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.     

The anthelmintic efficacy of ivermectin in experimentally infected cattle

The 22,23-dihydro derivative (ivermectin) of the major B₁ components of the avermectins was evaluated, by oral and parenteral routes, for anthelmintic activity in cattle experimentally infected with 7 gastro-intestinal nematode species and lungworm. Treatment at three dosage levels was administered when the parasites were in the L₄ developmental stage or when adult. The dosage expected to remove 95% of the nematodes (ED₉₅) by each route of administration against the various developmental stages of each species of parasite was calculated by methods of linear regression to choose dose levels to be used in subsequent developmental field trials. Parenterally, efficacy against L₄ stages of Haemonchus placei, Ostertagia ostertagi, Trichostrongylus axei, T. colubriformis, Cooperia oncophora, C. punctata, Oesophagostomum radiatum and Dictyocaulus viviparus exceeded 99% with treatment at 0.2 mg ivermectin/kg. Parenteral treatment of the adult parasites resulted in at least 98% reduction of worm burdens, except $\text{? 90\%}$ for T. colubriformis, with 0.2 mg/kg. Oral treatment of L₄ stages of development with 0.2 mg/kg resulted in at least a 95% reduction in worm burdens of each of the 8 species of parasite. Oral treatment of the adult worms with 0.1 mg ivermectin/kg produced at least a 98% decrease in each parasite species, except for T. colubriformis (90% reduction) for which dose-response calculations suggest 95% efficacy with oral treatment at 0.2 mg/kg.     

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.     

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.     

An Intradermal Test to Detect Latent Warble (Hypoderma spp.) Infection in Cattle

An intradermal test was developed to screen cattle for infection with the first-instar larvae of the warble flies Hypoderma lineatum and H. bovis. The diagnostic antigen, prepared from the first-instar larvae of H. lineatum, produced a distinct dermal reaction in cattle infected with the first-instar larvae of either species, but not in cattle in which the infection could not be confirmed later either on necropsy or by the appearance of warbles in the back. The reaction to the antigen was unpredictable in cattle with warbles in their backs. The diagnostic property of the antigen was also demonstrated in rabbits and guinea pigs artificially sensitized to the antigen.     

Culture requirements of Ascaris suum larvae using a stationary multi-well system: Increased survival,development and growth with cholesterol

Ascaris suum third-stage larvae (L₃) were converted from infected rabbits and cultured in a stationary multi-well plate system. Several different culture media including Mediuim 199, NCTC-135, Minimum Essential Medium-Eagle, Dulbecco's Modified Eagle's Medium, RPMI 1640, McCoy's 5A and Neuman-Tytell mdium were tested to determine which was best for overall larval survival, development and growth. Larvae developed from L₃ to fourth-stage (L₄) in all media tested, but larval survival and the yield and growth of L. were superior in RPMI 1640. Pyruvate is an important medium component because its addition to RPMI 1640 enhanced the yield and growth of L₄ while its removal from DMEM reduced larval survival and the yield and growth of L₄. Cholesterol markedly enhanced larval survival and the yield and growth of L₄ when added to RPMI 1640 as either a soluble supplement in Tween 80 or to a lesser degree, as a component of liposomes. The multi-well culture plate system is a convenient method for determining the effects of different media and changes in media composition on larvae in vitro.     

Antigenic differences between the life cycle stages of Cooperia oncophora

The antigenic differences among the life cycle stages of Cooperia oncophora were studied by SDS-gel electrophoresis and Western blotting. Sodium dodecylsulphate polyacrylamide gel electrophoresis of the different life cycle stages of C oncophora revealed a complex protein pattern with a decreasing number of protein bands towards the adult stages. Several bands of the fourth stage larvae were in common with both the third stage and the adult nematode. Western blotting with sera from C oncophora monoinfected calves showed that the antigens of the fourth stage larvae were recognised predominantly and the presence of stage specific antigens in all stages. Strong cross reactivity was demonstrated when serum from Ostertagia ostertagi infected calves was used.     

