Ascaris suum Infection in Calves I. Clinical Signs

Clinical signs consistent with those of atypical interstitial pneumonia (AIP) were induced in calves sensitized with infective Ascaris suum eggs at seven to 20 weeks of age and challenged at three-week intervals one or more times. These signs usually appeared on the sixth or seventh day postinfection and reached maximum severity between the tenth and 13th days following infection. Prominent signs were: dyspnea, often with expiratory grunt, coughing, mouth breathing and emphysema as well as increased respiration and heart rates. In general, the intensity of signs was dependent upon dose size, although a single small dose resulted in acute signs and death in one calf. Intermittent coughing and vesicular sounds were induced in calves given A. suum eggs continually over prolonged periods. No respiratory abnormalities resulted from challenge with Toxocara canis after sensitization with A. suum. Antihistamine therapy did not alter the clinical signs in A. suum infected calves.     

Effects of Salmonellosis on subsequent infections with Ascris suum in swine

Weanling pigs were exposed to Salmonella cholerae-suis var. kunzendorf and sixteen days later were inoculated with infective Ascaris suum eggs. Necropsy at seventeen days post-Ascaris-infection revealed fewer lesions of hepatic fibrosis and a lesser degree of tissue eosinophilia in the hepatic interstitium in pigs receiving inoculations of Salmonella and Ascaris compared to those receiving Ascaris inoculation only. The numbers of juvenile nematodes recovered from the small intestines of both groups were variable. The comparatively less-intensive hepatic lesions may have been caused by reduction of the number of penetrating larvae at the intestinal level. Alternatively, the decreased tissue eosinophilia and fibrosis of the liver may be due to the previous stress of the Salmonella infection.     

Experimental Ascaris suum infection in the pig. Population kinetics following low and high levels of primary infection in piglets

Groups of piglets 20 to 28 days of age were dosed with 50 or 10 000 Ascaris suum eggs. The pigs were sacrificed from day 16 post infection onwards and worms present in the intestine were counted. Large numbers of immature parasites were recovered initially following the 10 000 egg dose, but the numbers decreased rapidly with the and worms were not found in pigs examined in the late prepatent phase. All pigletts infected with 50 eggs were found to harbour worms and the infection reached patency. The high level infections resulted in an eosinophil response which reached maximum values during the second week after infection. The low level infections which resulted in patency were not followed by any significant eosinophil response.     

The protective effect of developmental stages in an Ascaris suum infection in the guinea pig

The capacity of various developmental stages of Ascaris suum to induce a protective immune response in the guinea pig model was evaluated. Larvated eggs, second, third or fourth stage larvae were injected into guinea pigs by either the suncutaneous, intramuscular, ear vein or mesenteric vein route. Animals were challenged with a mesenteric vein injection of artificially hatched infective larvae of A. suum. Second stage larvae produced the highest degree of immunity and fourth stage larvae produced the least protection comparing all routes of administration. The most effective route of immunization for all the developmental stages was the mesenteric vein. Antibody titer as assessed by indirect hemagglutination was not correlated with the degree of protection.     

Development of immune responsiveness to Ascarissuum antigens in pigs vaccinated with ultraviolet-attenuated eggs

Pigs fed $\text{Ascaris}$$\text{suum}$ eggs attenuated by short wave ultraviolet-radiation developed resistance to challenge infection. Per os inoculation of pigs on three successive weeks with 10,000 eggs irradiated to total exposures of 150,100 and 75 μW-min/cm², respectively, resulted in an 88% reduction in the number of larvae recovered from the lungs 7 days after challenge with 10,000 infective eggs. Peripheral blood lymphocytes (PBL) from vaccinated pigs were specifically stimulated in vitro to incorporate tritiated thymidine by egg hatching fluid (Ea) and by excretory-secretory products obtained from cultures in defined-media of second-stage larvae (L2) developing to third-stage (L2–3) and from cultures of third-stage larvae developing to fourth-stage (L3–4). PBL were also specifically stimulated by living L2. Ea and L2 stimulated pig PBL significantly at 7 days after the first inoculation; responses to L2–3 and L3–4 developed 7 days after a second inoculation. The antigen-responsive cells in the PBL population were non-immunoglobulin-bearing lymphocytes. Antibodies to Ea and L2–3 were not detected in the serum of vaccinated pigs, and only 3 of 7 pigs had low concentrations of serum antibodies to L3–4.     

