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.     

Influence of an experimental infection of Ascaris suum on performance of pigs

Thirty-two pigs (average 26.6 kg live weight) were individually housed and fed to study the effect of an infection of Ascaris suum (either 0, 600, 6,000 or 60,000 A. suum eggs/pig) on performance of growing-finishing pigs. Increasing the level of A. suum infection produced linear (P less than .07) and quadratic (P less than .09) effects on final weight, weight gain and average daily gain. Feed to gain ratio and number of A. suum worms recovered from the intestines of pigs at slaughter increased linearly (P less than .01) with increasing doses of A. suum eggs. Pigs receiving 60,000 A. suum eggs were 13% less (P less than .01) efficient than the noninfected controls. In each of two trials, eight crossbred barrows (15.7 kg in trial 1 and 16.1 kg body weight in trial 2) were examined for the effects of two levels of A. suum infection (0 and 20,000 eggs/pig) on digestibility coefficients for dry matter, crude protein and gross energy. The infection did not affect (P greater than .05) digestibility coefficients during the first two collection periods (d 6 through 10 and 19 through 23). However, digestion coefficients for dry matter, crude protein and gross energy obtained from the total collection period on d 33 through 37 postinfection were greater (P less than .01) for control pigs than for pigs given 20,000 A. suum eggs each. Also, N retention was greater (P less than .05) for control pigs than for infected pigs.     

Effects of multiple dose infections with Ascaris suum on blood gastrointestinal hormone levels in pigs

Ten consecutive daily doses of infective Ascaris suum eggs were administered to pigs in two experiments and the levels of gastrointestinal hormones in their blood were measured. The piglets in each experiment were divided into low-dose (LDI) and high-dose (HDI) infections and control groups. Infected pigs had lower feed consumption, lower weight gains, and lower feed efficiency than control pigs. Serum gastrin levels in infected pigs were significantly lower than the controls from Days 7 to 17 post first inoculation (PFI), and so were their serum glucagon levels from Days 12 to 24 PFI. Serum insulin levels in infected animals were sometimes lower than those in controls. These differences were usually more intense in the LDI pigs than in HDI pigs. The plasma cholecystokinin (CCK) levels in the LDI group were significantly higher than those in controls from Day 10 PFI to the end of the experiment, while the CCK levels in the HDI group did not differ significantly from the controls. Increased plasma CCK levels could be a satiety factor in A. suum infection since the time of occurrence of high levels of CCK matched the period of reduced feed consumption.     

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.     

Comparative studies of experimental oral inoculation of pigs with Ascaris suum eggs or third stage larvae

This experimental study was designed to compare the acquired resistance in pigs to Ascaris suum eggs following 4-weekly oral immunizations with either 200 A. suum infective eggs or 50 A. suum third stage larvae (L3). The two immunized groups (n = 7) together with an unimmunized control group (n = 7) of pigs were challenged orally with 50 infective A. suum eggs per kilogram bodyweight on day 19 after the last immunization. Seven days post-challenge the group immunized with eggs showed signs of resistance as evidenced by reduced lung larval counts compared with the challenge control group. Such significant resistance was not observed in the L3-immunized group. However, a markedly increased inflammatory liver reaction and white spot formation was demonstrated in the L3-immunized pigs after challenge compared with both control animals and egg-immunized pigs. On the day of challenge only the egg-immunized pigs mounted an anti-Ascaris antibody response both in serum and in lung lavage fluid. Ascaris-antigen induced increased histamine release from peripheral leucocytes following both immunization and challenge could only be demonstrated in the egg-immunized pigs. On day 7 post-challenge local IgA-anti-Ascaris antibodies were further demonstrated in bile of the egg-immunized group and in the small intestine of both immunized groups. In conclusion, oral A. suum egg immunization of pigs induced a significant reduction in lung larval counts upon challenge. In contrast, oral L3 immunization seemed to prime the pigs as observed by the presence of stunted lung larval growth and increased liver reaction post-challenge with A. suum eggs.     

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.     