In vitro evaluation of the ovistatic and ovicidal effect of the cosmopolitan filamentous fungi isolated from soil on Ascaris suum eggs

The ovicidal activity of seven fungal strains: Acremonium alabamense, Alternaria chlamydospora, Cladosporium herbarum, Fusarium solani, Paecilomyces variotii, Paecilomyces viridis and Penicillium verruculosum isolated from urban soil samples from Poland was determined in vitro. The fungal mycelium was co-cultured with Ascaris suum eggs on plates with 2% water–agar for 28 days. Eggs exposed and unexposed (control) to fungal mycelium were observed weekly by light microscopy and the percentage of malformed eggs were determined. The eggs were classified according to following parameters: type 1 – biochemical and physiological effect without morphological damage to the eggshell; type 2 – lytic effect with morphological alteration of the eggshell and embryo; type 3 – lytic effect with morphological alteration of eggshell and embryo with hyphal penetration and internal egg colonization. All examined species of fungi extended embryogenesis, but the retardation of embryonic development was varied and depended on the species. A. alabamense, A. chlamydospora and P. verruculosum exhibited very high inhibitory activity on A. suum egg development. The fungus-exposed eggs revealed morphological alternations in all stages of embryogenesis. Isolates of F. solani, P. variotii and P. viridis showed hyphal penetration and internal colonization of A. suum eggs (type 3 effect). No appressoria were produced and simple hyphal penetrations were most commonly observed. A. alabamense and P. verruculosum demonstrated morphological destruction, with eggshell destruction. The remaining fungi showed type 1 effect. The results demonstrated that examined strains of F. solani, P. variotii and P. viridis may be considered to be potential limiting factors of parasitic geohelminth populations.     

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.     

In vitro development of cyathostomin larvae from the third stage larvae to the fourth stage: morphologic characterization, effects of refrigeration, and species-specific patterns

A mixed population of equine cyathostomin (Nematoda, Strongyloidea) infective third stage larvae (L3) was cultured in vitro using a cell-free medium. Some L3 were cultured immediately after Baermann collection from fecal cultures, while others were kept in water at 4 °C for 7 days before initiating the in vitro cultures. Cultures were examined daily for viability. At days 2, 7, 14 and 21 larvae were collected for identification of developmental stage and morphological changes, using both light and scanning electron microscopy. Larvae were classified as early L3 (EL3), developing L3 (DL3), late L3 (LL3) and fourth stage larvae (L4) on the basis of morphological features. Viability remained high throughout the entire study period in cultures of both non-refrigerated (84.7%) and refrigerated (77.4%) larvae. However, viability of the non-refrigerated was significantly greater from 7 through 21 days of culture. Significant differences were also observed in the percentage of DL3 between the non-refrigerated and refrigerated larval cultures by day 7. The highest percentage of DL3 larvae (22.5%) was reached at the end of study in those larvae that were not previously refrigerated. The data suggests that prior refrigeration decreases viability and slows L3 development. At day 21 LL3 larvae were only a small percentage of the DL3: 6.9 and 5% in non-refrigerated and refrigerated cultures, respectively. Few of these larvae freed themselves from the L3 cuticle and moulted to L4 stage. Characteristics of individual species in vitro developmental patterns were determined by the molecular identification of individual larvae in pools of larvae randomly collected at days 0 and 21. Seven species (Coronocyclus coronatus, Cylicostephanus goldi, Cylicostephanus longibursatus, Cyathostomum catinatum, Cylicocyclus nassatus, Cylicocyclus ashworthi, Petrovinema poculatum) were identified in the day 0 pool. The greatest tendency to develop in vitro was shown by the genus Cylicostephanus with the species C. goldi and C. longibursatus that developed to the LL3-L4 stages. C. nassatus, C. ashworthi and C. coronatus did not progress in their development beyond the EL3 stage, while no apparent signs of development were registered for C. catinatum.     