A Comparison of the Susceptibility of Growing Mukota and Large White Pigs to Infection with Ascaris suum

The influence of A. suum infection on the haematology, liver-related serum enzymes, blood urea and live weight gain in Mukota and Large White (LW) weaner pigs was compared. Six pigs of each genotype were infected with a single dose of 4000 A. suum eggs per pig and another six were not. The pigs were kept for 100 days. Blood was collected daily for the first 7 days and also after 100 days. In the infected pigs, there was an increase (p<0.05) in alanine aminotransferase (ALT) activity in the LW but not in the Mukota pigs. Although the alkaline phosphatase (ALP) activity rose (p<0.05) in both infected and non-infected LW pigs from day 1 to day 3, the activity in the non-infected LW pigs then decreased, while that of the infected LW pigs remained elevated. The infected LW pigs had higher (p<0.05) levels of ALT, ALP and aspartate aminotransferase than their non-infected counterparts. Non-infected LW pigs tended to have higher (p<0.05) haematological parameters, daily weight gain and urea concentrations than infected LW pigs, but these differences were not significant. These preliminary findings suggest that more A. suum larvae reached the livers in the LW than in the Mukota pigs and that the latter may be more resistant to A. suum infection.     

A comparison of the enzyme linked immunosorbent assay and the indirect haemagglutination technique applied to sera from cattle experimentally infected with Taenia saginata (Goeze, 1782)

The serological response of 6 calves to experimental oral infection with between 60,000 and 100,000 Taenia saginata eggs at 3–12 months of age was monitored by the enzyme-linked immunosorbent assay (ELISA) and the tanned cell indirect haemagglutination technique (IDH). A serum antibody response was detected by both techniques by 2–3 weeks post infection, rising to a plateau about 4–6 weeks post infection. The serum antibody levels began to decline by about 30 weeks post infection. Two uninfected control cattle gave negative reactions.In addition, the serological response of 5 calves which had received a dose of 10,000 T. saginata eggs at 2–3 days of age and then weekly serial doses of 500 eggs for 12 months thereafter, was compared with a similar group of 5 calves, which had received the single infection of 10,000 eggs at 2–3 days of age only. Calves in both groups developed an antibody response detectable by the ELISA technique whereas those in a group of 5 control calves did not show such a response. When studied individually however there was marked variation in the serum antibody levels of these young cattle, as although some calves gave a relatively strong serological response, others hardly varied from the controls.     

Biological control of <Emphasis Type="Italic">Ascaris suum</Emphasis> eggs by <Emphasis Type="Italic">Pochonia chlamydosporia</Emphasis> fungus

Ascaris suum is a gastrointestinal nematode parasite of swines. The aim of this study was to observe Pochonia chlamydosporia fungus on biological control of A. suum eggs after fungus passage through swines gastrointestinal tract. Eighteen pigs, previously dewormed, were randomly divided into three groups: group 1, treated with the fungus isolate VC4; group 2, treated with the fungus isolate VC1 and group 3 did not receive fungus (control). In the treated groups, each animal received a 9 g single dose of mycelium mass containing P. chlamydosporia (VC1 or VC4). Thereafter, animal fecal samples were collected at the following intervals: 8, 12, 24, 36, 48, 72 and 96 h after treatment beginning and these were poured in Petri dishes containing 2% water-agar culture medium. Then, 1,000 A. suum eggs were poured into each dish and kept in an incubator at 26°C and in the dark for 30 days. After this period, approximately 100 eggs were removed from each Petri dish and morphologically analyzed under light microscopy following the ovicidal activity parameters. The higher percentage observed for isolated VC4 eggs destruction was 57.5% (36 h) after fungus administration and for isolate VC1 this percentage was 45.8% (24 h and 72 h) (p > 0.01). P. chlamydosporia remained viable after passing through the gastrointestinal tract of swines, maintaining its ability of destroying A. suum eggs.     

