Natural nodular worm infection in goats induces eosinophil extracellular DNA trap (EET) formation.

Extracellular DNA Trap (ET) formation by granulocyte is a strong innate immune machinery that plays crucial roles in trapping and killing of pathogens. Here, we show Eosinophil Extracellular DNA Trap (EET) formation in goats naturally infected with nodular worms (Oesophagostomum columbianum, Strongyloidae: Nematoda). By a slaughterhouse based survey, we found that 60% goats were infected with nodular worms. We detected numerous, hard and pale yellow to dark black nodules of variable sizes (0.25-2 cm) in the large intestine and the number of nodules were significantly (p < .05) higher in the cecum (21.7 ± 17.9) than in the colon (10.1 ± 9.9). Histologically, pink colored circumscribed caseous mass was surrounded by a dense zone of infiltration and fibrous proliferation along with massive infiltration of eosinophils in and around the necrotic mass. DAPI staining revealed huge accumulation of extracellular DNA, which formed wide ridge like structure surrounding the necrotic zone. Massive release of eosinophils cationic proteins (ECP), a helmintho-toxic substance, was found into the lesions. Collectively, our results suggest that nodular worm infection induces EETosis and ECP release, and is one of the major parasitic problem affecting Black Bengal goats that causes distortion of normal architecture of the gut wall.

Excretion in feces and mucosal persistence of Salmonella ser. Typhimurium in pigs subclinically infected with Oesophagostomum spp.

OBJECTIVE: To determine interactions between Oesophagostomum spp and Salmonella ser. Typhimurium in pigs. ANIMALS: 30 healthy 5- to 6-week-old pigs. PROCEDURE: Pigs were allotted to 3 groups (n = 10 pigs/group) and treated as follows: group A was given Oesophagostomum dentatum and O quadrispinulatum; group B was given O dentatum, O quadrispinulatum, and S Typhimurium; and group C was given S Typhimurium only. Pigs in groups A and B were trickle infected with Oesophagostomum spp 3 times weekly throughout the study. After 19 days, groups B and C were inoculated once with S Typhimurium. One pig from each group was euthanatized on the day of Salmonella exposure and 2 and 4 days after Salmonella exposure. The remaining pigs were euthanatized on days 16 and 17 after Salmonella exposure. RESULTS: Pigs with dual infections of nematodes and bacteria (group B) excreted significantly higher amounts of S Typhimurium in feces, compared with nematode-free pigs (group C). In addition, group-B pigs excreted S Typhimurium on more days than pigs in group C. Salmonella Typhimurium was detected in the cecum and colon in the majority of pigs in group B, whereas S Typhimurium was only detected in the colon in pigs in group C. Immunohistochemical examination detected S Typhimurium in 7 of 9 pigs in group B but only 2 of 9 pigs in group C. CONCLUSIONS AND CLINICAL RELEVANCE: Interactions between intestinal nematodes and bacteria may play an important role in the dynamics of S Typhimurium infections.

Development and infectivity of eggs and larvae derived from pigs trickle-infected with Oesophagostomum dentatum at different dose levels.

We examined the impact of different Oesophagostomum dentatum dose levels and durations of infection on the development and infectivity of the following generation. Pigs were trickle-infected with 200, 2,000 or 20,000 L3/week over 20 weeks. Egg hatch assays were performed at monthly intervals; however, no consistent differences were found between any of the dose groups in the development of eggs into first-stage larvae. To compare larval infectivity, larvae were derived from faecal cultures set up from the low- and the high-dose groups in the early and the late part of the experiment, and were inoculated into helminth-free pigs (5,000 L3/pig). Worm establishments were significantly higher (P<0.05) in the group of pigs receiving larvae derived early in the experiment from the low-dose group compared with the two groups receiving larvae from high-dose groups, thus indicating an adverse effect of high doses of trickle infection on the later infectivity of L3 larvae derived from excreted eggs.

Electro- and transportphysiological changes in pig proximal colon during parasitic infection.

Oesophagostomum dentatum, one of the most common nematodes in pigs, causes the formation of subepithelial granuloma in the large intestine. To investigate possible changes in epithelial function or response during the infection we incubated epithelia of pig proximal colon in Ussing chambers at different days post infectionem (p.i.). Transepithelial conductance, gt, and the Cl flux from serosal to mucosal, JsmCl, were increased on day 2 p.i., when the nematodes penetrate the epithelium of the large intestine, and declined toward control levels thereafter. Histamine, PGE2 and carbachol caused transient increases in short circuit current, Isc, and conductance that could partly be attributed to a higher JsmCl. The Isc responses were highest on the days of nematode penetration in or out of the epithelium (days 2 and 14 p.i.) and did decline on day 7 p.i. during the histotropic development of the parasite. This reduced epithelial reaction on day 7 p.i. might be an adaptation to secretory stimuli released from the inflammatory cells in the intestinal wall or might reflect modulation by the parasite and could be responsible for the absence of marked clinical signs during the infection.

