Production and distribution of immunoglobulin-bearing cells in the intestine of young pigs infected with Ascaris suum

Pigs of 10 days-1 month old were orally infected with eggs of Ascaris suum at different rates and inoculation schedules. Histological sections from various parts of the small intestines were prepared to observe the production and localization of immunoglobulin-bearing cells. Fluorescent antibody and immunoperoxidase staining methods were used to determine the number of IgM-, IgA- and IgG-producing plasma cells in the intestinal lamina propria. Significant increases in immunoglobulin-bearing cells were observed in those pigs which received single inoculations of A. suum eggs. Pigs infected every 2,4,8 and 10 days with 10,000-20,000 embryonated eggs showed numerical increases in IgM-bearing cells. Increases in IgA-bearing cells were noted in pigs which received the higher number of eggs every 8-10 days. Higher concentrations of IgA- and IgM-bearing cells were observed in the jejunal mucosa of infected pigs as compared to those in the duodenum and ileum.     

Immunohistochemical distribution of viral antigens in pigs naturally infected with porcine teschovirus

A distribution of porcine teschovirus (PTV) antigens in pigs naturally infected with PTV is presented using the method of immunohistochemical examination. In the nervous system, PTV antigens were found in the cytoplasm of neuronal cells and glial cells distributed in the spinal ventral horn and brain stem, and also in the cytoplasm of ganglion cells in the spinal ganglion. No antigens were seen in the cerebral hemisphere. In the nervous system, the distribution of PTV antigens was consistent with lesions characteristic of nonsuppurative encephalomyelitis. In the other examined organ, PTV antigens were observed in bronchiolar epithelial cells in the lung, hepatocytes in the liver, epithelial cells in the tonsils and the myenteric nerve plexus in the small and large intestine.     

Tumor necrosis factor-like cytotoxicity and anorexia in Ascaris suum infected pigs

Tumor Necrosis Factor-Like Cytotoxicity (TNF-LC) was examined in sera from 12 pigs experimentally infected with Ascaris suum. The difference of TNF-LC levels between eight infected and four uninfected controls was not significant. When an endotoxin challenge was intravenously administered 1 month after the first dose of A. suum, the levels of TNF-LC in the sera of infected pigs were one-third that of the controls 125 min post-challenge (PC). In a more detailed study on four infected and two uninfected control pigs, TNF-LC was monitored every 10-15 min until 125 min after endotoxin challenge. The TNF-LC levels in these pigs increased at 40 min PC, reached maximum in another 10-25 min and then decreased. This pattern was seen in all except one infected pig. The infected pigs showed milder shock symptoms and their serum TNF-LC levels returned to pre-challenge levels 30 min earlier than controls.     

Presence of immunoglobulins and antigens in serum, lung and small intestine in Ascaris suum infected and immunised pigs

The immunodetection of local Ascaris suum antigens and local and systemic antibodies were analysed in pigs reinfected with eggs or immunized with the 14, 42 and 97 kilodalton (kDa) fractions from A. suum. Twenty-one Iberian pigs were divided in 7 groups of 3 pigs. Groups 1 and 2 were uninfected and challenge control groups, respectively. Groups 3 and 4 were infected weekly with increasing doses of A. suum eggs and Group 4 was additionally treated with pyrantel pamoate. Groups 5, 6 and 7 were immunised with the 14, 42 or 97 kDa fractions from adult worms, respectively. Groups 2-7 were challenged with 10,000 infective eggs. Animals of Groups 3 and 4 showed a pulmonary granulomatous reaction with moderate number of eosinophils and leukocytes, while Groups 5-7 presented higher number of cells, especially in animals immunized with the 42 kDa fraction. These immunized groups presented abundant deposition of Ascaris body fluid (BF) and body wall (BW) antigens as well as the 14 and 42 kDa fractions in the pulmonary and intestinal tissues, while lower deposition of antigens was observed in animals of Groups 3 and 4. The immunized pigs of Groups 5 and 6 showed the highest systemic IgG titres in serum and these antibodies were negatively correlated with the number of larvae recovered in the lungs, suggesting that the IgG response may have a protective function against the ascariosis. The highest concentrations of IgA-bearing cells were observed in animals of Groups 3 and 4 compared to the immunised pigs (Groups 5-7), suggesting that local IgA production may be involved in the protection against migrating larvae. The main localisations of IgA-bearing cells were the bronchial and peribronchial areas of lungs and the lamina propia of duodenum. Low numbers of local IgG-bearing cells were observed in all animals and no IgM-bearing cells were detected in the local tissues.     

