Bert A.P. Urlings

Role of Volatile Fatty Acids in Development of the Cecal Microflora in Broiler Chickens during Growth

ABSTRACT It is known that volatile fatty acids can inhibit growth of species of the family Enterobacteriaceae in vitro. However, whether these volatile fatty acids affect bacterial populations in the ceca of chickens is unknown. Therefore, a study was conducted to investigate if changes in volatile fatty acids in ceca of broiler chickens during growth affect bacterial populations. Results showed that members of theEnterobacteriaceae and enterococci are present in large numbers in 3-day-old broilers and start to decrease when broilers grow older. Lactobacilli are present in large numbers as well in 3-day-old broilers, but they remain stable during the growth of broilers. Acetate, butyrate, and propionate increase from undetectable levels in 1-day-old broilers to high concentrations in 15-day-old broilers, after which they stabilize. Significant negative correlations could be calculated between numbers of Enterobacteriaceae and concentrations of undissociated acetate, propionate, and butyrate. Furthermore, pure cultures of Enterobacteriaceae isolated from the ceca were grown in the presence of volatile fatty acids. Growth rates and maximal optical density decreased when these strains grew in the presence of increasing volatile fatty acid concentrations. It is concluded that volatile fatty acids are responsible for the reduction in numbers of Enterobacteriaceae in the ceca of broiler chickens during growth.

Effect of Fermented Feed on the Microbial Population of the Gastrointestinal Tracts of Pigs

ABSTRACT An in vivo experiment was performed with pigs to study the inhibitory effect of fermented feed on the bacterial population of the gastrointestinal tract. Results demonstrated a significant positive correlation between pH and lactobacilli in the stomach contents of pigs in dry feed as well as in the stomach contents of pigs fed fermented feed. Furthermore, a significant positive correlation between the pH and the numbers of bacteria in the familyEnterobacteriaceae in the contents of the stomach of pigs fed dry feed was found. In the stomach contents of pigs fed fermented feed, a significant negative correlation was found between the concentration of the undissociated form of lactic acid and the numbers of Enterobacteriaceae. The numbers ofEnterobacteriaceae in the contents of the stomach, ileum, cecum, colon, and rectum of pigs fed fermented feed were significantly lower compared with the contents of the stomach, ileum, caecum, colon, and rectum of pigs fed dry feed. The numbers of total lactobacilli were significantly higher in the stomach contents of pigs fed fermented feed and in the ileum contents of one pig group fed fermented feed compared with the contents of pigs fed dry feed. However, the influence of lactobacilli on numbers of Enterobacteriaceae could not be demonstrated. It was concluded that fermented feed influences the bacterial ecology of the gastrointestinal tract and reduces the levels of Enterobacteriaceae in the different parts of the gastrointestinal tract.

Mechanism of Salmonella reduction in fermented pig feed

To protect consumers from Salmonella infection acquired through the consumption of pork meat, it is necessary to eradicate Salmonella from pork. In order to achieve this, the whole pork production chain should be free from Salmonella, including the pigs at the farm. In epidemiological studies it was concluded that the use of fermented feed plays a significant role in the reduction of Salmonella prevalence in pig farms. However, the mechanism of Salmonella reduction in fermented feed is not known. A controlled feed fermentation was performed using a pure culture of Lactobacillus plantarum, pH reduction, organic acid profiles and bacterial counts were determined. In L plantarum-fermented feed, lactic acid and acetic acid were produced and the pH dropped to a value below 4.0. Antimicrobial products (bacteriocins) could not be detected. The results showed that the produced lactic and acetic acid and the pH in the feed are responsible for Salmonella reduction in fermented feed. L plantarum did not show any other antimicrobial effect on Salmonella

Livestock-associated MRSA ST398 carriage in pig slaughterhouse workers related to quantitative environmental exposure

