Campylobacter in broiler slaughter samples assessed by direct count on mCCDA and Campy-Cefex agar

ABSTRACT Campylobacter spp. cause foodborne illnesses in humans primarily through the consumption of contaminated chicken. The aim of this study was to evaluate the United States Department of Agriculture's (USDA) recommended methodology, protocol MLG 41.02, for the isolation, identification and direct plate counting of Campylobacter jejuni and C. coli samples from the broiler slaughtering process. A plating method using both mCCDA and Campy-Cefex agars is recommended to recover Campylobacter cells. It is also possible to use this method in different matrices (cloacal swabs and water samples). Cloacal swabs, samples from pre-chiller and post-chiller carcasses and samples of pre-chiller, chiller and direct supply water were collected each week for four weeks from the same flock at a slaughterhouse located in an abattoir in southern Brazil. Samples were analyzed to directly count Campylobacter spp., and the results showed a high frequency of Campylobacter spp. on Campy-Cefex agar. For the isolated species, 72% were identified as Campylobacter jejuni and 38% as Campylobacter coli. It was possible to count Campylobacter jejuni and Campylobacter coli from different samples, including the water supply samples, using the two-agar method. These results suggest that slaughterhouses can use direct counting methods with both agars and different matrices as a monitoring tool to assess the presence of Campylobacter bacteria in their products.

Detecção de resistência às fluoroquinolonas em Campylobacter isolados de frangos de criação orgânica / Detection of fluoroquinolone resistance in Campylobacter strains from organic poultry

Estudos têm revelado que a resistência às quinolonas em cepas de Campylobacter está relacionada à presença da mutação Treonina-86 para Isoleucina. Com o objetivo de investigar a presença dessa mutação em cepas de Campylobacter sensíveis e resistentes à ciprofloxacina e enrofloxacina, o conteúdo cecal de 80 frangos de corte de criação orgânica, abatidos sob Serviço de Inspeção Estadual (S.I.E.) do Estado do Rio de Janeiro, foram coletados e investigados para a presença de Campylobacter. A determinação da resistência à ciprofloxacina e enrofloxacina foi feita pela técnica de difusão em disco e de diluição em ágar para determinação da Concentração Inibitória Mínima (CIM). A detecção da mutação na Região Determinante de Resistencia às Quinolonas (RDRQ) no gene gyrA foi realizada através de sequenciamento. Campylobacter foi isolado a partir de 100% das amostras avaliadas, sendo 68,75% correspondente à C. jejuni e 31,25% à C. coli. No teste de difusão em disco, 100% das cepas foram resistentes à ciprofloxacina e 56,25% das cepas foram resistentes à enrofloxacina. No teste de diluição em ágar, todas as cepas foram resistentes à ciprofloxacina apresentando CIM variando de ≥ 16-64μg/mL, e resistência ou resistência intermediaria à enrofloxacina foi detectada em 42,50% (CIM ≥ 4-32μg/mL) e 38,75% (CIM = 2μg/mL) das cepas, respectivamente. A mutação Tre-86-Ile, foi observada em 100% das cepas analisadas. Além dessa mutação, foram observadas outras mutações não silenciosas (Val-73-Glu, Ser-114-Leu, Val-88-Asp, Ala-75-Asp, Ser-119-Gli, Arg-79-Lis) e mutações silenciosas (His-81-His, Ser-119-Ser, Ala-120-Ala, Fen-99-Fen, Ala-122-Ala, Gli-74-Gli, Ile-77-Ile, Ala-91-Ala, Leu-92-Leu, Val-93-Val, Ile-106-Ile, Tre-107-Tre, Gli-113-Gli, Ile-115-Ile, Gli-110-Gli). A observação de que cepas sensíveis à enrofloxacina pelos testes fenotípicos apresentavam a substituição Tre-86 para Ile sugere que outros mecanismos podem contribuir para a resistência à enrofloxacina em Campylobacter...