Evidence of interaction between Ascaris suum and Metastrongylus apri in experimentally infected pigs

A study has been carried out with the aim to determine possible interactions between Ascaris suum and Metastrongylus apri under experimentally infected pigs. Twenty-eight Iberian pigs were allocated into four groups. Group 1 was inoculated with 5000 infective A. suum eggs; group 2 received concurrently 5000 infective A. suum eggs and 5000 infective M. apri larvae; group 3 received 5000 infective M. apri larvae; group 4 served as uninfected controls. In each group, pigs were necropsied on day 7 (n = 4) and day 28 (n = 3) post-infection (p.i.). Pigs with single M. apri infections showed earlier and more severe respiratory symptoms compared to pigs with mixed infection, while no clinical signs were observed in pigs single infected with A. suum. Mean burdens of immature A. suum and immature and adult M. apri were reduced in pigs with concomitant infection both on day 7 and 28 p.i., respectively. In contrast, the number of white spots was significantly increased on day 7 in pigs with mixed infection. In addition, pigs of group 1 showed the highest eosinophil levels in blood compared to pigs in groups 2 (intermediate levels) and 3 (moderate levels). The results suggest an antagonistic interaction between A. suum and M. apri in concomitantly infected pigs.     

Persistent activity of doramectin and ivermectin against Ascaris suum in experimentally infected pigs.

A study was conducted to investigate the persistent nematocidal activity of two avermectins against experimentally-induced infections of Ascaris suum in swine. Seventy-two nematode-free cross-bred pigs of similar bodyweight were randomly allotted to nine treatment groups of eight pigs each. Eight of the groups were treated with injectable solutions containing 300 microg of doramectin/kg (IM) or 300 microg of ivermectin/kg (SC) either 0 (same day), 7, 14, or 21 days prior to an oral challenge of 50000 embryonated A. suum eggs. The ninth group (control) was challenged in parallel without any avermectin treatment. At 41 or 42 days after challenge, pigs were euthanatized and adult and larval stages of A. suum were collected from the gastrointestinal tract of each pig and counted. Both avermectins significantly (P < 0.0002) reduced nematode counts when given on the day of challenge (0 days prior), and the efficacy was 100% and 97.5% for doramectin and ivermectin, respectively. Doramectin given 7 days prior to challenge significantly (P < 0.0001) reduced nematode counts, and the efficacy was 98.4%. For all other avermectin-treatment groups, nematode counts were not significantly reduced compared to those in control pigs. These data indicated that anthelmintic activity of ivermectin against A. suum persisted for less than 7 days and the activity of doramectin persisted for more than 7, but less than 14 days.     

Efficacy of piperazine dihydrochloride against Ascaris suum and Oesophagostomum species in naturally infected pigs

A controlled trial was performed to evaluate the efficacy of piperazine dihydrochloride in a new granular formulation (Ascarex D) against naturally occurring infections with Ascaris suum, Oesophagostomum dentatum and O quadrispinulatum. Treatment effects were estimated on the basis of parasites recoverable from the intestinal contents. Given orally at 200 mg per kg body weight the compound showed an efficacy of 99 to 100 per cent against A suum and the nodular worms. Egg excretion of the respective species was reduced by 98 per cent and 100 per cent six days after treatment. No adverse reactions were observed after the treatment.     

