Institute for Milk Hygiene

Vienna, Austria

Institute for Milk Hygiene

Vienna, Austria

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Schoder D.,Institute for Milk Hygiene | Skandamis P.,Agricultural University of Athens | Wagner M.,Institute for Milk Hygiene
International Journal of Food Microbiology | Year: 2013

A cluster of 34 cases of listeriosis was traced to consumption of quargel cheese, a sour milk specialty, in Austria, Germany and Czech Republic between 2009 and 2010. After recall from the retail market all soft cheese batches (n=18) were sent for investigation and ISO 11290 based microbiological analysis revealed all red smear-ripened batches (16/18) to be positive for Listeria monocytogenes whereas mold ripened cheeses were negative. The 16 positive batches were grouped into three categories: those having exceeded shelf-life (G1), those around shelf-life (±4days, G2) and those within shelf-life (G3). Tracing the contamination levels as measured after recall (CLR) to the theoretical contamination level after processing (CL0) was considered to provide an estimate as to whether the in-house monitoring system would have been capable of unraveling the contamination scenario. Growth simulations starting from various hypothetical initial contamination levels of cheese at the plant and considering the potential variability in growth of L. monocytogenes due to model parameters and storage conditions suggested that a very low initial contamination level (e.g., <1CFU/g or 5CFU/100g) could justify the levels of L. monocytogenes enumerated in recalled samples of G1 and G2 lots. This in turn, may have resulted in low detection probability using ISO 11290:1996. In lots of G3 group, however, high initial contamination levels or temperature abuse at retail are inferred, based on simulated outputs. © 2013 Elsevier B.V.


Jobstl M.,Austrian Agency for Health and Food Safety AGES | Heuberger S.,IMED AGES Graz | Indra A.,IMED AGES Vienna | Nepf R.,Austrian Agency for Health and Food Safety AGES | And 2 more authors.
International Journal of Food Microbiology | Year: 2010

Prevalence of Clostridium difficile was examined in Austrian ground meat samples and bactofugates, following an evaluation of enrichment broths. Bactofugation is a centrifugation procedure used at sensitive dairies to lower the concentration of spores in raw milk before heat treatment. Among the five enrichment broths tested, C. difficile moxalactam norfloxacin boullion (CDMN) was the only one that allowed recovery of C. difficile from artificially spiked meat samples. Use of Tween 80 as a detergent in the enrichment of artificially contaminated bactofugates improved recovery of C. difficile. Following the enrichment procedures (meat without the use of TWEEN 80), one hundred ground meat samples and fifty bactofugates were enriched for 10-15 days in CDMN and presumed positive colonies were isolated and identified by Gram staining, observation of colony fluorescence and ID 32 A ribotyping. Subsequently PCR ribotyping, PCR-based identification of toxin genes (tcdA, tcdB) and antimicrobial drug susceptibility testing to metronidazole, vancomycin, clindamycin and moxifloxacin were performed. C. difficile was isolated from three (3%) of the one hundred retail ground meat samples. Two C. difficile isolates of the same rare ribotype AI-57 were toxin gene-negative and sensitive to all antibiotics tested. One isolate was assignable to one of the most prevalent clinical ribotypes isolated in Austria and harboured the tcdA and tcdB genes. This isolate was also resistant to clindamycin and moxifloxacin. None of the fifty bactofugates tested were positive for C. difficile. The presence of an isolate of human origin could indicate contamination by human shedders during food processing rather than evidencing zoonotic potential. Bactofugates, although constituting concentrated spore suspensions, were not contaminated with C. difficile spores. This finding excludes raw milk as a major source of food contamination. In conclusion, C. difficile recovery rates found in our study were lower than expected from the literature. Sources other than zoonotic origin must be considered when studying the epidemiology of community acquired infections with this bacterium. © 2010 Elsevier B.V. All rights reserved.


