RESUMEN

Campylobacter is gram-negative bacteria considered the predominant genera isolated from poultry samples and associated with gastroenteritis. Due to the problems in conventional cultural methods of time-consuming and technically demanding requirements, a rapid and feasible method for their identification and discrimination of the closely related spp. Including Campylobacter coli, Campylobacter fetus, and Campylobacter jejuni is needed. This study analyzes the chicken and sheep meats samples (n = 125) using culture and pre-enrichment-based Quadraplex real-time PCR by targeting OrfA, CstA, HipO, and 16 S rRNA genes of C. coli, C. fetus, C. jejuni and Campylobacter spp. Respectively. The analysis of 125 chicken and sheep meat samples by culture and real-time PCR showed high concordance between the results of the two methods. The present study show high prevalence of Campylobacter species (35% and 32% from chicken and meat respectively) of which C. jejuni were the most abundant. Reaction efficiencies were between 90 and 110%, and detect as low as 8.9 fg in C. jejuni. The need for quick detection and discrimination methods in sheep and chicken meat can be met using the described Quadraplex real-time PCR methodology.

Asunto(s)

Campylobacter coli , Campylobacter jejuni , Pollos , Carne , Reacción en Cadena en Tiempo Real de la Polimerasa , Animales , Pollos/microbiología , Ovinos/microbiología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Campylobacter coli/genética , Campylobacter coli/aislamiento & purificación , Campylobacter coli/clasificación , Campylobacter jejuni/genética , Campylobacter jejuni/aislamiento & purificación , Campylobacter jejuni/clasificación , Carne/microbiología , Campylobacter fetus/genética , Campylobacter fetus/aislamiento & purificación , Campylobacter fetus/clasificación , Campylobacter/genética , Campylobacter/aislamiento & purificación , Campylobacter/clasificación , Microbiología de Alimentos , ADN Bacteriano/genética

RESUMEN

Campylobacter spp. are considered the most frequent cause of acute gastroenteritis worldwide. However, outside high-income countries, its burden is poorly understood. Limited published data suggest that Campylobacter prevalence in low- and middle-income countries is high, but their reservoirs and age distribution are different. Culturing Campylobacter is expensive due to laboratory equipment and supplies needed to grow the bacterium (e.g., selective culture media, microaerophilic atmosphere, and a 42°C incubator). These requirements limit the diagnostic capacity of clinical laboratories in many resource-poor regions, leading to significant underdiagnosis and underreporting of isolation of the pathogen. CAMPYAIR, a newly developed selective differential medium, permits Campylobacter isolation without the need for microaerophilic incubation. The medium is supplemented with antibiotics to allow Campylobacter isolation in complex matrices such as human feces. The present study aims to evaluate the ability of the medium to recover Campylobacter from routine clinical samples. A total of 191 human stool samples were used to compare the ability of CAMPYAIR (aerobic incubation) and a commercial Campylobacter medium (CASA, microaerophilic incubation) to recover Campylobacter. All Campylobacter isolates were then identified by MALDI-TOF MS. CAMPYAIR showed sensitivity and specificity values of 87.5% (95% CI 47.4%-99.7%) and 100% (95% CI 98%-100%), respectively. The positive predictive value of CAMPYAIR was 100% and its negative predictive value was 99.5% (95% CI 96.7%-99.9%); Kappa Cohen coefficient was 0.93 (95% CI 0.79-1.0). The high diagnostic performance and low technical requirements of the CAMPYAIR medium could permit Campylobacter culture in countries with limited resources.

Asunto(s)

Infecciones por Campylobacter , Campylobacter , Medios de Cultivo , Técnicas Microbiológicas , Medios de Cultivo/normas , Aerobiosis , Campylobacter/clasificación , Campylobacter/crecimiento & desarrollo , Campylobacter/aislamiento & purificación , Infecciones por Campylobacter/diagnóstico , Infecciones por Campylobacter/microbiología , Heces/microbiología , Valor Predictivo de las Pruebas , Técnicas Microbiológicas/métodos , Técnicas Microbiológicas/normas

