Helminth parasites in black rats (Rattus rattus) and brown rats (Rattus norvegicus) from different environments in the Netherlands.

BACKGROUND: Rattus norvegicus (brown rat) and Rattus rattus (black rat) are known carriers of bacteria, viruses, and parasites of zoonotic and veterinary importance. Moreover, rats may play a role in the transmission of muscle larvae of the zoonotic nematode Trichinella spiralis to farm animals. We aimed to study the intestinal and intramuscular helminths in wild rats from three different environments to assess the relevance of rats as carrier of zoonotic parasites for public health. MATERIALS AND METHODS: Wild brown rats (117 individuals) and black rats (44 individuals) were captured at farms, in suburban and in rural environments in the Netherlands. Intestinal helminths were isolated and identified morphologically. Artificial digestion was used to isolate muscle larvae. RESULTS AND DISCUSSION: Morphological analysis of rat intestinal contents yielded six nematode species (Syphacia muris, Heterakis spumosa, Aonchotheca murissylvatici, Trichuris muris, Nippostrongylus brasiliensis, and Strongyloides sp.), three cestode species (Hymenolepis diminuta, H. nana and Hymenolepis (=Rodentolepis) fraterna), and four trematode species (Plagiorchis muris, Plagiorchis proximus, Echinostoma chloropodis, and Notocotylus imbricatus).Black rats at farms displayed the lowest intestinal helminth species variation (six species) and carried overall on average 0.93 species simultaneously. In comparison, brown rats at farms carried seven helminth species and 1.91 species simultaneously. Brown rats from suburban environments displayed the highest species variation (11 species) at 1.82 simultaneous helminth species. Absence of trematodes from rats at farms may suggest limited exchange of rats between farms and surrounding wet rural environments. We report four species of veterinary (Syphacia muris) or zoonotic relevance (Hymenolepis diminuta, Hymenolepis nana and Plagiorchis muris). We did not find Trichinella muscle larvae, consistent with long-term prevalence in Dutch wild rats.

Bayesian estimation of diagnostic accuracy of a new bead-based antibody detection test to reveal Toxoplasma gondii infections in pig populations.

The success of a Toxoplasma gondii surveillance program in European pig production systems depends partly on the quality of the test to detect infection in the population. The test accuracy of a recently developed serological bead-based assay (BBA) was investigated earlier using sera from experimentally infected animals. In this study, the accuracy of the BBA was determined by the use of sera from animals from two field subpopulations. As no T. gondii infection information of these animals was available, test accuracy was determined through a Bayesian approach allowing for conditional dependency between BBA and an ELISA test. The priors for prevalence were based on available information from literature, whereas for specificity vague non-informative priors were used. Priors for sensitivity were based either on available information or specified as non-informative. Posterior estimates for BBA sensitivity and specificity were (mode) 0.855 (Bayesian 95% credibility interval (bCI) 0.702-0.960) and 0.913 (bCI 0.893-0.931), respectively. Comparing the results of BBA and ELISA, sensitivity was higher for the BBA while specificity was higher for ELISA. Alternative priors for the sensitivity affected posterior estimates for sensitivity of both BBA and ELISA, but not for specificity. Because the difference in prevalence between the two subpopulations is small, and the number of infected animals is small as well, the precision of the posterior estimates for sensitivity may be less accurate in comparison to the estimates for specificity. The estimated value for specificity of BBA is at least optimally defined for testing pigs from conventional and organic Dutch farms.

Veterinary and public health aspects of Toxocara spp.

Pet dogs and cats can play an important role in the transmission of zoonotic nematodes such as Toxocara canis and Toxocara cati, by excreting eggs directly into the human environment, without the involvement of vectors or intermediate hosts. Human toxocarosis remains a hazard despite the availability of highly effective anthelmintics for dogs and cats. A good understanding of the biology and epidemiology of these parasites, and the risk factors that lead to their transmission to humans is required for effective prevention strategies. In this respect, the maintenance of high quality continuing education for veterinarians and the provision of suitably presented information to pet owners are of priority importance. A closer collaboration between veterinary and public health professionals within the 'One Health' concept is also required.

[Is being licked by dogs not dirty?]. / Is likken door hondentong minder vies?

