Studies on Haemonchus contortus. XII. Effect of Trichostrongylus axei in Dorper lambs on natural pasture lightly infested with H. contortus

Weaned Dorper lambs on natural pasture were predosed with 40 000 infective larvae (L3) of Trichostrongylus axei, irradiated (0,3 kGy) L3 of T. axei or closantel at 10 mg/kg either in September or November 1978 and were compared with Merino yearlings predosed with 40 000 L3 of T. axei in November 1977. The following summer (December-March) only 178,6 mm of rain fell and very few H. contortus were present on pastures. Artificial challenge with 20 000 L3 of Haemonchus contortus with the local strain from the University of Pretoria's experimental farm was given 6-7 months after predosing with T. axei. When compared with the controls, significant reductions occurred only in Group 11 (T. axei irradiated at 0,3 kGy on Day +63) (P = 0,025); Group 2 (T. axei on Day 0) (P = 0,003) and Group 4 (T. axei and closantel on Day 0) (P = 0,049). We concluded that predosing with T. axei was unsuccessful in Dorpers and Merinos artificially challenged 6-7 months later with H. contortus.     

Effect of sericea lespedeza (Lespedeza cuneata) fed as hay, on natural and experimental Haemonchus contortus infections in lambs

Parasitic infection is one of the leading economic constraints in small ruminant production. This problem is exacerbated as the resistance of nematode populations to chemical treatment (anthelmintics) becomes increasingly more common. Condensed tannin (CT) containing plants are being investigated as alternative solutions to address these problems. This investigation was conducted to evaluate the effect of the CT containing forage, Lespedeza cuneata (sericea lespedeza, SL) fed as hay, on Haemonchus contortus infection in sheep. Naturally and experimentally infected lambs were fed either SL or bermudagrass (BG, control) hay for 49 days. All lambs were fed BG hay for an additional 14 days. SL hay effectively reduced (67-98%) fecal egg count (FEC) during the time of feeding for both infection groups. FEC increased in both infection groups after SL feeding was stopped which indicated an effect on fecundity. SL hay feeding also reduced worm numbers, with more of an effect on reducing naturally infected worm burdens (67.2%) than on establishment of incoming larvae (26.1%). SL fed as hay may be more useful to remove existing worms than establishing worms. The decrease in FEC would have the benefit of reduced pasture contamination.     

Resistance of St. Croix lambs to Haemonchus contortus in experimentally and naturally acquired infections.

Parasitological and immunological parameters of experimental or naturally acquired infections with Haemonchus contortus were compared in St. Croix and Dorset lambs. In experimental infections, St. Croix lambs developed significantly greater levels of resistance to H. contortus, following primary exposure, as compared with Dorset lambs. This resistance was influenced both by age and by prior exposure to parasites. In grazing experiments on H. contortus-infected pasture, St. Croix lambs shed significantly fewer eggs as early as 5 weeks following initial exposure. Further, St. Croix lambs had more than 99% fewer worms in the abomasum at necropsy compared with age-matched Dorset lambs. Lymphoproliferative assays using peripheral blood mononuclear cells and antigen-specific serological tests demonstrated only minor differences in immune responsiveness between the two breeds despite the dramatic parasitological differences. Similarly, abomasal mucus from both breeds had elevated levels of parasite-specific antibodies and contained substances mediating larval paralysis. In contrast, St. Croix lambs which had become resistant to nematode infection had dramatically higher numbers of globule leukocytes in the abomasal mucosa compared with Dorset lambs.     

Age group interaction of Haemonchus contortus in premunized lambs.

Lambs premunized with a relatively less pathogenic (RLP) isolate of Haemonchus contortus were challenge exposed each day with 500 3rd-stage larvae (L3) of a normally pathogenic isolate of the same worm, starting on day 9 of the premunition-induced infection. In 6-month-old lambs, there was a fourfold decrease in egg production by the premunizing RLP isolate with challenge exposure compared with egg production in lambs given only the premunizing RLP isolate and not challenge exposed. Premunized lambs were able to carry larger worm burdens with less hemoglobin loss than were nonpremunized challenge-exposed control lambs. Although challenge exposure had affected parasitic egg production by the 9-day-old premunizing RLP worms, the same young infective isolates were able to survive and protect lambs against the additional parasitism.     

