Airborne pathogens can cause infections within parrot (Psittaciformes) and pigeon (Columbiformes) holdings and, in the case of zoonoses, can even spread to humans. Air sampling is a useful, noninvasive method which can enhance the common sampling methods for detection of microorganisms in bird flocks. In this study, fecal and air samples were taken from four parrot holdings. Additionally, cloacal and oropharyngeal swabs as well as air samples were taken from 15 racing pigeon holdings. Parrots were examined for psittacine beak and feather disease virus (PBFDV), proventricular dilatation disease virus (PDDV), adenoviruses (AdVs), avian paramyxovirus type-1 (APMV-1), avian influenza virus (AIV), Chlamydia psittaci (CP), and Mycobacterium avium complex (MAC). MAC and AdVs were detected in three parrot holdings, CP was detected in two parrot holdings, and PBFDV and PDDV were each detected in one parrot holding. Pigeons were examined for the pigeon circovirus (PiCV), AdVs, and CP; PiCV and AdVs were detected in all investigated pigeon holdings and CP was detected in five pigeon holdings.
Background: Clostridioides difficile is an important human and animal intestinal pathogen. Because of increasing indications of an association between C. difficile and food, in 2015, the Administration of the Republic of Slovenia for Food Safety, Veterinary Sector and Plant Protection (UVHVVR) included C. difficile in its national food surveillance. Aim: We aim to report the results and experience with a nationwide and longterm testing of food for C. difficile as a part of a regular national food surveillance programme. Methods: Retail minced meat and meat preparations (beef, pork and poultry) were sampled within a three-year period, 2015 to 2017. Selected raw retail vegetables, leaf salads and root vegetables, and ready-to-eat salads were only sampled during 2016 and 2017. Seafood was only sampled in 2017. Results: Altogether, 434 samples were tested, with 12 of 336 (3.6%) meat samples and 6 of 98 (6.1%) raw vegetables contaminated with C. difficile. Twelve of 18 recovered food isolates were toxigenic (toxinotypes 0, III, V, XII). The isolates belonged to 13 different PCR ribotypes, 001 being most common (5 isolates). Several food types with an increased potential of being contaminated with C. difficile were detected by surveillance. Conclusion: The three-year C. difficile testing within the national food surveillance revealed a low proportion of C. difficile-contaminated food and high genotype variability. Because the risk of C. difficile infection associated with C. difficile-contaminated food is unknown, no measures were recommended in the case of positive results.
There is no recommended protocol for detecting and isolating Clostridium difficile present in food samples. Here, we have evaluated the recovery of C. difficile in meat samples after incubating them in various enrichment broths. The media were as follows: cycloserine-cefoxitin fructose broth supplemented with taurocholic acid, d-cycloserine, cefoxitin, and lysozyme; cycloserine-cefoxitin mannitol broth with taurocholate and lysozyme; and cycloserine-cefoxitin fructose broth supplemented with taurocholic acid, C. difficile moxalactam norfloxacin selective supplement, and lysozyme. Samples were inoculated with various strains and quantities of C. difficile and then enriched in the different broths for 1, 4, and 7 days. C. difficile was isolated on agar plates and detected with quantitative real-time PCR (qPCR). The procedure using enrichment in cycloserine-cefoxitin fructose broth supplemented with taurocholic acid, d-cycloserine, cefoxitin, and lysozyme and incubation for 4 days for qPCR detection and 7 days for isolation (plating on C. difficile agar base with added C. difficile selective supplement and 7% [v/v] defibrinated horse blood after alcoholic shock and centrifugation) was validated. Samples of different kinds of meat and meat preparation were contaminated and used for validation of the chosen protocol. The sensitivity of detection with qPCR was 100%, and the sensitivity of the isolation method was 96%.
Purpose The purpose of this paper is to study the microbiological quality of raw milk delivered by 17 vending machines (VM) owned by different Slovenian milk producers. Design/methodology/approach For the determination of hygiene-technical conditions of VM, an observation list that included criteria for estimation of hygiene-technical suitability was made. A total of 51 milk samples were collected in three different seasons. The swabs and the cleaning liquid (eluates) of dispensing nozzles and chambers were also sampled. The main groups of microorganisms were determined by colony count technique according to international standards in all collected samples. Findings The aerobic colony count was higher than 100,000 CFU/mL in 20 (39.2 per cent) of milk samples. Its mean value was 4.8 log10 CFU/mL. The mean values of Enterobacteriaceae, psychrotrophic microorganisms, lipolytes, proteolytes, yeasts and moulds together, coagulase-positive staphylococci and somatic cell count were 3.3 log10 CFU/mL, 4.1 log10 CFU/mL, 3.2 log10 CFU/mL, 3.9 log10 CFU/mL, 2.2 log10 CFU/mL, 2.8 log10 CFU/mL and 5.3 log10 cells/mL, respectively. E. coli was found in 33.3 per cent of milk samples, while Listeria monocytogenes and antibiotics were not detected. The inner surface contamination of the dispensing nozzles and chambers was estimated in the range from 1.8 log10 CFU to 6.0 log10 CFU/cm2. The presence of detergents and disinfectants in supply valve eluates was determined in more than one-third of the samples. The hygienic-technical conditions of observed VM show some deviations from specified hygienic-technical requirements which could influence the safety of raw milk. Research limitations/implications The data about construction and the cleaning practice of VM, included in the experiment, were not available during the inspection facility. Originality/value In the paper the pathogenic and also the spoilage microorganisms in milk in the combination with hygienic conditions of inside surfaces of VM were studied.
The aim of this study was to determine the prevalence of Vibrio parahaemolyticus in shellfish samples harvested along the Slovenian coast. Shellfish samples of Mediterranean mussels (Mytilus galloprovincialis) were collected along the Slovenian coast at four locations (Seča, Piran, Strunjan and Debeli Rtič) between 2006 and 2008. Samples were examined and analysed for the presence of V. parahaemolyticus by conventional and molecular methods. The presence of Vibrio in the samples was examined by conventional methods on plate grown bacterial cells before and after enrichment in alkaline saline peptone water (ASPW). PCR methods were used for the detection of V. parahaemolyticus-specific toxR and tlh genes and of the virulence-associated tdh and trh genes. Out of 168 samples examined, 24 were positive for toxR and tlh genes by PCR from enrichment broth. Five out of 62 (8.1%), 4 out of 32 (12.5%) and 15 out of 74 (20.2%) samples were positive in 2006, 2007 and 2008, respectively. Colonies of V. parahaemolyticus were isolated from only one sample positive for V. parahaemolyticus by PCR.
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