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PubMed1999

Structural characterisation and pharmacology of KHEYLRFamide (AF2) and KSAYMRFamide (PF3/AF8) from Haemonchus contortus.

The FMRFamide-related peptides (FaRPs), KHEYLRFamide (AF2) and KSAYMRFamide (PF3) were structurally characterised from the parasitic nematode of sheep, Haemonchus contortus (MH isolate). Both peptides were sequenced in a single gas-phase sequencing run and their structure confirmed by mass spectrometry which identified peptides of 920 Da (C-terminally amidated AF2) and 902/918 Da (C-terminally amidated non-oxidised/oxidised PF3, respectively). AF2 had inhibitory effects on H. contortus muscle and inhibited acetylcholine (ACh, 10 microM)-induced contractions, with a threshold for activity of 1 microM. PF3 induced concentration-dependent contractions of H. contortus (activity threshold, 10 nM) and enhanced ACh contractions. Compared with the MH isolate, an isolate of H. contortus which has reduced sensitivity to cholinergic drugs (Lawes isolate) was less sensitive to the effects of PF3. The concentration-response curves for the cholinergic compounds ACh and levamisole (LEV), and PF3, but not a control, KPNFIRFamide (PF4), showed a statistically similar shift. This study implicates PF3 in the modulation of cholinergic function in H. contortus.

Amino Acid Sequence↗
PubMed2000

Multiple lethal effects induced by a benzimidazole anthelmintic in the anterior intestine of the nematode Haemonchus contortus.

A mechanism of benzimidazole efficacy against parasitic nematodes is postulated to involve inhibition of intestinal secretory vesicle transport via depolymerization of microtubules. We show that fenbendazole (FBZ) treatment of lambs causes pathology localized to the anterior intestine in the parasitic nematode Haemonchus contortus. The pathology included gross disintegration of the anterior intestine, DNA fragmentation in anterior intestinal nuclei with characteristics of an apoptosis-like process, and inhibition of host erythrocyte digestion. These lethal effects were associated with inhibited transport of apical secretory vesicles in the anterior intestine, and then generalized dispersal of these vesicles contents throughout the intestinal cytoplasm and worm body. Cytoplasmic accumulation of secretory vesicles and undigested erythrocytes preceded DNA fragmentation and vesicle content dispersal. Both DNA fragmentation and vesicle content dispersal were detected in disintegrated intestine and intestine that had not yet undergone disintegration. These pathologic effects in the anterior intestine appear sufficient to explain the efficacy of FBZ against adult H. contortus. The results implicate mechanisms in the anterior intestine that govern dispersal of apical secretory vesicle contents, DNA fragmentation and tissue disintegration as effectors of this pathology.

Animals↗
PubMed1998

Sustained-release delivery systems and their application for endoparasite control in animals.

A solid formulation of a potent anthelmintic macrocyclic lactone, moxidectin, was administered using a non-degradable delivery device to discharge the agent into the subcutaneous tissues of sheep. In vivo release was monitored in sheep indirectly using faecal egg counts. Using a dose of 0.2 mg moxidectin/kg body weight when applied in the form of a solid pellet, protection of sheep against Haemonchus contortus challenge was conferred to a level greater than that of sheep which received Cydectin, the commercial liquid injectable form delivered at the same dosage. The anthelmintic efficacy of the solid formulation was assessed at four dosage levels in sheep and it was demonstrated that the dosage of anthelmintic agent could be reduced to 1/6 of the present recommended injectable dose. When two pellets containing the recommended dose of moxidectin were loaded into a non-degradable delivery device, the period of H. contortus control was extended from 42 to 183 days. Antibody levels of sheep receiving repeated infections of H. contortus L3 larvae and treated with moxidectin-loaded devices were reduced significantly compared to the levels observed in sheep treated with Cydectin (p < 0.0005). This implies that the group treated with the moxidectin-loaded devices was exposed to a reduced antigenic load compared to sheep treated with placebo devices, and sheep treated with Cydectin. The antibody levels generated in the sheep treated with placebo devices were no different to those treated with Cydectin. Application of this sustained release device may allow the control of nematode diseases in livestock throughout an entire season with a single administration.

Animals↗
PubMed1999

The development of vaccines against gastrointestinal nematode parasites, particularly Haemonchus contortus.

Many parasitic nematodes are developing resistance to chemical treatment, and the search is on to produce commercially viable molecular vaccines. Much progress has been made with highly protective 'hidden antigens', especially for Haemonchus contortus, and recent work with new 'natural antigens' has yielded promising results. Here, Sue Newton and Ed Munn review the most recent advances in these two main approaches to this problem.

