Résumé
This study was aimed to understand the current status of the antimicrobial resistance and molecular distribution of Campylobacter in various poultry in Jiaodong area,to provide a basis for effective prevention and control of the Campy-lobacter risk to poultry products and human health.Campylobacter was isolated and identified from 565 cloacal samples collect-ed in the Jiaodong area from August to October 2021 through conventional bacterial isolation and culture,mass spectrometry,microbroth dilution and multilocus sequence typing(MLST).The drug resistance and molecular typing of 131 representative strains(67 Campylobacter jejuni and 64 Campylobacter coli)were studied separately.Antimicrobial resistance analysis indica-ted that 131 isolates were highly resistant to ciprofloxacin,nalixic acid and tetracycline,with resistance rates of 96.21%,96.21%and 95.45%,respectively.Except for 2 strains,62 strains of C.coli were completely resistant to these three drugs(100%).A total of 65 strains of 131 strains were multidrug re-sistant,and the overall multidrug resistance rate was 49.62%,among which 11 strains(16.42%)of C.jejuni were resistance to 3-5 antibiotics,and 54 strains(84.38%)of C.coli were re-sistance to 3-6 antibiotics.Among the isolates from different poultry sources,waterfowl isolates were the most resistant,fol-lowed by broiler isolates.The MLST typing results revealed 72 alleles and 35 sequence types obtained from 67 strains of C.je-juni,and the distribution was relatively dispersed,without a dominant ST type and homologous complex.A total of 27 alleles and 19 sequence types were obtained from 64 strains of C.coli.Moreover,59.38%(38/64)strains were homologous complex CC-828,in which the ST-1586 sequence type was most frequent,followed by ST-825.ST-1586,ST-9944 and ST-3735 were the main sources of C.coli in broilers,and ST-825 and ST-1586 were the main sources of C.coli in waterfowl.Differences in C.jejuni and C.coli carriage were observed among poultry in the Jiaodong area.Carriage of the two bacteria was more common in laying hens than in broilers and waterfowl.C.jejuni from poultry in the Jiaodong area was highly resistant to ciprofloxacin,nalixic acid and tetracycline,but had good sensitivity to other drugs.C.coli was highly resistant to a variety of antibiotics,and multiple drug resistance was common.St-type dispersal of C.jejuni showed high genetic diversity.C.coli was cloned and transmitted mainly by ST-1586 in broiler chickens and waterfowl.Poultry carry C.jejuni,which can cause serious diseases in humans.Therefore,dynamic monitoring of Campylobacter from poultry should be strengthened.
Résumé
Resumen El objetivo de este estudio fue evaluar la capacidad inhibitoria de Lactiplantibacillus plantarum LP5 frente a Campylobacter coli en ensayos de formación de biopelículas in vitro y exclusión competitiva. La formación de biopelículas por C. coli NCTC11366, C. coli DSPV458, C. coli DSPV541 y C. coli DSPV570 fue evaluada mediante medición de DO. La capacidad inhibitoria de L. plantarum LP5 frente a C. coli fue evaluada sobre discos de vidrio, nailon y aluminio. Sobre una biopelícula de L. plantarum se adicionó C. coli para cuantificar el efecto inhibidor de L. plantarum LP5 sobre el patógeno. Las cuatro cepas de C. coli fueron clasificadas como moderadas formadoras de biopelículas. El ensayo de exclusión competitiva mostró que la formación de biopelículas de las cepas de C. coli en todos los materiales fue significativamente mayor que la formación de biopelículas de cada patógeno en presencia de biopelículas de L. plantarum LP5. Si bien es necesario realizar más pruebas para confirmar la capacidad de supervivencia de C. coli en ambientes hostiles hasta llegar al huésped, este estudio permitiría avanzar en el esclarecimiento de su comportamiento mediante la formación de biopelículas.