The effect of fasting on Ascaris suum and Oesophagostomum spp. in growing pigs.

Experiments were conducted to study the possible influence of fasting on Ascaris suum and Oesophagostomum spp. In growing pigs. Forty young crossbred pigs naturally infected with A. suum and Oesophagostomum spp. were used. In one experiment 10 pigs were fasted and offered water ad libitum for 6 days, in another experiment for 10 days. Subsequently, these pigs, together with 10 non-fasted control pigs per experiment were slaughtered, and worm numbers, worm location, sex, developmental stage and female worm fecundity were determined. Pigs fasted for 10 but not for 6 days had decreased numbers of A. suum and Oesophagostomum spp. at slaughter vs controls, and worms were found in more distal locations in the gastrointestinal tract. Fasting for both 6 and 10 days significantly lowered the fecundity of both worm species.

Prepatent periods of different Oesophagostomum spp. isolates in experimentally infected pigs.

To define prepatent periods of different Oesophagostomum spp. isolates we carried out two separate experiments, one using two monospecific laboratory isolates and another using laboratory isolates as well as isolates obtained from pig herds having different management systems and with different anthelmintic treatment histories. Pigs were inoculated with 1,000-2,000 infective larvae. Fecal samples were collected daily beginning on days 15 and 16 postinoculation (p.i.). Fecal cultures were set up at different times to yield larvae that could be identified by DNA analyses. All pigs started to excrete eggs on days 18-24 p.i. The mean prepatent period was 20.2 +/- 1.4 days, with no significant difference being observed between species and isolates. Prepatent periods of 17-19 days were found for the monospecific laboratory isolates of O. dentatum and O. quadrispinulatum. These findings conflict with parasitology textbooks; therefore, suggestions as to the possible reasons for the observed short prepatent periods are given.

A possible influence of diet composition on the establishment of nematodes in the pig.

Two groups of 20 pigs weighing approx. 26 kg were offered either ground barley plus protein supplement (GBP) or commercial pelleted feed (CPF), two diets consisting of nearly the same amounts of carbohydrates, proteins and micronutrients. All pigs were experimentally infected with 600 infective Ascaris suum eggs and 6000 infective Oesophagostomum spp. larvae and followed coprologically for approx. 12 weeks post infection, whereafter they were slaughtered. The number of Oesophagostomum spp. and A. suum eggs was significantly higher in the GBP-fed pigs compared with the CPF-fed pigs (P < 0.0001). The mean worm burden of adult Oesophagostomum spp. was higher in the GBP-pigs than in the CPF-pigs (P < 0.05), while the mean worm burden of adult A. suum in the two groups was not significantly different. The results indicate that two diets, both fulfilling the requirements of modern pig raising, may result in different establishment rates of natural adult gastrointestinal helminths in swine.

Experimental Oesophagostomum dentatum infections in the pig: worm populations resulting from trickle infections with three dose levels of larvae.

This paper describes the worm populations in pigs experimentally infected by trickle infections with different dose levels of the nodular worm, Oesophagostomum dentatum. Four groups each of 20 helminth naïve pigs, 10-12 weeks old, were inoculated with 0 (group 1), 100 (group 2), 1000 (group 3), or 10,000 (group 4) infective larvae twice weekly, and the pigs were killed after 10-13 weeks. No overt clinical signs were observed, and only group 4 had slightly lower food conversion rate (P < 0.05) than the controls. Faecal egg counts revealed that the nodular worms in pigs of groups 2 and 3 had a short prepatent period (3-4 1/2 weeks) and a fairly stable egg output, while the worms in the pigs of group 4 had prepatent periods of 3-10 weeks and low, unstable egg excretion. The mean worm burdens increased with the dose rate (group 2: 929 worms; group 3: 7467 worms; group 4: 19,847 worms), but detailed analyses of the worm populations from 10 pigs from each of the infected groups revealed a clear dose-dependency in worm recovery, percentage adult worms, worm lengths and female fecundity, as all these measures declined significantly with increasing dose level. The adult worms seemed to be shorter and less fertile when they were located posteriorly to their predilection site, and especially in group 4 many stunted infertile adults measuring only 2-5 mm were found in the posterior half of the colon, but there were no indications of worm expulsion. Superimposed on the main experiment was a cohort study in which 4 pigs of group 3 were given a single dose of 1000 pyrantel resistant larvae at day 56 (all other larvae were pyrantel sensitive), treated with 28 mg pyrantel per kg body weight at day 85 and killed at day 90. Appropriate control groups were included. The mean establishment of the cohort was similar to previously uninfected controls, but between-animal variation was much higher in the trickle infected group.

Growth and fecundity of Oesophagostomum dentatum in high-level infections and after transplantation into naive pigs.