Allerberger F.,Austrian Agency for Health and Food Safety AGES | Wagner M.,Institute for Milk Hygiene
Clinical Microbiology and Infection | Year: 2010

Listeria monocytogenes is the causative agent of human listeriosis, a potentially fatal foodborne infection. Clinical manifestations range from febrile gastroenteritis to more severe invasive forms, including sepsis, meningitis, rhombencephalitis, perinatal infections, and abortions. In recent years, an increasing rate of listeriosis has been reported in several European countries. These increases primarily reflect a higher rate of bacteraemic listeriosis in those ≥65 years of age, and are not otherwise correlated with geography, gender, ethnicity, socioeconomic factors or infectious serotypes. In the late 1980s, an upsurge in listeriosis rates was due to the contamination of a small number of food products. However, a restricted range of strains was responsible for most of the additional cases at that time, and no evidence exists for such a pattern since 2001. From a clinical perspective, the importance of isolating the pathogen as a prerequisite for an accurate epidemiological investigation and ultimately stopping transmission cannot be overemphasized. © 2009 The Authors. Journal Compilation © 2009 European Society of Clinical Microbiology and Infectious Diseases.


Wetzels S.U.,Institute of Animal Nutrition and Functional Plant Compounds | Wetzels S.U.,Institute for Milk Hygiene | Wetzels S.U.,Research Cluster Animal Gut Health | Mann E.,Institute for Milk Hygiene | And 11 more authors.
Journal of Dairy Science | Year: 2015

Ecological balance in the rumen is highly sensitive to concentrate-rich diets. Yet the effects of these feeding practices on the caprine bacterial epimural microbiome (CBEM), a microbial community with putative important physiological functions in the rumen, are largely unexplored. This study aimed to investigate the effect of dietary concentrate amount on ruminal CBEM. Seventeen growing goats were fed diets with 0 [n = 5; 6.2. MJ of metabolizable energy (ME)/d], 30 (n = 6; 7.3. MJ of /d), or 60% (n = 6; 10.2. MJ of ME/d) concentrate for 6 wk. Two hours after their last feeding, goats were euthanized and tissue samples of the ventral rumen wall were collected, washed in phosphate-buffered saline to detach loosely attached bacteria, and stored at -20°C for further processing. Genomic DNA was isolated from thawed rumen mucosa samples and used for Roche/454 Life Science (Branford, CT) 16S rRNA gene amplicon pyrosequencing yielding 122,458 reads. Pyrosequencing data were clustered into 1,879 operational taxonomic units (OTU; 0.03 distance level). Pyrosequencing revealed Proteobacteria, Bacteroidetes, Firmicutes, and Spirochaetes as the most abundant phyla (97.7%). Compared with the 30% group, both the 60 and 0% concentrate groups harbored significantly more Firmicutes and SR1, respectively. On an OTU level, a Bergeriella-related OTU was most abundant in the CBEM, followed by 2 Campylobacter OTU, which responded differently to diets: 1 OTU was significantly increased whereas the other significantly decreased with highest concentrate amount in the diet. At the genus level, the 0% concentrate group harbored increased Kingella-like sequences compared with the other feeding groups. Furthermore, the 0% concentrate group tended to have more Bergeriella than the 30 and 60% concentrate groups. The genus Bergeriella was significantly decreased in the 60% feeding group compared with the other diets. In conclusion, this is the first report of CBEM using deep-sequencing methods on the genus and OTU level, and our study revealed major shifts in the CBEM in response to concentrate-rich diets with potential health relevance in goats. © 2015 American Dairy Science Association.


Reindl A.,Institute for Milk Hygiene | Reindl A.,Federal Institute for Alpine Dairying | Dzieciol M.,Institute for Milk Hygiene | Hein I.,Institute for Milk Hygiene | And 2 more authors.
Journal of Dairy Science | Year: 2014