Establishment of Ascaris suum in the pig: development of immunity following a single primary infection

Development of immunity after a single primary infection of Ascaris suum in pigs was investigated with regard to the worm population dynamics of a superimposed A. suum infection, host immune response and gross liver pathological changes. Group A was given a primary infection of 60,000 infective A. suum eggs and group B was left uninfected. Four weeks later both groups A and B were inoculated with 1,000 A. suum eggs, and subgroups were slaughtered 7, 14 and 21 days post challenge infection (p.c.i.). An uninfected control group C was slaughtered on day 21 p.c.i. The challenge worm recovery in group A was reduced compared to group B by 12%, 50% and 75% on day 7, 14 and 21 days p.c.i., respectively. In both groups was the expulsion of worms initiated between day 14 and 21 p.c.i. However, in group A the worms were recovered more posteriorly in the small intestine and 21 days p.c.i. the mean worm length was significantly shorter than in group B (p = 0.01). The results above were associated with significantly higher (p < 0.05) antibody response and higher eosinophil counts in group A compared to group B. The present results suggest that the larval growth and survival of a challenge infection are decreased, probably due to higher antibody and eosinophil attack during the migratory phase.     

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.     

Survival of Salmonellas and Ascaris Suum Eggs in a Thermophilic Biogas Plant

In a continuous biogas plant, receiving manure from 200 dairy cows and 400 calves and young stock, survival of salmonellas and Ascaris suum eggs was studied. The bacteria and parasite eggs were kept in filter sacs in the manure that had a temperature of 55°C. No viable salmonellas or Ascaris suum eggs could be found after 24h in the digester.Survival of salmonellas and Ascaris suum eggs was also studied in the manure pit where the manure was stored after digestion. The temperature in the manure pit varied between 22–27°C. Salmonellas survived 35 but not 42 days. On day 56, when the experiments had to be stopped, 60% of the Ascaris eggs were viable.     

Application of a real-time PCR assay for the detection of Ascaris suum DNA in the liver of experimentally infected chickens

This study aimed to evaluate the sampling method for the detection of Ascaris suum larval DNA in chicken livers using real-time PCR. Chickens were inoculated with A. suum eggs of a single dose (Group A) or repeatedly low doses (Group B). White spots (WSs) were continuously observed on liver from day 3 after the last infection in Group B and day 14 in Group A. In Group A, larval DNA was detected in WS lesions (78.6%) at a significantly higher rate than in the remaining tissue samples (31.3%). In conclusion, applying WS lesions to the assay improved the detection rate of A. suum DNA in chicken livers, especially in the case of a single infection.     

Ascaris suum in Saskatchewan Pigs: An Abattoir Survey of Prevalence and Intensity of Infection

In a survey of 2500 market weight pigs in a Saskatchewan abattoir, 37% were infected with adult Ascaris suum and in 46% there were milkspot hepatic lesions. A total of 60% of the pigs examined had some evidence of A. suum infection. Most infected pigs contained less than 50 adult ascarids.

In a second abattoir survey of a further 2500 market weight pigs, 44% of the animals had milk-spot hepatic lesions. In approximately 12% of the affected pigs these lesions were very severe and a very similar proportion of pigs' livers were condemned or sent for use as animal feed.

The results of these surveys are discussed with particular reference to the significance of A. suum infection in growing pigs in Saskatchewan.