RESUMEN

Campylobacteriosis is an important contributor to the global burden of acute gastroenteritis (AGE). In Nicaragua, the burden, risk factors, and species diversity for infant campylobacteriosis are unknown. Between June 2017 and December 2018, we enrolled 444 infants from León, Nicaragua, in a population-based birth cohort, conducting weekly household AGE surveillance. First, we described clinical characteristics of symptomatic Campylobacter infections, and then compared clinical characteristics between Campylobacter jejuni/coli and non-jejuni/coli infections. Next, we conducted a nested case-control analysis to examine campylobacteriosis risk factors. Finally, we estimated the population attributable fraction of campylobacteriosis among infants experiencing AGE. Of 296 AGE episodes in the first year of life, Campylobacter was detected in 59 (20%), 39 were C. jejuni/coli, and 20 were non-jejuni/coli species, including the first report of Campylobacter vulpis infection in humans. Acute gastroenteritis symptoms associated with C. jejuni/coli lasted longer than those attributed to other Campylobacter species. In a conditional logistic regression model, chickens in the home (odds ratio [OR]: 3.8, 95% CI: 1.4-9.8), a prior AGE episode (OR: 3.3; 95% CI: 1.4-7.8), and poverty (OR: 0.4; 95% CI: 0.2-0.9) were independently associated with campylobacteriosis. Comparing 90 infants experiencing AGE with 90 healthy controls, 22.4% (95% CI: 11.2-32.1) of AGE episodes in the first year of life could be attributed to Campylobacter infection. Campylobacter infections contribute substantially to infant AGE in León, Nicaragua, with non-jejuni/coli species frequently detected. Reducing contact with poultry in the home and interventions to prevent all-cause AGE may reduce campylobacteriosis in this setting.

Asunto(s)

Infecciones por Campylobacter/epidemiología , Campylobacter/genética , Cohorte de Nacimiento , Campylobacter/clasificación , Campylobacter/patogenicidad , Infecciones por Campylobacter/etiología , Estudios de Casos y Controles , Preescolar , Heces/microbiología , Femenino , Gastroenteritis/epidemiología , Gastroenteritis/microbiología , Humanos , Lactante , Recién Nacido , Masculino , Oportunidad Relativa , Factores de Riesgo

RESUMEN

Campylobacter is among the four main causes of gastroenteritis worldwide and has increased in both developed and developing countries over the last 10 years. The vast majority of reported Campylobacter infections are caused by Campylobacter jejuni and, to a lesser extent, C. coli; however, the increasing recognition of other emerging Campylobacter pathogens is urgently demanding a better understanding of how these underestimated species cause disease, transmit, and evolve. In parallel to the enhanced clinical awareness of campylobacteriosis due to improved diagnostic protocols, the application of high-throughput sequencing has increased the number of whole-genome sequences available to dozens of strains of many emerging campylobacters. This has allowed for comprehensive comparative pathogenomic analyses for several species, such as C. fetus and C. concisus These studies have started to reveal the evolutionary forces shaping their genomes and have brought to light many genomic features related to pathogenicity in these neglected species, promoting the development of new tools and approaches relevant for clinical microbiology. Despite the need for additional characterization of genomic diversity in emerging campylobacters, the increasing body of literature describing pathogenomic studies on these species deserves to be discussed from an integrative perspective. This review compiles the current knowledge and highlights future work toward deepening our understanding about genome dynamics and the mechanisms governing the evolution of pathogenicity in emerging Campylobacter species, which is urgently needed to develop strategies to prevent or control the spread of these pathogens.

Asunto(s)

Infecciones por Campylobacter/microbiología , Campylobacter/genética , Enfermedades Transmisibles Emergentes/microbiología , Genoma Bacteriano/genética , Evolución Biológica , Campylobacter/clasificación , Campylobacter/patogenicidad , Genómica , Humanos

RESUMEN

A study using sentinel broiler chickens was performed to address Campylobacter persistence in litter that was reused for successive flocks. Cloacal swabs, litter, drag swabs, darkling beetles, feed, and drinking water were weekly sampled and analyzed by standard microbiological procedures. Thermotolerant Campylobacter isolated strains were confirmed by polymerase chain reaction and subtyped by pulsed-field gel electrophoresis analysis. Campylobacter was not detected in samples collected immediately after downtime between broiler flocks. However, Campylobacter-positive samples were first detected at 21 d. After Campylobacter was initially isolated from the cloacal swabs, reused litter, drag swabs, or darkling beetles, these samples remained Campylobacter positive in the following weeks until the end of the rearing period. Campylobacter-positive cloacal swabs obtained from sentinel broilers ranged from 97.3% to 100% at 42 d. All isolated strains were identified as Campylobacter jejuni. Among the subtypes identified, an indistinguishable C. jejuni strain was predominant in sentinel broilers and was also detected in the other environmental samples analyzed, suggesting a common and persistent contamination source within the flocks. Sentinel broilers may have contributed to amplify the Campylobacter level, maintaining flock and broiler house contamination until the end of the production cycle.