Serum IgG response of Manchego lambs to infections with Haemonchus contortus and preliminary characterization of adult antigens

Manchego lambs (16-18 weeks old) were infected with 2500 infective larvae (L3) of Haemonchus contortus and challenged 2 months later with 5000 L3. The serum IgG anti-Haemonchus response was estimated by enzyme-linked immunosorbent assay (ELISA) using soluble proteins from adults and L3. Previously infected Manchego lambs failed to mount a protective immune response against challenge, at least as assessed by faecal egg counts and pre-patency periods. Primary infection did not provoke any rise in specific anti-parasite serum antibodies, whereas a weak but significant rise was observed in challenged 6.5-month-old lambs which was very similar in both infected and non-infected lambs. The serum IgG anti-parasite response was higher against larval antigens than adult soluble proteins. Preliminary characterization of adult and larval soluble proteins by electrophoresis under reducing and denaturing conditions and Western blotting showed high cross-reactivity of both extracts. Immunoblots of adult H. contortus probed with infected and challenged lambs' sera did not yield conclusive results, although some low molecular weight peptides were recognized.     

Evaluation of immune response to artificial infections of Haemonchus contortus in Gulf Coast Native compared with Suffolk lambs

Due to their critical functions in worm physiology, glutathione peroxidases in Haemonchus contortus are potential candidates for vaccine to control haemonchosis. However, information on the protection potential of these molecules is lacking. In this study, recombinant HC29 antigen was tested for protection against experimental H. contortus infections in goats. Fifteen animals were allocated into three trial groups. The animals of HC29 group was vaccinated with a recombinant HC29 DNA vaccine twice at day 0 and day 14, and then challenged with 5000 infective H. contortus L3 (third larval stage) on the 28th day. An unvaccinated positive control group was challenged with L3 at the same time. An unvaccinated negative control group was not challenged with L3. The results indicated HC29 DNA vaccine was transcribed at local injection sites and expressed in vivo post immunizations, respectively. Following L3 challenge, the mean eggs per gram feces and worm burdens of HC29 group were reduced by 36.1% and 35.6% respectively compared to the positive control group. After immunization with the DNA vaccine, significantly high levels of serum IgG, serum IgA, mucosal IgA and CD4(+) T lymphocytes were detected. These results suggest that recombinant H. contortus HC29 glutathione peroxidase DNA vaccine induced a partial immune response and has protective potentials against caprine haemonchosis.     

The effect of transfer factor treatment on two challenge infections of Haemonchus contortus in immuno-competent 7-month-old lambs

A number of immuno-competent Blackface lambs were infected with 10,000 Haemonchus contortus third stage larvae and were later challenged with a second dose of 10,000 H. contortus third stage larvae. One group of lambs was treated with dialysed transfer factor prior to the second challenge dose. Histological changes in the abomasal wall, reductions in abomasal worm burdens at slaughter and faecal egg counts monitored throughout the experiment suggested the presence in some animals of an innate resistance, a "self curing" effect, an enhancement of this "self curing" effect in the transfer factor treated group and a response in one transfer factor treated animal comparable to "classical self cure".     

