Comparison of innate and Th1-type host immune responses in Oesophagostomum dentatum and Trichuris suis infections in pigs.

This study investigated the details of the innate and Th1/Treg-type-associated host immune responses in Trichuris suis and Oesophagostomum dentatum mono- and co-infected pigs and in vitro in stimulated porcine dendritic cell cultures. Forty-eight pigs were allocated into a 2-factorial design with two groups trickle-inoculated with 10 T. suis eggs/kg/day (Group T) or 20 O. dentatum L3/kg/day (O). Another group (OT) was infected with both parasites. Group C remained uninfected. Expression of innate and Th1/Treg-cell-associated genes in gut mucosa and associated lymph nodes was determined by qPCR at necropsy day 35 and 71. Gene expression showed suppressed/inhibited Th1 and Treg-type immune reactions, in accordance with previous findings of a predominant Th2-type immune response to both nematodes. The in vitro part examined the production of TNF-α in porcine dendritic cells (DC) exposed to T. suis and/or O. dentatum excretory/secretory (E/S) products. Further, binding capacity and structure of E/S products were characterized. Glycan and lectin-binding capacity were generally lower in O. dentatum E/S products compared to T. suis which may explain the earlier found weaker Th2 response to the former. Surprisingly, O. dentatum E/S products induced a significant (P < 0·0001) increase in TNF-α DC production, potentially indicating a new mode of helminth-host immune response interaction.

Gastrointestinal parasites of cats in Denmark assessed by necropsy and concentration McMaster technique.

The large population of feral cats in Denmark may potentially transmit pathogens to household cats and zoonotic parasites to humans. A total of 99 euthanized cats; feral cats (n=92) and household cats with outdoor access (n=7), were collected from March to May 2014 from the Zealand region, Denmark. The sedimentation and counting technique (SCT) was used to isolate helminths and coproscopy was done by concentration McMaster technique (c-McMaster). Overall, 90.1% of the cats were infected and a total of 10 species were recorded by SCT: 5 nematode species: Toxocara cati (84.8%), Ollulanus tricuspis (13.1%), Aonchotheca putorii (7.1%), Paersonema spp. (3.0%), Strongyloides spp. (1.0%); 3 cestodes: Hydatigera taeniaeformis (36.4%), Mesocestoides sp. (3.0%), Dipylidium caninum (1.0%); and 2 trematodes: Cryptocotyle spp. (5.1%) and Pseudamphistomum truncatum (1.0%). O. tricuspis was the second most common gastrointestinal nematode of cats but had the highest intensity of infection. For T. cati, prevalence and worm burden were significantly higher in feral than household cats. No juvenile cats were infected with H. taeniaeformis, and age thus had a significant effect on prevalence and worm burdens of this species. Rural cats had a higher prevalence and worm burden of A. putorii than urban cats. By c-McMaster, ascarid, capillarid, strongylid or taeniid type eggs were found in 77.9% of the cats while Cystoisospora felis was found in 2.1%. The sensitivity of the c-McMaster was 82.5% for T. cati but 26.5% for taeniid eggs, using the SCT as gold standard. A positive correlation between faecal egg counts and worm burdens was seen for T. cati, but not for taeniid eggs (assumed to be H. taeniaeformis). Coprological examination also detected the eggs of extraintestinal Capillariidae species including Eucoleus aerophilus and Eucoleus boehmi, but further necropsy studies are needed to confirm these findings.

Detection of a quantitative trait locus associated with resistance to infection with Trichuris suis in pigs.