Abstract The objective of this study was to evaluate the inhibitory capacity of Lactiplantibacillus plantarum LP5 against Campylobacter coli in in vitro biofilm formation and competitive exclusion assays. Biofilm formation by C. coli NCTC11366, C. coli DSPV458, C. coli DSPV541 and C. coli DSPV570 was evaluated by OD measurement. The inhibitory capacity of L. plantarum LP5 against C. coli was evaluated on glass, nylon and aluminium discs, added with L. plantarum and incubated at 37°C for 72 h. C. coli was added to each washed well. The plates were incubated at 42°C for 72 h in microaerophilic conditions and the biofilms were detached for quantification. The four strains of C. coli were classified as moderate biofilm former. The competitive exclusion test showed that the biofilm formation of the C. coli strains in all materials was significantly higher than the biofilm formation of each pathogen in the presence of L. plantarum LP5 biofilms. Although it is necessary to carry out more tests to confirm the ability of C. coli to survive in hostile environments until reaching the host, this study would allow progress in the elucidation of its behaviour through the formation of biofilms.
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Campylobacter spp. is a bacterial agent that causes gastroenteritis in humans and may trigger Guillain-Barré Syndrome (GBS) and is also considered one of the main foodborne diseases in developed countries. Poultry and pigs are considered reservoirs of these microorganisms, as well as raw or undercooked by-products are often incriminated as a source of human infection. Treatment in human cases is with macrolide, such erythromycin, that inhibits the protein synthesis of the microorganism. This study aimed to isolate Campylobacter jejuni and Campylobacter coli from intestinal content samples of broiler chickens (n=20) and swine (n=30) to characterize the erythromycin resistance profile of the strains and to detect molecular mechanisms involved in this resistance. The minimum inhibitory concentration was determined by agar dilution. The Mismatch Amplification Mutation Assay-Polymerase Chain Reaction (MAMA-PCR) was performed to detect mutations at positions 2074 and 2075 of 23S rRNA region, in addition to PCR test to detect the erm(B) gene. From the intestinal content of broiler chickens, 18 strains of C. jejuni and two strains of C. coli were isolated, whereas, from swine samples, no C. jejuni strain and 14 strains of C. coli were isolated. All C. coli strains were resistant, and three C. jejuni strains from broilers chickens were characterized with intermediate resistance to erythromycin. The MIC of the strains ranged from ≤0.5mg/μL to ≥128mg/μL. All resistant strains had the A2075G mutation, and one strain with intermediate resistance had the A2075G mutation. However, the A2074C mutation and the erm(B) gene were not detected. High resistance levels were detected in C. coli strains isolated from swine. The MAMA-PCR is a practical tool for detecting the erythromycin resistance in Campylobacter strains.(AU)
Campylobacter spp. é um agente bacteriano causador de gastroenterite em humanos e associado à síndrome de Guillain-Barré, sendo a campilobacteriose considerada uma das principais enfermidades de origem alimentar. Aves e suínos são importantes reservatórios desses microrganismos e seus produtos derivados crus ou mal cozidos são muitas vezes incriminados como fonte de infecção humana. A primeira escolha para o tratamento em casos humanos são os antimicrobianos da classe dos macrolídeos como à eritromicina. Dentro desse contexto, o objetivo deste estudo foi isolar Campylobacter jejuni e C. coli a partir de 20 amostras de conteúdo intestinal de frangos de corte e de 30 de suínos ao abate e investigar a resistência à eritromicina das estirpes obtidas e os possíveis mecanismos moleculares envolvidos nesta resistência. A concentração inibitória mínima foi determinada pela diluição em ágar e a técnica MAMA-PCR foi utilizada para detecção de mutações nas posições 2074 e 2075 da região 23s rRNA, foi pesquisado também a presença do gene erm(B) pela PCR. A partir do conteúdo intestinal de frangos de corte foram isoladas 18 estirpes de C. jejuni e duas de C. coli, enquanto de suínos foram obtidas 14 estirpes de C. coli e nenhuma estirpe de C. jejuni. Todas as estirpes de C. coli de suínos foram identificadas como resistentes e três estirpes de C. jejuni de frangos foram caracterizadas com resistência intermediária. A CIM das estirpes variou de ≤0,5mg/μL a ≥128mg/μL. Todas as estirpes resistentes tinham a mutação A2075G e uma cepa com resistência intermediária também apresentou a mutação A2075G. Não foi detectada a mutação A2074C ou a presença do gene erm(B) em nenhuma das estirpes obtidas. Os resultados revelam um alto nível de resistência em estirpes de C. coli isoladas de suínos frente a eritromicina. A técnica MAMA PCR utilizada se constitui em uma ferramenta prática para detecção da resistência à eritromicina em estirpes de C. jejuni e C. coli.(AU)