Asunto(s)

Crianza de Animales Domésticos/instrumentación , Campylobacter/clasificación , Campylobacter/crecimiento & desarrollo , Pollos/microbiología , Vivienda para Animales , Termotolerancia , Crianza de Animales Domésticos/métodos , Animales , Brasil , Campylobacter/aislamiento & purificación , Campylobacter jejuni/genética , Campylobacter jejuni/crecimiento & desarrollo , Campylobacter jejuni/aislamiento & purificación , Cloaca/microbiología , Escarabajos/microbiología , ADN Bacteriano/análisis , ADN Bacteriano/aislamiento & purificación , Masculino

RESUMEN

The objective of this meta-analysis was to summarize available information on the prevalence of thermotolerant Campylobacter (TC) in different food-producing animals worldwide. Databases (i.e., PubMed, ScienceDirect, Scopus) were searched from 1980 to 2017 unrestricted by language. The inclusion criteria were as follows: prevalence or incidence studies, published in peer-reviewed journals, and they must have reported the total number of animal samples studied and the number of samples that were positive for the presence of TC. When the identification of Campylobacter species was available, this information was included in the analysis. Multilevel random-effect meta-analysis models were fitted to estimate mean occurrence rate of TC and to compare them among different factors potentially associated with the outcome. The mean occurrence rate of TC in food-producing animals was 0.424 (95% CI: 0.394-0.455), and the mean occurrence rate of Campylobacter jejuni and Campylobacter coli were 0.214 and 0.133, respectively. Pigs and poultry showed the highest prevalence of TC; however, there were differences in the prevalence of each Campylobacter species. Campylobacter jejuni was observed in broilers (0.322; 95% CI: 0.273-0.377) and hens (0.395; 95% CI: 0.265-0.542), while C. coli was restricted essentially in pigs (0.553; 95% CI: 0.541-0.650). The prevalence of C. jejuni in intensively bred cattle was higher (0.302; 95% CI: 0.227-0.389) than the prevalence in extensively bred cattle (0.172; 95% CI: 0.119-0.242) while the prevalence of C. coli was similar (0.051; 95% CI: 0.028-0.091 vs. 0.050; 95% CI: 0.027-0.091) in both production systems. Agar with or without blood used for the isolation of TC did not affect the prevalence observed. The method of species identification did not seem to generate differences in the prevalence of Campylobacter species. The prevalence of Campylobacter in primary food production has a strong impact on the entire agri-food chain. National authorities must monitor the situation with the aim to establish the appropriate risk management measures.

Asunto(s)

Infecciones por Campylobacter/veterinaria , Campylobacter/aislamiento & purificación , Microbiología de Alimentos , Animales , Campylobacter/clasificación , Infecciones por Campylobacter/epidemiología , Campylobacter coli/aislamiento & purificación , Campylobacter jejuni/aislamiento & purificación , Bovinos/microbiología , Pollos/microbiología , Aves de Corral/microbiología , Enfermedades de las Aves de Corral/epidemiología , Prevalencia , Porcinos/microbiología

RESUMEN

INTRODUCTION: Microbes such as Salmonella, Campylobacter and S. aureus have been implicated in Foodborne disease outbreaks (FBDOs) worldwide, yet information on their occurrence in Barbados is scanty. The purpose of this study was to determine the aetiological agents, food vehicles, locations and peak seasons of FBDOs in Barbados; assess the quality of epidemiological investigations; and identify deficiencies in food production practices and laboratory detection. METHODOLOGY: A search of FBDOs occurring in Barbados between 1998-2009 was conducted among published and unpublished literature sources and reports. The search terms included the keywords "foodborne disease," "outbreaks" and "Barbados". RESULTS: During the period 1998 to 2009, there were 24 foodborne outbreaks, 215 cases of illness, one hospitalisation and no deaths. Overall, 37.5% of outbreaks were associated with hotels/resorts. Salmonella Enteritidis phage type 8 was most commonly implicated with eggs and poultry being the primary vehicles. Three outbreak reports were available for assessment and revealed that there were deficiencies in the outbreak investigations. These reports also recorded high levels of food contamination with indicator organisms, suggesting that improvements in food hygiene and production practices were required. CONCLUSIONS: The number of FBDOs is low in comparison to developed countries. However, the data was likely affected by under-reporting and inadequacies in the outbreak investigations and laboratory detection. Improvements in these areas would lead to not only better detection and characterisation of FBDOs in Barbados but improved food safety control measures.

Asunto(s)

Campylobacter/aislamiento & purificación , Brotes de Enfermedades , Enfermedades Transmitidas por los Alimentos/epidemiología , Enfermedades Transmitidas por los Alimentos/etiología , Salmonella/aislamiento & purificación , Staphylococcus aureus/aislamiento & purificación , Barbados/epidemiología , Campylobacter/clasificación , Contaminación de Alimentos , Humanos , Salmonella/clasificación , Estaciones del Año , Staphylococcus aureus/clasificación

