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.     

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.     

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.     

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     

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.     

Reinfection of Yearling Calves with Ascaris suum

Naturally occuring outbreaks of Ascaris suum infection in calves have usually beeen found in animals nine to 12 months of age. The circumstances surrounding these outbreaks suggest that yearling calves are either particularly susceptible to a primary exposure to A. suum or react strongly to A. summ after sensitization early in life to this or some related ascarid. To determine the effect of reinfection with A. suum nine to 12 months after varying levels of exposure to this nematode, six calves were inoculated with 200,000 to 9,000,000 eggs. Neither death nor, in general, severe clinical signs resulted from reinfection. All calves were examined 15 days after reinfection with pathological changes noted only in the lungs and consisting of emphysema, alveolar wall thickening as well as accumulations of fibrin, eosinophils and hemorrhage in the lumina of alveoli. The findings suggested that exposure to A. suum early in life is not a factor in the development of disease in calves infected at one year of age. It was also found that the eosinophilia that develops following a primary infection with A. suum evidently persists for at least one year.     

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.     

Pulmonary bacterial deposition and clearance during ascarid larval migration in weanling pigs.

Pulmonary deposition and clearance of bacteria were measured in weanling pigs, half of which had been inoculated at age 31 days with larvated Ascaris suum ova. Seven days later, when breathing signs of larval migration were pronounced, all pigs were exposed to aerosolized Escherichia coli (strain B). Then, either immediately after aerosol exposure (for deposition assessment) or immediately after a 120 minute period in filtered air (for clearance), bacteria in the pigs' lungs were counted. Ascarid ova-inoculated pigs did not differ significantly from control pigs for number of bacteria in the lungs after aerosol exposure, but after the 120 minute clearance period they had 7.2 times more than did the control pigs. Thus, in weanling pigs, the breathing-pattern changes that were evident during ascarid-larval migration did not affect pulmonary deposition of inhaled bacteria significantly, but the presence of ascarid larvae in the lungs was associated with impaired pulmonary bacterial clearance.     

Effectiveness of antigens from various developmental stages of Ascaris suum in the deomonstration of the migratory phase of experimental ascariasis in pigs]

Tests of the efficiency of antigens prepared from different developmental stages of Ascaris suum in indirect haemagglutination test in the course of proving the migration phase of experimental ascariasis in pigs show that the antigens prepared by ultrasound from the invasive larval stage of A. suum in comparison with antigens of sexually mature stages have higher serological activity. By using this antigen it is possible to prove specific antibodies in experimentally invaded pigs from 6 to 120 days after invasion as opposed to the other tested antigens (the detectability of antibodies from the 7th-8th day to the 64th day after invasion).     

Immunization of sheep against the larval stage of Taenia multiceps

A trial with 200 ewes and 127 lambs showed that vaccination of lambs with 2 doses of Oncosphere Secretory Antigen (OSA) 4 weeks apart, at either 4-8 and 8-12 weeks or at 12-16 and 16-20 weeks of age, was equally effective in protecting them against infestation with the larval stage of Taenia multiceps. In this trial, the lambs of ewes, vaccinated with OSA when they were 90 and again 120 days pregnant, were as susceptible to infestation as the lambs of untreated control ewes. Another trial to provoke passive immunity in lambs gave inconclusive results. In a 3rd trial 2 doses of regular OSA and 2 doses of freeze-dried OSA protected all the lambs in each group, while a single dose of regular OSA protected 9 out of 10 lambs against cerebral lesions.     

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.