A membrane filtration technique developed for counting butyric acid bacteria in cow milk was further developed for analysis of goat milk. Reduction of the sample volume, prolongation of incubation time after addition of proteolytic enzyme and detergent, and a novel step of ultrasonic treatment during incubation allowed filtration of goat milk even in the case of somatic cell counts (SCC) exceeding 106/mL. However, filterability was impaired in milk from goats in late lactation. In total, spore counts were assessed in 329 farm bulk goat milk samples. Membrane filtration technique counts were lower than numbers revealed by the classic most probable number technique. Thus, method-specific thresholds for milk to evaluate the risk of late blowing have to be set. As expected, the spore counts of milk samples from suppliers not feeding silage were significantly lower than the spore counts of milk samples from suppliers using silage feeds. Not only were counts different, the clostridial spore population also varied significantly. By using 16S rRNA gene PCR and gene sequencing, 342 strains from 15 clostridial species were identified. The most common Clostridium species were Clostridium tyrobutyricum (40.4%), Clostridium sporogenes (38.3%), Clostridium bifermentans (7.6%), and Clostridium perfringens (5.3%). The 2 most frequently occurring species C. tyrobutyricum and C. sporogenes accounted for 84.7% of the isolates derived from samples of suppliers feeding silage (n=288). In contrast, in samples from suppliers without silage feeding (n=55), these species were detected in only 45.5% of the isolates. © 2014 American Dairy Science Association.


PubMed | Institute for Milk Hygiene
Type: Journal Article | Journal: International journal of food microbiology | Year: 2013

A cluster of 34 cases of listeriosis was traced to consumption of quargel cheese, a sour milk specialty, in Austria, Germany and Czech Republic between 2009 and 2010. After recall from the retail market all soft cheese batches (n = 18) were sent for investigation and ISO 11290 based microbiological analysis revealed all red smear-ripened batches (16/18) to be positive for Listeria monocytogenes whereas mold ripened cheeses were negative. The 16 positive batches were grouped into three categories: those having exceeded shelf-life (G1), those around shelf-life ( 4 days, G2) and those within shelf-life (G3). Tracing the contamination levels as measured after recall (CLR) to the theoretical contamination level after processing (CL0) was considered to provide an estimate as to whether the in-house monitoring system would have been capable of unraveling the contamination scenario. Growth simulations starting from various hypothetical initial contamination levels of cheese at the plant and considering the potential variability in growth of L. monocytogenes due to model parameters and storage conditions suggested that a very low initial contamination level (e.g., <1 CFU/g or 5 CFU/100g) could justify the levels of L. monocytogenes enumerated in recalled samples of G1 and G2 lots. This in turn, may have resulted in low detection probability using ISO 11290:1996. In lots of G3 group, however, high initial contamination levels or temperature abuse at retail are inferred, based on simulated outputs.


PubMed | University of Veterinary Medicine Vienna, Institute for Veterinary Public Health, Institute for Milk Hygiene and Institute of Animal Nutrition and Functional Plant Compounds
Type: | Journal: Journal of dairy science | Year: 2017

Subacute ruminal acidosis (SARA) is a prevalent metabolic disorder in cattle, characterized by intermittent drops in ruminal pH. This study investigated the effect of a gradual adaptation and continuously induced long-term SARA challenge diet on the epimural bacterial community structure in the rumen of cows. Eight rumen-cannulated nonlactating Holstein cows were transitioned over 1 wk from a forage-based baseline feeding diet (grass silage-hay mix) to a SARA challenge diet, which they were fed for 4 wk. The SARA challenge diet consisted of 60% concentrates (dry matter basis) and 40% grass silage-hay mix. Rumen papillae biopsies were taken at the baseline, on the last day of the 1-wk adaptation, and on the last day of the 4-wk SARA challenge period; ruminal pH was measured using wireless sensors. We isolated DNA from papillae samples for 16S rRNA gene amplicon sequencing using Illumina MiSeq. Sequencing results of most abundant key phylotypes were confirmed by quantitative PCR. Although they were fed similar amounts of concentrate, cows responded differently in terms of ruminal pH during the SARA feeding challenge. Cows were therefore classified as responders (n = 4) and nonresponders (n = 4): only responders met the SARA criterion of a ruminal pH drop below 5.8 for longer than 330 min/d. Data showed that Proteobacteria, Firmicutes, and Bacteroidetes were the most abundant phyla, and at genus level, Campylobacter and Kingella showed highest relative abundance, at 15.5 and 7.8%, respectively. Diversity analyses revealed a significant increase of diversity after the 1-wk adaptation but a decrease of diversity and species richness after the 4-wk SARA feeding challenge, although without distinction between responders and nonresponders. At the level of the operational taxonomic unit, we detected diet-specific shifts in epimural community structure, but in the overall epimural bacterial community structure, we found no differences between responders and nonresponders. Correlation analysis revealed significant associations between grain intake and operational taxonomic unit abundance. The study revealed major shifts in the 3 dominating phyla and, most importantly, a loss of diversity in the epimural bacterial communities during a long-term SARA diet challenge, in which 60% concentrate supply for 4 wk was instrumental rather than the magnitude of the drop of ruminal pH below 5.8.