RESUMEN

BACKGROUND: Campylobacter is one of the main causes of gastroenteritis worldwide. Most of the current knowledge about the epidemiology of this food-borne infection concerns two species, C. coli and C. jejuni. Recent studies conducted in developing countries and using novel diagnostic techniques have generated evidence of the increasing burden and importance of other Campylobacter species, i.e. non-C. coli/jejuni. We performed a nested case-control study to compare the prevalence of C. coli/jejuni and other Campylobacter in children with clinical dysentery and severe diarrhea as well as without diarrhea to better understand the clinical importance of infections with Campylobacter species other than C. coli/jejuni. METHODOLOGY/PRINCIPAL FINDINGS: Our nested case-control study of 439 stool samples included dysenteric stools, stools collected during severe diarrhea episodes, and asymptomatic stools which were systematically selected to be representative of clinical phenotypes from 9,160 stools collected during a birth cohort study of 201 children followed until two years of age. Other Campylobacter accounted for 76.4% of the 216 Campylobacter detections by qPCR and were more prevalent than C. coli/jejuni across all clinical groups. Other Campylobacter were also more prevalent than C. coli/jejuni across all age groups, with older children bearing a higher burden of other Campylobacter. Biomarkers of intestinal inflammation and injury (methylene blue, fecal occult test, myeloperoxidase or MPO) showed a strong association with dysentery, but mixed results with infection. MPO levels were generally higher among children infected with C. coli/jejuni, but Shigella-infected children suffering from dysentery recorded the highest levels (26,224 ng/mL); the lowest levels (10,625 ng/mL) were among asymptomatic children infected with other Campylobacter. Adjusting for age, sex, and Shigella infection, dysentery was significantly associated with C. coli/jejuni but not with other Campylobacter, whereas severe diarrhea was significantly associated with both C. coli/jejuni and other Campylobacter. Compared to asymptomatic children, children suffering from dysentery had a 14.6 odds of C. coli/jejuni infection (p-value < 0.001, 95% CI 5.5-38.7) but were equally likely to have other Campylobacter infections-odds ratio of 1.3 (0.434, 0.7-2.4). Children suffering from severe diarrhea were more likely than asymptomatic children to test positive for both C. coli/jejuni and other Campylobacter-OR of 2.8 (0.034, 1.1-7.1) and 1.9 (0.018, 1.1-3.1), respectively. Compared to the Campylobacter-free group, the odds of all diarrhea given C. coli/jejuni infection and other Campylobacter infection were 8.8 (<0.001, 3.0-25.7) and 2.4 (0.002, 1.4-4.2), respectively. Eliminating other Campylobacter in this population would eliminate 24.9% of the diarrhea cases, which is almost twice the population attributable fraction of 15.1% due to C. coli/jejuni. CONCLUSIONS/SIGNIFICANCE: Eighty-seven percent of the dysentery and 59.5% of the severe diarrhea samples were positive for Campylobacter, Shigella, or both, emphasizing the importance of targeting these pathogens to limit the impact of dysentery and severe diarrhea in children. Notably, the higher prevalence of other Campylobacter compared to C. coli/jejuni, their increasing burden during early childhood, and their association with severe diarrhea highlight the importance of these non-C. coli/jejuni Campylobacter species and suggest a need to clarify their importance in the etiology of clinical disease across different epidemiological contexts.

Asunto(s)

Infecciones por Campylobacter/epidemiología , Infecciones por Campylobacter/microbiología , Campylobacter/patogenicidad , Diarrea/epidemiología , Diarrea/microbiología , Disentería/epidemiología , Disentería/microbiología , Biomarcadores/análisis , Campylobacter/clasificación , Campylobacter/genética , Campylobacter/aislamiento & purificación , Infecciones por Campylobacter/diagnóstico , Campylobacter coli/patogenicidad , Campylobacter jejuni/patogenicidad , Estudios de Casos y Controles , Preescolar , Estudios de Cohortes , Coinfección/diagnóstico , Coinfección/microbiología , ADN Bacteriano/análisis , Disentería Bacilar/diagnóstico , Disentería Bacilar/epidemiología , Heces/microbiología , Femenino , Humanos , Lactante , Recién Nacido , Intestinos/lesiones , Intestinos/microbiología , Masculino , Oportunidad Relativa , Perú/epidemiología , Pobreza , Prevalencia , ARN Ribosómico 16S/genética , Shigella/genética , Shigella/aislamiento & purificación , Shigella/patogenicidad