     

Yeast-expressed classical swine fever virus glycoprotein E2 induces a protective immune response

Classical swine fever (CSF) is an economically important swine disease worldwide. The glycoprotein E2 of classical swine fever virus (CSFV) is a viral antigen that can induce a protective immune response against CSF. A recombinant E2 protein was constructed using the yeast Pichia pastoris expression system and evaluated for its vaccine efficacy. The yeast-expressed E2 (yE2) was shown to have N-linked glycosylation and to form homodimer molecules. Four 6-week-old specified-pathogen-free (SPF) piglets were intramuscularly immunized with yE2 twice at 3-week intervals. All yE2-vaccinated pigs could mount an anamnestic response after booster vaccination with neutralizing antibody titers ranging from 1:96 to 1:768. Neutralizing antibody titers at 10 weeks post booster vaccination ranged from 1:16 to 1:64. At this time, the pigs were subjected to challenge infection with a dose of 1 × 10⁵ TCID₅₀ (50% tissue culture infective dose) virulent CSFV strain. At 1 week post challenge infection, all of the yE2-immunized pigs were alive and without symptoms or signs of CSF. Neutralizing antibody titers at this time ranged from 1:4,800 to 1:12,800 and even to 1:51,200 one week later. In contrast, the control pigs continuously exhibited signs of CSF and had to be euthanized because of severe clinical symptoms at 6 days post challenge infection. All of the yE2-vaccinated pigs were E^rns antibody negative and had seroconverted against E^rns by post challenge day 11, suggesting that yE2 is a potential DIVA (differentiating infected from vaccinated animals) vaccine. The yeast-expressed E2 protein retains correct immunogenicity and is able to induce a protective immune response against CSFV infection.     

Heterologous helminth induced resistance to Ascaris suum in guinea pigs

The protection inducing capacity of Toxocara canis, Ancylostoma caninum, Haemonchus contortus, Caenorhabditis briggsae and Turbatrix aceti given via the mesenteric vein to a challenge infection with Ascaris suum was evaluated. Embryonated eggs and second stage larvae of T. canis and infective larvae of A. caninum were able to induce a statistically significant level of protective immunity, while the other non-related helminths were unable to protect against a challenge infection.     

Turbinate atrophy in gnotobiotic pigs intranasally inoculated with protein toxin isolated from type D Pasteurella multocida

Three doses (75 micrograms, 25 micrograms, and 25 micrograms) of purified toxin isolated from a toxigenic strain of type D Pasteurella multocida were given (by atomizer) into the right nasal cavities of each of 10 gnotobiotic pigs on the 21st, 24th, and 27th days of age, respectively. Inoculated pigs (usually 2) and 1 noninoculated control pig each were necropsied on 3, 6, 9, 12, and 15 days after inoculations were given. Severe bilateral atrophy of turbinates occurred in all toxin challenge-exposed pigs. Atrophy was more severe in the inoculated nasal cavity than that in the noninoculated side in 2 of the 10 pigs. Microscopic changes in turbinates of toxin challenge-exposed pigs were more severe in pigs killed at later dates. Dominant changes included degeneration and necrosis of osteoblasts, markedly accelerated osteoclastic osteolysis, replacement of the osseous core by a highly cellular mesenchymal stroma, and multifocal atrophy of submucosal glands. Seemingly, a protein toxin isolated from toxigenic type D strains of P multocida produced rapid atrophy of turbinates and may be a contributing factor in development of clinical progressive atrophic rhinitis in swine.     

Does Porcine Reproductive and Respiratory Syndrome Virus Potentiate Classical Swine Fever Virus Infection in Weaner Pigs?

Fifteen 6-week-old crossbred weaners weighing about 12 kg each were randomly divided into three groups of five animals each. One group of pigs was inoculated first with porcine reproductive and respiratory syndrome (PRRS) virus and then 3 days later with CSF virus. The second group received classical swine fever (CSF) virus, while the third group was inoculated with PRRS virus only. The aim of the experiment was to determine whether a primary PRRS virus infection influences the clinical outcome of experimentally induced CSF in young pigs. The PRRS virus infected weaners developed mild respiratory symptoms and recovered completely. All five weaners which were inoculated with CSF virus only showed severe clinical signs typical of the acute form of CSF. One pig had to be killed 15 days post-inoculation (p.i.); the remaining four died between the 18th and 22nd day p.i. The clinical course of the animals inoculated with both viruses was slightly different from that of the pigs that received only CSF virus. Four out of five pigs from the PRRS/CSF group became febrile and viraemic earlier than the animals which received CSF virus only. These pigs had to be killed 15–17 days post CSF virus inoculation. One animal in this group survived the acute phase of CSF and recovered completely. It was concluded that the observed divergences of the clinical courses would not have been noticed under field conditions. Therefore these findings cast doubt on the relevance of PRRS virus infection potentiating significantly the clinical outcome of CSF in young pigs.     

