F. van Knapen

International Commission on Trichinellosis: Recommendations on methods for the control of Trichinella in domestic and wild animals intended for human consumption.

This document provides a uniform set of recommendations for the control of Trichinella at all levels (on the farm, at slaughter and in processed meats). These recommendations are based on the best scientific information available and represent the official position of the International Commission on Trichinellosis regarding acceptable control methods. These recommendations are subject to change as new scientific information becomes available.

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.

Salmonella in slaughter pigs: prevalence, serotypes and critical control points during slaughter in two slaughterhouses

The purpose of this study was to show the distribution of Salmonella in slaughtered pigs and the environment of the slaughterhouse. 1,114 samples of slaughtered pigs (six different samples for Salmonella isolation and one serum sample for ELISA on antibodies per pig) and 477 samples of the slaughterhouse environment were collected in two slaughterhouses on two sampling days per slaughterhouse. Salmonella was isolated from one or more samples of 47% of the pigs. The highest prevalence of Salmonella was observed in rectal content samples (25.6%), whereas the lowest prevalence of Salmonella was observed on the carcasses (1.4%). The prevalence of Salmonella in other samples was: 19.6% in tonsils, 9.3% on livers, 9.3% on tongues, and 9.3% in mesenterial lymphnodes. The prevalence of Salmonella in environmental samples was high in the drain water samples in both slaughterhouses (61%) and on the carcass splitter in one slaughterhouse (33%). Salmonella typhimurium was the most frequently isolated serotype in pig samples and environmental samples in both slaughterhouses: 43% of the Salmonella isolates from pigs and 33% of the Salmonella isolates from the environment was S. typhimurium. The results of this study show that Salmonella prevalences in pigs differ a lot, depending on which part of the pig is sampled. Not all different samples of the pig will become available for human consumption, but collecting more than one sample per pig showed that Salmonella can be found in almost the whole pig. The result of surface samples of carcass and liver gives information about hygiene during the slaughter process; the result of tonsils, lymphnodes and rectal contents, combined with the serological result, gives information about infection of the pig before the slaughter process (on the farm, during transport or in lairage). It can be concluded that results of Salmonella isolation of slaughter pigs should always be carefully interpreted, depending on the type of sample that has been collected.

Survival and Resuscitation of Ten Strains of Campylobacter jejuni and Campylobacter coli under Acid Conditions

ABSTRACT The culturability of 10 strains of Campylobacter jejuni and Campylobacter coli was studied after the bacteria were exposed to acid conditions for various periods of time. Campylobacter cells could not survive 2 h under acid conditions (formic acid at pH 4). The 10 Campylobacter strains could not be recovered, even when enrichment media were used. Viable cells, however, could be detected by a double-staining (5-cyano-2,3-ditolyl tetrazolium chloride [CTC]-4′,6′-diamidino-2-phenylindole [DAPI]) technique, demonstrating that the treated bacteria changed into a viable but nonculturable (VBNC) form; the number of VBNC forms decreased over time. Moreover, some VBNC forms of Campylobacter could be successfully resuscitated in specific-free-pathogen fertilized eggs via two routes, amniotic and yolk sac injecting.

Salmonella in slaughter pigs: the effect of logistic slaughter procedures of pigs on the prevalence of Salmonella in pork.