Sujets)
Animaux , Infections à Campylobacter/médecine vétérinaire , Érythromycine , Campylobacter jejuni/effets des médicaments et des substances chimiques , Campylobacter coli/effets des médicaments et des substances chimiques , Résistance bactérienne aux médicaments , Poulets , Sus scrofaRésumé
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)
Sujets)
Humains , Animaux , Zoonoses/étiologie , Campylobacter jejuni/isolement et purification , Campylobacter coli/isolement et purification , Poulets/virologie , Facteurs de virulence , Réaction de polymérisation en chaine en temps réel/médecine vétérinaire , TranscriptomeRésumé
Foodborne Campylobacter is recognized as the leading causes of the bacterial diarrheal illness in both developing and developed countries. C. jejuni and C. coli caused 95% of the human campylobacterisosis. Bacteria culture has been recognized as the "Gold standard" for the diagnosis of the Campylobacter infection. The National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention coordinated the experienced researchers from China National Center for Food Safety Risk Assessment and other local Centers for Disease Control and Prevention to write up the standards for entitled Isolation and Identification of Campylobacter jejuni and Campylobacter coli (T/CPMA 006-2019). The standard is drafted with principles of emphasizing the scientific, normative, applicability and feasible nature. This group standard recommended the procedures and steps for the isolation and identification of C. jejuni and C.coli from variant samples. The standard aims to improve the capacity for Campylobacter identification in China.
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The objective of this study was to investigate a clonal relationship among thermo-tolerant Campylobacter spp. isolates from different stages of the poultry meat supply chain in Argentina. A total of 128 thermotolerant Campylobacter spp. (89 C. jejuni and 39 C. coli) isolates from six poultry meat chains were examined. These isolates were from: a) hens from breeder flocks, b) chickens on the farm (at ages 1 wk and 5 wk), c) chicken carcasses in the slaughterhouse, and d) chicken carcasses in the retail market. Chickens sampled along each food chain were from the same batch. Campylobacter spp. isolates were analyzed using pulsed-field gel electrophoresis to compare different profiles according to the source. Clustering of C. jejuni isolates resulted in 17 profiles, with four predominant genotypes and many small profiles with just a few isolates or unique patterns, showing a very high degree of heterogeneity among the C. jejuni isolates. Some clusters included isolates from different stages within the same chain, which would indicate a spread of strains along the same poultry meat chain. Moreover, twenty-two strains of C. coli clustered in seven groups and the remaining 17 isolates exhibited unique profiles. Evidence for transmission of thermotolerant Campylobacter spp. through the food chain and cross contamination in the slaughterhouses were obtained. This collective evidence should be considered as the scientific basis to implement risk management measures to protect the public health.
El objetivo de este trabajo fue investigar la relación clonal entre aislamientos de Campylobacter spp. termotolerantes obtenidos de diferentes etapas de la cadena cárnica aviar en Argentina. En total se examinaron 128 aislamientos de Campylobacter spp. (89 de Campylobacter jejuni y 39 de Campylobacter coli) obtenidos de 6 cadenas cárnicas muestreadas en los siguientes puntos del circuito productivo: a) gallinas reproductoras; b) pollos en las granjas (de una y 5 semanas de edad); c) carcasas de pollo en frigorífico, y d) carcasas de pollo en puntos de venta final. Las muestras de pollos fueron obtenidas a lo largo de las cadenas cárnicas siguiendo el mismo lote. Los aislamientos de Campylobacter spp. fueron analizados mediante electroforesis de campos pulsados y se compararon los diferentes perfiles. Los aislamientos de C. jejuni se agruparon en 17 perfiles, 4 de ellos predominantes y el resto en perfiles que agruparon pocos aislamientos o patrones únicos, lo que ilustra una gran heterogeneidad. Algunos agrupamientos incluyeron aislamientos obtenidos de diferentes etapas de una misma cadena cárnica, lo cual indicaría una dispersión de cepas a lo largo de las cadenas cárnicas. Por otra parte, 22 aislamientos de C. coli se agruparon en 7 grupos y otros 17 aislamientos presentaron perfiles únicos. Se obtuvieron evidencias de transmisión de Campylobacter spp. termotolerante en la cadena cárnica aviary contaminación cruzada en frigoríficos. La evidencia reunida debería servir como base científica para implementar estrategias de manejo del riesgo, destinadas a proteger la salud de los consumidores de carne aviar.