Animals↗
PubMed1999

Pharmacology of anthelmintic resistance.

Anthelmintic resistance has grown from a curiosity to an important economic problem in several animal industries and is now set to threaten the control of human parasites. The pharmacology of anthelmintics and anthelmintic resistance has been studied most extensively in the nematode parasites of sheep. Here, Nick Sangster and Jenny Gill discuss this veterinary experience, summarizing the progress made in understanding anthelmintic resistance and highlighting the tools available for research.

Animals↗
PubMed1999

Vaccination against the gastrointestinal nematode, Haemonchus contortus, using a purified larval surface antigen.

Sheep were immunized with a purified antigen (Hc-sL3) expressed on the surface of L3 larvae of the gastro-intestinal parasite, Haemonchus contortus, using different adjuvant and immunization routes. In the first experiment, intradermal immunization of sheep with Hc-sL3 and QuilA did not result in reductions in faecal egg counts after subsequent challenge infection while significant reductions were obtained when aluminium hydroxide (AH) was used as the adjuvant. Significant protection with Hc-sL3 absorbed on AH was confirmed in a second experiment and this protection was maintained when dextran sulphate was added to the Hc-sL3/AH mixture while the addition of pertussis toxin abrogated the protective effect. Significant levels of protection, as determined by reductions in both faecal egg counts and worm burdens, were also obtained when the Hc-sL3/AH mixture was injected into the rectal mucosa or the Hc-sL3 antigen was deposited on the surface of the rectal mucosa with cholera toxin. No correlations with antibody levels or isotype and protection were observed.

Animals↗
PubMed2000

Attempts to control haemonchosis in grazing ewes by vaccination with gut membrane proteins of the parasite.

A vaccination trial was conducted to evaluate the potential benefit of Haemonchus contortus gut membrane proteins as vaccine antigens under field conditions in Louisiana. The trial was conducted in the summer of 1996 in a flock of ewes grazing pasture naturally infected with H. contortus. Ewes were randomly assigned to three treatment groups (vaccine, adjuvant only, and saline) and fecal egg counts (FEC, measured as eggs per gram of feces), packed cell volumes (PCV), and antibody levels were monitored fortnightly for 12 weeks. It was shown by FEC that there were large individual variations in susceptibility to H. contortus in both vaccinated and non-vaccinated sheep, a finding which could have masked differences between treatments when analyzed by conventional statistical methods. Based on their egg counts before the period when the vaccination could have had an effect, all ewes were categorized as 'susceptible' or 'relatively resistant'. The significance of differences between FEC, PCV and antibody responses of vaccinated and control sheep were tested separately for the 'susceptible' and 'relatively resistant' category. The 'susceptible' vaccinates shed 65% fewer worm eggs during the period when the vaccine could have had an effect, but the difference was only significant on Week 6 post-vaccination. In these experiments, it was difficult to completely exclude the confounding effect of having 'relatively resistant' sheep in the control group. More studies are needed to further evaluate H11 and H-gal-GP antigens under field conditions.

Animals↗
PubMed2000

Interaction between Trypanosoma evansi and Haemonchus contortus infection in goats.

An experimental study on interaction between Trypanosoma evansi and Haemonchus contortus infections was conducted in 42 male Barbari goats of 6-9 months age. Parasitological observations like prepatent period, faecal egg count, worm burden and mortality were made in animals infected with H. contortus followed 1 week later with T. evansi and vice versa. These parameters were also examined in animals with single infection with each of these parasites. It was revealed that T. evansi infection in goats lowered the normal resistance to H. contortus. The prepatent period was markedly reduced to 16-18 days in cases where T. evansi preceded H. contortus as compared to 21-25 and 21-23 days in single H. contortus and in a H. contortus preceding T. evansi infection, respectively. The rate of L(3) establishment was significantly enhanced in infection where T. evansi preceded H. contortus. A higher rate of mortality and more pronounced pathological effects were observed in combined infections than in single ones.

Animals↗
PubMed2000

Uptake of albendazole and albendazole sulphoxide by Haemonchus contortus and Fasciola hepatica in sheep.

