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The great public health success in prevention of human trichinellosis has resulted in a decades-long decline in the incidence and health impact of this zoonosis. The important elements of this achievement include improvement in animal husbandry practices, meat inspection, consumer education, and medical care. Most outbreaks now involve consumption of infected game. However, domestic pork still accounts for many outbreaks, mostly in Eastern Europe and Argentina, where traditional small, "backyard" rearing of pigs for household and local use often involve high risk rearing practices, especially the feeding of food waste. Although commercially produced pork under controlled management now accounts for about half of the world's pork production the demand by consumers, especially in Europe and North America, for free-range pork is increasing. Because of the varying degrees of outdoor exposure in free-range systems, there is concern that such exposure increases the risk of spillover of Trichinella spiralis, (also Trichinella britovi and Trichinella pseudospiralis in Europe) from wild animal reservoirs. This review examines the knowledge gaps hindering the risk assessments needed to provide the producers with guidelines for achieving the food safety assurance demanded by consumers.

Humans suffer from numerous parasitic foodborne zoonoses, many of which are caused by helminths (‘worms’). The parasites discussed in this chapter include some of those which are transmitted by ingestion of raw or improperly prepared foods, such as meat, fish, and plants. While many people in developed countries are commonly aware of meat-borne parasites such as Trichinella ( trichinosis or trichinellosis), they are less familiar with those transmitted by fish, invertebrates and plants. The parasites selected for this chapter were chosen because they represent these different food hazards. In the discussion of these parasites basic information on the biology and epidemiology of the parasitic zoonosis is presented, along with information on the major human pathogenic and clinical problems caused by the accidental infection with them. Diagnsotic challenges and treatments for these parasites are also descrbed, however, emphasis is placed on measures that should be taken by people to prevent exposure to them. The information presented in this chapter is an overview, therefore, for a more detailed understanding of these zoonoses the reader is encouraged to consult the sources listed in the Citation list.

Fish-borne zoonotic trematode parasites (FZT) pose a food safety and public health problem in Vietnam. The transmission cycle is complex as domestic animals, especially dogs, cats, fish-eating birds and pigs together with humans serve as reservoir hosts and contribute to FZT egg contamination of aquaculture ponds and the environment. This intervention trial was conducted to determine the effectiveness of various on-farm interventions, including reduction in FZT egg contamination through treatment of infected people and domestic animals, reduction in snail density through mud removal from aquaculture ponds prior to fish stocking, and various other measures in reducing FZT infection in juvenile striped catfish (Pangasianodon hypophthalmus) and giant gourami (Osphronemus goramy). Interventions were implemented on 5 farms for each fish species during production cycles in 2009 and 2010 while 5 similar farms for each species served as control. For both fish species, both prevalence and intensity of infection did not differ significantly between intervention and non-intervention farms prior to the interventions. The interventions significantly reduced both prevalence and intensity of FZT infection in the juvenile fish compared to control ponds. For giant gourami, odds of infection in intervention ponds was 0.13 (95% CL: 0.09-0.20; p

Infection with fish-borne zoonotic trematodes (FZT) is an important public health problem in many parts of Southeast Asia. People become infected with FZT when eating raw or undercooked fish that contain the infective stage (metacercariae) of FZT. The parasites require specific freshwater snails as first intermediate host and a variety of fish species, both wild caught and cultured, as second intermediate host. Aquaculture production has grown almost exponentially in SE Asia and in order to produce fish free from FZT metacercariae, it is important to mitigate factors promoting transmission to fish. Here we report results from a cross-sectional study to look at the association between pond depth and infection with FZT in giant gourami nursery ponds. Density of intermediate host snails was positively associated with pond depth (count ratio associated with a 1m increase in pond depth was 10.4 (95% C.L.: 1.61-67.1, p0.5)) and this may partly explain the higher prevalence and intensity of FZT infection in juvenile fish. High fry stocking density (200 fry m(-3)) was associated with lower host snail density (count ratio=0.15) than low stocking density (100 fry m(3)). Ponds stocked with 100-200 fry m(-3) had snail counts 0.76 (95% C.L.: 0.33-1.75, p n.s.) of those in ponds stocked with fry density of100 fry m(-3). Since density of intermediate snail hosts was associated with FZT transmission to fish, effort should be taken to reduce snail density prior to stocking the fry, but focus should also be on habitats surrounding ponds as transmission may occur through cercariae produced outside ponds and carried into ponds with water pumped into ponds.

Fish-borne zoonotic trematodes (FZT) are a major public health problem. It is estimated that in Vietnam 26,366 “disability-adjusted life years” (DALYs) are lost due to FZT. Fish from aquaculture are a main source of protein and of great economic importance in both rural and urban areas. The objective of this study was to compare the impact on transmission of FZT to juvenile fish of drug treatment intervention of final hosts, and pond management interventions on fingerlings produced in nurseries producing Rohu carp in Northern Vietnam. The study design was a parallel group design of nurseries with two intervention groups; a drug treatment of human and animal groups and a farm management group (control of snail vectors and fecal pollution of pond). A third group (non-intervention) served as control. Fish were examined for FZT metacercariae prevalence and intensity before and after interventions, along with density of potential host pond snails. Fecal exams for FZT infections in farm household humans, dogs and cats were also performed. The results showed that although FZT prevalence in fish was not significantly reduced, the intensity of metacercariae in fish from ponds receiving management interventions was reduced 91.7% compared to before interventions. The intensity of FZT was also significantly lower in the pond management group, compared to the drug treatment group and the control group after interventions. The results demonstrate that improving farm and pond management practices can be effective in reducing FZT infections without using drug treatment. This study also highlighted certain obstacles to attaining higher levels of protection for which more research is needed. A major challenge is the ability to completely isolate (or bio-secure) the pond from its surrounding environment, especially surrounding water sources which may harbor snails and cercariae that can contaminate the pond through water resupply or runoff. Another is the unknown risk from piscivorous birds, host for intestinal FZT. The strategies employed in this study now need long-term evaluation on a regional basis, the outcome of which could be the development of good management practices crucial to ensuring a good quality product that benefits farmers and the consumers.