Sujets)
Animaux , Femelle , Variation génétique , Campylobacter , Microbiologie alimentaire , Argentine , Volaille , Campylobacter/isolement et purification , Campylobacter/génétique , Infections à Campylobacter , Poulets/microbiologie , Viande/microbiologieRésumé
To investigate the contamination conditions of Campylobacter spp.in duck production chain and its antimicrobial resistance,virulence gene distribution,samples were collected at the duck slaughterhouse according to GB 4789.9-2014.Triplex PCR assay was applied to identify the species of Campylobacter and the recovered Campylobacter strains were tested for the antimicrobial susceptibility against 8 kinds of antimicrobial agents using a broth microdilution method,the susceptibility results were determined according to the NARMS criteria (2011).Subsequently,4 virulence genes were detected by PCR method.The result showed that 187 Campylobacter isolates were obtained from 489 samples,including 160 C.jejuni,130 C.coli and 10 unidentified Campylobacter isolates.The total isolation rate of Campylobacter was 38.24%.The prevalence of Camnpytobacter before slaughtering,at depilation stage,evisceration stage and duck products was 76.33%,5.62%,24.00%,and 0%,respectively.The Campylobacter isolates were most frequently resistant to tetracycline (95.72 %),followed by resistance to clindamycin(90.91%),the resistance rate of azithromycin (63.64%) was in the middle,the resistance rates of ciprofloxacin(31.02%),gentamicin(34.76%),nalidixic acid (37.43 %),erythromycin (41.18 %),chloramphenicol (41.18%) were relatively low.The multi-drug resistance was common among Campylobacter isolates with a rate of 72.19%.The prevalence of adhesion-associated gene cadF,flagellin gene flaA,invasion associated protein gene iamA,toxin regulation gene cdtB was 100%,80.75 %,71.12% and 94.65%,respectively.The results indicated that the Campylobacter contamination occurred in the slaughtering procedures of duck,and the antimicrobial resistance of Campylobacter isolates was relatively serious.In addition,the virulence-associated genes were detected widely among Campylobacter isolates.Therefore,the supervision of antimicrobial agents using at rearing stage should be strengthened,along with health management in duck production chain.
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Doença bacteriana zoonótica, a campilobacteriose é responsável mundialmente por frequentes casos de gastroenterite humana. Campylobacter spp. apresenta fator de virulência associado à diarreia, denominado toxina citoletal distensiva (CDT), sendo codificado pelos genes do complexo cdt. Os objetivos do presente estudo foram: 1) isolar e identificar estirpes de Campylobacter spp. de 102 suabes de carcaças e 102 suabes retais de ovinos (Ovis aries) e de sete amostras de água dos efluentes, antes e depois do tratamento de desinfecção de abatedouro localizado no estado de São Paulo; e 2) detectar, pela técnica de multiplex-PCR, a presença do complexo de genes cdt. Foram isoladas e identificadas, por métodos fenotípicos e genotípicos, sete estirpes de Campylobacter coli provenientes de 4/102 (3,92%) das amostras de suabes retais, 1/102 (0,98%) de suabes de carcaças e 2/7 (28,5%) das águas dos efluentes. Dos isolados de suabes retais, em 2/7 (28,6%) estirpes foi detectada a presença dos genes cdt. Trata-se do primeiro relato de isolamento de estirpes de Campylobacter coli provenientes de abatedouro de ovinos e das estirpes portadoras do complexo de genes cdt nessa espécie animal no Brasil.