This experiment was designed to examine the growth, proportion of stages, and fecundity of an Oesophagostomum dentatum population by transplantation of a known small number of worms from a high-density population into helminth-naive recipient pigs. Approximately 1,500 4-week-old worms [69% fourth-stage larvae (L4), 31% adult worms] were transplanted into each of 5 recipient pigs (group B), and these pigs, along with a group of 5 high-level-infection control pigs (group C), were killed at 4 weeks after transplantation to determine and compare the worm burdens. By 2 weeks after transplantation and throughout the experiment, fecal egg counts of group B exceeded those of group C and the fecundity of the worms was higher, though not statistically significantly so, in the transplanted worms. In the recipient pigs, all worms (approx. 70% establishment) had developed to the adult stage and were significantly longer than worms recovered from the group C pigs.

Development of the free-living stages of Hyostrongylus rubidus and Oesophagostomum spp. at different temperatures and humidities.

Faeces containing a mixture of Hyostrongylus rubidus and Oesophagostomum spp. eggs were mixed with vermiculite and water and set up at combinations of different relative humidities (65.5%, 79.5% 90.0%, and 100%) and temperatures (5 degrees C, 10 degrees C, 15 degrees C, 20 degrees C, and 25 degrees C) in order to study the rate of egg hatching and larval development. The study established that the development from egg to infective larva showed similar patterns for the two parasites. Optimum development and survival was in the temperature range from 15 degrees C to 20 degrees C and at humidities from 79.5 to 95.5%.

[Digestibility of nutrients in a feed mixture in certain swine helminthiases]. / Stravitelnost zivin krmné smesi pri vybraných helmintózách prasat.

The average digestibility coefficients of feed mixture nutrients in pigs infected medium severely to severely by the helminths Ascaris suum and Oesophagostomum dentatum were lower in comparison with the post-dehelminthization condition (dry matter by 3.1%, crude protein by 6.7%, fat by 8.9%, nitrogen-free extracts and fibre by 2.2%, organic matter by 3.1%). The content of total digestible nutrients made 679 g in infected pigs and 708 g in dehelminthized pigs. In the period following the experimental infection of pigs by the helminths Ascaris suum, characterized by the formation of antibodies and adaptation after heavy injury of the organism, the digestibility coefficients of feed mixture nutrients were in comparison with the state prior to infection influenced positively. In two metabolism trials, on the 9th to 13th day after infection the digestibility coefficients increased as follows: of dry matter by 4.8% and 3.6%, crude protein by 8.2 and 4.2%, nitrogen-free extracts and fibre by 2.0 and 3.5% and organic matter by 3.0 and 3.3%. The decrease in fat digestibility coefficients was observed in both cases, by 12.4% and 9.9%. Prior to infection the content of total digestible nutrients was 712 g and 682 g, after infection 731 g and 707 g.

Influence of an experimental infection of nodular worms (Oesophagostomum spp.) on performance of pigs.

Four trials were conducted to determine the effect of different levels of nodular worm (Oesophagostomum spp.) infection on the performance of growing-finishing swine and on digestion and absorption of nutrients by the growing pig. In each of two growing-finishing trials, 48 crossbred pigs (average body weight 26.6 kg in trial 1 and 24.4 kg in trial 2) were individually housed and given a single infection of nodular worms (0, 550, 1,100 or 1,650 infective larvae/kg body weight in trial 1 and 0, 1,500, 3,000 or 4,500 infective larvae/kg body weight in trial 2). At 21 days postinfection, pigs given either 1,100 or 1,650 nodular worm larvae/kg body weight required more feed (P less than .05) per unit gain than pigs not infected, but for the 77-day test period, as a whole, nodular worms did not significantly affect performance. in trial 2, pigs given either 3,000 or 4,500 nodular worm larvae/kg body weight were lighter (P less than .05) and gained weight slower (P less than .05) for the first 21 days postinfection than pigs not infected, while pigs given 4,500 larvae gained slower (P less than .05) than pigs on the other three treatments. During the first 21 days after infection, pigs given 4,500 larvae/kg body weight consumed less feed (P less than .05 than pigs given either 0 or 1,500 larvae. Even at the higher levels of infection, nodular worms did not significantly affect performance over the entire 77-day test period. In each of the two other trials, eight crossbred barrows (average body weight 28.8 kg in trial 1 and 28.4 kg body weight in trials 2) were examined for the effects of two levels of nodular worm infection (0 and 4,950 larvae/kg body weight) on digestion and absorption of nutrients and on N balance. Pigs experimentally infected had a lower digestion coefficient for dry matter (P less than .01), ash (P less than .05), crude fiber (P less than .01), N free extract (P less than .05) and N (P less than .05) than noninfected pigs. Pigs not infected with nodular worms had a higher (P less than .05) N balance.