PubMed | Federal Institute for Alpine Dairying and Institute for Milk Hygiene
Type: Journal Article | Journal: Journal of dairy science | Year: 2014

A membrane filtration technique developed for counting butyric acid bacteria in cow milk was further developed for analysis of goat milk. Reduction of the sample volume, prolongation of incubation time after addition of proteolytic enzyme and detergent, and a novel step of ultrasonic treatment during incubation allowed filtration of goat milk even in the case of somatic cell counts (SCC) exceeding 10(6)/mL. However, filterability was impaired in milk from goats in late lactation. In total, spore counts were assessed in 329 farm bulk goat milk samples. Membrane filtration technique counts were lower than numbers revealed by the classic most probable number technique. Thus, method-specific thresholds for milk to evaluate the risk of late blowing have to be set. As expected, the spore counts of milk samples from suppliers not feeding silage were significantly lower than the spore counts of milk samples from suppliers using silage feeds. Not only were counts different, the clostridial spore population also varied significantly. By using 16S rRNA gene PCR and gene sequencing, 342 strains from 15 clostridial species were identified. The most common Clostridium species were Clostridium tyrobutyricum (40.4%), Clostridium sporogenes (38.3%), Clostridium bifermentans (7.6%), and Clostridium perfringens (5.3%). The 2 most frequently occurring species C. tyrobutyricum and C. sporogenes accounted for 84.7% of the isolates derived from samples of suppliers feeding silage (n=288). In contrast, in samples from suppliers without silage feeding (n=55), these species were detected in only 45.5% of the isolates.


PubMed | University of Veterinary Medicine Vienna, Institute for Milk Hygiene and Institute of Animal Nutrition and Functional Plant Compounds
Type: Journal Article | Journal: Journal of dairy science | Year: 2015

Ecological balance in the rumen is highly sensitive to concentrate-rich diets. Yet the effects of these feeding practices on the caprine bacterial epimural microbiome (CBEM), a microbial community with putative important physiological functions in the rumen, are largely unexplored. This study aimed to investigate the effect of dietary concentrate amount on ruminal CBEM. Seventeen growing goats were fed diets with 0 [n=5; 6.2MJ of metabolizable energy (ME)/d], 30 (n=6; 7.3MJ of /d), or 60% (n=6; 10.2MJ of ME/d) concentrate for 6 wk. Two hours after their last feeding, goats were euthanized and tissue samples of the ventral rumen wall were collected, washed in phosphate-buffered saline to detach loosely attached bacteria, and stored at -20C for further processing. Genomic DNA was isolated from thawed rumen mucosa samples and used for Roche/454 Life Science (Branford, CT) 16S rRNA gene amplicon pyrosequencing yielding 122,458 reads. Pyrosequencing data were clustered into 1,879 operational taxonomic units (OTU; 0.03 distance level). Pyrosequencing revealed Proteobacteria, Bacteroidetes, Firmicutes, and Spirochaetes as the most abundant phyla (97.7%). Compared with the 30% group, both the 60 and 0% concentrate groups harbored significantly more Firmicutes and SR1, respectively. On an OTU level, a Bergeriella-related OTU was most abundant in the CBEM, followed by 2 Campylobacter OTU, which responded differently to diets: 1 OTU was significantly increased whereas the other significantly decreased with highest concentrate amount in the diet. At the genus level, the 0% concentrate group harbored increased Kingella-like sequences compared with the other feeding groups. Furthermore, the 0% concentrate group tended to have more Bergeriella than the 30 and 60% concentrate groups. The genus Bergeriella was significantly decreased in the 60% feeding group compared with the other diets. In conclusion, this is the first report of CBEM using deep-sequencing methods on the genus and OTU level, and our study revealed major shifts in the CBEM in response to concentrate-rich diets with potential health relevance in goats.

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