Whipworms (Trichuris spp.) infect a variety of hosts, including domestic animals and humans. Of considerable interest is the porcine whipworm, T. suis, which is particularly prevalent in outdoor production systems. High infection levels may cause growth retardation, anaemia and haemorrhagic diarrhoea. A significant proportion of the variation in Trichuris faecal egg count (FEC) has been attributed to the host's genetic make-up. The aim of the present study was to identify genetic loci associated with resistance to T. suis in pigs. We used single nucleotide polymorphism (SNP) markers to perform a whole-genome scan of an F1 resource population (n = 195) trickle-infected with T. suis. A measured genotype analysis revealed a putative quantitative trait locus (QTL) for T. suis FEC on chromosome 13 covering ∼ 4.5 Mbp, although none of the SNPs reached genome-wide significance. We tested the hypothesis that this region of SSC13 harboured genes with effects on T. suis burden by genotyping three SNPs within the putative QTL in unrelated pigs exposed to either experimental or natural T. suis infections and from which we had FEC (n = 113) or worm counts (n = 178). In these studies, two of the SNPs (rs55618716, ST) were associated with FEC (P < 0.01), thus confirming our initial findings. However, we did not find any of the SNPs to be associated with T. suis worm burden. In conclusion, our study demonstrates that genetic markers for resistance to T. suis as indicated by low FEC can be identified in pigs.

Third-stage nematode larvae of Contracaecum osculatum from Baltic cod (Gadus morhua) elicit eosinophilic granulomatous reactions when penetrating the stomach mucosa of pigs.

Third-stage larvae of the anisakid nematode Contracaecum osculatum were recovered from livers of Atlantic cod (Gadus morhua) caught in the Baltic Sea (June 2014) and used for experimental infection of two pigs (one male and one female). Each pig received 215 larvae by oral infection (feeding with minced cod liver containing live nematode larvae). Pigs were euthanized after 5 days, necropsied, and subjected to parasitological investigation. A total of 12 larvae were found penetrating the mucosa of the ventricle (7 in the female pig and 5 in the male pig) eliciting a granulomatous reaction at the penetration site. Four non-attached larvae were found in the female pig stomach and one in the male pig. Petechial bleeding was observed at several locations in the ventricular mucosa where larvae were located. Histological examination of the stomach mucosa revealed a massive cellular infiltration (giant cells, lymphocytes, macrophages, granulocytes, and fibroblast like cells) around the penetrating larva. Mononuclear and polymorphonuclear cells containing eosinophilic granulae were particularly prominent in the granulomas. Reactions correspond to reactions in pigs following experimental infection with the human pathogenic anisakid larvae Anisakis sp. and Pseudoterranova sp. which suggests that C. osculatum might have a zoonotic potential as well.

Helminth parasites in pigs: new challenges in pig production and current research highlights.

Helminths in pigs have generally received little attention from veterinary parasitologists, despite Ascaris suum, Trichuris suis, and Oesophagostomum sp. being common worldwide. The present paper presents challenges and current research highlights connected with these parasites. In Danish swine herds, new indoor production systems may favour helminth transmission and growing knowledge on pasture survival and infectivity of A. suum and T. suis eggs indicates that they may constitute a serious threat to outdoor pig production. Furthermore, it is now evident that A. suum is zoonotic and the same may be true for T. suis. With these 'new' challenges and the economic impact of the infections, further research is warranted. Better understanding of host-parasite relationships and A. suum and T. suis egg ecology may also improve the understanding and control of human A. lumbricoides and T. trichiura infections. The population dynamics of the three parasites are well documented and may be used to study phenomena, such as predisposition and worm aggregation. Furthermore, better methods to recover larvae have provided tools for quantifying parasite transmission. Thus, an on-going study using helminth naïve tracer pigs has surprisingly demonstrated that soil infectivity with A. suum and T. suis increases during the first 2-3 years after pasture contamination. Though all three helminth species stimulate the Th2 arm of the immune system, Oesophagostomum seems weakly immunogenic, perhaps via specific modulation of the host immune system. A. suum and T. suis potently modulate the host immune response, up-regulating Th2 and down-regulating Th1. As a consequence, A. suum may compromise the efficacy of certain bacterial vaccines, whereas T. suis, which establish only short-term in humans, is a favourite candidate for down-regulating autoimmune Th1-related diseases in man. Some basic research findings have offered new possibilities for future sustainable control measures. For example, the heredity of host resistance to A. suum and T. suis is so high that breeding for resistant pigs may be a possibility. Experimental studies have demonstrated that fermentable dietary carbohydrates have an antagonistic effect on Oesophagostomum and to a lesser extent on T. suis and A. suum, whereas egg-destroying microfungi may be used to inactivate the hard-shelled A. suum and T. suis eggs in the environment. Helminth control in Denmark has previously relied solely on anthelmintic treatment in herds with low helminth transmission. When indoor transmission rates increase, or in outdoor herds with high pasture contamination levels, medication may advantageously be combined with sustainable control measures, such as selected pig genomes, bioactive forages, and egg-destroying microfungi.