Evaluation of Cymbopogon schoenanthus essential oil in lambs experimentally infected with Haemonchus contortus

Hematophagous gastrointestinal parasites cause significant economic losses in small ruminant grazing systems. The growing reports of multi-drug resistant parasites call for intensive research on alternative treatments for anthelmintics to help small ruminants cope with these parasites. Two-month-old lambs with mean body weight (BW) of 22.5 kg were experimentally infected with a multidrug-resistant Haemonchus contortus strain. Infected animals were dosed orally with Cymbopogon schoenanthus essential oil to evaluate its anthelmintic potential. Eighteen animals were allocated into three groups of six animals, and each received one of the following treatments: Group 1 - control (10 mL of water), Group 2 - C. schoenanthus essential oil (180 mg/kg BW); and Group 3 - C. schoenanthus essential oil (360 mg/kg BW). Animals received the oil once a day for 3 consecutive days. Lambs were evaluated clinically for blood biochemistry before, at 1, 5, 10, 15 and 20 days after treatment, and then were euthanized to assess the total worm burden. No statistically significant reduction in fecal egg count, packed cell volume or total worm count was observed after treatments. Also, no statistical difference among group means for blood levels of urea, creatinine, albumin, alkaline phosphatase, aspartate aminotransferase and gamma glutamyl transferase was found. Larval development assay (LDA) and egg hatch assay (EHA) were performed from feces of treated animals at 1, 5, 10 and 15 days after essential oil administration. An inhibition in LDA was observed 1 day after the 3-day treatment in larvae from feces of animals treated with 360 mg/kg essential oil. In conclusion, the essential oil at the doses of 180 mg/kg and 360 mg/kg was safe to sheep, but failed as an anthelmintic treatment when applied to young sheep artificially infected with a multidrug-resistant H. contortus strain.     

Activity of febantel on natural infections of gastrointestinal helminths in lambs in a controlled test

The efficacy of febantel paste formulation (6 and 12 mg/kg) against natural infections of gastrointestinal helminths in lambs (n = 33) in Kentucky was evaluated in a controlled test. For the test, 23 lambs were treated orally (17 at 6 mg/kg and 6 at 12 mg/kg) and 10 lambs were not treated. Removals at both dosages in treated lambs were 95% to 100% for species of immature and mature Haemonchus, Trichostrongylus, and Cooperia; and mature Ostertagia females, Nematodirus, and Strongyloides. For immature Nematodirus, removals were 92% and 77% at the dosages of 6 and 12 mg/kg, respectively. Only a few specimens (av less than 100) of some other species or stages were found in the nontreated group and removal of them (at both dosages) were 94% to 100% for Ostertagia (immature and males), Strongyloides (immature), Oesophagostomum (immature), and Moniezia (mature); and 61% (at 6 mg/kg) and 100% (at 12 mg/kg) for Capillaria (mature), 0% for Trichuris (mature, at both dosages), and 67% (at 6 mg/kg) and 100% (at 12 mg/kg) for Oesophagostomum (mature).     

Effects of dietary molybdenum on nematode and host during Haemonchus contortus infection in lambs.

The addition of molybdenum (0.05 mmol kg-1 dry-matter) to the diet of lambs given a trickle infection of Haemonchus contortus larvae (500 third stage larvae d-1 over six weeks) reduced mean faecal egg counts (epg) from 3952 to 2312 +/- 402 by 32 days (P less than 0.02) and greatly reduced the mean number of worms recovered from the abomasum 14 days after infection ceased (907 compared with 4167: P less than 0.01). Infection reduced haemoglobin concentrations less in lambs given molybdenum although the difference was small relative to the reduction in worm burden. Lambs not given molybdenum had low intraepithelial mast cell counts in the abomasal mucosa and less abomasal hypertrophy than expected from abomasal parasitism. Molybdenum did not consistently reduce the copper status of the host or the parasite. Previous exposure to molybdenum greatly reduced protein but not proteinase activity in, or secreted by, adult worms cultured for eight hours. It is suggested that molybdenum either increased the inflammatory response which preceded worm rejection or that it indirectly enhanced that reaction by reducing the effectiveness of copper-dependent, anti-inflammatory enzymes in the gastrointestinal mucosa.     