Studies have shown that resistance to quinolones in Campylobacter strains is related with Threonine-86-Isoleucine mutation. In order to investigate the presence of this mutation in sensitive and resistant Campylobacter strains to ciprofloxacin and enrofloxacin, the cecal contents of 80 broilers from organic raising chickens, slaughtered under State Inspection Service (S.I.S) of the State of Rio de Janeiro, were collected and tested for the presence of Campylobacter. The determination of ciprofloxacin and enrofloxacin susceptibility was done by disk diffusion and agar dilution methods for determining the Minimum Inhibitory Concentration (MIC). The detection of mutation in Quinolone Resistance Determinant Region (QRDR) in gyrA gene was done by sequencing. Campylobacter was isolated from 100% of the samples, being 68.75% C. jejuni and 31.25% C. coli. By the disk diffusion method, resistance to ciprofloxacin was observed in all isolates and 56.25% of the strains were resistant to enrofloxacin. By agar dilution method, all strains were resistant to ciprofloxacin (MIC ≥ 16μg/mL to ≥ 64μg/mL) and full and intermediate resistance to enrofloxacin was detected in 42.50% (MIC ≥ 4-32μg/mL) and 38.75% (MIC =2μg/mL) of the strains, respectively. Mutation Thr-86-Ile was observed in 100% of the isolates investigated. In addition to this mutation, others no silent mutations (Val-73-Glu, Ser-114-Leu, Val-88-Asp, Ala-75-Asp, Gly-119-Ser, Arg-79-Lys) and silent mutations (His-81-His, Ser-119-Ser, Ala-120-Ala, Phe-99-Phe, Ala-122-Ala, Gly-74-Gly, Ile-77-Ile, Ala-91-Ala, Leu-92-Leu, Val-93-Val, Ile-106-Ile, Thr-107-Thr, Gly-113-Gly, Ile-115-Ile, Gly-110-Gly) were detected. All the enrofloxacin-sensitive strains by the phenotypic methods had the Thr-86 to Ile substitution, which suggests other mechanisms contributing to enrofloxacin resistance in Campylobacter...

Circulation of Campylobacter spp. in rhesus monkeys (Macaca mulatta) held in captivity: a longitudinal study

Campylobacteriosis is an extremely important zoonosis, circulating freely in the environment. In nonhuman primates kept in open facilities and bred for experimental purposes, the presence of Campylobacter spp. could cause severe damage to the production and interfere with the results of scientific research. In this paper, we assessed the circulation of Campylobacter spp. in a colony of clinically healthy rhesus monkeys (Macaca mulatta) destined to research. The analysis was carried out during seven non-consecutive years. Data showed that despite several changes made in animal management along the studied years in order to control this zoonosis, reduction of bacterial charge did not occur. Significant differences among the age groups and sex were observed. Infants showed higher susceptibility than adult animals. In general males were more infected than females. Modifications adopted in the handling techniques need to be reviewed with the intent of improving the production, reducing bacterial infection of the stock and avoiding undesirable cross reactions in the research carried out with these animals. Therefore, this paper alerts professionals that work directly with captive rhesus monkeys about the risks of Campylobacter spp. infection and possible interference on the experimental procedures.

Application of biochemical and polymerase chain reaction assays for identification of Campylobacter isolates from non-human primates

A multiplex polymerase chain reaction (PCR) assay was performed on 167 thermophilic campylobacters isolated from non-human primates. Samples were first identified by phenotypic methods resulting in 64 Campylobacter jejuni and 103 C. coli strains. Four strains identified biochemically as C. coli, were then determined to be C. jejuni by PCR. Comparison of methodologies showed that the main discrepancies were attributed to the hippurate hydrolysis test and sensitivity to cephalothin and nalidixic acid. Analysis of data showed that the application of phenotypic methods should be supplemented by a molecular method to offer a more reliable Campylobacter identification.

Molecular characterization of Campylobacter isolated from chickens and humans in northern Thailand.

A study was conducted in northern Thailand to characterize Campylobacter spp isolated from chickens at farms, slaughterhouse, and chicken meat at fresh markets, and stools from chicken farmers as well as neighboring crop farmers who served as controls. The Campylobacter isolates were collected during the rainy months of the years 2000, 2001, and 2002. Standard methods were used for primary isolation and identification of Camplobacter and the resulting isolates were frozen and stored in 30% glycerol with Mueller-Hinton broth at -70 degrees C until used in 2003. A multiplex PCR assay was used for differentiation of the Campylobacter spp. A total of 415 Campylobacter spp were isolated from 849 (48.9%) samples from chickens at the farm, slaughter house and fresh chicken meat market. Campylobacter spp were isolated from 5 of the 129 (3.9%) chicken farmers but none from the 100 neighboring crop farmers. C. jejuni was the most prevalent (42.5%) at the farm, followed by C. coli (39.1%) and other species (8.0%). In contrast, C. coli was the most prevalent at the slaughter house (72.4%) while C. jejuni was only 17.2% and others 3.4%. Similarly, at the fresh chicken market, C. coli was the most prevalent (54.4%) while C. jejuni was 26.5% and others were 13.2%. Campylobacter spp isolated from the chicken farmers were predominantly (75%) C. coli and the rest (25%) were C. jejuni. The results of the study show that both C. coli and C. jejuni are highly prevalent in chickens, along the chicken production system and in chicken farmers in northern Thailand. Critical control points for exposure and contamination of the chicken meat supply should be identified so that methods can be developed to protect human exposure to Campylobacter spp.