The direct influence of intracaecal infusion of short-chain fatty acids (SCFA) and lactic acids (LA) on already established Oesophagostomum dentatum infection in cannulated pigs was investigated. We tested the hypothesis that the previously discovered anti-parasitic effect of inulin is mediated through its metabolic products SCFA and LA by infusing into cannulated pigs these compounds in amounts approximating to those produced in the pigs large intestine and caecum during the metabolism of inulin. The experiment comprised of 18 pigs--2 groups of 9 pigs in each. The normal diet used in the experiment was based on barley flour with insoluble fibre from oat husk with added soybean meal, vitamins and minerals. After 2 weeks of adaptation to the diet all the pigs were inoculated with 6,000 infective larvae of O. dentatum. Six weeks later, surgery on all pigs was performed to install cannulas into caeci. At 7 weeks post-infection (p.i.) the SCFA and LA infusion was initiated in Group 1 (experimental) pigs; at the same time pigs in Group 2 (controls) were infused with saline. At week 10 p.i., all pigs were killed and their worm burdens determined. SCFA and LA infused pigs exhibited markedly reduced fecal egg counts and worm recoveries (98 and 92% reduction, respectively, compared to saline controls). The results from this study demonstrate that SCFA and LA have a significant negative influence on established O. dentatum infection in growing pigs. The results also show that the type of dietary carbohydrates fed and its intestinal degradation can yield metabolic by products that profoundly influence helminth survival.

The present study was designed to assess the role of inulin and sugar beet fibres (SBF) on adult O. dentatum in growing pigs. Four experimental diets were formulated based on barley flour with added insoluble fibre from oat husk (Diet 1), a pure carbohydrate source inulin (Diet 2), soluble fibre from sugar beet fibre (SBF) with a high proportion of soluble fibre components (Diet 3) or inulin plus SBF (Diet 4). Thirty-two 10-week-old pigs were divided randomly into 4 groups each of 8 pigs. After 3 weeks adaptation on Diet 1 all pigs were infected with a single dose of 6000 L 3 O. dentatum . At week 7 p.i. one group was switched to Diet 2, another group to Diet 3 and another group to Diet 4. The remaining 8 pigs continued on Diet 1 until the end of the experiment and served as controls. At week 13, all pigs were necropsied and their worm burdens determined. The worm recoveries from the pigs on the inulin supplemented diet (Diet 2) were reduced by 97% compared to the controls (Diet 1). Further, the inulin-fed pigs exhibited markedly reduced faecal egg counts. The pigs on inulin plus SBF diet (Diet 4) and on SBF diet (Diet 3) had 86% and 70% adult worm reductions compared with the controls, respectively. The results from this study indicate that highly degradable and rapidly fermentable carbohydrates such as dietary inulin have a profound deworming effect on O. dentatum infection.

Trichinellosis is a re-emerging zoonosis and more clinical awareness is needed. In particular, the description of new Trichinella species such as T papuae and T murrelli and the occurrence of human cases caused by T pseudospiralis, until very recently thought to occur only in animals, requires changes in our handling of clinical trichinellosis, because existing knowledge is based mostly on cases due to classical T spiralis infection. The aim of the present review is to integrate the experiences derived from different outbreaks around the world, caused by different Trichinella species, in order to provide a more comprehensive approach to diagnosis and treatment.

A study was conducted to determine the distribution and transmission rate of Ascaris suum eggs and Oesophagostomum dentatum larvae in a pasture/pig house facility, which during the preceding summer was contaminated with helminth eggs by infected pigs. In May, four groups of 10 helminth naïve tracer pigs were exposed to fenced sections of the facility for 7 days and necropsied for parasite recovery 9-10 days later (trial 1). The highest rate of A. suum transmission (201 eggs per day) occurred in the pig house (A). On the pasture, egg transmission decreased with the distance from the house: 8 eggs per day in the feeding/dunging area (B); 1 egg per day on the nearest pasture (C);1 egg per day on the distant pasture (D). Only a few O. dentatum infections were detected, indicating a poor ability of the infective larvae to overwinter. Soil analyses revealed that the highest percentage (5.8%) of embryonated A. suum eggs were in the house (A). Subsequently, the facility was recontaminated with A. suum eggs by infected pigs. A replicate trial 2 was conducted in the following May. A major finding was the complete reversal of egg distribution between the 2 years (trials 1 and 2). In contrast to previous results, the highest rates of transmission (569 and 480 eggs per day) occurred in pasture sections C and D, and the lowest transmission rates (192 and 64 eggs per day) were associated with the feeding/dunging sections and the house (B and A). Soil analyses again supported the tracer pig results, as the pasture sections had the highest concentrations of embryonated eggs. Detailed soil analysis also revealed a non-random, aggregated egg distribution pattern. The different results of the two trials may be due to the seasonal timing of egg deposition and tracer pig exposure. Many eggs deposited during the summer prior to trial 1 may have died rapidly due to high temperatures and dessication, especially when they were not protected by the house, while deposition in the autumn may have favored egg survival through lower temperatures, more moisture, and greater sequestration of eggs in the soil by rain and earthworms. The latter eggs may, however, not have become embryonated until turnout the next year. The results demonstrate that yearly rotations may not be sufficient in the control of parasites with long-lived eggs, such as A. suum, and that a pasture rotation scheme must include all areas, including housing.