The pattern of in vivo uptake of albendazole (ABZ) and its major metabolite, ABZ-sulphoxide (ABZSO), by Haemonchus contortus and Fasciola hepatica recovered from ABZ-treated sheep, was investigated. Concentration profiles of both compounds were simultaneously measured in target tissues/fluids from the same infected sheep. In addition, the proportion of the (+) and (-) ABZSO enantiomers was determined in plasma, bile and F. hepatica recovered from treated sheep. Sheep naturally infected with H. contortus were intraruminally (i.r.) treated with ABZ (micronized suspension, 7. 5mg/kg) and the plasma concentrations of ABZSO and ABZ-sulphone (ABZSO(2)) determined in addition to the concentration of ABZ and ABZSO in H. contortus, abomasal mucosa and fluid content samples. In addition, F. hepatica artificially infected sheep were treated i.r. with the same ABZ suspension (7.5mg/kg), and samples of blood, bile, liver tissue and adult flukes were collected and analysed by HPLC to determine the concentrations of ABZ and both enantiomers of ABZSO. ABZSO and ABZSO(2) were the analytes recovered in plasma with ABZ and ABZSO present in H. contortus. ABZ was the analyte recovered at the highest concentration in H. contortus and abomasal mucosa, whereas higher concentrations of ABZSO were measured in abomasal fluid content. Only low concentrations of ABZ were detected in F. hepatica and bile, but markedly higher concentrations of ABZ were measured in liver tissue. ABZSO was the main molecule recovered in F. hepatica, plasma and bile samples collected from ABZ-treated sheep. The (+) enantiomer of ABZSO was recovered at a higher proportion in plasma (75%), bile (78%) and F. hepatica (74%) after ABZ administration to infected sheep.

Abomasum↗
PubMed2001

Experimental cross-infections of Haemonchus placei (Place, 1893) in sheep and cattle.

To examine effects on biology and morphology, Haemonchus placei infections were propagated in cattle or sheep and infective larvae were introduced into the same or opposite host. Ovine source larvae had a geometric mean (GM) prepatent period of 26.5 days regardless of host species, compared to 30.6 days for bovine source larvae regardless of host species. Similarly, ovine source H. placei had a higher GM percentage establishment versus bovine source larvae (9.6% versus 3.4%) regardless of host species. The patent daily egg count for ovine source versus bovine source H. placei was 109.1 versus 50.0 FEC regardless of host species. Mean spicule length for ovine source H. placei was 492.5 microm while bovine source measured 496.5 microm. Mean female tail length for ovine source H. placei was 586.5 and 589.5 microm for bovine source H. placei. Using synlophe morphology, all worms that were measured were identified as H. placei. Sixty-three percent of females examined for vulvar flap morphology had knob-like vulvar flaps while the remaining 37% had linguiform vulvar flaps.

Animals↗
PubMed2001

Preliminary observations on the potential of gut membrane proteins of Haemonchus contortus as candidate vaccine antigens in sheep on naturally infected pasture.

In a trial lasting 11 months in South Africa, faecal egg counts and haematocrits of sheep vaccinated with gut membrane proteins of adult Haemonchus contortus were compared with unvaccinated controls grazing pasture contaminated with the parasite. Vaccination reduced egg output by >82% on average during one 4 month period of the trial and simultaneously significantly reduced the degree of anaemia and deaths due to haemonchosis. Although vaccine immunity was not sufficiently long lasting to prevent a surge in egg output which occurred after the onset of a period of irrigation, re-vaccinating the sheep at this point cleared their newly acquired infection and rapidly restored protection to approximately the level observed beforehand. It was clear that a vaccine based on parasite gut membrane proteins could offer substantial benefits in the control of natural haemonchosis.

Animals↗
PubMed2001

Anthelmintic resistance on goat farms in Georgia: efficacy of anthelmintics against gastrointestinal nematodes in two selected goat herds.