Being licked by pet dogs is frequently a common advice in articles for the uninitiated. An overview is given about the special antibacterial and wound healing properties of human and canine saliva. New developments in the human area are presumably assigned to dog saliva. Because of the presence of a quite different mouth flora including various potential zoonotic pathogens, it is strictly not advised to let dogs lick the wounds or face of the human.

A novel bead-based assay to detect specific antibody responses against Toxoplasma gondii and Trichinella spiralis simultaneously in sera of experimentally infected swine.

BACKGROUND: A novel, bead-based flow cytometric assay was developed for simultaneous determination of antibody responses against Toxoplasma gondii and Trichinella spiralis in pig serum. This high throughput screening assay could be an alternative for well known indirect tests like ELISA. One of the advantages of a bead-based assay over ELISA is the possibility to determine multiple specific antibody responses per single sample run facilitated by a series of antigens coupled to identifiable bead-levels. Furthermore, inclusion of a non-coupled bead-level in the same run facilitates the determination of, and correction for non-specific binding. The performance of this bead-based assay was compared to one T. spiralis and three T. gondii ELISAs. For this purpose, sera from T. gondii and T. spiralis experimentally infected pigs were used. With the experimental infection status as gold standard, the area under the curve, Youden Index, sensitivity and specificity were determined through receiver operator curve analysis. Marginal homogeneity and inter-rater agreement between bead-based assay and ELISAs were evaluated using McNemar's Test and Cohen's kappa, respectively. RESULTS: Results indicated that the areas under the curve of the bead-based assay were 0.911 and 0.885 for T. gondii and T. spiralis, respectively, while that of the T. gondii ELISAs ranged between 0.837 and 0.930 and the T. spiralis ELISA was 0.879. Bead-based T. gondii assay had a sensitivity of 86% and specificity of 96%, while the ELISAs ranged between 64-84% and 93-99%, respectively. The bead-based T. spiralis assay had a sensitivity of 68% and specificity of 100% while the ELISA scored 72% and 95%, respectively. Marginal homogeneity was found between the T. gondii bead-based test and one of the T. gondii ELISAs. Moreover, in this test combination and between T. spiralis bead-based assay and respective ELISA, an excellent inter-rater agreement was found. When results of samples before expected seroconversion were removed from evaluation, notably higher test specifications were found. CONCLUSIONS: This new bead-based test, which detects T. gondii and T. spiralis antibodies simultaneously within each sample, can replace two indirect tests for the determination of respective antibodies separately, while performing equally well or better.

Role of pet dogs and cats in the transmission of helminthic zoonoses in Europe, with a focus on echinococcosis and toxocarosis.

The close emotional tie between people and companion animals is a beneficial relation known as the human-animal bond. However, pet dogs and cats can play an important role in the transmission of helminthic zoonotic agents such as the tapeworms Echinococcus and the roundworms Toxocara which are directly transmitted from pets to the human environment without the involvement of vectors or intermediate hosts. In humans, echinococcosis has emerged in Europe and toxocarosis is still persisting in large endemic areas despite the availability of highly efficient anthelminthics for dogs and cats. Ecological changes significantly contributed to these trends: the high wild fox populations and the high density of freely roaming dogs and cats maintain a permanent infection pressure of these and other parasites. Further, the establishment of urban recreational environments closer to natural ecological systems boosted vole populations that represent urban reservoirs for zoonotic helminths. A good understanding of the parasites' biology and epidemiology including the transmission to humans is required for planning and implementing effective prevention strategies. The continuous education of veterinarians and the information of the pet owners by providing uniform recommendations are of priority importance. A close collaboration between veterinary and public health professionals in a 'One Health' concept is required.

[Burying family pets: legislation, risks, and recommendations]. / Begraven van (gezelschaps)dieren; wetgeving, mogelijke risico's en aanbevelingen.

At one point or other all pet owners are confronted with the death of their pet. To safeguard public health and to prevent the transmission of disease, there are regulations concerning the burial of family pets. This article focuses on relevant regulations and potential risks and provides recommendations for the burial of family pets.

Zoonotic parasites in fecal samples and fur from dogs and cats in The Netherlands.