A comparison of the efficacy of two ivermectin formulations against larval and adult Ascaris suum and Oesophagostomum dentatum in experimentally infected pigs

A study was conducted to evaluate and compare the efficacy of two injectable formulations of ivermectin (IVM-1 and IVM-2) at a dose rate of 0.3 mg/kg bodyweight versus placebo in the treatment and control of larval and adult stages of Ascaris suum and Oesophagostomum spp. in experimentally infected pigs. Seventy helminth free pigs were allocated on a liveweight basis to 7 groups each comprising 10 pigs (A-G). Group A served as an untreated control group. Groups B and C were used to investigate the efficacy of both formulations against adult stages of A. suum and Oesophagostomum spp., Groups D and E for efficacy against larval stages of A. suum and Groups F and G for efficacy against larval stages of Oesophagostomum spp. Pigs of groups A, B, C, D and E were infected on Day-0 with 1000 infective A. suum eggs each. Infective larvae of Oesophagostomum spp. (10,000/pig) were given on Day-0 to pigs of Groups F and G and on Day-21 to pigs of Groups A, B and C. Treatment was given to pigs of Group A (saline as placebo) on Day-7 and -28, IVM-1 to pigs of Group F on Day-7, pigs of Group D on Day-14 and pigs of Group B on Day-49. IVM-2 was given to pigs of Group G on Day-7, Group E on Day-28 and Group C on Day-49. Pigs of Groups F and G were sacrificed on Day-28, pigs of Groups A, D and E on Day-49 and pigs of Groups B and C on Day-56. Post mortem worm counts showed the following efficacies: (IVM-1) against larval A. suum 100%, against adult A. suum 94.4%, against larval Oesophagostomum spp. 52.0% and against adult Oesophagostomum spp. 83.0%. (IVM-2) against larval A. suum 100%, against adult A. suum 90.3%, against larval Oesophagostomum spp. 94.0% and against adult Oesophagostomum spp. 94.7%.     

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.     

猪蛔虫幼虫琼脂胶移行法的建立

本实验首次在国内建立起了线虫的琼脂胶移行法 (Agar- gel Migration Assay,AMA)。通过对影响幼虫活力的某些理化因素 ,如 :琼脂胶的温度、每孔加入的琼脂胶和幼虫混合液的体积、琼脂胶的浓度及幼虫在琼脂胶内移行的时间等研究发现 :加胶的温度和加胶的量对幼虫的移出率有较大的影响。幼虫的移出率随加胶温度的升高而降低 ,当温度达到 70℃时 ,则几乎无虫体从琼脂胶内移出 (移出率仅为 0 .5 % ) ;幼虫的移出率随加入琼脂胶量的增多而减小 ,当每孔加入 10 0 0μl时 ,幼虫的移出率为 16 .6 8% ,当加入 4 0 0μl时幼虫的移出率则能达到2 5 .17%。琼脂胶的浓度对幼虫移出的影响并不明显 ,采用 SAS软件 (for Windows V6 .12 )分析发现 1.2 %、1.0 %、0 .8%、0 .4 %的结果间无显著差异 (P>0 .0 5 ) ,但琼脂胶的浓度能影响胶液凝固的时间和凝固后的韧性。实验表明 :最适宜的加胶量为 4 0 0μl /孔 ,最适宜的加胶温度为 5 3℃ (Agar- gel,sigm a) ,在琼脂胶中最佳培养移行时间为 2 4 h,琼脂胶液的最适实验浓度为 1.5 %琼脂胶     

Susceptibility of fourth-stage Ascaris suum larvae to fenbendazole and to host response in the pig

Fenbendazole given at the rate of 2.5 g/kg of feed for 3 days had 100% efficacy against 4th-stage Ascaris suum larvae in 8 pigs. Eight control pigs had a total of 108 A suum. In 6 pigs infected 3 times with 3rd-stage A suum larvae and treated with fenbendazole after the larvae molted to the 4th stage, the challenge exposure-derived population was decreased by 64%. Similar sequential infections in 6 pigs similarly infected, but not treated with fenbendazole, decreased the challenge exposure-derived population by 98%; however, developing and/or adult worms from the vaccinating infections were present.     

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.     