Objectives To assess livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) carriage among workers in pig slaughterhouses and assess associated risk factors, including occupational exposure to LA-MRSA. Methods A cross-sectional study in three Dutch pig slaughterhouses was undertaken. Nasal swabs of participants were taken. Nasal swabs and surface wipes, air and glove samples were screened for presence of methicillin-resistant Staphylococcus aureus (MRSA). MRSA was quantitatively determined on gloves and in air samples by culturing and real-time PCR. Results 11 of 341 (3.2%) participants were identified as nasal MRSA carriers. MRSA-positive workers were predominantly found at the start of the slaughter process. Major risk factors for carriage were working in the lairage and working in the scalding and dehairing area. Most nasal isolates (73%) belonged to the LA-MRSA clone ST398. MRSA ST398-positive environmental samples were found throughout the slaughter process. A clear decrease was seen along the slaughterline in the number of MRSA-positive samples and in the MRSA amount per sample. Conclusions This study showed that working in the lairage area or scalding and dehairing area were the major risk factors for MRSA carriage in pig slaughterhouse workers, while the overall prevalence of MRSA carriage is low. Occupational exposure to MRSA decreased along the slaughterline, and the risk of carriage showed a parallel decrease.

Effect of fermented feed on shedding of Enterobacteriaceae by fattening pigs

Epidemiological studies showed that the use of fermented feed could significantly reduce Salmonella prevalence in pigs compared to the use of normal feed. Experimental challenge experiments with Salmonella have however never been conducted to reveal the efficacy of fermented feed in reducing Salmonella shedding and/or reducing the number of Salmonella-positive pigs. A longitudinal study was conducted to measure the effect of fermented feed, in particular of its components lactic acid and Lactobacillus plantarum, on gastrointestinal bacterial ecology (Salmonella, Enterobacteriaceae, lactobacilli, volatile fatty acids (VFAs), pH). Seeder pigs were used as a mode for Salmonella transmission within a pig herd. Bacteriological measurements were performed in faeces of the pigs. The results showed that fermented feed affected/reduced the Enterobacteriaceae population in faeces of the pigs. No differences were found in the number of positive pigs infected or in the number of shedding with Salmonella serovar Typhimurium fed fermented feed and between the normal feed group. S. serovar Goldcoast could not establish an infection in the seeder pigs in the fermented feed group as well as in the normal feed group. The pH of the faeces in the fermented feed groups was significantly higher than the pH of the faeces of the normal feed groups. The role of the undissociated form of the faecal VFAs on the significantly lower Enterobacteriaceae number in faeces of the pigs of the fermented feed groups could not be demonstrated because of the significant higher pH in the faeces of the pigs fed fermented feed.

Effect of fermented feed on the susceptibility for Campylobacter jejuni colonisation in broiler chickens with and without concurrent inoculation of Salmonella enteritidis

Fermented liquid feed (FLF) protects broiler chickens against colonisation with Salmonella. While Campylobacter causes more disease cases in humans than Salmonella, the effect of FLF on Campylobacter was assessed. The fermented liquid feed is a moistened feed with a high number of lactobacilli, a high concentration of lactic acid, and a pH of 4. In three experiments Campylobacter was orally applied to individually housed 9-day-old broiler chickens. A significant reduction of susceptibility, as determined by cloacal swabs, was observed. At any moment where an animal has not started to shed Campylobacter yet, the probability to start shedding Campylobacter in a subsequent small time interval was nine times as high for the control chickens than for the animals that were fed FLF. FLF did not consistently change the Campylobacter colonisation level in the caeca. It was concluded that FLF could reduce the probability of introduction of Campylobacter in broiler flocks. In an experiment where some chickens were simultaneously inoculated with Salmonella enteritidis and Campylobacter, no interaction on susceptibility or caecal colonisation level was observed.

