Molecular screening of gastric Helicobacter pullorum recovered from different avian species in Egypt.

Helicobacter pullorum (H. pullorum) is a bacterium that colonizes the intestines of poultry and causes gastroenteritis. Because these species are known as human and/or animal pathogens, identification of H. pullorum is becoming increasingly necessary. The bacterium has been linked to colitis and hepatitis in humans after being transmitted by infected meat consumption. Misdiagnosis of other enteric zoonotic pathogens such as Campylobacter and other Helicobacter species makes the diagnosis of H. pullorum extremely difficult. This study focused on the molecular detection of H. pullorum from the stomach (proventriculus and gizzard) of different avian species as new target organs for detection and transmission between avian species. Proventriculus and gizzards were obtained from 40 freshly dead chickens and resident wild birds (n=40). Diarrhea was found in the farms that were surveyed. DNA was extracted from all collected samples to conduct PCR amplification. The samples were screened for Helicobacter genus-specific 16s using C97 and C05 primers. To confirm the existence of H. pullorum, the positive samples were sequenced. H. pullorum was recorded in two out of 40 chicken samples. In addition, H. pullorum was recorded in one out of 40 resident wild birds. The 16S rRNA gene sequence for Helicobacter genus-specific in poultry and wild birds showed a 100% homology. In conclusion, broiler chickens and resident wild birds are possible reservoirs for H. pullorum, according to this report, and possibly act as a source of infection for humans via the food supply.

Rat as an overlooked reservoir for Coxiella burnetii: A public health implication.

Q fever is a zoonosis with a mounting public health concern throughout the world. Rodents have been assumed to be a potential reservoir for Coxiella burnetii, a bacterium which causes Q fever. The current study was carried out to investigate the possible role of rats in the epidemiology of such disease. For this purpose, fecal samples were collected from 75 rats (55 Rattus norvegicus and 20 Rattus rattus) trapped from Giza governorate, Egypt. DNAs were extracted and samples were examined for the presence of C. burnetii using nested PCR technique. Out of examined rats, 5 yielded C. burnetii in their feces with an overall prevalence 6.7%, whereas the prevalence rates among R. norvegicus and R. rattus were (2/55) 3.6% and (3/20) 15% respectively. In addition, the phylogenetic analysis of three selected amplicons (2 R. rattus and one R. norvegicus) revealed that these sequences were highly related to each others and to those detected among humans. In conclusion, the results of the current study point out the role of rats as a potential reservoir for C. burnetii.

Epidemiological studies on Clostridium perfringens food poisoning in retail foods.

Clostridium perfringens is an important anaerobic pathogen causing food-borne gastrointestinal (GI) diseases in humans and animals. Meat and meat products are the most common vehicles of C. perfringens type A food poisoning. Contamination of meat by the intestinal contents of slaughtered animals may serve as an important source of this pathogen to the food supply. One hundred and fifty-five non-outbreak food samples were obtained from meat and retail food and examined for the presence of C. perfringens. Multiplex polymerase chain reaction assay to determine the toxin genotype of C. perfringens isolates, and extraction and purification of C. perfringens enterotoxin from enterotoxin gene (cpe)-positive isolates were carried out. The homogeneity of the purified enterotoxin was demonstrated by polyacrylamide gel electrophoresis. In addition, stool samples were collected from 150 persons who had been in contact with animals, and enzyme-linked immunosorbent assays were carried out for the qualitative determination of C. perfringens enterotoxin in the stool samples. The results demonstrated that approximately 2.6% of the tested meat and retail meat samples were contaminated with cpe-positive C. perfringens. The recommended laboratory criteria used to implicate C. perfringens in food-borne disease should involve the detection of C. perfringens enterotoxin production or the presence of the cpe gene in foods or faeces, or in the suspected C. perfringens isolates. In the present study some isolates such as tuna contained the enterotoxin gene although they had a low count of C. perfringens.

Clostridium perfringens est une bactérie anaérobie majeure responsable de toxi-infections alimentaires à symptomatologie gastro-intestinale chez l'homme comme chez l'animal. La viande et les produits carnés contaminés sont les sources les plus fréquentes des toxi-infections dues aux C. perfringens de type A. La contamination de la viande par le contenu intestinal des animaux abattus est une voie de pénétration majeure de la bactérie dans la chaîne alimentaire. Lors d'une étude visant à déterminer la présence de C. perfringens en dehors des épisodes de toxi-infection, 155 échantillons de viande et de denrées alimentaires vendues au détail ont été analysés. Les échantillons ont été soumis à une amplification en chaîne par polymérase multiplex afin de caractériser les toxinotypes des C. perfringens isolées ; l'entérotoxine de C. perfringens a été extraite et purifiée à partir des isolats possédant le gène de l'entérotoxine (cpe). L'homogénéité de l'entérotoxine purifiée a été mise en évidence par électrophorèse sur gel de polyacrylamide. En outre, 150 échantillons de selles provenant de personnes en contact avec des animaux ont été collectés et soumis à une épreuve immuno-enzymatique en vue d'une évaluation qualitative de la présence d'entérotoxines produites par C. perfringens dans ces échantillons. Il ressort de l'étude qu'environ 2,6 % des échantillons de viande et de denrées alimentaires vendues au détail étaient contaminés par des C. perfringens possédant le gène cpe. Les procédures de laboratoire recommandées pour incriminer C. perfringens en cas de toxi-infection alimentaire devraient porter sur la mise en évidence de la production d'entérotoxines par C. perfringens ou sur la détection du gène cpe dans les aliments ou les fèces ou dans les isolats présumés de C. perfringens. Dans l'étude présentée par les auteurs, certains échantillons (notamment de thon) contenaient le gène de l'entérotoxine malgré un faible taux de contamination par C. perfringens.

Clostridium perfringens es un importante patógeno anaeróbico, causante de enfermedades gastrointestinales de transmisión alimentaria en el hombre y los animales. La carne y los productos cárnicos son el vehículo más frecuente de las intoxicaciones alimentarias por C. perfringens de tipo A. La contaminación de la carne por el contenido intestinal de los animales sacrificados puede ser una importante vía de entrada de este patógeno en el suministro alimentario. A partir de carne y alimentos vendidos al por menor se obtuvieron 155 muestras alimentarias no relacionadas con brote alguno, que fueron analizadas para detectar la eventual presencia de C. perfringens. Para determinar el genotipo toxínico de los C. perfringens aislados se empleó una reacción en cadena de la polimerasa (PCR) múltiple. Después se procedió a extraer y purificar la enterotoxina (cpe) a partir de los C. perfringens positivos para el gen de que la codifica. Por electroforesis en gel de poliacrilamida se comprobó la homogeneidad de la enterotoxina purificada. Por otro lado, se obtuvieron muestras fecales de 150 personas que estaban en contacto con los animales y se practicaron ensayos inmunoenzimáticos para determinar, cualitativamente, la presencia de la enterotoxina de C. perfringens en dichas muestras. Los resultados pusieron de manifiesto que alrededor de un 2,6% de la carne analizada y de la carne vendida al por menor estaba contaminada por organismos C. perfringens cpe-positivos. Para discernir la intervención de C. perfringens en una enfermedad de transmisión alimentaria, los criterios de laboratorio recomendados pasan por detectar la producción de la enterotoxina de C. perfringens o la presencia del gen cpe en heces, alimentos o muestras sospechosas de contener C. perfringens. En el estudio descrito por los autores algunas muestras, como las de atún, contenían el gen de la enterotoxina, aunque presentaban un bajo recuento de organismos C. perfringens.