RESUMEN

Foodborne diseases are a major public health issue but their overall incidence is underestimated due to insufficient report. The present study aimed to investigate the presence of Brucella spp., Campylobacter spp. and Listeria monocytogenes in raw milk and cheese of uninspected production obtained from cattle bred on the polluted banks of the Tietê River. Generally, milk from these animals is used to prepare fresh cheese, which is then commercialized by the producers themselves or in local markets. We analyzed 81 samples consisting of 38 samples of cheeses, 15 samples of raw milk and 28 samples of water collected from the Tietê River. These samples were evaluated for the presence of the three pathogens using bacteriological methods and the conventional polymerase chain reaction (PCR), with primers specific for each bacterial genus. In the bacteriological examination, all samples were negative for Brucella spp., Campylobacter spp. and Listeria monocytogenes. In the PCR test, Brucella spp. was detected in 5/38 (13.16%) cheese samples. Campylobacter spp. was present in 18/38 (47.37%) cheese samples, 1/15 (6.66%) raw milk samples and in 12/28 (42.86%) water samples. Listeria monocytogenes was not detected by PCR. The detection of Brucella spp. DNA in cheese and Campylobacter spp. DNA in cheese, milk and water may reflect inadequate animal sanitary management and deficiencies in goodmanufacturing practices. The presence of these pathogens in the food and water may pose a threat to thehealth of the consumer and increase the incidence of zoonosis.(AU)

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As doenças transmitidas por alimentos (DTA) ainda são subnotificadas, sendo difícil estimar a incidência global. O presente estudo objetivou detectar a presença de Brucella spp., Campylobacter spp. e Listeria monocytogenes em queijo de produção informal e leite cru, provenientes de bovinos criados às margens poluídas do Rio Tietê. O leite desses animais era utilizado na preparação de queijos tipo frescal, comercializados pelos próprios produtores ou no comércio local. No total, 81 amostras foram analisadas: 38 de queijos, 15 de leite cru e 28 alíquotas de água do Rio Tietê. Os materiais foram processados por métodos bacteriológicos e pela reação da polimerase em cadeia (PCR) convencional, específicos para cada gênero. Todas as amostras foram negativas no exame bacteriológico para Brucella spp., Campylobacter spp. e Listeria monocytogenes. Pela PCR, Brucella spp. foi detectada em 5/38 (13,16%) queijos de produção informal; Campylobacter spp. em 18/38 (47,37%) queijos, 1/15 (6,66%) leite cru e em 12/28 (42,86%) amostras das águas analisadas. Listeria monocytogenes não foi detectada pela PCR. A detecção da presença de DNA de Brucella spp. no queijo e de DNA de Campylobacter spp. no queijo, leite e água, podem indicar manejo sanitário inadequado dos animais e deficiência nas boas práticas de fabricação, com potencial risco para a saúde do consumidor e, aumentando assim, o risco da disseminação de zoonoses.(AU)

Asunto(s)

Leche/microbiología , Brucella/clasificación , Campylobacter/clasificación , Listeria monocytogenes/clasificación , Queso/microbiología

RESUMEN

Infections caused by Campylobacter species pose a severe threat to public health worldwide. However, in Grenada, the occurrence and characteristics of Campylobacter in food animals, including pigs, remain mostly unknown. In this study, we identified the sequence types (STs) of Campylobacter from young healthy pigs in Grenada and compared the results with previous studies in Grenada and other countries. Antimicrobial resistance patterns and diversity of the Campylobacter clones were evaluated. Ninety-nine Campylobacter isolates (97 Campylobacter coli and 2 Campylobacter jejuni) were analyzed by multilocus sequence typing. Eighteen previously reported STs and 13 novel STs were identified. Of the 18 previously reported STs, eight STs (ST-854, -887, -1068, -1096, -1445, -1446, 1556, and -1579) have been associated with human gastroenteritis in different geographical regions. Among these 18 previously reported STs, ST-1428, -1096, -1450, and -1058 predominated and accounted for 18.2%, 14.1%, 11.1%, and 8.1% of all isolates, respectively. Of the 13 novel STs, ST-7675 predominated and accounted for 20% (4 of 20 isolates), followed by ST-7678, -7682, and -7691, each accounting for 10% (2 of 20 isolates). Antimicrobial resistance testing using Epsilometer test revealed a low resistance rate (1-3%) of all C. coli/jejuni STs to all antimicrobials except for tetracycline (1-10.1%). Some of the C. coli STs (13 STs, 24/99 isolates, 24.2%) were resistant to multiple antimicrobials. This is the first report on antimicrobial resistance and multidrug resistance patterns associated with Campylobacter STs recovered from swine in Grenada. This study showed that pigs in Grenada are not major reservoirs for STs of C. coli and C. jejuni that are associated with human gastroenteritis worldwide.