An experiment was undertaken to study the effect of dietary carbohydrates with different digestibility on the populations of Oesophagostomum dentatum in the intestinal tract of pigs. Sixty-four worm-free pigs from a specific pathogen-free farm were randomly divided into 8 equal groups. The animals in 4 groups were assigned to a diet with partially undegradable carbohydrates (diet 1), while the pigs in the 4 remaining groups were given a diet with fermentable carbohydrates (diet 2). Diet 1 was comprised of barley flour, oat husk meal, soybean meal, vitamins and minerals and diet 2 of barley flour, inulin and sugar beet fibre, soybean meal, vitamins and minerals. The pigs in 6 of the groups (n = 48) were inoculated with 6000 infective larvae of O. dentatum. To determine O. dentatum populations at the early stage of infection, 16 pigs were slaughtered 3 weeks p.i., while the remaining 4 groups continued on the diets for a further 9 weeks after which they were slaughtered. In a diet cross-over experiment 6 weeks after inoculation, 8 pigs changed from diet 1 to diet 2 (diet 1>diet 2), and 8 pigs from diet 2 to diet 1 (diet 2>diet 1). The results showed that partially undegradable carbohydrates provided favourable conditions not only for parasite establishment and sustainability, but also for already established O. dentatum infection while, in contrast, the diet composed of highly degradable carbohydrates decreased worm establishment, size and female fecundity. The implications for pastured pigs or pigs fed different complex carbohydrate diets is discussed.

The authors review the major biological, biochemical, and molecular characters that are used to distinguish the seven Trichinella species (T. spiralis, T. nativa, T. britovi, T. pseudospiralis, T. murrelli, T. nelsoni, T. papuae) and three genotypes whose taxonomic status is yet uncertain (T-6, T-8, T-9). A comparison of host specificity, morphology, reproductive abilities, nurse cell development and freeze resistance is presented, along with useful biochemical and molecular markers. Finally, this information is used to construct a diagnostic key for the species. A phylogenetic classification of the species is needed.

The infectivity of Trichinella spiralis, T. nativa, and T. britovi was experimentally compared in pigs. Blood sampling was performed weekly, and muscle juices were obtained at slaughter 10 weeks after inoculation. Muscle larvae were found in all of four pigs inoculated with T. spiralis [mean 190 larvae per gram (lpg)] and in three of four pigs inoculated with T. britovi (mean 7 lpg). No larvae were found in pigs inoculated with T. nativa. For T. spiralis and T. britovi, the neck muscle (m. splenius) appears to be a predilection site in addition to the tongue, the diaphragm, and the jaw. High antibody responses were found in all experimental groups, independent of the antigen used, and even in pigs in which no muscle larvae were recovered. The strong and consistent antibody response found with meat juice indicates the usefulness of this material where a blood sample is not obtainable, e.g. meat samples from wild animals. Immunoblotting (Western blots) on slaughter sera revealed no species specificity when comparing homologous versus heterologous staining.

A multitude of parasites have been reported in fish, but only a few species are capable of infecting humans. The most important of the helminths acquired by humans from fish are the anisakid nematodes (particularly Anisakis simplex and Pseudoterranova decipiens), cestodes of the genus Diphyllobothrium and digenetic trematodes of the families Heterophyidae, Opisthorchiidae and Nanophyetidae. Seafood-associated infections by acanthocephalans are rarely reported in humans. All of the helminths mentioned above are associated with social-cultural and behavioural factors, in particular the consumption of raw or undercooked seafood. Measures can be taken during harvesting, processing or post-processing (e.g., by the consumer) to mitigate the risks of infection. The seafood industry and government authorities can apply various programmes to reduce these risks, including good manufacturing practices (GMPs) and hazard analysis and critical control point (HACCP) systems. Such measures may include avoiding particular harvest areas, sizes of fish, or even particular species of fish. The method of capture, handling and storage of the catch can directly affect the quality of the seafood with regard to the presence and numbers of parasites. The extent of processing--including heading and gutting, candling and trimming--and the type of product derived (fresh, frozen, salted or pickled) can all contribute to the control of the risks posed by helminths. The most effective means of killing the parasites are either freezing or heat inactivation.

In Denmark, alternative outdoor production systems for pigs are becoming more frequent, and information on the transmission of Ascaris suum and Trichuris suis under continuously grazed pasture conditions is needed. A group of pigs was turned out on a pasture in May 1993 (Year 1 of the study), inoculated with 200 eggs of A. suum and 1000 eggs of T. suis , and followed parasitologically. A non-experimentally infected group of pigs was similarly turned out on the same pasture the following year (Year 2) and again followed parasitologically. Pasture infectivity was measured using helminth-naive tracer pigs. During the summer of Year 1, A. suum eggs became infective within 4–6 weeks on the pasture. However, transmission was moderate until August of Year 2, when a pronounced increase in transmission occurred. After 2 months on the pasture, the continuously exposed pigs in summer seasons of both Years 1 and 2 harboured small overdispersed populations of adult A. suum , moderate numbers of liver white spots, and high specific IgG responses. These parasitological measures on chronically exposed pigs did not, however, correlate well with pasture infectivity or with each other. In contrast, the liver inflammation and specific IgG responses (but not the intestinal A. suum burdens) of the tracer pigs were highly correlated ( P = 0.0001) and appeared to better reflect pasture infectivity. The inoculated pigs excreted T. suis eggs by the late summer of Year 1, but no transmission took place before August of Year 2. Thus, the T. suis population of infective eggs built up very slowly. The results indicate that T. suis eggs may survive for a considerable time, however. The study results reveal that A. suum and T. suis eggs are much more resistant to environmental factors than free-living infective larvae of pig parasites such as Oesophagostomum dentatum and Hyostrongylus rubidus . Control of these parasites in outdoor systems will present more difficult challenges than that for parasites transmitted by free-living larvae.