Influence of dietary protein supply on resistance to experimental infections with Haemonchus contortus in Ile de France and Santa Ines lambs

The effect of Haemonchus contortus infection in sheep fed with a moderate and high protein content diet was evaluated in two breeds of sheep. Forty-eight Ile de France and Santa Ines lambs were maintained indoors since birth, in worm-free conditions. The lambs were allocated after weaning in four groups of six animals per breed, which were either infected or remain uninfected and given access to either a moderately or highly metabolizable protein diet. The moderately and highly metabolizable protein diets were calculated to supply 75 and 129 g metabolizable protein per kg of dry matter (MP/kg DM), respectively. The infection consisted of a trickle infection with 300 infective larvae, three times a week, for 12 weeks. Significant differences were observed for mast cell, globule leukocyte and eosinophil counts in the abomasal mucosa of the infected groups compared to the control of both breeds (P<0.05), regardless of the diet supplied. Significantly higher IgA anti-L5 antibody was detected in the infected Santa Ines groups than in the infected Ile de France groups (P<0.05). Increased metabolizable protein supply resulted in larger body weight gain and higher packed cell volumes for both breeds (P<0.05). Both breeds showed an increased ability to withstand the pathophysiological effects of H. contortus infection when given access to the highly metabolizable protein diet. However, increased metabolizable protein supply resulted in reduced worm burdens in Santa Ines lambs but not in the Ile de France lambs (P<0.05). The present results show that the increase in protein content in growing lamb diets may benefit resistance and resilience to gastrointestinal parasites but that these benefits may vary among breeds.     

Lymphocyte reactivity to antigens of Haemonchus contortus in antigen-inoculated and H contortus-naive lambs.

Lambs were inoculated during either fetal or early neonatal life with antigens of Haemonchus contortus (HcAg) third-stage larvae, or ovalbumin (OVA), or were left untreated. By the time lambs were 4 weeks old, blood lymphocytes from all of them responded to HcAg. This was evident in all lambs, regardless of the previous inoculation regimen, although higher response was seen in HcAg-inoculated lambs, compared with naive lambs. Only lambs inoculated with OVA had OVA-specific lymphocyte activity. In HcAg-naive lambs, the maximal HcAg lymphocyte response was observed between the ages of 5 and 25 weeks, with gradual decrease thereafter. In vitro detection of HcAg-reactive lymphocytes was affected by antigen concentration and presence of plasma. Plasma inhibited HcAg-reactive lymphocytes from all lambs at high in vitro antigen concentration and completely abrogated the response in H contortus antigen-naive lambs.     

Anthelmintic activity of Jatropha curcas L. seeds on Haemonchus contortus

The aim of this study was to evaluate the anthelmintic activity of hexane (HE), ethyl acetate (EA) and ethanol (EE) extracts obtained from the seeds of Jatropha curcas using the egg hatch inhibition assay (EHA) and the artificial larval exsheathment inhibition assay (LEIA). For the egg hatch assay, HE, EA and EE were used in concentrations of 3.12, 6.25, 12.5, 25 and 50 mg ml(-1), accompanied by a negative control (5% Tween 80) and a positive control (0.025 g ml(-1) thiabendazole). In LEIA, the extracts were tested at a concentration of 1000 μg ml(-1), accompanied by a negative control (PBS). To evaluate the effect of tannins, the extract with the greatest effect was incubated with polyvinyl polypyrrolidone (PVPP). The EE (50 mg ml(-1)) inhibited 99.8% of egg hatching. After the addition of PVPP, the ovicidal effectiveness of EE was reduced to 91.9%. Using the HE and EA, inhibition of egg hatching was 15.3% and 32.2%, respectively. In the LEIA, 18.9% of L3 incubated with EE were exsheathed (p<0.01). The addition of PVPP to EE reversed the inhibitory effect on larval exsheathment. The percentage of exsheathment of L3 incubated with HE (99.6%) and EA (97.8%) did not differ from the control group (p>0.05). The results show that the effects of EE on eggs are not solely due to the tannins. However, these secondary metabolites are implicated in blocking the larval exsheathment.     