Asunto(s)

Infecciones por Campylobacter/epidemiología , Campylobacter/aislamiento & purificación , Gastroenteritis/epidemiología , Animales , Antibacterianos/farmacología , Campylobacter/clasificación , Campylobacter/efectos de los fármacos , Infecciones por Campylobacter/tratamiento farmacológico , Infecciones por Campylobacter/veterinaria , Campylobacter coli/efectos de los fármacos , Campylobacter coli/aislamiento & purificación , Campylobacter jejuni/efectos de los fármacos , Campylobacter jejuni/aislamiento & purificación , Clonación Molecular , ADN Bacteriano/aislamiento & purificación , Pruebas Antimicrobianas de Difusión por Disco , Farmacorresistencia Bacteriana Múltiple/genética , Gastroenteritis/tratamiento farmacológico , Gastroenteritis/microbiología , Genotipo , Grenada/epidemiología , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , Análisis de Secuencia de ADN , Porcinos , Tetraciclina/farmacología

RESUMEN

During a study on the prevalence and diversity of campylobacteria in wild birds faecal samples from the city of Valdivia (southern Chile) 17 Gram-stain-negative, curved-rod-shaped isolates, were initially identified as Campylobacter lari by PCR-RFLP. Further identification by 16S rRNA sequence analysis revealed that they formed a distinct group in the genus Campylobacter. This unique position was confirmed by the results of analysis of rpoB, atpA and cpn60 gene sequences. The average nucleotide identity between the representative strain WBE38T and the type strain of the most closely related taxon C. larisubsp.concheus (LMG 11760) was 90.8 %. The oxidase and urease activity of the novel isolates enabled them to be phenotypically differentiated from species of the genus Campylobacter with validly published names. Therefore, on the basis of phenotypic, genetic and genomic characterizations, the results of this study clearly indicate that these strains represent a novel species within the genus Campylobacter, for which the name Campylobacter ornithocola sp. nov. is proposed, with the type strain WBE38T (=CECT 9147T=LMG 29815T).

Asunto(s)

Aves/microbiología , Campylobacter/clasificación , Heces/microbiología , Filogenia , Animales , Animales Salvajes/microbiología , Técnicas de Tipificación Bacteriana , Composición de Base , Campylobacter/genética , Campylobacter/aislamiento & purificación , Chile , ADN Bacteriano/genética , Genes Bacterianos , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN

RESUMEN

Foodborne diseases are a major public health issue but their overall incidence is underestimated due to insufficient report. The present study aimed to investigate the presence of Brucella spp., Campylobacter spp. and Listeria monocytogenes in raw milk and cheese of uninspected production obtained from cattle bred on the polluted banks of the Tietê River. Generally, milk from these animals is used to prepare fresh cheese, which is then commercialized by the producers themselves or in local markets. We analyzed 81 samples consisting of 38 samples of cheeses, 15 samples of raw milk and 28 samples of water collected from the Tietê River. These samples were evaluated for the presence of the three pathogens using bacteriological methods and the conventional polymerase chain reaction (PCR), with primers specific for each bacterial genus. In the bacteriological examination, all samples were negative for Brucella spp., Campylobacter spp. and Listeria monocytogenes. In the PCR test, Brucella spp. was detected in 5/38 (13.16%) cheese samples. Campylobacter spp. was present in 18/38 (47.37%) cheese samples, 1/15 (6.66%) raw milk samples and in 12/28 (42.86%) water samples. Listeria monocytogenes was not detected by PCR. The detection of Brucella spp. DNA in cheese and Campylobacter spp. DNA in cheese, milk and water may reflect inadequate animal sanitary management and deficiencies in goodmanufacturing practices. The presence of these pathogens in the food and water may pose a threat to thehealth of the consumer and increase the incidence of zoonosis.

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As doenças transmitidas por alimentos (DTA) ainda são subnotificadas, sendo difícil estimar a incidência global. O presente estudo objetivou detectar a presença de Brucella spp., Campylobacter spp. e Listeria monocytogenes em queijo de produção informal e leite cru, provenientes de bovinos criados às margens poluídas do Rio Tietê. O leite desses animais era utilizado na preparação de queijos tipo frescal, comercializados pelos próprios produtores ou no comércio local. No total, 81 amostras foram analisadas: 38 de queijos, 15 de leite cru e 28 alíquotas de água do Rio Tietê. Os materiais foram processados por métodos bacteriológicos e pela reação da polimerase em cadeia (PCR) convencional, específicos para cada gênero. Todas as amostras foram negativas no exame bacteriológico para Brucella spp., Campylobacter spp. e Listeria monocytogenes. Pela PCR, Brucella spp. foi detectada em 5/38 (13,16%) queijos de produção informal; Campylobacter spp. em 18/38 (47,37%) queijos, 1/15 (6,66%) leite cru e em 12/28 (42,86%) amostras das águas analisadas. Listeria monocytogenes não foi detectada pela PCR. A detecção da presença de DNA de Brucella spp. no queijo e de DNA de Campylobacter spp. no queijo, leite e água, podem indicar manejo sanitário inadequado dos animais e deficiência nas boas práticas de fabricação, com potencial risco para a saúde do consumidor e, aumentando assim, o risco da disseminação de zoonoses.