A substantial part of the finishing pigs in the Netherlands is infected with Salmonella. Infection of pigs with Salmonella can occur already on the farm. Pigs can also get infected or contaminated during transport, lairage or slaughter. The aim of this study was to evaluate the effect of separating pigs from Salmonella-infected farms from pigs from Salmonella-free farms during transport, lairage and slaughter on the prevalence of Salmonella on pork after slaughter. Two experiments were carried out. In the first experiment, farms were selected to participate, based on serology of the pigs (Dutch Salmonella ELISA). The pigs were slaughtered at the beginning of the day: firstly, sero-negative herds, secondly, sero-positive herds and thirdly, again sero-negative herds. The latter were slaughtered to investigate the effect of a contaminated slaughterline due to a previously slaughtered positive herd. In the second experiment, farms were selected to participate, based on both serology and bacteriology of the pigs on the farm. Two hundred pigs from Salmonella-free farms were slaughtered after 200 pigs from Salmonella-infected farms. Results showed that the prevalence of Salmonella in pork samples of sero-negative herds was lower than in samples of sero-positive herds. Results also showed that Salmonella contamination of carcasses after slaughter was partially caused by Salmonella-infected herds that were slaughtered before, and partially by residential flora of the slaughterhouse. It is concluded that separate slaughter of sero-negative pig herds can be useful to decrease the prevalence of Salmonella-contaminated pork after slaughter. To avoid cross-contamination by residential flora from trucks, lairage and slaughterline, cleaning and disinfection have to be improved.

Monitoring of transmission of Salmonella enterica serovars in pigs using bacteriological and serological detection methods

The standard method to detect Salmonella positive pigs is bacteriological examination of the faeces, but in recent years the use of Salmonella-ELISAs have become available to screen pigs for serological evidence of infection. This study was conducted to monitor the transmission of five different Salmonella enterica serovars (S. Typhimurium, S. Brandenburg, S. Panama, S. Livingstone, and S. Goldcoast) in fattening pigs and to test the feasibility of Salmonella-ELISA, using seeder pigs as a mode of transmission. Serovar dependence in transmission was observed. The Salmonella-ELISA proved to be useful to detect S. Typhimurium and S. Brandenburg in herds but was of limited value to demonstrate S. Livingstone, S. Goldcoast, and S. Panama.

Antagonistic activities of several bacteria on in vitro growth of 10 strains of Campylobacter jejuni/coli

Chicken meat contaminated with Campylobacter jejuni can be the source of human enteritis. To decrease the risk of human infection, Campylobacter should be controlled at farm levels. Orally given probiotic bacteria could prevent colonisation of chicken with pathogenic bacteria like Campylobacter. The aim of this study was to investigate the effect of different bacteria on Campylobacter growth. Our results demonstrated that bacteria isolated from conventional chicken had potential inhibitory activities against Campylobacter. Other bacteria not isolated from chickens but with known antagonistic capacities, e.g. Enterococcus (56 strains) and Escherichia coli (20 strains), did not show any negative effect on Campylobacter. Interestingly, one Lactobacillus (P93) strain isolated from the chicken gut showed bactericidal activity against all tested Campylobacter. The bactericidal effect was characterised as the production of organic acids in combination with probably production of an anti-Campylobacter protein. In a co-culture study of Campylobacter and Lactobacillus (P93), the culturability of Campylobacter was under the detection limit after 48 h of incubation. A chicken experiment is needed to further evaluate the effect of the promising probiotic bacteria against Campylobacter colonisation in chicken.

Quantitative assessment of infection risk from exposure to waterborne pathogens in urban floodwater

Flooding and heavy rainfall have been associated with waterborne infectious disease outbreaks, however, it is unclear to which extent they pose a risk for public health. Here, risks of infection from exposure to urban floodwater were assessed using quantitative microbial risk assessment (QMRA). To that aim, urban floodwaters were sampled in the Netherlands during 23 events in 2011 and 2012. The water contained Campylobacter jejuni (prevalence 61%, range 14- >10(3) MPN/l), Giardia spp. (35%, 0.1-142 cysts/l), Cryptosporidium (30%, 0.1-9.8 oocysts/l), noroviruses (29%, 10(2)-10(4) pdu/l) and enteroviruses (35%, 10(3)-10(4) pdu/l). Exposure data collected by questionnaire, revealed that children swallowed 1.7 ml (mean, 95% Confidence Interval 0-4.6 ml) per exposure event and adults swallowed 0.016 ml (mean, 95% CI 0-0.068 ml) due to hand-mouth contact. The mean risk of infection per event for children, who were exposed to floodwater originating from combined sewers, storm sewers and rainfall generated surface runoff was 33%, 23% and 3.5%, respectively, and for adults it was 3.9%, 0.58% and 0.039%. The annual risk of infection was calculated to compare flooding from different urban drainage systems. An exposure frequency of once every 10 years to flooding originating from combined sewers resulted in an annual risk of infection of 8%, which was equal to the risk of infection of flooding originating from rainfall generated surface runoff 2.3 times per year. However, these annual infection risks will increase with a higher frequency of urban flooding due to heavy rainfall as foreseen in climate change projections.