Guillain-Barre syndrome and Campylobacter infection.

Campylobacter infection of the gastrointestinal tract has been observed as an antecedent illness in some patients with Guillain-Barre syndrome (GBS); these patients have been reported to have poor prognosis. We investigated 29 patients with GBS, admitted to our hospital from January 1996 to December 1999 for recent Campylobacter enteritis by culture of their stool specimens. Campylobacter upsaliensis and C. jejuni were isolated from stools of one patient each with acute motor axonal neuropathy (AMAN) and acute inflammatory demyelinating polyradiculoneuropathy (AIDP) respectively. The patient with C. upsaliensis infection was a 7 year-old male child who developed features of AMAN, 7 days after onset of diarrhea. He recovered gradually within 24 days with residual deficit in the form of foot drop. This deficit has persisted for last three and half years. The other patient with C. jejuni infection was a 9 year-old boy, who developed AIDP after 9 days of acute diarrhea. This patient recovered completely within 28 days of illness without any deficit. None of the patients had relapse of GBS. The present findings indicate the need of planned systematic studies to explore the role of C. upsaliensis and other campylobacters as agents of antecedent diarrhea in patients of GBS with different clinical presentations and prognosis.

Campylobacter species associated with diarrhoea in patients from a tertiary care centre of north India.

BACKGROUND & OBJECTIVES: Most laboratories do not routinely distinguish the various Campylobacter species, though almost all Campylobacter species have been isolated from human faeces. The epidemiological and clinical aspects of its infection and the species involved in genesis of diarrhoea are least understood in the developing countries. The aim of the present study was to find out frequency of Campylobacter species isolated from patients with diarrhoea over a 12-year period and to analyse their features. METHODS: Campylobacter strains isolated from stool samples of patients with diarrhoea were identified to the species level on appropriate media at 42 degrees C micro-aerobically. Patients' demography and clinical data were analyzed retrospectively; 25 Campylobacter jejuni strains were tested for toxin production and 23 strains were typed by Penner scheme. RESULTS: A total of 62 strains were isolated from 59 patients and the various species were C. jejuni 51 (82.3%), C. coli 8 (12.9%), C. lari 2 (3.2%), and C. upsaliensis 1 (1.6%). Children < 5 yr of age were most affected (34/59; 57.6%), followed by patients in 15-30 yr of age (12/59; 20.3%). Presentation of watery diarrhoea was significantly more common than inflammatory diarrhoea (50/59, 84.7% vs 9/59, 15.3%; P < 0.001). Recurrence occurred in 3 (5.1%) patients. Majority of the infections resolved within one week; one HIV-positive patient had chronic diarrhoea. Two patients developed Guillain-Barré syndrome following Campylobacter infection. Twenty (80%) of 25 strains were toxigenic and 20 (87%) of 23 strains could be typed by Penner scheme. INTERPRETATION & CONCLUSION: In our patients, 4 different Campylobacter species and various C. jejuni serotypes were involved in gastroenteritis. Majority of the infections were watery diarrhoea and in children < 5 yr of age. There is a need of a population-based systematic study to know the epidemiology of whole spectrum of campylobacters in India.

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.

Carriage of the classical thermotolerant campylobacters in health domestic animals from Earstern Peru

Com o objetivo de conhecer a importancia dos animais domesticos como reservatorios naturais dos classicos campylobacters termotolerantes, amostras de fezes foram obtidas de mamiferos e aves do leste do Peru e imediatamente colocadas num meio de enriquecimento. Tecnicas convencionais foram utilizadas para identificar C. jejuni ssp. jejuni, C. coli e C. lari. Campylobacter foi isolado em 26,5 por cento dos animais estudados, sendo C. jejuni ssp. jejuni biovar I o mais frequente (8,9 por cento). O frango foi o reservatorio mais importante destes microorganismos (54,0 por cento).