A zoonosis and bacterial disease, campylobacteriosis is responsible for frequent cases of human gastroenteritis worldwide. Campylobacter spp. presents the virulence factor called cytolethal distensive toxine (CDT), responsible for diarrhea and codified by the cdt gene. The aims of this study were: 1) to isolate and identify Campylobacter spp. strains from 102 carcass swabs and 102 rectal swabs of sheep (Ovis aries) and seven samples of wastewater, before and after the disinfection treatment, collected from the abattoir of the state of São Paulo; and 2) to detect the presence of cdt gene complex by Multiplex-PCR in strains of Campylobacter spp. Seven strains of Campylobacter coli were isolated and identified by phenotypic and genotypic methods: 4/102 (3.92%) from rectal swabs, 1/102 (0.98%) from carcass swabs and 2/7 (28.5%) from wastewater. From the rectal swab samples 2/7 (28.6%) strains were detected with the cdt gene. This is the first report on the isolation of Campylobacter coli from sheep abattoir, and of strains carrying the cdt gene complex in this animal species in Brazil.
Sujets)
Animaux , Abattoirs , Campylobacter coli , Effluents Industriels , Ovis , Désinfection de l'Eau , Infections bactériennes , Gastroentérite/épidémiologie , Réaction de polymérisation en chaine multiplex/médecine vétérinaire , ZoonosesRésumé
The antibacterial effects of a combination of Coptidis rhizoma and Galla rhois extracts (CGE) were evaluated in piglets. The minimum bactericidal concentration of CGE was 2.0 mg/mL. Thirty 5-week-old piglets were challenged with Campylobacter (C.) coli after allocation to three different groups, a control and two treatment groups fed with CGE at 2.0 or 4.0 g/kg feed for 7 days. On day 7, C. coli in the feces of the CGE-treated groups were significantly lower than in the control (p < 0.01). These results suggest that CGE can be used to control C. coli in piglets.
Sujets)
Campylobacter coli , Campylobacter , FècesRésumé
Purpose: To study the prevalence and the antimicrobial resistance of campylobacter species isolated from children suffering from gastroenteritis. Materials and Methods: A total of 125 stool samples were collected from children with gastroenteritis. The identification of isolates was performed with conventional methods as well as with molecular methods based on 16SrRNA species‑specific gene amplification by PCR and product analysis. Resistance pattern of the isolated strains was determined using agar dilution method. Results: Conventional methods including sodium hippurate hydrolysis revealed that 12 (9.6%) samples were positive for Campylobacter species. Ten out of 12 Campylobacter spp. were identified as Campylobacter jejuni and 2 as Campylobacter coli but PCR assay revealed that five samples only were positive for Campylobacter and all were C. jejuni. Antimicrobial susceptibility to 10 antimicrobials was performed and all isolates (five isolates of C. jejuni) were susceptible to chloramphenicol, gentamicin and amikacin but all were resistant to ceftriaxone. Conclusion: PCR assay method allows reliable detection of C. jejuni. C. jejuni was the most prevalent Campylobacter species. Gentamicin, amikacin and chloramphenicol were the most effective antibiotic.
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Isolaram-se estirpes de Campylobacter spp. em amostras de carcaças (n=65), fezes (n=65) e linfonodos mesentéricos (n=65) de suínos abatidos em frigoríficos do estado de São Paulo e detectaram, pela técnica da Multiplex-PCR, a presença do complexo de genes cdt, responsáveis pela expressão do fator de virulência da toxina CDT. Do total de 195 amostras de origem suína, Campylobacter spp. foi isolado de 31 (15,9%), sendo 29 (93,6%) de amostras de suabe retal, 1/65 (3,2%) de suabe de carcaça e um (3,2%) de linfonodo. Vinte e oito estirpes de C. coli foram positivas para a detecção dos genes cdt, e três estirpes de C. jejuni foram negativas para a detecção desses genes. Foi detectada, pela primeira vez no estado de São Paulo, a presença dos genes cdt em 100% das estirpes de Campylobacter coli provenientes de suínos abatidos em frigoríficos.