Does porcine reproductive and respiratory syndrome virus potentiate classical swine fever virus infection in weaner pigs?

Fifteen 6-week-old crossbred weaners weighing about 12 kg each were randomly divided into three groups of five animals each. One group of pigs was inoculated first with porcine reproductive and respiratory syndrome (PRRS) virus and then 3 days later with CSF virus. The second group received classical swine fever (CSF) virus, while the third group was inoculated with PRRS virus only. The aim of the experiment was to determine whether a primary PRRS virus infection influences the clinical outcome of experimentally induced CSF in young pigs. The PRRS virus infected weaners developed mild respiratory symptoms and recovered completely. All five weaners which were inoculated with CSF virus only showed severe clinical signs typical of the acute form of CSF. One pig had to be killed 15 days post-inoculation (p.i.); the remaining four died between the 18th and 22nd day p.i. The clinical course of the animals inoculated with both viruses was slightly different from that of the pigs that received only CSF virus. Four out of five pigs from the PRRS/CSF group became febrile and viraemic earlier than the animals which received CSF virus only. These pigs had to be killed 15-17 days post CSF virus inoculation. One animal in this group survived the acute phase of CSF and recovered completely. It was concluded that the observed divergences of the clinical courses would not have been noticed under field conditions. Therefore these findings cast doubt on the relevance of PRRS virus infection potentiating significantly the clinical outcome of CSF in young pigs.     

Antigens in perienteric fluid of Ascaris suum as detected through antibodies in pigs orally inoculated with fully embryonated eggs

Perienteric fluid (Pf) of adult Ascaris suum was fractionated by ammonium sulfate precipitation, gel filtration or DEAE-cellulose chromatography, and sucrose density gradient centrifugation. Anti-sera (anti-EE) from pigs which were inoculated orally with fully embryonated eggs (EE) of A. suum were used in an indirect radioimmunoassay (IRIA) to determine which fractions of Pf reacted positively in the analyses at the lowest protein concentration. These fractions were considered to contain more potent antigens. Comparative IRIA were performed employing antisera (anti-Pf) produced by injecting Pf into pigs. Six out of 35 fractions reacted positively at ? 0.2 μg protein when anti-EE was used in the IRIA. Twenty-two out of 35 fractions reacted positively at ? 0.2 μg protein when anti-Pf was used. Five of the 6 fractions reacting positively with anti-EE also reacted with anti-Pf. A 76 000 dalton component appears to be one of the major proteins in the 6 fractions which react positively with anti-EE while components from 10 000–138 000 daltons were present in the various fractions reacting positively with anti-Pf at ? 0.2 μg protein.     

Ascaris suum Infection in Calves II. Circulating and Marrow Eosinophil Responses

An increment in the number of circulating eosinophils occurred between the 11th and 14th days after infection with Ascaris suum and this increase was generally greater after a challenge infection. Continual infection with small numbers of A. suum eggs over prolonged periods resulted in circulating eosinophil levels which fluctuated and were dose-dependent. The per cent marrow eosinophils always increased after a primary infection and a greater increase usually followed a challenge infection. The maximum increment of marrow eosinophils occurred between the tenth and 12th days and preceded the rise in circulating eosinophils by 36 hours. Antihistamine therapy did not alter eosinophil responses to A. suum. Circulating eosinophilia was not usually reflected by drastic changes in differential white cell counts. However, an increase in total white cell count often followed infection with A. suum and frequently parallelled changes in eosinophil counts. Hemoglobin and P.C.V. values remained within normal limits in A. summ infected calves.