Mature Ascaris suum in naturally infected calves

In two small farms in Sweden young calves were found to be naturally infected with Ascaris suum. One of the calves expelled mature worms with the faeces, one had a great number of worms in the ductus choledochus and two others had worms in the intestine. Most of the worms were mature and at the egg-producing stage. The morphology of the eggs and the adult worms indicated that these parasites were A. suum and that Toxocara (syn. Neoascaris) vitulorum could be excluded.     

Concurrent Ascaris suum and Oesophagostomum dentatum infections in pigs.

The aim of this study was to examine interactions between Ascaris suum and Oesophagostomum dentatum infections in pigs with regard to population dynamics of the worms such as recovery, location and length; and host reactions such as weight gain, pathological changes in the liver and immune response. Seventy-two helminth-na?ve pigs were allocated into four groups. Group A was inoculated twice weekly with 10000 O. dentatum larvae for 8 weeks and subsequently challenge-infected with 1000 A. suum eggs, while Group B was infected with only 1000 A. suum eggs; Group C was inoculated twice weekly with 500 A. suum eggs for 8 weeks and subsequently challenge-infected with 5000 O. dentatum larvae, whereas Group D was given only 5000 O. dentatum larvae. All trickle infections continued until slaughter. Twelve pigs from Group A and B were slaughtered 10 days post challenge infection (p.c.i.) and the remaining 12 pigs from the each of the four groups were slaughtered 28 days p.c.i.. No clinical signs of parasitism were observed. The total worm burdens and the distributions of the challenge infection species were not influenced by previous primary trickle-infections with the heterologous species. Until day 10 p.c.i. the ELISA response between A. suum antigen and sera from the O. dentatum trickle infected pigs (Group A) pigs were significantly higher compared to the uninfected Group B. This was correlated with a significantly higher number of white spots on the liver surface both on Day 10 and 28 p.c.i. in Group A compared to Group B. The mean length of the adult O. dentatum worms was significantly reduced in the A. suum trickle infected group compared to the control group. These results indicate low level of interaction between the two parasite species investigated.     

Serum levels of gastrin, insulin and glucagon as possible factors of anorexia in pigs infected once with Ascaris suum

In order to determine possible mediators for development of anorexia in pigs infected with Ascaris suum, serum levels of gastrin, insulin and glucagon were measured. After a single high oral dose of 100,000-200,000 embryonated eggs the serum levels of gastrin and insulin in the infected pigs did not significantly differ from those in controls. Serum glucagon levels in the infected groups, however, were lower than those in controls and the difference was more evident 24 days postinoculation and later.     

Effect of fenbendazole and pyrantel tartrate on the induction of protective immunity in pigs naturally or experimentally infected with Ascaris suum

An experiment was conducted with 96 weanling pigs (avg initial wt 18.5 kg) divided into six treatment with two replicates of eight pigs each. Pigs in Treatments 1, 2 and 3 were penned in outside pens with dirt lots that previously were contaminated with A. suum ova to induce a natural ascaris infection. Pigs in Treatments 4, 5 and 6 were penned in an open-front building with solid concrete floors and were experimentally infected with 2,000 embryonated A. suum. ova on d 1, 15 and 29 of the experiment. Pigs in Treatments 1 and 4 were medicated with fenbendazole (FBZ, 3 mg/[kg BW.d]) for three consecutive days during three consecutive time periods. Pigs in Treatments 2 and 5 were medicated with pyrantel tartrate (PT, 106 mg/kg feed) for 28 d. Pigs in Treatments 3 and 6 served as infected, unmedicated controls. All pigs were challenged with 100 A. suum eggs 7 d after termination of the final FBZ treatment. All pigs were killed 66 d after challenge and worms were recovered. Fenbendazole treatment resulted in greater (P less than .07) average daily gain than PT treatment in pigs penned outside. Among inside pigs, FBZ treatment resulted in better (P less than .02) feed utilization than in controls. The FBZ and PT treatments reduced (P less than .03) the total number of A. suum, the length and weight of female ascarids and the length of male ascarids compared with controls. A natural continual infection with A. suum was less effective than experimental infection in inducing protective immunity in pigs.     