The Tenth International Conference on trichinellosis, organized by the International Commission on Trichinellosis, gathered together 147 scientists from 34 countries. The 173 presentations ranged over most aspects of this parasitic zoonosis: history, epidemiology, speciation, biology, immunology, biochemistry, human and animal pathology, new diagnostic methods and control.

The re-emergence of the zoonotic importance of trichinellosis reminds us that this parasite's unique biological features serve it well in helping it to persist in the face of 150 years of efforts to eliminate it from the food chain. Because of Trichinella's re-emergence as an important public health risk, and the consequent stimulation of research, our knowledge of the parasite has grown, especially on the complexity of its biology and epidemiology. This overview will highlight the major features of this parasite's adaptive traits and the human factors that account for its re-emergence in many regions, along with the actions of the International Trichinellosis Commission to address these issues. The attractiveness of Trichinella as a research subject for study of the parasitic lifestyle will also be emphasized, including areas for future research.

A novel lateral flow card (TS-Card pork) test was developed for the serological detection of Trichinella infected pigs. Based on extensive studies performed in Romania during 1 999-2000 this test proved to be highly specific, sensitive, rapid (3-12 minutes) and easy to use (no need for laboratory facilities). It can be used both for the detection of Trichinella infection in carcasses and for epizootiological studies using a variety of samples including whole or dried blood, serum, or tissue fluids. The TS-Card pork test, used as a screening test, can be the foundation of an on-farm or field based inspection system to significantly improve food safety in countries with a high prevalence of Trichinella in pigs or other food animal species. The results presented are also promising for application of the test in an on-line laboratory based inspection system since the speed of the test allows sufficient time to rail out suspected hog carcasses during the slaughter process.

Many helminth infections of humans are recognised as zoonoses and the route of infection invariably involves one of the pathogen’s natural hosts having become a human food item. Some of these zoonotic infections are deadly while others cause chronic ill health. This chapter presents a selection of foodborne species of trematode, cestode and nematode parasites, their distribution and abundance, and their public health and economic importance. The helminths include those transmitted by infected meat, fish, invertebrates and plants. Finally, a selection of approaches to the prevention and control of foodborne helminth infections is presented.

Zoonotic helminths that invade host tissues, particularly as larvae, generally have low prevalence rates in most areas of the world. Taken as an aggregate, however, the likelihood that a human will be exposed to a member of this group of parasites is not insignificant. This article covers the specific agents or parasites involved in these zoonotic infections, their epidemiology, the clinical aspects of the disease, and strategies for preventing or controlling infection. Although not all-inclusive, the zoonoses discussed are trichinellosis, anisakiasis, angiostongyliasis, gnathostomiasis, and infection with Baylisascaris. Except perhaps for the latter parasite, the rest have a common feature: transmission through consumption of raw or undercooked food. For humans, this food behavioral trait is the most important risk factor, and a factor that is very difficult, from a public health perspective, to alter. In fact, the demand for raw or lightly cooked food, especially fish, is an increasing global phenomenon, and this will ensure that risk of transmission of these parasites persists unless new approaches to control are introduced.

SUMMARYThe aim of this study was to investigate the intra-litter infection dynamics ofIsospora suisunder natural conditions, and to study any association between parasite transmission and the contamination level of the farrowing pen by applying different interventions in order to reduce the transmission ofI. suisinfection within the litter. The study was divided in 2 trials including in total 22 litters (254 piglets). The first trial included 4 litters (where standard procedures practiced routinely on the farm piglets were applied) and the piglets were followed coprologically from farrowing until 2 weeks after weaning. The sows of those litters were also examined at various intervals before and after farrowing. The second trial included the application of 3 different management procedures: (A) standard farm hygiene and management procedures, (B) standard farm hygiene and management procedures+the first piglets found to excreteI. suisoocysts in each pen were removed from the pen, and (C) reduced cleaning. Each procedure was studied in 2 litters. This was replicated 3 times to yield a total of 18 litters. The results suggested that (i) the sow does not play an important role in transmission ofI. suisin the farrowing pen; (ii) in natural infections, both the age of the piglet age at onset of oocyst excretion and the oocyst excretion patterns may vary considerably; (iii) the course of oocyst excretion or development of diarrhoea is related to the time of initial infection and (iii) piglets, which are heavy at birth, are more prone to acquireI. suisinfection. Moreover, it was demonstrated that cleaning could be an effective means of restricting the spread of the parasite within the litter and thus the development of diarrhoea.