Effect of Haemonchus contortus infection on the clearance of antipyrine, sulfobromophthalein, chloramphenicol, and sulfathiazole in lambs

A study was made to determine the effect of Haemonchus contortus parasitic infection in lambs on the clearance of several IV administered drugs. Clearance of sulfobromophthalein or sulfathiazole from the plasma of lambs was unaffected by infection with H contortus. Clearance of antipyrine was enhanced by the infection, and thiabendazole treatment did not alter this effect. Clearance of chloramphenicol (CAP), administered as the succinate ester (CAPS), was not changed by the infection, but it was increased after treatment with thiabendazole. Changes in the mean body residence time and initial plasma concentration of CAPS and CAP after treatment with thiabendazole indicate that hydrolysis of CAPS to CAP was reduced. High concentrations of CAPS apparently enhanced its own elimination directly rather than via the expected sequence involving hydrolysis, glucuronidation, and excretion of CAP-glucuronide. Enhanced clearance of antipyrine following infection of lambs with H contortus can be explained in at least 2 ways. First, it is possible that the lambs did not have mature amounts of hepatic drug metabolizing enzyme activity as reported by other investigators, which may be explained by breed differences or animal husbandry practices. Second, infection of lambs by H contortus may have triggered an inductive response in hepatic cytochrome P-450-mediated activities, which might result via a generalized enhancement in hepatic protein synthesis associated with the physiologic response to replace plasma proteins and other blood components lost through gastrointestinal hemorrhage caused by the active feeding of adult worms. Other phase-II reactions such as acetylation, glucuronidation, and glutathione-S-transferase apparently were not affected.     

The effects of transfer factor, parasitic antigen and precipitated immunoglobulin treatments in lambs infected with Haemonchus contortus

Significant reductions in adult worm burdens in lambs less than six months old infected with 5000 third stage Haemonchus contortus larvae were achieved following the combination of a single transfer factor treatment with multiple parenteral treatments of adult parasitic antigen and precipitated immunoglobulin. Specific cellular responses in the abomasa associated with individual treatments were discernible in all the experimental groups and quantitative differences in these responses reflected the varying treatment levels and treatment combinations. The reactions associated with transfer factor were similar to immediate type hypersensitivity responses and their role as possible effector mechanisms in correcting partial immune incompetency in lambs is discussed.     

Dose titration of sericea lespedeza leaf meal on Haemonchus contortus infection in lambs and kids

The objective of three experiments was to determine the impact of supplementing sericea lespedeza (Lespedeza cuneata; SL) in three concentrations in a loose or pelleted diet on gastrointestinal nematodes (GIN) in small ruminants. Experiments on lambs were conducted at the USDA, Agricultural Research Service in Booneville, AR (Exp. 1) and at Louisiana State University in Baton Rouge, LA (Exp. 2); an experiment on goat kids occurred at University of Maryland-Eastern Shore (Exp. 3). Exp. 1 used crossbred hair sheep lambs naturally infected with GIN that were randomly allocated to diets containing 0, 25, 50, and 75% SL diets (n=11 or 12/diet). Exp. 2 consisted of Haemonchus contortus-inoculated crossbred wool breed lambs that were blocked by gender and FEC and randomly assigned to 0, 25, 50, or 75% SL diet (n=8/diet). Fecal egg counts (FEC) and blood packed cell volume (PCV) were not influenced by SL supplementation in Exp. 1 and 2. Exp. 3 consisted of naturally GIN infected Boer crossbred goat kids in individual pens. Kids were blocked by FEC and randomly allotted to treatments of 0, 20, 40, or 60% SL with 9-13 goats/diet. The more SL fed, the greater the reduction in FEC (P<0.001). There was an increase in PCV in SL fed goats (P<0.001). Larval speciation at the end of the experiment indicated that feces from control animals produced 43% H. contortus larva while 20, 40 and 60% SL resulted in 39%, 35% and 31% H. contortus larvae, respectively. Feeding dried SL may be less effective in lambs than kids, though concurrent studies must be conducted to confirm this.