter species that might be potential pathogens in the future.