This research was carried out to determine the time/temperature exposure of Trichinella spiralis to freezing conditions necessary to destroy the infectivity of the trichinae. Experimentally infected pork was subjected to temperatures of -1 to -193°C for one sec to 182 d and the treated pork samples, which contained about 1000 larvae per gram of tissue, were subjected to rat bioassay to determine infectivity of the larvae. A linear regression equation, log10t = 5.98 + 0.40T where t = required inactivation time in hours and T = temperature in degrees Celsius, described the exposure necessary to destroy the trichinae. The correlation for that relationship was r = 0.942. The predicted thermal death times (+7 min) at -20, -15, and -10°C were 8 min, 64 min, and 4.0 d, respectively. The predicted upper confidence limits (99%) for the thermal death times (+7 min) for exposure at -20, -15, and - 10°C, were 48 min, 63 h, and 266 d, respectively. These data provide a continuum of definitive times and temperatures necessary to destroy T. spiralis by freezing and are of value to the meat industry and the regulatory agencies.

A heretofore undescribed host-mediated reactivity against encapsulated muscle larvae (ML) of the nematode Trichinella spiralis is reported. Inbred miniature swine (NIH minipigs) of three independent SLA phenotypes, which received a primary oral dose of 300 T. spiralis ML, successfully resisted a secondary infection of 10,000 ML; however, only pigs of the SLAa/a phenotype exhibited an unusual and highly significant reduction in the numbers of encysted ML from the primary infection (P less than 0.0003). This initial anti-encysted ML reactivity was confirmed in subsequent trials by comparing the prechallenge ML burdens with the reduced ML numbers in primary-infected aa pigs after challenge. Analyses of inbred strains of mice, selected for major histocompatibility type and for resistance or susceptibility to infection with T. spiralis, showed no such anti-encysted ML response. Because elimination of encysted T. spiralis ML had been accomplished previously only through selected drug regimens, our demonstration of a nonpharmacological, host-mediated reactivity against this stage of the parasite in swine highlights the importance of MHC genes in regulating disease resistance in a livestock species.

The objective of this experiment was to investigate the potential influence of inulin on the establishment of new and patent infections ofTrichuris suisin growing pigs. Two experimental diets were formulated based on barley flour with either added insoluble fibre from oat husk (Diet 1) or a pure inulin (16%) supplementation (Diet 2). Twenty-eight 10-week-old pigs were divided randomly into 4 groups (Groups 1–4) each of 7 pigs. After 3 weeks adaptation to the experimental diets all pigs were infected with a single dose of 2000 infectiveT. suiseggs. Group 1 was fed Diet 1 until 7 weeks post-infection (p.i.) and Group 3 until 9 weeks p.i., Group 2 was fed Diet 2 until 7 weeks p.i., Group 4 was fed Diet 1 until week 7 p.i. and was switched-over from Diet 1 to Diet 2 until week 9 p.i. Seven weeks p.i. pigs in Groups 1 and 2 were slaughtered, and pigs in Groups 3 and 4 were slaughtered at 9 weeks p.i.Trichuris suisworm burdens were determined for all pigs. Inulin-fed pigs (Group 2) exhibited an 87% reduction in EPG, compared to the pigs on standard diet (Group 1) (PPPT. suisestablishment, egg excretion, and female worm fecundity and can be used as a treatment for patent infections.

In the Balkan countries, where trichinellosis is a re-emerging zoonosis, it is of great importance to determine Trichinella infection prevalence among the major hosts, including horses. One method for monitoring prevalence is serological surveillance; however, the validity of serological methods in horses is not well understood. The dynamics of anti-Trichinella IgG production and circulating excretory/secretory (ES) antigens were investigated in three horses experimentally-infected with Trichinella spiralis. Horses were slaughtered at 32 week post infection (p.i.). Low worm burdens were found in all three animals. Anti-Trichinella IgG was detected up to 32 weeks p.i. by an indirect immunofluorescence assay (IFA) and by Western blot (Wb), but not by ELISA. The ELISA test detected antibodies for only a short period of time (up to 18 weeks p.i. using ES antigen or up to 20 weeks p.i. using tyvelose-BSA antigen). The presence of circulating muscle larvae ES antigen in sera of infected horses was observed by dot blot from the 4th week p.i. up to the 32nd week p.i.

A discovery in 2002 of a Trichinella spiralis-infected horse in Serbia offered an opportunity to conduct needed epidemiological studies on how horses, considered herbivores, acquire a meat-borne parasite. This enigma has persisted since the first human outbreaks from infected horse meat occurred in then 1970s. The trace back of the infected horse to a farm owner was carried out. Interviews and investigations on the farm led to the conclusion that the owner had fed the horse food waste in order to condition the horse prior to sale. Further investigations were then carried out to determine the frequency of such practices among horse owners. Based on interviews of horse producers at local horse markets, it was revealed that the feeding of animal products to horses was a common practice. Further, it was alleged that many horses, particularly those in poor nutritional condition would readily consume meat. A subsequent series of trials involving the experimental feeding of 219 horses demonstrated that 32% would consume meat patties. To confirm that horses would eat infected meat under normal farm conditions, three horses were offered infected ground pork balls containing 1100 larvae. All three became infected, and at necropsy at 32 weeks later, were still positive by indirect IFA testing, but not by ELISA using an excretory-secretory (ES) antigen. This result indicates that further study is needed on the nature of the antigen(s) used for potential serological monitoring and surveillance of horse trichinellosis, especially the importance of antigenic diversity. The experimentally-infected horses also had very low infection levels (larvae per gram of muscle) at 32 weeks of infection, and although the public health consequences are unknown, the question of whether current recommended inspection procedures based on pepsin digestion of selected muscle samples require sufficient quantities of muscle should be addressed. It is concluded that horses are more willing to consume meat than realized and that the intentional feeding of animal products and kitchen waste is a common occurrence among horse owners in Serbia (and elsewhere?). This is a high risk practice which demands closer scrutiny by veterinary and food safety authorities, including the implementation of rules and procedures to ensure that such feeds are rendered safe for horses, as is now required for feeding to swine.