猪蛔虫幼虫琼脂胶移行法的建立

本实验首次在国内建立起了线虫的琼脂胶移行法(Agar-gelMigrationAssay,AMA).通过对影响幼虫活力的某些理化因素,如琼脂胶的温度、每孔加入的琼脂胶和幼虫混合液的体积、琼脂胶的浓度及幼虫在琼脂胶内移行的时间等研究发现加胶的温度和加胶的量对幼虫的移行率有较大的影响.幼虫的移行率随加胶温度的升高而降低,当温度达到70℃时,则几乎无虫体从琼脂胶内移出(移行率仅为0.5%);幼虫的移行率随加入琼脂胶量的增多而减小,当每孔加入1000μl时,幼虫的移行率为16.68%,当加入400μl时幼虫的移行率则能达到25.17%.琼脂胶的浓度对幼虫移出的影响并不明显,采用SAS软件(forWindowsV6.12)分析发现1.2%、1.0%、0.8%、0.4%的结果间无显著差异(P＞0.05),但琼脂胶的浓度能影响胶液凝固的时间和凝固后的韧性.实验表明最适宜的加胶量为400μl/孔,最适宜的加胶温度为53℃(Agar-gel,Sigma),在琼脂胶中最佳培养移行时间为24h,琼脂胶液的最适实验浓度为1.5%琼脂胶.     

Immunohistochemical distribution of antigens in liver of infected and immunized pigs with Ascaris suum

In the present work, we carry out an immunopathological study of the swine ascariosis, under different conditions (control, infection and immunization). Twenty-one Iberian pigs were used and divided in seven groups. Groups 1 and 2 were the uninfected and challenged controls, respectively. Groups 3 and 4 were weakly infected with increasing doses of Ascaris suum eggs and treated with pyrantel (Group 4). Groups 5-7 were immunized with 14, 42 and 97 kDa proteins from the parasite, respectively. Groups 2-7 were challenged with 10,000 infective eggs 7 days before the sacrifice. The focal parasitic granulomata with eosinophils and lymphocytes were the main histopathological lesions in the liver of reinfected pigs, while more marked cellular infiltrate and abundant connective tissue were seen in the livers of immunized animals. There were important deposits of antigens in the livers of immunized and infected pigs. Antigens were mainly located in the connective tissue, with positive staining detection of the somatic larvae antigen, the body wall from the adult worms and the 14-, 42- and 97-kDa proteins. However, cholangiols, biliary ducts and macrophages presented an immunohistochemical positive stain against excretory-secretory and somatic antigens from the larvae and the body fluid antigen from the adult parasite. The detection of A. suum antigens in the liver of infected pigs improves the diagnosis of swine ascariosis. It may be possible to apply these procedures for diagnosis of human ascariosis in liver biopsies since A. suum from swine have been previously used as a substitute for the study of the human parasite Ascaris lumbricoides.     

Ecological influences on transmission rates of Ascaris suum to pigs on pastures

A study was conducted to determine the distribution and transmission rate of Ascaris suum eggs and Oesophagostomum dentatum larvae in a pasture/pig house facility, which during the preceding summer was contaminated with helminth eggs by infected pigs. In May, four groups of 10 helminth na?ve tracer pigs were exposed to fenced sections of the facility for 7 days and necropsied for parasite recovery 9-10 days later (trial 1). The highest rate of A. suum transmission (201 eggs per day) occurred in the pig house (A). On the pasture, egg transmission decreased with the distance from the house: 8 eggs per day in the feeding/dunging area (B); 1 egg per day on the nearest pasture (C); <1 egg per day on the distant pasture (D). Only a few O. dentatum infections were detected, indicating a poor ability of the infective larvae to overwinter. Soil analyses revealed that the highest percentage (5.8%) of embryonated A. suum eggs were in the house (A). Subsequently, the facility was recontaminated with A. suum eggs by infected pigs. A replicate trial 2 was conducted in the following May. A major finding was the complete reversal of egg distribution between the 2 years (trials 1 and 2). In contrast to previous results, the highest rates of transmission (569 and 480 eggs per day) occurred in pasture sections C and D, and the lowest transmission rates (192 and 64 eggs per day) were associated with the feeding/dunging sections and the house (B and A). Soil analyses again supported the tracer pig results, as the pasture sections had the highest concentrations of embryonated eggs. Detailed soil analysis also revealed a non-random, aggregated egg distribution pattern. The different results of the two trials may be due to the seasonal timing of egg deposition and tracer pig exposure. Many eggs deposited during the summer prior to trial 1 may have died rapidly due to high temperatures and dessication, especially when they were not protected by the house, while deposition in the autumn may have favored egg survival through lower temperatures, more moisture, and greater sequestration of eggs in the soil by rain and earthworms. The latter eggs may, however, not have become embryonated until turnout the next year. The results demonstrate that yearly rotations may not be sufficient in the control of parasites with long-lived eggs, such as A. suum, and that a pasture rotation scheme must include all areas, including housing.