Quantification of horizontal transmission of Salmonella enterica serovar Enteritidis bacteria in pair-housed groups of laying hens.

ABSTRACT An important source of human salmonellosis is the consumption of table eggs contaminated with Salmonella enterica serovar Enteritidis. Optimization of the various surveillance programs currently implemented to reduce human exposure requires knowledge of the dynamics of S. Enteritidis infection within flocks. The aim of this study was to provide parameter estimates for a transmission model of S. Enteritidis in laying-type chicken flocks. An experiment was carried out with 60 pairs of laying hens. Per pair, one hen was inoculated with S. Enteritidis and the other was contact exposed. After inoculation, cloacal swab samples from all hens were collected over 18 days and tested for the presence of S. Enteritidis. On the basis of this test, it was determined if and when each contact-exposed hen became colonized. A transmission model including a latency period of 1 day and a slowly declining infectivity level was fitted. The mean initial transmission rate was estimated to be 0.47 (95% confidence interval [CI], 0.30 to 0.72) per day. The reproduction number R0, the average number of hens infected by one colonized hen in a susceptible population, was estimated to be 2.8 (95% CI, 1.9 to 4.2). The generation time, the average time between colonization of a “primary” hen and colonization of contact-exposed hens, was estimated to be 7.0 days (95% CI, 5.0 to 11.6 days). Simulations using these parameters showed that a flock of 20,000 hens would reach a maximum colonization level of 92% within 80 days after colonization of the first hen. These results can be used, for example, to evaluate the effectiveness of control and surveillance programs and to optimize these programs in a cost-benefit analysis.

Prevalence of antibodies to toxoplasma in farm animals in the Netherlands and its implication for meat inspection

Summary A serological survey on toxoplasmosis was carried out amongst horses, sheep, cattle, pigs, and poultry in the Netherlands. Sera were obtained from slaughter animals and the results were compared with those of previous studies of this kind in the Netherlands. In horses and cattle the percentage of seropositive animals remained the same in the past 20 years. In sheep and particularly in pigs, however, a remarkable decline of seropositive animals was found. This is probably due to the age of the animals examined and a change in present day methods of farming. In poultry it was shown that only free scratching hens were seropositive in up to 30 per cent of the animals tested, whereas broilers and battery animals were seronegative. Since as yet no individual control measures in the slaughterhouse exist with regard to toxoplasmosis it is recommended to carry out periodic epizootiological surveys amongst farm animals to be able to follow trends of the infection.A serological survey on toxoplasmosis was carried out amongst horses, sheep, cattle, pigs and poultry in the Netherlands. Sera were obtained from slaughter animals and the results were compared with those of previous studies of this kind in the Netherlands. In horses and cattle the percentage of seropositive animals remained the same in the past 20 years. In sheep and particularly in pigs, however, a remarkable decline of seropositive animals was found. This is probably due to the age of the animals examined and a change in present day methods of farming. In poultry it was shown that only free scratching hens were seropositive in up to 30 per cent of the animals tested, whereas broilers and battery animals were seronegative. Since as yet no individual control measures in the slaughterhouse exist with regard to toxoplasmosis it is recommended to carry out periodic epizootiological surveys amongst farm animals to be able to follow trends of the infection.