The population dynamics of Toxocara canis in pigs, and their immune response to a primary and a challenge infection, were studied by parasitological, haematological and serological parameters. Seventy pigs were divided into 4 groups; 35 pigs received a primary infection (group A), 15 pigs received both a primary and a challenge infection (group B), 15 pigs received the challenge infection only (group C), and 5 pigs served as helminth-free controls (group NC). A dose of 50 000 eggs was administered for the primary infection (day 0) and a dose of 10 000 eggs was given for the challenge infection (day 28). On days 7, 14, 21 and 28 p.i., 5 pigs of group A, and on days 35, 42 and 49 p.i., 5 pigs from each of groups A, B and C were necropsied. Numbers of recovered larvae varied widely among the 5 pigs of each group on all days of necropsy. Toxocara canis larvae were recovered predominantly from the lungs; migration of larvae to other organs or tissues from the lungs was restricted. In group A, the larval burden in the lungs peaked on day 14 p.i., and the larval densities decreased significantly over time. Thereafter, the majority of larvae were recovered from the lungs until the end of the experiment (day 49 p.i.). A few larvae were found in the muscles and brain until day 42 p.i., and 2 larvae were found in the eyes of 2 pigs on day 35 p.i. There was little evidence of protective immunity to a challenge infection in this experiment. The eosinophil levels tended not to increase in pigs receiving a challenge infection, in contrast to the challenge control pigs. The fact that T. canis larvae migrate and persist in the tissues of pigs for more than 1 month suggests a zoonotic risk in infected pigs. The relevance of these data to the population biology and immunology of porcine and human toxocarosis is discussed.

The effects of Oesophagostomum dentatum infection and dietary carbohydrates on the morphology and epithelial cell proliferation in the gastrointestinal tract of pigs were investigated experimentally. Thirty-two worm-free pigs (n=32) from a specific pathogen-free farm were randomly divided into four groups (A-D), of eight animals each. Pigs in groups A (control) and B (infected) were fed Diet 1, and pigs in groups C (control) and D (infected) were fed Diet 2. The two diets were formulated: Diet 1 (%) contained barley flour, oat husk meal plus soya bean meal (55:21:24) and Diet 2 (%) contained barley flour, inulin and sugar beet fibre (SBF) (80.1:7:12.9) plus soya bean meal (3:1) to contain carbohydrates from inulin and sugar beet fibre (SBF) that were readily fermentable in the large intestine. The two infected pig groups (16 pigs total) were inoculated with 6000 infective larvae of O. dentatum and all pigs, including the controls, were slaughtered 12 weeks p.i. The combination of O. dentatum infection and highly fermentable dietary carbohydrates affected the mucosal architecture, the epithelial cell proliferation and mucin secretion of the large intestine. Infection had a significant influence on the crypt volume, height and density, and on muscularis externa at the proximal and middle colon. The changes in the affected gut sections were proportional to the number of worms present. However, these parameters appeared unaffected by those diets alone. In pigs without infection non-digestible dietary carbohydrates significantly influenced the tissue weight of colon.

Experiments were conducted to investigate possible alternative routes of extraintestinal migration of Ascaris suum larvae in the pig. Pigs were infected with A. suum via injection of newly hatched larvae into cecal veins (i.v.), into cecal lymph nodes (LN), or intraperitoneally (i.p.), and control animals were inoculated orally with infective eggs (p.o.). Two pigs per inoculation route were necropsied on days 1, 4, and 13 postinoculation. The numbers of liver lesions and the percentage of larvae recovered was considerably greater in pigs inoculated i.v. or p.o. on each necropsy day. However, irrespective of inoculation route, at least a proportion of larvae passed through the livers and were able to complete migration to the small intestine by day 13. The results indicate that larval penetration of the intestinal wall is not necessary for liver-lung migration and that passage through the liver may be favorable for migrating A. suum larvae, although a delayed arrival in the small intestine cannot be ruled out for larvae following alternative routes.

Trichinellosis, is normally not included among those regarded as emerging zoonoses because it has been a public health threat for more than 150 years. However, its dramatic re-emergence in many areas around the world over the past 10-20 years, inspite of a century of veterinary public health efforts to control and eradicate it, justifies it being included in this group. The reasons for this re-emergence are diverse, and include human pertubation and manipulation of ecosystems, war and political turmoil, rapidly changing food distribution and marketing systems, and even, surprisingly, rising affluence in developing countries. These influences, and their impact on the epidemiology of both domestic and sylvatic trichinellosis, are discussed, along with recommendations for confronting this altered status as a public health threat.

Fragmentary data indicate that zoonotic parasites cause human illnesses with medical costs and productivity and disability losses totalling billions of dollars annually. Food is an important vehicle for some of these parasitic diseases. The cost to public health is not reflected in the priorities given to these parasitic diseases in either research or public health planning. In this article, Tanya Roberts, Darwin Murrell and Suzanne Marks discuss the cost of toxoplasmosis, taeniasis, cysticercosis, trichinellosis and other foodborne parasitic diseases.