Gastrointestinal nematode (GIN) parasitism is a major constraint to production of goats in the southeastern United States. The conventional method of control used by producers in this region is frequent use of anthelmintics during the warm season. Overuse of anthelmintics has led to an increase in the incidence of anthelmintic resistance in many parts of the world, but data on prevalence of anthelmintic resistance in GIN of goats in the southeastern United States are very limited. To address this issue, anthelmintic efficacy was determined in goat herds at the Fort Valley State University, Agricultural Research Station (FVSU-ARS) and the University of Georgia, College of Veterinary Medicine (UGA-CVM) using fecal egg count reduction (FECR) tests and DrenchRite((R)) larval development assays (LDA). At FVSU-ARS, 2-year-old Spanish goat does were randomly allocated to one of nine different treatment groups (n = 10): albendazole (ABZ; 20mg/kg body weight (BW)), fenbendazole (FBZ; 20mg/kg BW), ivermectin (IVM; 0.4 mg/kg BW), doramectin (DRM; 0.4 mg/kg BW), moxidectin (MOX; 0.4 mg/kg BW), levamisole (LEV; 12 mg/kg BW), morantel tartrate (MOR; 10mg/kg BW), a combination of IVM (0.4 mg/kg BW) and ABZ (20 mg/kg BW), and untreated controls. At UGA-CVM, goats were randomly allocated to one of five different treatment groups (n = 8): ABZ (20 mg/kg BW), IVM (0.4 mg/kg BW), MOX (0.4 mg/kg BW), LEV (12 mg/kg BW), and untreated controls. All drugs in both experiments were administered orally. Anthelmintic efficacy was calculated by comparing 14-day post-treatment FEC of treated and control animals, and percent reductions were interpreted using the World Association for the Advancement of Veterinary Parasitology guidelines for resistance. For the LDA, nematode eggs were isolated from pooled fecal samples of untreated control goats in each herd and used to perform DrenchRite((R)) assays. In the FVSU-ARS herd, MOX, LEV, the combination of IVM and ABZ, IVM, DRM, ABZ, MOR, and FBZ reduced FEC by 100, 91, 88, 78, 76, 62, 48, and 10%, respectively. In the UGA-CVM herd, MOX, LEV, ABZ and IVM, reduced FEC by 100, 94, 87, and 0%, respectively. In both herds moxidectin was the only drug tested that was fully effective. Results of the LDA were in agreement with results of the FECR tests for both herds. These data demonstrate the presence of GINs resistant to all three major anthelmintic classes in both goat herds.

Animals↗
PubMed2001

Testing for clinical anaemia caused by Haemonchus spp. in goats farmed under resource-poor conditions in South Africa using an eye colour chart developed for sheep.

A novel clinical assay for the assessment and subsequent treatment of Haemonchus infection in sheep to slow down the development of anthelmintic resistance--the FAMACHA system--has been developed, tested and validated in South Africa. The system is based on a colour chart with five colour categories depicting varying degrees of anaemia that are compared with the colour of the mucous membranes of the eyes of sheep. The animal is then scored from severely anaemic (pale) through anaemic to non-anaemic (red) and those animals considered in danger of succumbing to the effects of haemonchosis are treated. This method was tested in goats farmed under resource-poor conditions in South Africa. Analyses in goats performed during the summers of 1998/1999 and 1999/2000 show a test sensitivity of 76 and 85%, respectively, meaning that the system may be used to identify correctly 76-85% of those animals in need of treatment with an anthelmintic. However, the test specificity remains low at 52-55%. This means that a large proportion of those animals that would not require treatment would in fact be treated. However, when the use of the FAMACHA system is compared with conventional dosing practices where all the animals are treated, using the FAMACHA system would result in a large proportion of the animals being left untreated. The untreated animals are then able to deposit the eggs of anthelmintic-susceptible worms on the pasture, while the treated ones should pass very few ova, given an effective anthelmintic. This maintains a reservoir of susceptible larvae in refugia, and should slow down the development of anthelmintic resistance. The validation of the FAMACHA system for goats for use by resource-poor farmers, which this paper describes, may have wide application in the tropics and subtropics of sub-Saharan Africa and elsewhere.

Anemia↗
PubMed2001

Vaccination against gastrointestinal nematode parasites of ruminants using gut-expressed antigens.

To date, proteins isolated from the surface of the gut of gastrointestinal nematodes, particularly Haemonchus contortus, have generally proved to be useful protective antigens and several are being progressed towards recombinant protein-based vaccines. This paper describes the properties of some of the most promising antigens and summarises their performance in laboratory and field based trials. The antigens described include contortin, H11, H-gal-GP, GPI and cysteine proteinases. In addition, the discussion addresses the utility of selected antigens to protect against co-infecting nematode species such as Teladorsagia circumcincta and against related nematode infections such as Ostertagia ostertagi in cattle.

Animals↗
PubMed2001

The potential of nematophagous fungi to control the free-living stages of nematode parasites of sheep: towards the development of a fungal controlled release device.

Studies showed that chlamydospores of the nematophagous fungus, Duddingtonia flagrans, are capable of surviving pressures of several tonnes when incorporated into matrices and pressed into tablets for the manufacture of prototype intraruminal controlled release devices (CRDs). They remain viable in this tabletted form for at least 9 months when stored at 4 degrees C. In vitro studies demonstrated that there was no effect on spore viability of prolonged exposure to either room or elevated temperature (40 degrees C) in air, or under an atmosphere of either of the major ruminal gases, carbon dioxide and methane. In vivo, studies showed that viable chlamydospores could be detected at the erosion surface of prototype CRDs recovered from the rumen and also in faeces of fistulated sheep, for up to 3 weeks after administration. Further studies have shown that chlamydospores released from such devices can substantially reduce the number of infective larvae that develop in cultures of faeces collected from sheep infected with the nematode parasite, Haemonchus contortus. This work demonstrates, in principle, that the deployment of chlamydospores of D. flagrans in intraruminal CRDs, is another possibility in the development of a range of methods for the biological control of parasites in livestock.