A novel technique for identification of Ascaris suum cohorts in pigs

The objective of the present study was to develop a fast, cheap and reliable technique for identifying different cohorts of the swine parasite, Ascaris suum. A polymerase chain reaction linked restriction fragment length polymorphism (PCR-RFLP) technique on mt-DNA was used to identify unique haplotypes of four gravid A. suum females on agarose gels after eggs were recovered from each of the worms. Each of four pigs was inoculated with 2000 embryonated eggs originating from one of the four identified Ascaris haplotypes, respectively. Ascaris larvae were isolated from the small intestine at day 14 post-infection using an agar technique. Single larvae from each pig were transferred to 96-well PCR plates and a simple DNA extraction using a worm lysis buffer was carried out and followed by the PCR-RFLP analysis. More than 100 larvae from each of the four pigs were analysed and all were found to have the same haplotype as the parental female. We conclude that unique haplotypes of female A. suum and offspring can be identified by means of PCR-RFLP on mt-DNA and suggest that this method can be used in future research on Ascaris population biology using cohorts with distinct mt-DNA profile.     

Pharmacokinetics of ampicillin and sulfadimidine in pigs infected experimentally with Streptococcus suum

Twenty-three hybrid pigs (23 ± 3 kg body wt) were assigned to three groups to investigate the pharmacokinetics of ampicillin (APC, 10 mg/kg) administered intravenously (i.v.) and intramuscularly (i.m.), and sulfadimidine (SDM, 50 mg/kg) administered intravenously as a bolus injection. In the first series of experiments the animals remained healthy. Subsequently, the pigs were infected with Streptococcus suum by subcutaneous (s.c.) inoculation and the experiments were repeated. The total apparent distribution volume of APC given intravenously was increased from 0.512 ± 0.026 L/kg in uninfected pigs to 0.68 ± 0.06 L/kg (P < 0.01) in infected pigs, whereas there were no significant changes in the same parameter for SDM (P > 0.05). The clearance of APC was increased markedly from 0.52 ± 0.07 L/kg/h in uninfected pigs to 0.62 ± 0.10 L/kg/h in infected pigs. In contrast, SDM clearance was decreased markedly from 0.023 ± 0.003 L/kg/h to 0.017 ± 0.003 L/kg/h (P < 0.05). As a result, the biological half-lives of the drugs were altered to varying degrees in infected pigs. The half-life of SDM was increased from 15.0 ± 3.0 h in uninfected pigs to 20 ± 7h in infected pigs (P < 0.05), but differences in APC half-lives between uninfected and infected animals were not observed (P > 0.05). There were no statistically significant differences in pharmacokinetic parameters of APC administered by intramuscular injection between the healthy and the diseased status, although its half-life was shortened from 0.76 ± 0.22 h in the healthy to 0.57 ± 0.23 h in the diseased. The results suggest that blood concentrations of APC and SDM are affected differently by the same disease due to its specific effects on their distribution and elimination.     

Blocking of nonspecific IgG in indirect radioimmunoassay for detecting Ascaris suum antigens

The sensitivity of an indirect radioimmunoassay (IRIA) used for detecting larval body wall (LBW) antigens of Ascaris suum was enhanced by using various blocking agents which prevented nonspecific binding of immunoglobulin G (IgG) or free ¹²⁵I without preventing binding of specific antibodies to the antigen. The use of blocking agents reduced counts for both positive and negative sera, resulting in an increase in calculated binding ratios (BR) and deltas and, thus, in the sensitivity of the assay. The relative effectiveness of blocking agents, in decreasing order, were turkey serum (TS), rabbit gamma globulin (RGG), rabbit whole serum (RS), bovine serum albumin (BSA), rabbit IgG (RIgG), bovine alpha globulin (BAG), bovine gamma globulin (BGG) and Tween-20.