Allozyme analysis was carried out on 152 Trichinella isolates from synanthropic and wild animals and from humans; the isolates were collected from 5 continents. The analysis, involving 27 enzymes, revealed the presence of 8 distinct gene pools, termed T1-T8. Four of the genetic groups represent the 4 previously proposed species: Trichinella spiralis sensu stricto (T1), Trichinella nativa (T2), Trichinella nelsoni (T7), and Trichinella pseudospiralis (T4). The other 4, T3, T5, T6, and T8 are distinct from previously described species. The absence of allozymic hybrid patterns among even sympatric groups indicates a lack of gene flow among the groups. Principal component analysis and the unweighted pair group method of analysis were used to assemble allozyme patterns of the 152 isolates into discrete groups and to show their relative relationships. Both analyses indicated the presence of 8 primary clusters that correlated with the gene pools revealed by direct allozyme profile analysis. The absence of evidence of gene flow among the gene pools and the high level of allozymic differentiation between the cluster groups support the concept that the genus Trichinella is composed of several sibling species.

Increasing concern over food safety has focused greater attention on foodborne parasitic diseases. This review highlights instances in which the application of biotechnological tools is facilitating the development of new strategies to control foodborne parasites.

A study was made of the early migratory pattern of Ascaris suum in mice. Mice were each inoculated orally with a single dose of 2,500 infective eggs and then killed at 2, 4, 6, 8, 10, and 12 hr postinoculation (PI). At necropsy, it was observed that the larvae had penetrated the mouse cecum and colon and had reached the liver by 4 hr PI. This migratory behavior closely mimics what is observed in the pig and suggests that the mouse may serve as an experimental model for intestinal immunity in the early phase of A. suum infection.

During the course of carrying out studies on the role of intestinal immunity in blocking the migration of larval Ascaris suum in the pig, it was discovered that the prevailing understanding of larval penetration sites was at variance with our observations. Therefore, a detailed investigation of the migration of A. suum 1.2 larvae through the intestine was initiated. The results demonstrate that the 1.2 larvae invade almost exclusively the wall of the pig cecum and colon and not the small intestine as is generally believed. The larvae were recovered from the mucosa of the cecum and colon as early as 3 hr postinoculation (PI) with infective eggs and were recovered from the liver by 6 hr PI. The maximal recovery of larvae (total larvae and larval/g of mucosa) from the intestinal mucosa occurred between 6 and 12 hr PI; by 24 hr PI, virtually all of the larvae had disappeared from the mucosa. These observations correct a common misunderstanding of this aspect of the life cycle of A. suum in the pig, and they raise 2 issues related to the biology and pathogenesis of Ascaris in humans. What is the actual migratory and development behavior of Ascaris lumbricoides and A. suum in humans and the potential risk for liver lesions? Most authors, in describing the life cycle of A. lumbricoides, either ignore or discount a possible obligatory liver stage of development, and, consequently, the potential for lesion formation similar to that which occurs in pigs infected with A. suum. This issue takes on added importance with the growing evidence that A. suum is an important zoonoses.

In the course of a comprehensive study on the population biology of Ascaris suum, it became necessary to determine quantitatively the migration of the larvae (L 2 ) through the pig intestine. Because no satisfactory methods for recovering these larvae from the intestinal mucosa were apparently available in the literature, we undertook the development of such a procedure. Direct Baermannization of the intestine proved inadequate, so a series of studies was undertaken to find a method to solubilize the mucosa and free the larvae. Both pepsin digestion and mucus reduction with N-acetyl cysteine were evaluated. The highest recovery rates were obtained with short-term pepsin digestion (45-90 min), followed by a specific settling and washing procedure. A third alternative, migration of larvae out of agar-gel, was also tested. Although the recovery efficiency was low (10% of pepsin digestion), the larvae recovered were clean, with good motility. The agar-gel procedure could be useful when L2 larvae are needed for other purposes, such as antigen preparation or cultivation.

The analysis of genetic, biochemical, and biological data on about 300 Trichinella isolates, reported in the literature, allows a taxonomic revision of this genus. We propose the recognition of 5 sibling species, Trichinella spiralis (Owen, 1835) sensu stricto; Trichinella nativa Britov and Boev, 1972; Trichinella pseudospiralis Garkavi, 1972; Trichinella nelsoni Britov and Boev, 1972 sensu stricto; and Trichinella britovi n. sp., on the basis of biochemical and biological characteristics. Trichinella britovi n. sp. is characterized by distribution in the Palaearctic Region; newborn larvae (NBL) production in vitro of 35-55 NBL/72 hr; nurse cell development time (NC d.t.) between 24 and 42 days postinfection (d.p.i.); low reproductive capacity index (RCI) in mice, rats, and pigs; low resistance to freezing; 1 unique marker allozyme; and moderate pathogenicity for humans. The new species is most similar to Trichinella nativa but differs from it in 4 allozymes, in having less resistance to freezing, in having a different pattern of major ribosomal DNA fragments after endonuclease digestion, and in distribution area. Trichinella nativa is characterized by a holarctic distribution; hosts that are sylvatic mammals; NBL production in vitro 28-54/72 hr; NC d.t. between 20 and 30 d.p.i.; low RCI in mice, rats, and pigs; high resistance to freezing; 2 unique marker allozymes; and moderate to severe pathogenicity for humans. Trichinella spiralis sensu stricto is characterized by a cosmopolitan distribution in domestic pigs, associated wildlife, and humans; high NBL production in vitro (greater than 90 NBL/72 hr); NC d.t. between 16 and 37 d.p.i.; high RCI in mice, rats, and pigs; no resistance to freezing; 6 unique marker allozymes; and high pathogenicity for humans.(ABSTRACT TRUNCATED AT 250 WORDS)