Efficacy of a single high oxfendazole dose against gastrointestinal nematodes in naturally infected pigs

The goal of the current experiment was to assess the clinical efficacy of oxfendazole (OFZ) administered as a single oral dose (30 mg/kg) to pigs naturally parasitized with Ascaris suum, Oesophagostomum spp., Metastrongylus spp. and Trichuris suis. Thirty-six local ecotype piglets were divided into three independent experiments, named I, II and III (n = 12 each), respectively. Each experiment involved two different groups (n = 6): Untreated Control and OFZ treated. Animals were naturally parasitized with A. suum (Experiments I, II and III), Oesophagostomum spp. (Experiments I and II), T. suis (Experiments II and III) and Metastrongylus spp. (Experiment I). Pigs in the treated group received OFZ (Synanthic^®, Merial Ltd., 9.06% suspension) orally at 30 mg/kg dose. At five (5) days post-treatment, animals were sacrificed and the clinical efficacy of the OFZ treatment was established following the currently available WAAVP guidelines for a controlled efficacy test. None of the animals involved in this experiment showed any adverse events during the study. OFZ treatment given as a single 30 mg/kg oral dose showed a 100% efficacy against all the nematode parasites present in the three experiments. In conclusion, under the current experimental conditions, OFZ orally administered to naturally parasitized piglets at a single dose of 30 mg/kg was safe and highly efficacious (100%) against adult stages of A. suum, Oesophagostomum spp., T. suis and Metastrongylus spp.     

Inhibited development of Obeliscoides cuniculi in rabbits: the effects of active and passive immunization and resumption of larval development

Rabbits were either actively or passively immunized against Obeliscoides cuniculi and challenged with 5 000 infective larvae (IL). Worm development was inhibited in actively immunized animals and worm egg production was impaired by administration of immune serum. Inhibited fourth stage larvae (L₄) from high-level infections (200 000 IL) and normal L₄ from low-level infections (4 000 IL) were transferred to actively immunized and normal rabbits. In both instances, large numbers of L₄ developed to maturity; however, a significant number of inhibited L₄ failed to resume development. Inhibition of larvae resulted in the development of fewer adult male than female worms, and the change in sex ratio was further accentuated in actively immunized recipients. Worm egg production was also reduced in actively immunized rabbits. These data indicate that host immune responses contribute to inhibition of O. cuniculi in rabbits and that worm egg production is also responsive to immunologic control.     

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.     

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.     

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.

     

Can carotenoids mediate the potentially harmful effects of ultraviolet light in Silurana (Xenopus) tropicalis larvae?

Amphibians have recently experienced unprecedented declines in the wild, the causes of which are often difficult to mitigate. This has increased the importance of ex situ conservation; however, long‐term maintenance and breeding of amphibians in captivity often has limited success. In vertebrates, vitamin D is required for calcium homeostasis and is produced endogenously in skin exposed to ultraviolet (UV) light; however, UV light can be harmful to amphibians in some instances. Carotenoids are dietary pigments that may provide protection from UV light. The aim of this study was to assess the protective capability of carotenoids against the potentially harmful effects of UV light in Silurana (=Xenopus) tropicalis larvae raised in an enhanced or limited UV environment. Tadpole survival and the size and developmental stage reached by the end of the study period were measured. Carotenoids had a significantly positive effect on developmental rate in both UV‐limited and UV‐enhanced environments. Larvae in an enhanced UV environment were significantly larger than those raised under a limited UV environment, irrespective of diet. Carotenoid‐fed larvae tended to have increased survival in relation to those raised without carotenoids, but only in a limited UV environment. Carotenoids appear to provide little protection against UV light in this case. The role of carotenoids in amphibian health has not previously been studied. We show that carotenoid availability significantly influences development and may increase survival in S. (X.) tropicalis larvae. This finding may have important implications for recommendations made on the nutrition of amphibians in captivity.     

