ABSTRACT
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)
Subject(s)
Animals , Campylobacter Infections/veterinary , Erythromycin , Campylobacter jejuni/drug effects , Campylobacter coli/drug effects , Drug Resistance, Bacterial , Chickens , Sus scrofaABSTRACT
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.
Subject(s)
Humans , Infant , Child, Preschool , Child , Campylobacter Infections/microbiology , Campylobacter Infections/epidemiology , Campylobacter jejuni/isolation & purification , Campylobacter jejuni/drug effects , Campylobacter coli/isolation & purification , Campylobacter coli/drug effects , Drug Resistance, Multiple, Bacterial , Diarrhea/microbiology , Anti-Bacterial Agents/pharmacology , Campylobacter Infections/drug therapy , Prevalence , Ecuador/epidemiology , Feces/microbiology , Anti-Bacterial Agents/classificationABSTRACT
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.
Subject(s)
Adolescent , Animals , Child , Child, Preschool , Humans , Infant , Infant, Newborn , Anti-Bacterial Agents/pharmacology , Campylobacter coli/drug effects , Campylobacter jejuni/drug effects , Feces/microbiology , Poultry/microbiology , Acute Disease , Campylobacter coli/genetics , Campylobacter coli/isolation & purification , Campylobacter jejuni/genetics , Campylobacter jejuni/isolation & purification , Diarrhea/microbiology , Genotype , Random Amplified Polymorphic DNA TechniqueABSTRACT
A total of 495 Campylobacterjejuni and 122 C. coli isolated from Thai children were screened for macrolide (erythromycin and azithromycin) resistance by disk diffusion assay. Minimum inhibitory concentrations for erythromycin, azithromycin, nalidixic acid, ciprofloxacin, tetracycline, streptomycin, gentamicin and chloramphenicol were further determined for these macrolide-resistant Campylobacter isolates. Presence of known point mutations resulting in reduced susceptibility to macrolides was investigated by PCR and DNA sequencing. Seventeen percent (23/122) of C. coli and 2.4% (12/495) of C. jejuni isolates were resistant to macrolides. By sequencing domain V of the 23S ribosomal DNA from all 35 macrolide-resistant isolates, a known point mutation of 23S rRNA associated with reduced susceptibility to macrolides was detected in all isolates except one. Among the macrolide-resistant isolates, all were multiply resistant to nalidixic acid and ciprofloxacin, of which the latter is the preferred antimicrobial used for diarrheal treatment in Thailand. Furthermore, most macrolide-resistant isolates were also resistant to tetracycline and streptomycin. The spread of macrolide and quinolone resistant Campylobacter should be monitored closely in Thailand and elsewhere as these antimicrobials are preferred drugs for treatment of diarrhea.