Pets may carry zoonotic pathogens for which owners are at risk. The aim of the study is to investigate whether healthy pets harbour zoonotic parasitic infections and to make an inventory of the interactions between pet-owners and their companion animals in The Netherlands. Fecal and hair samples were collected from healthy household dogs and cats in Dutch veterinary practices. Owners were interviewed about interaction with their pets. The samples were investigated by microscopy, ELISA, and PCR. From 159 households, 152 dogs (D) and 60 cats (C), information and samples were collected and examination for several zoonotic parasites was performed. Toxocara eggs were found in 4.4% (D) and 4.6% (C) of the fecal samples and in 12.2% (D) and 3.4% (C) of the fur samples. The median epg in the fur was 17 (D) and 28 (C) and none of these eggs were viable. From 15.2% of the dog and 13.6% of the cat feces Giardia was isolated. One canine and one feline Giardia isolate was a zoonotic assemblage A (12%). Cryptosporidium sp. were present in 8.7% (D) and 4.6% (C) of the feces. Fifty percent of the owners allow the pet to lick their faces. Sixty percent of the pets visit the bedroom; 45-60% (D-C) are allowed on the bed, and 18-30% (D-C) sleep with the owner in bed. Six percent of the pets always sleep in the bedroom. Of the cats, 45% are allowed to jump onto the kitchen sink. Nearly 39% of the dog owners never clean up the feces of their dog. Fifteen percent of the dog owners and 8% of the cat owners always wash their hands after contact with the animals. Close physical contact between owners and their pets is common and poses an increased risk of transmission of zoonotic pathogens. Education of owners by the vet, specifically about hygiene and potential risks, is required.

Risk analysis-based food safety policy: scientific factors versus socio-cultural factors.

The purpose of this article is to illustrate the importance of socio-cultural factors in risk management and the need to incorporate these factors in a standard, internationally recognized (WTO) framework. This was achieved by analysing the relevance of these factors in three cases. It can be concluded that the pre-eminent role of science in food-related regulatory decisions is debatable. At a risk management level, other factors, such as cultural, social, or economic issues, are often more important than scientific advice in determining policy. There is a need for transparency at an international level as trade barriers are gradually being removed and these other factors are becoming more apparent. Therefore it is important that all the factors implicated in the food safety policy-making process are recognized in a standard framework.

Inhibition of Salmonella enterica serotype Enteritidis on agar and raw chicken by carvacrol vapour.

Carvacrol is known to inhibit a number of food borne pathogens. The activity of carvacrol vapour was evaluated against S. enterica serotype Enteritidis on tryptone soya agar and on pieces of raw chicken. On agar, the size of the inhibition zone increased with decreasing volume of agar, increasing vapour exposure period, increasing temperature, increasing volume of carvacrol used and with decreasing bacterial density. Inhibition was equally effective under aerobic and anaerobic conditions. On chicken pieces (10 x 10 x 5 mm, UV-sterilized and inoculated with approx. 5 x 10(3) cfu) carvacrol vapour significantly reduced viable numbers of salmonellae at 4, 20 and 37 degrees C and all viable cells were eliminated by a minimum of 3 h at 37 degrees C (p<0.05). A minimum concentration of 20% carvacrol v/v in ethanol was required to achieve a significant reduction and from 40% v/v no viable cells were recovered (p<0.05). In conclusion, carvacrol vapour is effective at inhibiting the growth of S. enterica serotype Enteritidis on agar and at inhibiting and eliminating these bacteria on the surface of raw chicken.

Carvacrol induces heat shock protein 60 and inhibits synthesis of flagellin in Escherichia coli O157:H7.

The essential oils of oregano and thyme are active against a number of food-borne pathogens, such as Escherichia coli O157:H7. Carvacrol is one of the major antibacterial components of these oils, and p-cymene is thought to be its precursor in the plant. The effects of carvacrol and p-cymene on protein synthesis in E. coli O157:H7 ATCC 43895 cells were investigated. Bacteria were grown overnight in Mueller-Hinton broth with a sublethal concentration of carvacrol or p-cymene, and their protein compositions were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and confirmed by Western blotting. The presence of 1 mM carvacrol during overnight incubation caused E. coli O157:H7 to produce significant amounts of heat shock protein 60 (HSP60) (GroEL) (P < 0.05) and inhibited the synthesis of flagellin highly significantly (P < 0.001), causing cells to be aflagellate and therefore nonmotile. The amounts of HSP70 (DnaK) were not significantly affected. p-Cymene at 1 mM or 10 mM did not induce HSP60 or HSP70 in significant amounts and did not have a significant effect on flagellar synthesis. Neither carvacrol (0.3, 0.5, 0.8, or 1 mM) nor p-cymene (0.3, 0.5, or 0.8 mM) treatment of cells in the mid-exponential growth phase induced significant amounts of HSP60 or HSP70 within 3 h, although numerical increases of HSP60 were observed. Motility decreased with increasing concentrations of both compounds, but existing flagella were not shed. This study is the first to demonstrate that essential oil components induce HSP60 in bacteria and that overnight incubation with carvacrol prevents the development of flagella in E. coli O157:H7.