In vitro cultivation of Ostertagia ostertagi, the medium stomach worm of cattle. II. Effect of insect-growth-disrupting amines and amides on development

Thirteen secondary and tertiary amides and amines that disrupt growth and development in certain insects were tested at concentrations of 0.25–5.0 p.p.m. for their effects on development of Ostertagia ostertagi, from infective larvae to egg-laying adults, in a two-step roller culture system. These compounds did not affect exsheathment of infective larvae. However, some of these compounds inhibited development or killed 100% of the exsheathed larvae at concentrations of 1.0–2.5 p.p.m. Minimum inhibitive concentrations of several of the amines and amides affected the nematodes in one or more of the following ways: decreased motility or paralysis; reduced survival; delayed or blocked third or fourth ecdysis; lowered yields of advanced stages; decreased production of fertile and non-fertile eggs. The results also indicated that, as in insects, these chemicals exerted their lethal effects against O. ostertagi at the time of molt.     

Pilot project to investigate over-wintering of free-living gastrointestinal nematode larvae of sheep in Ontario,Canada

This study investigated the overwintering survival and infectivity of free-living gastrointestinal nematode (GIN) stages on pasture. The presence of GIN larvae was assessed on 3 sheep farms in Ontario with a reported history of clinical haemonchosis, by collecting monthly pasture samples over the winter months of 2009/2010. The infectivity of GIN larvae on spring pastures was evaluated using 16 tracer lambs. Air and soil temperature and moisture were recorded hourly. Free-living stages of Trichostrongylus spp. and Nematodirus spp. were isolated from herbage samples. Gastrointestinal nematodes were recovered from all tracer lambs on all farms; Teladorsagia sp. was the predominant species. Very low levels of Haemonchus contortus were recovered from 1 animal on 1 farm. The results suggest that Haemonchus larvae do not survive well on pasture, while Teladorsagia sp., Trichostrongylus spp. and Nematodirus spp. are able to overwinter on pasture in Ontario and are still infective for sheep in the spring.     

Sequential development of esophagogastric ulcers induced in swine by infections with Ascaris suum goeze, 1782

Twenty-five experimental and five control hysterectomy derived pigs were raised colostrum free under specific pathogen-free conditions. The experimental pigs were first inoculated with A. suum eggs at 8 weeks of age, and a second dose of infective eggs was given 15 days later. They were necropsied from 4 to 10 days post second infection.The ulcers found in the stomach of these pigs were confined to the esophagogastric area. A sequence of epithelial changes, erosions, and ulceration was seen in the stratified squamous epithelium. Edema and hemorrhage were noted in the lamina propria and submucosa. Vascular changes in the area were characterized by thickening of the wall and presence of thrombi in arteries as well as cellular degeneration and vacuolization of the media and intima of the veins.Changes in the muscularis were characterized by edema and degeneration of the muscle cells, which later were replaced by fibroblasts and heavy infiltration by eosinophils.Three of the control pigs received one dose of A. suum eggs, and were killed one on each of the 8th, 9th, and 10th days post infection. The remaining two pigs received no infective material and were killed when they were 11 weeks of age. Neither the uninoculated pigs nor the once-inoculated animals developed ulcerations in their stomachs.The possible mechanisms by which esophagogastric lesions are produced following A. suum larval migrations are discussed.     

Study on the pathogenicity of ostertagiosis due to Ostertagia circumcincta in sheep,in Iraq

The response of Awassi sheep to Ostertagia circumcincta, the most prevalent ovine strongylid species in Iraq, was studied.A dose of 100000 larvae of O. circumcincta (Iraqi strain) induced moderate clinical symptoms of ostertagiosis. These symptoms were correlated with elevated pH of the abomasal fluid and increased plasma pepsinogen levels. There was no evidence of larval inhibition since the majority of the fourth stage larvae (L₄) continued their development. No appreciable loss of worm population was observed and most of the parasites survived and exhibited a prolific egg-laying potential six moths after initial infection. Assessments of the pathophysiological changes were performed and correlated with parasitological and clinical observations.The Awassi breed of sheep, the most prevalent in the Arab Middle East, seems to be more susceptible to ostertagiosis than other breeds and there was no spontaneous self-cure in this breed.