Forty isolates of Trichinella collected from 5 continents were compared for 7 biological characters: newborn larvae produced per female worm cultured in vitro at the seventh, eighth, and ninth day postinfection, host muscle nurse cell development time, reproductive capacity index in rats and chickens, and resistance of muscle larvae to freezing. The isolates also were compared by analyses of an environmental character of the location from which they were isolated: the isotherms for January and July. By factorial analysis of correspondence of the biological and environmental data, the 40 isolates were grouped into 8 gene pools (T1-T8). The environmental temperature-related distribution was more evident for the sylvatic isolates (T2, T3, T5, T6, T7, T8), than for T1, which was isolated from domestic pigs, and for T4, a bird-adapted, nonencapsulating genetic type. The 8 biological groups correlated closely with the 8 gene pools previously identified on the basis of allozyme analysis. These results support the concept that the genus Trichinella is composed of at least 5 distinct gene pools or sibling species: Trichinella spiralis sensu stricto (T1), Trichinella nativa (T2), Trichinella sp. (T3), Trichinella pseudospiralis (T4), and Trichinella nelsoni (T7), and 3 other groups of uncertain taxonomic status (i.e., T5, T6, and T8).

The ability of domestic pigs to develop protective immunity to Trichinella spiralis in response to inoculation with different doses of muscle larvae was assessed. Adult worms developing from the inoculations of 112, 500, and 10,000 larvae were expelled from the intestine about 6 weeks after inoculation. Inoculation with 25,000 larvae, however, resulted in more rapid intestinal worms expulsion, indicating that gut expulsion is dose dependent. Secondary expulsion also tended to be dependent upon primary infection level. Pigs initially inoculated with 500 to 10,000 larvae expelled the challenge infection of adult worms after 22 to 25 days; in contrast, infection by inoculation of only 112 larvae failed to induce significant enchanced gut expulsion of the challenge infection intestinal worms. However, all primary infection levels, including inoculation with 112 larvae, induced nearly absolute resistance to the muscle establishment of larvae from challenge adult worms. The fecundity of female worms recovered from immune pigs was reduced 75% in comparison to controls. These results show that, in contrast to some host species, very rapid gut expulsion does not occur in domestic swine. Yet, immune responses at the gut level are important, perhaps responsible for much of the inhibition reflected as reduction in the establishment of muscle larvae.

Parameters of immunization of mice with /sup 60/Co-irradiated Schistosoma mansoni cercariae are described and related to protection against subsequent challenge infection. Such immunization was found to be dependent on dose of irradiation, number of immunizing cercariae, and number and time course of infections. Low levels of resistance were obtained with low irradiation doses, in contrast to previous studies in mice. In general, resistance increased with increasing irradiation doses, up to approximately 48 to 56 Kr. Maximal resistance (70 to 80%) was induced by a single exposure to 250 to 500 cercariae, irradiated at a dose rate of 2 Kr/minute to a total dose of 56 Kr, administered percutaneously 4 to 6 wk prior to challenge. Challenge could be delayed for at least 15 wk after immunization without a decrease in resistance. The resistance obtained was not attributable to a delayed migration of challenge worms.

Mice reinfected with Schistosoma mansoni 6--8 weeks after a primary infection largely or completely eliminated the second infection prior to the 7-week adult worm stage. In contrast, challenge worm counts were not lower than controls at the 6-day lung schistosomulum stage. At reinfection intervals of 12 or more weeks, worm counts were reduced at both stages. The reduction in lung schistosomulum count was proportional to the number of schistosome eggs present in the lungs, with no significant reduction being detected at any challenge time in mice free of lung eggs. Isolated schistosome eggs injected intravenously into the lungs of normal mice induced moderate to high levels of resistance to infection, while eggs injected subcutaneously or imtraperitoneally did not. It is concluded that the deposition of schistosome eggs in sites encountered by migrating schistosomula may be essential for mice to become resistant to reinfection with S. mansoni.

Center cut chops (longissimus dorsi), 2.5 cm in thickness, from 31 pigs experimentally infected with Trichinella spiralis larvae and containing 37 ± 5 larvae per gram were cooked to a final internal temperature of 66, 71, 77 or 82° C by one of eight methods to determine their efficacy in killing encysted larvae. The results indicate that with the time and temperatures used in this study, some rapid methods of cooking pork chops that involved the use of a microwave oven did not completely destroy T. spiralis larvae at 77 or 82° C. The data also showed that cooking pork chops to an internal temperature of 77° C in the conventional oven, convection oven, flat grill, charbroiler or deep fat gryer did inactivate encysted T. spiralis larvae in pork chops.

The filariform larvae of Strongyloides ransomi can infect their hosts by penetration through skin. In this report, homogenates of these organisms were prepared and their proteolytic enzymes examined. Homogenates prepared in 0.2 M citrate, pH 4.0, contain two thiol-dependent proteinases with molecular weights of approximately 32,000 and 28,000. These proteinases have an acidic pH optimum and show substrate preferences and inhibitor susceptibilities similar to the vertebrate acidic cysteinyl proteinases. Homogenates prepared in 0.1 M Tris, pH 7.5, contain multiple proteolytic enzymes, active against both Azocoll and synthetic substrates. These enzymes do not require thiols for activity and they have an alkaline pH optimum. The enzymes are inhibited by both chelating agents and heavy metals, but not by serine-proteinase inhibitors. Extracts prepared in 0.1 M Tris-HCl, pH 7.5, contain endogenous proteinase inhibitors.