The effect of a diet with fructan-rich chicory roots on intestinal helminths and microbiota with special focus on Bifidobacteria and Campylobacter in piglets around weaning.

The restrictions on the use of antibiotic and anthelmintic treatments in organic pig farming necessitate alternative non-medical control strategies. Therefore, the antibiotic and parasite-reducing effect of a fructan-rich (prebiotic) diet of dried chicory was investigated in free-ranging piglets. Approximately half of 67 piglets from nine litters were experimentally infected with Ascaris suum and Trichuris suis in the suckling period (1 to 7 weeks of age) and 58 of the piglets were challenged daily with Eschericia coli O138:F8 for 9 days after weaning to induce weaning diarrhoea. The litters were fed either chicory (30% dry matter) or a control diet. The effect of chicory on intestinal helminths, intestinal microbiota, especially Bifidobacteria and Campylobacter spp. and E. coli post-weaning diarrhoea was assessed. The weight gain of the piglets was not impaired significantly by chicory. The intestinal A. suum worm burden was reduced by 64% (P = 0.034) in the chicory-fed piglets, whereas these same piglets had 63% more T. suis worms (P = 0.016). Feeding with chicory elicited no changes among the main bacterial groups in ileum according to terminal restriction fragment length polymorphism analysis. However, the terminal-restriction fragment (T-RF) 208 bp, which may belong to Lachnospiraceae, was stimulated by the chicory feed (P = 0.03), and T-RF 370 bp that matches Enterobacter belonging to the Enterobacteria was reduced (P = 0.004). In addition, chicory increased the level of Bifidobacteria (P = 0.001) and the faecal Campylobacter excretion level was transitorily reduced in chicory-fed piglets at 7 weeks of age (P = 0.029). Unfortunately, it was not possible to assess the effect of chicory on post-weaning diarrhoea as it did not develop. In conclusion, feeding piglets chicory around the time of weaning caused complex changes of the microbiota and parasite communities within the intestinal tract, and feeding piglets chicory may therefore serve as an animal-friendly strategy to control pathogens.

Ascaris suum infections in pigs born and raised on contaminated paddocks.

The transmission of Ascaris suum was studied in outdoor reared pigs. From May to June 2001, 6 farrowing paddocks were naturally contaminated with A. suum using experimentally infected seeder pigs. Early July, 1 sow farrowed on each paddock. One piglet per litter was slaughtered every second week starting at week 3 post-partum (p.p.) for registration of liver white spots and recovery of A. suum from the lungs and the small intestine. The last pigs were slaughtered at week 19 p.p. Faeces was examined for parasite eggs and blood was analysed for A. suum-specific antibodies. Weaning took place at week 7 p.p. by removing the sow. Paddock infection levels were estimated by regular examination of soil samples and in late June and late November using parasite naïve tracer pigs. Paddock contamination was high but eggs developed slowly resulting in a low initial transmission to the experimental pigs. By week 5 p.p. transmission had increased and the numbers of infective eggs in the soil increased during the study. The results indicate a continuous uptake of infective eggs, but visceral larval migration was reduced with time, probably due to the development of a pre-hepatic barrier. Nevertheless, a rather large population of adult worms remained in the pigs throughout the study, and it may primarily have been eggs ingested in the early infection phase that gave rise to the patent infections. It is suggested that neonatal exposure may result in increased persistence and size of adult worm burden and that the higher 'life-time worm burden' may be of significant economic importance.

Oesophagostomum dentatum and Trichuris suis infections in pigs born and raised on contaminated paddocks.