The purposes of this study were to isolate and identify Campylobacter spp. strains from the carcasses (n=65), feces (n=65) and mesenteric lymph nodes (n=65) of swine slaughtered in abattoirs in the State of Sao Paulo and to detect the presence of the cdt gene complex - responsible for the expression of the virulence factor cytolethal distensive toxin - in these Campylobacter spp. strains through Multiplex-PCR. From 195 samples analyzed, Campylobacter spp. was isolated in 31 (15.9%): 29 (93,6%) samples of rectal swab, 1 (3.2%) carcass swab and 1 (3.2%) lymph node sample. The 28 strains of isolated C. coli were positive for CDT toxin genes and the three strains of isolated C. jejuni were negative for these genes. It was also the first time that the cdt gene cluster was detected in strains isolated from swine in the state of São Paulo. These findings indicate swine as a potential spreading source of virulent strains of Campylobacter coli, either for slaughterhouse staff or consumers of carcasses and sub products.
Sujets)
Animaux , Abattoirs , Campylobacter/virologie , Suidae , Réaction de polymérisation en chaine multiplexRésumé
Campylobacter coli is an important species involved in human cases of enteritis, and chickens are carriers of the pathogen mainly in developing country. The current study aimed to evaluate the transmission of C. coli and its pathogenic effects in chicken embryos. Breeder hens were inoculated intra-esophageally with C. coli isolated from chickens, and their eggs and embryos were analyzed for the presence of bacteria using real-time PCR and plate culture. The viability of embryos was verified. In parallel, SPF eggs were inoculated with C. coli in the air sac; after incubation, the embryos were submitted to the same analysis as the embryos from breeder hens. In embryos and fertile eggs from breeder hens, the bacterium was only identified by molecular methods; in the SPF eggs, however, the bacterium was detected by both techniques. The results showed no relationship between embryo mortality and positivity for C. coli in the embryos from breeder hens. However, the presence of bacteria is a cause of precocious mortality for SPF embryos. This study revealed that although the vertical transmission is a possible event, the bacteria can not grow in embryonic field samples.
Sujets)
Animaux , Embryon de poulet , Infections à Campylobacter , Poulets , Campylobacter/génétique , Campylobacter/isolement et purification , Techniques in vitro , Viabilité microbienne , Mortalité , Réaction de polymérisation en chaîne/méthodes , Embryon de poulet , Méthodes , VirulenceRésumé
The aim of this study is to develop the real-time PCR assays to detect and qualify C. coli from stool specimen. The primers and probe were designed based on the specific sequence of ceuE gene in C. coli using Primer 5.0 and Vector NTI Suite 6.0 e. The PCR assay was optimized with the reference C. coli strains. The standard curve based on the dilutions of genomic DNA showed a linear relationship between log CFU/mL and threshold cycles. The detectable limitation was 5.62 CFU/mL by using purified DNA from bacteria culture. The reproducibility of this assay was assessed by calculating the variation of the threshold cycle value and the slope from test repeats for the same strains and different strains. Our results indicate that the developed assay has high sensitivity and specificity for identification of C. coli.
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Swine is a common source of Campylobacter coli human gastroenteritis, for the treatment of which erythromycin and fluoroquinolones are recommended. The prevalence of antimicrobial-resistant C. coli differs significantly depending on countries. We investigated the prevalence of C. coli in swine from a farm in Buan-gun, Korea in 2010, and determined antimicrobial susceptibility of the isolates. Rectal swab specimens were used to inoculate Campylobacter Preston media and incubated microaerophilically at 42degrees C for 48 h. The species were identified by phenotypic tests and by detecting hipO and glyA genes. PCR was used to detect mutations of A2074C in 23S rRNA gene, and quinolone resistance-determining region (QRDR) of gyrA, which are associated with high level resistance to erythromycin, and with ciprofloxacin, respectively. Antimicrobial susceptibility was determined by the disk diffusion and agar dilution tests. Of the 100 specimens, 55 (55%) yielded C. coli, and 23 of them (41.8%) had A2074G mutation. A2074G mutated isolates showed the lowest MIC90 of imipenem, while those of ampicillin and clindamycin were relatively low. The majority of both A2074G mutation-positive and -negative isolate were susceptible to ampicillin, cefotaxime, and chloramphenicol. All isolates were resistant to ciprofloxacin, and had mutation in QRDR of gyrA. In conclusion, C. coli was detected in 55% of swine, and A2074G mutation was detected in 41.8% of the isolates. All isolates had gyrA mutation-mediated ciprofloxacin resistance.