Animals↗
PubMed2002

Selection for resistance to macrocyclic lactones by Haemonchus contortus in sheep.

An anthelmintic-sensitive Haemonchus contortus strain was selected for moxidectin and ivermectin resistance concurrently for 22 generations. Treatment with 0.002 mg moxidectin/kg BW or 0.02 mg ivermectin/kg BW produced >99% efficacy against the susceptible parent strain passaged for 22 generations without any anthelmintic exposure. However, to obtain similar efficacy the moxidectin-selected and the ivermectin-selected strains of H. contortus required 0.05 mg moxidectin/kg BW or 0.4 mg ivermectin/kg BW. These results indicate that development of resistance to one macrocyclic lactone, simultaneously results in resistance to another macrocyclic lactone. However, rates of resistance development differ between compounds and occurs more slowly with moxidectin than with ivermectin.

Animals↗
PubMed2002

Incidence of Haemonchus spp. and effect on haematocrit and eye colour in goats farmed under resource-poor conditions in South Africa.

The diversity and predominance of nematode genera in goats of resource-poor farmers at Rust de Winter, Gauteng Province, Impendle, KwaZulu-Natal Province, and Kraaipan, North-West Province, South Africa, was determined by means of a longitudinal study of the nematode faecal egg counts (FECs) and differential third-stage nematode larvae. The animals were bled for haematocrit determination and scored for pallor of ocular mucous membranes using the FAMACHA( Copyright) method, an assay for clinical evaluation of anaemia caused by Haemonchus spp. Animals considered to be in danger of dying from anaemia caused by haemonchosis were selectively treated with an anthelmintic. Lower haematocrit values were registered during periods of heavier Haemonchus infection, which occurred from December/January to March for Rust de Winter; from December to March/April for Impendle; and from November/December to February or April for Kraaipan. There was agreement too between the lower haematocrits and paler mucous membranes scored according to the FAMACHA( Copyright) method. The use of this system may be recommended as part of an integrated approach to worm control in goats kept in the resource-poor areas studied.

Anemia↗
PubMed2002

Evaluation of Duddingtonia flagrans in reducing infective larvae of Haemonchus contortus in feces of sheep.

Consequences of nematode infections due to Haemonchus contortus are a serious constraint for the sheep industry worldwide. Development of anthelmintic resistance and increasing concern about the impact of anthelmintic use dictate the need of alternative control. Such an alternative is using the nematode trapping fungus Duddingtonia flagrans to reduce infective larvae levels on pasture. Two trials were conducted to determine the effect of D. flagrans in reducing infective larvae (predominantly H. contortus) in feces. The first trial determined the dose effect of D. flagrans in reducing infective larvae in feces. Eighteen ewes were dewormed to remove existing infections and randomly assigned to six treatment groups: 5 x 10(4), 1 x 10(5), 2.5 x 10(5), 5 x 10(5), 1 x 10(6) or no (control) spores of D. flagrans per kg of body weight mixed in their feed for 7 days. Fecal samples were collected daily from these and from infected donor ewes. Feces from individual-treated ewes were mixed with equal amounts of donor ewe feces, theoretically approximating oral dose spore concentrations of 2.5 x 10(4), 5 x 10(4), 1.25 x 10(5), 2.5 x 10(5), 5 x 10(5) and no spores, and were cultured. Across dosages and during the 7 days of fungus feeding, percent reduction of infective larvae ranged from 76.6 to 100.0%. The second trial determined the effect of D. flagrans at the dose of 10(5) spores per kg body weight on reducing infective larvae in feces from naturally infected lambs. Twenty lambs were randomly assigned to either treatment or control groups based on fecal egg count. Treatment lambs were fed spores mixed in feed for 7 days. Feces were collected daily and cultured. During the 7 days of fungus feeding, the percent reduction of infective larvae ranged from 82.8 to 99.7%. Results of these trials demonstrated that the nematode trapping fungus D. flagrans was highly effective in reducing infective larvae in sheep feces and should be considered as a biological control agent for integrated nematode control programs.

Administration, Oral↗