Transmission of Oesophagostomum dentatum and Trichuris suis was studied in outdoor reared pigs. Six farrowing paddocks were naturally contaminated in May to mid-June 2001 by experimentally infected seeder pigs. In early July 1 sow farrowed on each paddock and starting at week 3 post-partum (p.p.) the offspring was slaughtered serially every 2 weeks for parasite recovery. Faeces were collected regularly for parasite egg counts and acid-insoluble ash (AIA) content as an indicator of geophagy. Weaning took place at week 7 p.p. by removing the sow. Paddock infection levels were estimated in mid-June (O. dentatum) and late November (O. dentatum and T. suis) using helminth-naïve tracer pigs. Soil and vegetation samples were collected regularly. Despite a high initial contamination by the seeder pigs, O. dentatum paddock infectivity was negligible to low throughout the raising of the experimental piglets, which had a slow accumulation of nodular worms ending with a mean of 422 worms/pig at week 19 p.p. As only few eggs developed to infectivity overall T. suis transmission was minimal. The first T. suis were recovered at week 11 p.p. and the highest mean burden of 21 worms/pig was recorded at week 19 p.p. The experimental pigs initially had a high faecal level of AIA although it decreased over time. The results are discussed in relation to the biological characteristics of the 2 parasites and their occurrence in organic pig production.

The influence of stocking rate on transmission of helminth parasites in pigs on permanent pasture during two consecutive summers.

This study was made to elucidate the transmission of nematode infections in outdoor pigs at different stocking rates during two consecutive seasons. Five pigs (Group 1A) inoculated with low doses of Oesophagostomum dentatum, Ascaris suum, and Trichuris suis and five helminth-naïve pigs (Group 1B) were turned out together in June 1996 on each of four pastures at stocking rates of 100, 240 (two pastures) and 576m(2) per pig, respectively. The pigs were slaughtered in early October, and pasture infectivity was subsequently measured using helminth-naïve tracer pigs (Tracer). In 1997, 10 helminth-naïve pigs were turned out on each pasture in May (Group 2) and again in August (Group 3), and allowed to graze for 12 weeks. The percentage of grass cover was reduced considerably at the high stocking rate in comparison to the other stocking rates. Transmission of all three helminths was observed on all pastures. In 1996, the O. dentatum faecal egg counts and worm burdens were significantly higher in pigs at the high stocking rate compared to pigs at the other stocking rates. O. dentatum did not survive the winter and pigs of Group 2 were inoculated with 3000 larvae each to reintroduce this parasite. Ascaris suum ELISA values and worm counts were highest at the high stocking rate in 1997 (Group 3). Transmission of T. suis was not significantly influenced by stocking rate. The results indicate that transmission of O. dentatum, and to some extent A. suum is influenced by stocking rate. However, both A. suum and T. suis eggs are still expected to constitute a high risk of infection on intensively used pastures where eggs may accumulate for years. The relationship between host density and helminth transmission seems more complex for grazing/rooting pigs than for grazing ruminants. This may be due to the differences in behaviour of the animals and the resulting differences in microclimate of the developing eggs/larvae.

Nose-rings and transmission of helminth parasites in outdoor pigs.

Five growing pigs experimentally infected with low doses of Oesophagostomum dentatum, Ascaris suum, and Trichuris suis were turned out with 5 helminth-naïve pigs on each of 3 pastures in June 1996 (Group 1). On one pasture all pigs received nose-rings. After slaughter of Group 1 in October, pasture infectivity was monitored using helminth-naïve, unringed tracer pigs. In 1997, helminth-naïve young pigs were turned out on the contaminated pastures in May (Group 2) and again in August (Group 3). Again all pigs on one pasture received nose-rings. All pigs and pastures were followed parasitologically and reduction in grass cover was monitored. Based on the acquisition of infection by the naïve pigs in Group 1, the estimated minimal embryonation times for eggs deposited on pasture were 23-25 days for O. dentatum, 5-6 weeks for A. suum and 9-10 weeks for T. suis. Results from tracer pigs and grass/soil samples indicated that pasture infectivity was light both years. Free-living stages of O. dentatum did not survive the winter. The nose-rings reduced rooting considerably, resulting in three-fold more grass cover on the nose-ring pasture compared to the control pastures by the end of the experiment. Nevertheless, the nose-rings did not significantly influence parasite transmission.