Asunto(s)

Brucella/clasificación , Campylobacter/clasificación , Leche/microbiología , Listeria monocytogenes/clasificación , Queso/microbiología

RESUMEN

The aim of this study was to evaluate the relationship among nutritional status, gingival health and the composition of oral microbiota in children of a public school from a very poor area of San Miguel de Tucuman. Forty-five children ranging in age from 6 to 14 years old, 13 males and 32 females were studied. Twenty of these children were undernourished (Lejarraga-Morasso Table) and twenty-five were eutrophic. A clinical study that included DMF and dmf indexes, Löe Silness Plaque Index and bleeding on probing was performed. For microbiological study, saliva samples without stimulation were taken; aliquots of them were immediately placed in TAE buffer pH 7.6, adding NaOH (N and keeping at -70 °C until processed by checkerboard DNA-DNA hybridization method to check the presence of 40 oral microorganism species. Positive bleeding on probing was present in more than 80% of children, without significant differences between eutrophic and undernourished groups. Same result were obtain for the other clinical indexes (p > 0.05, Two Way ANOVA). Significant differences were found for some oral microorganism species, with a higher percentage of undernourished children harboring them. That was the case of S. gordonii (p < 0.05), Capnocitophaga gingivalis and C. ochraceae (p < 0.01 and p < 0.10, respectively), F. nucleatum ss nucleatum (p < 0.05), P. nigrescens (p < 0.10), Campylobacter gracilis (p < 0,05), and T. denticola (p < 0.10, multiple logistic regression). Significant differences were also found between children groups for E. saborreum (p < 0.001), P. acnes (p < 0.10), G. morbillorum (p < 0.05) and L. buccalis (p < 0.10). Gingivitis and bleeding on probing would not be related to nutritional status in the groups of children studied. There were significant differences for the presence of some of the main periodontal pathogen species between eutrophic and undernourished children. It would be important to study the meaning of significant differences found for the other microorganisms more deeply.

Asunto(s)

ADN Bacteriano/genética , Encía/microbiología , Gingivitis/microbiología , Desnutrición/microbiología , Microbiota/genética , Adolescente , Aggregatibacter actinomycetemcomitans/clasificación , Aggregatibacter actinomycetemcomitans/genética , Aggregatibacter actinomycetemcomitans/aislamiento & purificación , Argentina , Bacteroides/clasificación , Bacteroides/genética , Bacteroides/aislamiento & purificación , Campylobacter/clasificación , Campylobacter/genética , Campylobacter/aislamiento & purificación , Capnocytophaga/clasificación , Capnocytophaga/genética , Capnocytophaga/aislamiento & purificación , Estudios de Casos y Controles , Niño , Femenino , Fusobacterium nucleatum/clasificación , Fusobacterium nucleatum/genética , Fusobacterium nucleatum/aislamiento & purificación , Gingivitis/fisiopatología , Humanos , Masculino , Desnutrición/fisiopatología , Hibridación de Ácido Nucleico , Peptostreptococcus/clasificación , Peptostreptococcus/genética , Peptostreptococcus/aislamiento & purificación , Porphyromonas gingivalis/clasificación , Porphyromonas gingivalis/genética , Porphyromonas gingivalis/aislamiento & purificación , Saliva/microbiología

RESUMEN

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.

Asunto(s)

Humanos , Animales , Campylobacter/aislamiento & purificación , Pollos/microbiología , Carga Bacteriana/métodos , Microbiología de Alimentos , Campylobacter/clasificación , Campylobacter/genética , Técnicas de Tipificación Bacteriana/métodos , Mataderos

RESUMEN

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.

Asunto(s)

Carga Bacteriana/métodos , Campylobacter/aislamiento & purificación , Pollos/microbiología , Microbiología de Alimentos , Mataderos , Animales , Técnicas de Tipificación Bacteriana/métodos , Campylobacter/clasificación , Campylobacter/genética , Humanos

RESUMEN

Rheumatoid arthritis (RA) and periodontitis (PD) are chronic inflammatory disorders that cause bone loss. PD tends to be more prevalent and severe in RA patients. Previous experimental studies demonstrated that RA triggers alveolar bone loss similarly to PD. The aim of this study was to investigate if arthritis-induced alveolar bone loss is associated with modification in the oral microbiota. Checkerboard DNA-DNA hybridization was employed to analyze forty oral bacterial species in 3 groups of C57BL/6 mice: control (n = 12; without any challenge); Y4 (n = 8; received oral inoculation of Aggregatibacter Actinomycetemcomitans strain FDC Y4) and AIA group (n = 12; chronic antigen-induced arthritis). The results showed that AIA and Y4 group exhibited similar patterns of bone loss. The AIA group exhibited higher counts of most bacterial species analyzed with predominance of Gram-negative species similarly to infection-induced PD. Prevotella nigrescens and Treponema denticola were detected only in the Y4 group whereas Campylobacter showae, Streptococcus mitis and Streptococcus oralis were only found in the AIA group. Counts of Parvimonas micra, Selenomonas Noxia and Veillonella parvula were greater in the AIA group whereas Actinomyces viscosus and Neisseira mucosa were in large proportion in Y4 group. In conclusion, AIA is associated with changes in the composition of the oral microbiota, which might account for the alveolar bone loss observed in AIA mice.