Prevalence of thermotolerant species of Campylobacter and their biotypes in children and domestic birds and dogs in southern Chile

Foi determinada, no Sul do Chile, a prevalencia de especies termotolerantes de Campylobacter e seus biotipos, em criancas normais e diarreicas, bem como em caes e aves domesticas. Campylobacter fa isolado em 34,5 por cento do total das amostras estudadas sendo sua frequencia de isolamento de 16,3 por cento e 6,4 por cento nas criancas com diarreia e normais, respectivamente.C.jejuni foi a especie mais frequente. Porem, C. coli foi isolado em 29 por cento das criancas com diarreia. Somente os biotipos I e II, tanto de C. jejuni como de C. coli, foram isolados das criancas, os quais tambem foram os mais frequentemente encontrados nos animais, sugerindo uma possivel associacao epidemiologica entre eles.

Biotipificación y serotipificación de campylobacter jejuni, acmpylobacter coli y campylobacter lari / Biotyping and sertyping of campylobacter jejuni, campylobacter coli and campylobacter lari

Se serotipificaron 26 cepas de Campylobacter termofílicos por el método de hemaglutinación pasiva y se biotipificaron 62 cepas por el método de Lior. Dos cepas correspondieron al serogrupo 13/50 y otras dos expresaron antígenos del serogrupo 4; el 78 por ciento restante se distribuyó entre diferentes serogrupos; 21,7 por ciento de las cepas fueron no tipificables. Entre los biotipos, 47 (75,8 por ciento) correspondieron a C. jejuni, 44 (71 por ciento) al biotipo I, 3 (4,8 por ciento) al biotipo II; 14 (22,6 por ciento) a C. coli, 11 (17,8 por ciento) al biotipo I, 3 (4,8 por ciento) al biotipo II y 1 (1,6 por ciento) a C. lari. Las cepas de C. jejuni y C. coli resistentes al ácido nalidíxico pueden complicar la identificación

Simplified method for the detection of DNA hydrolysis by enteric campylobacters.

A simplified medium was developed for the detection of DNase produced by enteric campylobacters. Sensitivity and reproducibility of the test were similar to that of the improved toluidine blue DNA agar method. Logistically, the simplified DNA hydrolysis test was cheaper (5.5 times) than the earlier medium. Based on this study we recommend the routine use of the simplified medium to perform the DNase test for biotyping enteric campylobacters.

Caracterización de Campylobacter, Helicobacter y bacterias curvadas asociadas con gastritis y úlceras pépticas / Campylobacter, Helicobacter, spiral shaped bacteria associated with gastritis and pepticulcers. A characterization

El género Campylobacter fue enmendado por Vandamme et al. 1991 y se propuso el nuevo género Arcobacter. Además, se revisó la taxonomía de Helicobacter. Al menos seis especies de este último género han sido descritas: H. pylori, en humanos y monos, H. mustelas en hurones, H. felis en gatos y perros y H. nemestrinae a partir de un mono (Macaca nemestrina), H. cinaedi y fennelliae (anteriormente C. cinaedi y C. fennelliae). Con excepción de las últimas dos especies, relacionadas con proctitis en hombres homoxesuales, este agente induce gastritis en sus respectivos hospederos. H. mustelae, una de las especies más estudiadas se ha propuesto como modelo para el estudio de la patogénesis dela gastritis. Además, se ha inoculado experimentalmente algunos animales como cerdos, perros y ratones con H. Pylori. Por otra parte, a partir de biopsias gáastricas de pacientes con molestias gástricas se ha observado, pero no cultivado, otras bacterias espirilares diferentes de Helicobacter, para las cuales se ha propuesto el nombre "Gastrospirillum hominis". Estos hallazgos sugieren que la mucosa gástrica de mamíferos está colonizada por bacterias espirilares adaptadas a ese medio. Por lo tanto, podrían darse las infecciones cruzadas entre humanos y otros animales

Ocorrência de Campylobacter termofílico em diferentes pontos de uma estaçäo de tratamento de esgotos na cidade do Rio de Janeiro, RJ / Occurrence of thermophilic Campylobacter in different points of a sawage treatment station in Rio de Janeiro, RJ