Detection of egg yolk antibodies reflecting Salmonella enteritidis infections using a surface plasmon resonance biosensor.

A surface plasmon resonance (SPR) biosensor assay was developed on the basis of a lipopolysaccharide antigen of Salmonella enterica serovar enteritidis (S. enterica serovar enteritidis) to detect egg yolk antibodies against S. enterica serovar enteritidis. This biosensor assay was compared to two commercial ELISA kits based on LPS antigen and flagellar antigen. A number of 163 egg yolk and combined egg white and yolk samples from chickens experimentally infected with S. enterica serovar enteritidis and 90 egg yolk and combined egg white and yolk samples from uninfected chickens were analyzed. Receiver operating characteristic analysis of the data calculated a diagnostic sensitivity of 82% and a diagnostic specificity of 100%. The within-day coefficient of variation of a positive internal-control egg yolk was 1%. The SPR biosensor assay was able to detect antibodies in a significantly higher percentage of known positive samples than the commercial ELISA's. The anticipated use of the SPR biosensor assay is to determine the S. enterica serovar enteritidis serostatus of non-vaccinated layer hens.

Surface plasmon resonance biosensors for detection of pathogenic microorganisms: strategies to secure food and environmental safety.

This review describes the exploitation of exclusively optical surface plasmon resonance (SPR) biosensors for the direct and indirect detection of pathogenic microorganisms in food chains and the environment. Direct detection is, in most cases, facilitated by the use of defined monoclonal or polyclonal antibodies raised against (a part of) the target pathogenic microorganisms. The antibodies were immobilized to a solid phase of the sensor to capture the microbe from the sample. Alternatively, antibodies were used in an inhibition-like assay involving incubation with the target organism prior to analysis of nonbound antibodies. The free immunoglobins were screened on a sensor surface coated with either purified antigens or with Fc or Fab binding antibodies. Discussed examples of these approaches are the determination of Escherichia coli O1 57:H7, Salmonella spp., and Listeria monocytogenes. Another direct detection strategy involved SPR analysis of polymerase chain reaction products of Shiga toxin-2 genes reporting the presence of E. coli O157:H7 in human stool. Metabolic products have been exploited as biomarkers for the presence of a microbial agent, such as enterotoxin B and a virulence factor for the occurrence of Staphylococcus aureus and Streptococcus suis, respectively. Indirect detection, on the other hand, is performed by analysis of a humoral immune response of the infected animal or human. By immobilization of specific antigenic structures, infections with Herpes simplex and human immunodeficiency viruses, Salmonella and Treponema pallidum bacteria, and Schistosoma spp. parasites were revealed using human, avian, and porcine sera and avian eggs. Bound antibodies were easily isotyped using an SPR biosensor to reveal the infection history of the individual. Discussed studies show the recent recognition of the suitability of this type of instrument for (rapid) detection of health-threatening microbes to food and environmental microbial safety.

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.

Immunochemical detection of Salmonella group B, D and E using an optical surface plasmon resonance biosensor.

A surface plasmon resonance biosensor (Biacore) was used to detect Salmonella through antibodies reacting with Salmonella group A, B, D and E (Kauffmann-White typing). In the assay designed, anti-Salmonella antibodies immobilized to the biosensor surface were allowed to bind injected bacteria followed by a pulse with soluble anti-Salmonella immunoglobulins to intensify the signal. No significant interference was found for (mixtures of) 30 non-Salmonella serovars at 10(9) CFU ml(-1). A total of 53 Salmonella serovars were successfully detected at 1 x 10(7) CFU ml(-1), except those of groups C, G, L and P, as expected. The cut-off point was determined with an equicellular mixture of Salmonella enteritidis and Salmonella typhimurium at a final amount of 1.7 x 10(3) CFU per test portion. Although further work is needed to cover the detection of all relevant Salmonella serovars in food-producing animals and food products, this work demonstrates the merits of this alternative biosensor approach in terms of automation, sensitivity, specificity, simple handling and limited hands-on time.