Asunto(s)

Pérdida de Hueso Alveolar/microbiología , Proceso Alveolar/microbiología , Artritis Experimental/microbiología , Maxilar/microbiología , Microbiota/genética , Periodontitis/microbiología , Aggregatibacter actinomycetemcomitans/clasificación , Aggregatibacter actinomycetemcomitans/genética , Aggregatibacter actinomycetemcomitans/aislamiento & purificación , Pérdida de Hueso Alveolar/patología , Proceso Alveolar/patología , Animales , Artritis Experimental/patología , Campylobacter/clasificación , Campylobacter/genética , Campylobacter/aislamiento & purificación , ADN Bacteriano/genética , Humanos , Masculino , Maxilar/patología , Ratones , Ratones Endogámicos C57BL , Boca/microbiología , Boca/patología , Periodontitis/patología , Prevotella nigrescens/clasificación , Prevotella nigrescens/genética , Prevotella nigrescens/aislamiento & purificación , Streptococcus mitis/clasificación , Streptococcus mitis/genética , Streptococcus mitis/aislamiento & purificación , Streptococcus oralis/clasificación , Streptococcus oralis/genética , Streptococcus oralis/aislamiento & purificación , Treponema denticola/clasificación , Treponema denticola/genética , Treponema denticola/aislamiento & purificación

RESUMEN

In 2006, the International Organization for Standardization (ISO) published two standard protocols for the detection and enumeration of Campylobacter spp. in foods: the ISO 10272-2(direct method) and the ISO 10272-1 (enrichment method). The aim of this study was to compare the efficiency of these two methods in the detection of Campylobacter spp. and propose a modification in the enrichment method. Compared with the enrichment method, the direct method yielded a higher number of positive results for Campylobacter spp. and, consequently, presented higher sensitivity percentage. The isolation of Campylobacter spp. was more difficult when 10 mL of rinse was used in the enrichment method, which is currently recommended by the ISO protocol. Therefore, different rinse volumes were tested (2.5 and 5 mL). The most efficient recovery of Campylobacter spp. occurred when 2.5 mL of rinse were used in the enrichment method, most likely due to a lower number of microbial contaminants than that present in the 5 or 10 mL rinses. The proposed modification of the enrichment method will contribute to the food analysis by improving the detection of Campylobacter spp. in chicken carcass.

Asunto(s)

Animales , Alimentos Fortificados , Alimentos Fortificados/análisis , Pollos , Campylobacter/clasificación , Campylobacter/química

RESUMEN

In 2006, the International Organization for Standardization (ISO) published two standard protocols for the detection and enumeration of Campylobacter spp. in foods: the ISO 10272-2(direct method) and the ISO 10272-1 (enrichment method). The aim of this study was to compare the efficiency of these two methods in the detection of Campylobacter spp. and propose a modification in the enrichment method. Compared with the enrichment method, the direct method yielded a higher number of positive results for Campylobacter spp. and, consequently, presented higher sensitivity percentage. The isolation of Campylobacter spp. was more difficult when 10 mL of rinse was used in the enrichment method, which is currently recommended by the ISO protocol. Therefore, different rinse volumes were tested (2.5 and 5 mL). The most efficient recovery of Campylobacter spp. occurred when 2.5 mL of rinse were used in the enrichment method, most likely due to a lower number of microbial contaminants than that present in the 5 or 10 mL rinses. The proposed modification of the enrichment method will contribute to the food analysis by improving the detection of Campylobacter spp. in chicken carcass.(AU)

Asunto(s)

Animales , Pollos , Alimentos Fortificados/análisis , Alimentos Fortificados , Campylobacter/química , Campylobacter/clasificación

RESUMEN

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...

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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...

Asunto(s)

Animales , Campylobacter/clasificación , Campylobacter/ultraestructura , Fluoroquinolonas/inmunología , Galliformes/inmunología , Mutación , Reacción en Cadena de la Polimerasa Multiplex/veterinaria , Resistencia a Medicamentos/inmunología

RESUMEN

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.(AU)

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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.(AU)

Asunto(s)

Animales , Campylobacter/clasificación , Campylobacter/ultraestructura , Galliformes/inmunología , Reacción en Cadena de la Polimerasa Multiplex/veterinaria , Resistencia a Medicamentos/inmunología , Mutación