Gary L. Zimmerman

MODULATION OF HOST IMMUNE RESPONSES BY FASCIOLA HEPATICA: RESPONSES OF PERIPHERAL LYMPHOCYTES TO MITOGENS DURING LIVER FLUKE INFECTIONS OF SHEEP

Fasciola hepatica infections of lambs (250 or 500 metacercariae) were shown to alter the proliferative responses of peripheral blood lymphocytes (whole blood culture) to mitogens at specific times postinfection (PI). Responses to concanavalin A (Con A) were significantly suppressed at weeks 4, 8, 10, and 11 PI whereas suppressed responses to phytohemagglutinin (PHA) occurred at weeks 4, 10, 11, and 16 PI. Only on weeks 4 and 6 PI were responses to pokeweed mitogen (PWM) suppressed. The fluke-induced modulation of responses appeared to be related more to specific phases of infection rather than to worm burdens.

Seroprevalence of Toxoplasma gondii in llamas (Lama glama) in the northwest USA

Serum samples from 283 llamas (Lama glama) from Oregon, Washington State and Idaho were tested for antibodies to Toxoplasma gondii using the modified agglutination test. Antibodies were found in 95 (33.5%) llamas. Percent seropositivity in serum dilutions of 1:25, 1:50, 1:500, and 1:5000 was 9.5%, 18.3%, 4.9%, and 0.7%, respectively.

Effect of ivermectin delivered from a sustained-release bolus on the productivity of beef cattle in Oregon.

The effect of ivermectin delivered from a sustained-release bolus (I-SRB) on the weight gain of beef cattle through a grazing season was evaluated using 20 yearling beef steer calves randomly divided into two groups of ten animals each. Calves in the control group each received a placebo bolus, while those in the treatment group each received an I-SRB designed to release 12 mg ivermectin day-1 for approximately 90 days. All animals were weighted and samples of feces were collected from the rectum at monthly intervals, beginning on Day 0, until trial termination (Day 148). Pasture larval counts were also conducted on herbage collected on each sampling date. On Day 119, two control and two treated calves were removed from pasture, housed in isolation for 3 weeks, then necropsied for recovery of gastrointestinal nematodes. Three sets of parasite-naive tracer calves were utilized to evaluate the initial, interim and final levels of pasture contamination by nematode larvae. A fourth set was used to evaluate the level of pasture contamination the following spring. The use of the I-SRB resulted in a greater than 99% reduction in fecal egg counts of trichostrongyles and numbers of gastrointestinal nematodes in the treated principals, as well as an average daily gain advantage of 0.114 kg over the 148 day period. A 67-98% reduction in pasture larval nematode contamination occurred on pastures grazed by the treated animals, as indicated by the parasite burden in tracer calves and pasture larval counts. The treatment effect was eventually lost by the following spring since tracer calves on the treated pasture had only 33% fewer nematodes than those on the control pasture.

Host influence on the banding profiles of whole-body protein and excretory-secretory product of Fasciola hepatica (trematoda) by isoelectric focusing

Various protein separation techniques have been used as alternative means of differentiating species and strains of parasites. Of these techniques, isoelectric focusing (IEF) has proved to be specific and reproducible with cestodes, protozoa and trematodes. As with the traditional morphological approach, there is, however, the potential for host influence. For a particular separation technique to be useful, it is important that banding profiles differentiating parasites be consistent, regardless of host species. To test for host influence, Fasciola hepatica were maintained in four definitive hosts. Soluble proteins of flukes from different hosts and their respective excretory-secretory product (ESP) were then subjected to IEF together with respective host tissues, and the resulting banding patterns were compared. Protein banding profiles of fluke body protein from llama, rat and mouse were very similar, while those of flukes from calves were distinct. Banding patterns of fluke ESP from different hosts were dissimilar. Overall banding patterns of body protein and ESP of flukes from different hosts were distinct from those of respective host tissues. The unique banding patterns of the body protein of flukes from calves indicate that there was host influence in their protein banding patterns, and the uniqueness of the calf fluke is discussed relative to hosts resistance to the fluke.

Spiculopteragia spiculoptera and S. asymmetrica (Nematoda: Trichostrongyloidea) from red deer (Cervus elaphus) in Texas.

Specimens of Spiculopteragia spiculoptera and S. asymmetrica were recovered from the abomasa of five of ten naturally infected red deer (Cervus elaphus) in Texas (USA). Female specimens of Spiculopteragia were present in all five animals. Male specimens of S. spiculoptera and S. asymmetrica were present in one of five and three of five red deer, respectively. Spiculopteragia spiculoptera has not previously been recognized in the United States and the present report constitutes the first records of Spiculopteragia spp. in red deer from North America. It is likely that species of Spiculopteragia have been introduced to North America with the import of exotic cervids on several occasions. Focal populations of these nematodes have been established in North America; however, distribution of the parasites likely coincides with areas of residence of introduced populations of red deer and fallow deer (Cervus dama) in the Nearctic.