Pathogenic micro-organisms in slaughterhouse sludge—a survey

During slaughtering of animals and subsequent meat processing the process water used becomes polluted with organic matter of animal origin (i.e. protein and fat). This organic sludge is, in principle, a product suitable for animal feeding. To investigate the microbiological contamination level of sludge, raw sludge was collected at pig (n = 8) and poultry (n = 5) slaughterhouses. Both flocculated and aerobically activated sludge was monitored. Slaughterhouse sludge was heavily contaminated with Enterobacteriaceae (6.3-10.0 in log10 N/gram dry matter) and enterococci (4.6-7.9). Clostridia were present in sludge at a level of 3.1-5.8 (in log10 N/g DM). Salmonella was present in the sludge from all slaughterhouses examined. Yersinia enterocolitica serotypes O:3 and O:9 were found in sludge from seven out of thirteen slaughterhouses. The prevalence of Campylobacter jejuni/coli was higher in flocculated poultry sludge than in both flocculated pig sludge and aerobically activated pig sludge. Obviously, decontamination of the sludge is mandatory when it is to be applied as a feed constituent, to prevent bacterial cycles from occurring in livestock, as well as the spread of human pathogenic zoonoses like campylobacter, salmonella and yersinia, to minimize loss of protein quality by the microbial breakdown of amino acids and the formation of possible toxic metabolites in sludge during storage.

Passage of Salmonella through the crop and gizzard of broiler chickens fed with fermented liquid feed.

In vivo experiments were conducted in order to investigate the passage and bacterial reduction of Salmonella in the crop and gizzard of chickens when fed two different feeds. The chickens were fed dry conventional feed and fermented liquid feed. The fermented feed contains a relatively high concentration of lactic and acetic acid and lactobacilli. One and three week old broiler chickens were necropsied at short intervals after inoculation with Salmonella Enteritidis. Counts of Salmonella from the crop, gizzard, duodenum, caecum and colon/rectum were obtained. This revealed a sharper decrease of Salmonella in the anterior parts of the gastro-intestinal tract in chickens fed with fermented feed than in chickens fed dry feed. It is therefore concluded that fermented feed improves the barrier formed by the crop and gizzard. The reduction of Salmonella is fully realised in the crop and gizzard. The lower intestinal compartment did not show a substantial effect on the reduction of Salmonella. The performed in vivo method appeared to be an appropriate way to study intervention strategies that aim to control Salmonella by improving the barrier function of the upper gastro-intestinal tract.

Tracing and analytical results of the dioxin contamination incident in 2008 originating from the Republic of Ireland

High levels of dioxins (PCDD/Fs) in pork were discovered in France and the Netherlands at the end of 2008. The contamination was rapidly traced back to a feed stock in the Republic of Ireland (RoI). Burning oil, used for the drying of bakery waste, appeared to be contaminated with PCBs. Consequently, very high levels up to 500 pg TEQ g−1 fat were found in pork. The congener pattern clearly pointed to PCB-oil as a source, but the ratio between the non-dioxin-like indicator PCBs (PCBs 28, 52, 101, 138, 152 and 180) and PCDD/Fs was much lower than observed during the Belgian incident, thereby limiting the suitability of indicator PCBs as a marker for the presence of dioxins and dioxin-like PCBs. This paper describes the tracking and tracing of the incident, the public–private cooperation, the surveillance activities and its results. A major lesson to be learned from this incident is the importance of good private food safety systems. In this incident, it was the private surveillance systems that identified the origin of contamination within 10 days after the first signal of increased dioxin levels in a product. On the other hand, retrospective analyses showed that signals were missed that could have led to an earlier detection of the incident and the source. Above all, the incident would not have occurred when food safety assurance systems had been effectively implemented in the involved feed chain. It is discussed that besides primary responsibility for effective private food safety systems, the competent authorities have to supervise whether the food safety procedures are capable of coping with these kinds of complex food safety issues, while private food companies need to implement the law, and public authorities should supervise and enforce them. Finally, it is discussed whether the health risks derived from consumption of the contaminated batches of meat may have been underestimated during the incident due to the unusually high intake of dioxins.

The Prevalence of Campylobacter in Pigs during Fattening

The aim of this study was to improve our knowledge of the epidemiology of campylobacters in pigs. This can help in understanding whether it is possible to reduce the prevalence of campylobacters in the pig population and, ultimately, to produce campylobacter-free pork. For this purpose the prevalence of campylobacters in porkers in 8 farms was monitored. A selection of campylobacter isolates was characterised by a genetic typing method known as RFLP (Restriction Fragment Length Polymorphism).