Sujets)
Humains , Agar-agar , Ampicilline , Campylobacter , Campylobacter coli , Céfotaxime , Chloramphénicol , Ciprofloxacine , Clindamycine , Diffusion , Érythromycine , Fluoroquinolones , Gastroentérite , Gènes d'ARN ribosomique , Imipénem , Corée , Réaction de polymérisation en chaîne , Prévalence , SuidaeRésumé
Nowadays salmonellosis and campylobacteriosis are the most prevalent diseases transmitted to humans through contaminated food of animal origin in developed countries. Propagation of Salmonella enterica serotype Enteritidis and Salmonella enterica serotype Typhimurium has increased since the second half of the twentieth century, derived from two changes in the epidemiology of salmonellosis that occurred worldwide: the emergence of human infections caused by Salmonella Enteritidis, and the multiple resistances against antibiotics of Salmonella Typhimurium strains. Most retrospective studies suggest an epidemiological relationship between human infections and poultry products. Modernization of poultry industry and exportation of progenitor stock birds have played an important roll disseminating Salmonella Enteritidis. Campylobacteriosis is the most frequent enteric bacterial diseases in developed countries. Campylobacter jejuni is the most frequent cause of acute infectious diarrheas, and the sources of infection are mainly poultry products. The present review includes relevant taxonomic and pathogenic aspects of these organisms; and stresses the problematic of diagnosis and detection, analyzing techniques that permit rapid detection of animal carriers. Finally, some preliminary findings, not yet published, suggest that a prevalence of salmonellosis and campylobacteriosis in Mexican poultry farms must be similar to that found in other countries with massive poultry production; therefore, epidemiological studies to determine the frequency of human infections, derived from poultry consumption, are recommended.
En la actualidad, la salmonelosis y la campilobacteriosis son las zoonosis de mayor prevalencia en países desarrollados, debido a la contaminación de alimentos de origen animal. La propagación de Salmonella enterica serotipo Enteritidis y Salmonella enterica serotipo Typhimurium va en aumento a partir de la segunda mitad del siglo XX, cuando ocurrieron dos cambios mundiales en la epidemiología de la salmonelosis: el surgimiento de infecciones en humanos por Salmonella Enteritidis, y la múltiple resistencia a los antibióticos de cepas de Salmonella Typhimurium. La mayoría de los análisis retrospectivos sugieren que existe relación epidemiológica entre las aves y los humanos: La modernización de la avicultura y las exportaciones de aves progenitoras son de importancia en la diseminación de Salmonella Enteritidis. La campilobacteriosis es la enfermedad entérica bacteriana más común en países desarrollados. Campylobacter jejuni es la causa más frecuente de diarreas infecciosas agudas, y las fuentes de esta infección son principalmente productos avícolas. En la presente revisión se incluyen aspectos relevantes de taxonomía y patogenia de estos organismos, y se hace hincapié en la problemática del diagnóstico y detección, analizando las técnicas para detectar rápidamente la existencia de animales portadores. Finalmente, se discuten hallazgos preliminares aún no publicados que sugieren que la prevalencia de salmonelosis y campilobacteriosis en granjas avícolas mexicanas debe ser semejante a la de otros países que tienen avicultura tecnificada, y se recomiendan los estudios epidemiológicos para determinar la frecuencia de estas infecciones en humanos, derivadas del consumo de alimentos de origen aviar.