Differentiation of Sarcocystis species in European sheep by isoelectric focusing

Isoelectric focusing was carried out on total proteins derived from cystozoites ofSarcocystis tenella, S. arieticanis andS. gigantea. The overall protein profile of each species was distinct and reproducible and enabled the unequivocal differentiation of the threeSarcocystis species. In total, approximately 60 bands could be counted forS. tenella andS. arieticanis and approximately 45 bands forS. gigantea cystozoites in the pH range of 3–9. Isoelectric points (pIs) were determined for dominant peaks of densitometric scans of gels in the pH range of 3–9 (cathodal and anodal sample application) and arbitrary identification numbers were assigned to 25 (5 and 20) peaks ofS. tenella, 25 (8 and 17) peaks ofS. arieticanis and 21 (7 and 14) peaks ofS. gigantea; similar assignments were also used for 11 peaks ofS. tenella, 18 peaks ofS. arieticanis and 12 peaks ofS. gigantea observed on densitometric scans of gels in the pH range of 4–6.5.

The anthelmintic efficacy of netobimin against experimental infections of Fasciola hepatica in sheep

Netobimin (coded SCH 32481, Schering Corporation), a new broad-spectrum anthelmintic having both fasciolicidal and nematocidal properties was evaluated for efficacy against mature Fasciola hepatica infections in sheep. The trial was conducted with 30 cross-bred spring lambs, each experimentally infected with 250 F. hepatica metacercariae. A single treatment of netobimin was administered at 17 weeks post-infection (PI) by oral drench at 7.5 or 20 mg kg-1 body weight while 10 animals remained as untreated controls. At necropsy (either 1 or 2 weeks post-treatment), the mean number of adult flukes recovered from the control, 7.5 and 20 mg kg-1 groups were 94.7, 35.9 and 8.8, respectively. The resulting efficacies were 62% (P less than or equal to 0.05) and 90.7% (P less than or equal to 0.01), respectively. No clinical signs of fascioliasis were noted in any sheep during the trial. No signs of toxicosis nor any adverse reactions to the drug were observed.

Influence of ivermectin and clorsulon treatment on productivity of a cow-calf herd on the southern Oregon coast☆

The reproductive performance of beef cows and the weight gain of their calves was evaluated after oral administration of clorsulon or clorsulon in combination with the subcutaneous administration of ivermectin. One hundred and fifty pregnant cows harboring infections of Fasciola hepatica were assigned to one of three treatment groups: Group 1 comprising 50 unmedicated controls; Group 2, 50 cows treated with clorsulon orally at 7 mg kg-1; Group 3, 50 cows treated with clorsulon orally at 7 mg kg-1 and ivermectin subcutaneously at 200 micrograms kg-1. Weights and body condition scores of the cows were measured and fecal and blood samples were taken at trial initiation and days 158 and 270. Pregnancy status was also determined at day 270. Weights and body condition scores were measured for the calves at days 158 and 270. Adjusted 205 day weaning weight of the calves was calculated and analyzed for differences between treatment groups. Four sets of tracer calves were used periodically throughout the trial to monitor the helminth challenge to the herd. Both gastro-intestinal nematodes and liver flukes were transmitted to the tracer calves during the entire trial. Even in the face of continual helminth challenge, beef cows treated with combined clorsulon/ivermectin conceived approximately 2 weeks earlier than their untreated counterparts and their calves had better body condition scores and weights than untreated calves.

The anthelmintic efficacy of netobimin against naturally acquired gastrointestinal nematodes in sheep

The broad-spectrum anthelmintic efficacy of netobimin (SCH 32481, Schering Corporation) was evaluated using 30 cross-bred spring lambs with naturally acquired infections of gastrointestinal nematodes. Three groups of 10 animals each were allotted into either control (given a tap water drench as a placebo) or 7.5 and 20 mg kg-1 dosage groups (given the netobimin as an oral drench). Seven to fourteen days post-treatment, animals were necropsied and nematodes recovered by standard techniques. Examination of fecal samples taken on dates of necropsy showed median egg production was reduced in treated animals (61.98% with 7.5 mg kg-1 and 100% with 20 mg kg-1). The compound was highly effective in removal of adult nematodes representing a number of genera and species of trichostrongyloids at the 7.5 and 20 mg kg-1 dose levels (shown, respectively, below). These included Ostertagia spp., with O. circumcincta, O. trifurcata, O. ostertagi and Teladorsagia davtiani (96.20%; 100%), Trichostrongylus spp., with T. axei, T. vitrinus and T. colubriformis (100%; 98.72%), Nematodirus spp., with N. spathiger, N. filicollis and N. battus (100% both levels) and Haemonchus contortus (100% both levels). High efficacies against other species of nematodes (at both dose levels) were not statistically significant (Cooperia spp., Chabertia ovina and Oesophagostomum venulosum). At 20 mg kg-1, netobimin significantly reduced populations of early and late fourth stage larvae of Ostertagia spp. by 100%. The overall efficacy (all life stages included) was 90.16% at 7.5 mg kg-1 and 98.77% at 20 mg kg-1 dose levels. No adverse reactions or signs of toxicosis were observed.

Banding patterns of Fasciola hepatica and Fasciola gigantica (Trematoda) by isoelectric focusing

Liver flukes Fasciola hepatica and Fasciola gigantica are polymorphic and vary morphologically depending upon the host being parasitized. It is known also that mixed infection occurs where both species are present. A technique involving protein separation was used to distinguish the 2 species. Isoelectric focusing of soluble proteins was performed on polyacrylamide gels using whole-body proteins from adult flukes. Although many bands appeared common to both species and some were shared with host tissues, the banding patterns could be used to distinguish 1 species from the other. Soluble protein isoelectric focusing is simple, reproducible, and has very good resolution. It seems well suited to the differentiation of the 2 fluke species.