Investigou-se durante o período de 1986 a 1988, a incidência de espécies termofílicas de Campylobacter em duzentas amostras de água colhidas de afluente, entrada e saída do tanque de aeraçäo e efluente da estaçäo de tratamento de esgotos, na ilha do Governador, R.J. No processo de isolamento, alguns esquemas foram experimentados, destacando-se a maior eficiência da semeadura em "Columbia Blood Agar Base" contendo 5% FBP, 0,5% de uma mistura de antibióticos e 0,4% de carväo ativado, após incubaçäo a 43-C por 48 horas em microaerofilia. Verificou-se a predominância de Campylobacter no afluente e entrada do tanque, representada por 40 estirpes ou 83% dos 48 isolamentos obtidos. A identificaçäo fenotípica caracterizou as seguintes espécies: C. jejuni (56,25%), C. coli (25,0%), C. laridis (6,25%) e Campylobacter sp. (12,5%), sendo que as duas primeiras näo foram detectadas nos efluentes examinados. Com base neste resultado, questiona-se a disseminaçäo de certas espécies termofílicas de Campylobacter, através de efluentes de estaçöes de tratamento de esgotos

Primer aislamiento de campylobacter jejuni en Misiones: importancia de su estudio en pacientes con gastroenteritis severas / First isolation of campylobacter jejuni in Misiones: its importance in patients with severe gastroenteritis

Informa del primer aislamiento de Campylobacter jejuni en un niño con diarrea aguda, internado en el Hospital Ramón Madariaga de la ciudad de Posadas, Misiones. Se trata de un paciente de dos meses de edad que en enero de 1987 presentó un cuadro de diarrea aguda, con temperatura elevada, deshidratación moderada, vómitos y 6 a 8 deposiciones diarias. El estudio microbiológico de sus heces resultó negativo para Escherichia coli enterotoxigénica (ECET), eschirichia coli entero invasiva (ECEI), escherichi coli enteropatógena (ECEP) escherichia coli enterhemorrágica (ECEH), salmonella, shigella, aeromona y rotavirus, aislándose como único agente Campylobacter jejuni. Luego del tratamiento el paciente evolucionó favorablemente

Serotypes & biotypes of Campylobacter jejuni & C. coli from diverse sources in Calcutta.

C. jejuni and C. coli isolates from diverse sources in Calcutta were serotyped and biotyped according to the Lior scheme. Of the 55 strains examined, 85.5 per cent reacted with one or another of the 73 antisera available. This included the formation of two new serogroups, LIO 67 and LIO 76. C. coli serogroup LIO 46 biotype II was the most frequently encountered strains (14.5%), followed by C. coli serogroup LIO 29, 55 biotype II (10.5%) and C. jejuni serogroup LIO 54, biotype I (5.5%). Serogroups recovered from animals and birds were also found to be prevalent in strains isolated from clinical sources, confirming the zoonotic implications of the disease.

Características bioquímicas de Campylobacter isolados de bezerros com e sem diarréia / Biochemical characters of Campylobacter sp, isolated from calves with and without diarrhea

Sessenta amostras de Campylobacter sp, isoladas de bezerros com e sem diarréia foram testadas frente a 11 testes bioquímicos. Dentro dos critérios propostos, Campylobacter sp, foi classificado, 53,3% como Campylobacter jejuni (biotipo 1 e biotipo 2); 8,4% como Campylobacter coli; 20% Campylobacter jejuni/coli; 15% como NARTC - grupo 3,3% como Campylobacter fetus subsp venerealis. Foram englobadas algumas novas cepas de Campylobacter sp, com determinadas características fenotípicas, obedecendo a nomenclatura aceita pela "Aproved List of Bacterial Names" (1980)

Campylobacter gastro-entéricos: bacteriología, epidemiología y patogenia / Gastro-enteric Campylobacter: bacteriology, epidemiology and pathogenesis

The genus Campylobacter compromises over ten species of Gram negative, curved or S shaped rods. Thermotolerant species (C. jejuni, C. coli and C. Laridis) are recognized as important agentes of human diarrhea, whreas C. fetus subsp. fetus is an apportunistic pathogen and C. pyloridis has been associated with gastritis and gastric ulcer. These bacteria can be isolated don selective media using a microaerobic atmosphere. Termotolerant species grow well at 43-C while the other species do not. Identification methods include morphological, cultural an biochemical test, that correlate well genetic characteristics. Enteric infections with Campylobacter spp. occur by ingestión of contaminated liquid or solid food, and through contact with infected animals. A large number of animal species are recognized as reservoirs of these zoonotic bacteria. Disease can be produced by means of invasiviness of by the production of toxins. Erythromycin is the drug of choice for the treatment of Campylobacter enteric infection. However, they are also susceptible to nitrofurans, chloramphenicol and gentamicin. Despite the great information existing about many aspects of Campylobacter infections, only the joint work of clinicians, epidemiologists and laboratory personnel could provide an oberview of the complex factor involved in the infections by Campylobacter spp