A comparison of two integrated approaches of controlling nematode parasites in small ruminants

Control of gastrointestinal nematodes (GIN) in small ruminants in regions of the world where anthelmintic resistance is prevalent must rely on more than just chemical deworming strategies. The objective of this experiment was to compare two integrated treatment protocols for control of GIN (primarily Haemonchus contortus in this region) using anthelmintics, copper oxide wire particles (COWP) and FAMACHA(?), compared to traditional anthelmintic use only. Three separate trials were conducted on mature ewes, weaned goats, and weaned lambs in which three deworming management strategies were applied: 1) all animals were dewormed with levamisole at four week intervals (LEV), 2) individual animals were dewormed with moxidectin when scored≥4 (ewes) or ≥3 (kids and lambs) using FAMACHA(?) (FAM/MOX), 3) all animals were dewormed with moxidectin initially and again with COWP (2g) when group mean FEC exceeded 500 (ewes), 3000 (kids), or 1000 (lambs) eggs/g (MOX/COWP). In this final group, during periods between group treatments, individual animals were dewormed with albendazole and levamisole according to FAMACHA(?) score. Fecal egg counts (FEC) and blood packed cell volume (PCV) were determined every 7 days and body weight every 28 days for 30, 20, or 16 weeks in ewes, goats, and lambs, respectively. Efficacy of levamisole was 83.4-86.4%, efficacy of moxidectin was 93.5-100%, and efficacy of COWP was 10.8-98.1% among the three trials. The mean number of deworming treatments per animal that occurred for the LEV, FAM/MOX, and MOX/COWP groups, respectively, was 7.9, 0.6, and 2.5±0.4 (P<0.001) for ewes, 5.2, 1.6, and 3.4±0.3 (P<0.001) for goats, and 4.0, 1.7, and 3.6±0.2 (P<0.001) for lambs. Production (body weight of lambs weaned from ewes or final body weight of kids and lambs) was similar among management strategy groups for ewes and kids, but FAM/MOX lambs were lighter by the end of the trial (P<0.003). While more time and labor was required to use the FAMACHA(?) system, a more conservative use of anthelmintics occurred in the FAM/MOX group.     

Anthelmintic resistance of gastrointestinal parasites in small ruminants

Consensus Statements of the American College of Veterinary Internal Medicine (ACVIM) provide veterinarians with guidelines regarding the pathophysiology, diagnosis, or treatment of animal diseases. The foundation of the Consensus Statement is evidence-based medicine, but if such evidence is conflicting or lacking, the panel provides interpretive recommendations on the basis of their collective expertise. The Consensus Statement is intended to be a guide for veterinarians, but it is not a statement of standard of care or a substitute for clinical judgment. Topics of statements and panel members to draft the statements are selected by the Board of Regents with input from the general membership. A draft prepared and input from Diplomates is solicited at the Forum and via the ACVIM Web site and incorporated in a final version. This Consensus Statement was approved by the Board of Regents of the ACVIM before publication.     

Anthelmintic resistance of nematode parasites of small ruminants in eastern Ethiopia: exploitation of refugia to restore anthelmintic efficacy

Faecal egg count reduction tests (FECRT) were conducted in May 2003 to determine the efficacy of anthelmintics used for treatment against nematode parasites in separately managed sheep and goat flocks at Alemaya University in eastern Ethiopia. These tests revealed high levels of anthelmintic resistance to albendazole, tetramisole, the combination of these two drugs, and to ivermectin in the goat flock (predominantly infected by Haemonchus contortus and Trichostrongylus spp.), whereas all drugs were highly efficacious in the sheep flock. A second FECRT confirmed these observations. Following this, a new management system was implemented on the goat flock for a period of 9 months (January-September 2004) in an attempt to restore the anthelmintic efficacy. This involved a combination of measures: eliminating the existing parasite infections in the goats, exclusion from the traditional goat pastures, and introducing communal grazing of the goats with the university sheep flock and livestock owned by neighbouring small-holder farmers. A second series of FECRTs (Tests 3 and 4) conducted 7 months after this change in management, showed high levels of efficacy to all three drugs (albendazole, tetramisole and ivermectin) in the goat flock. This is the first field study to demonstrate that anthelmintic efficacy in the control of nematode parasites of small ruminants can be restored by exploiting refugia.     

Prospects for biological control of nematode parasites of ruminants

Control of nematode parasites in ruminants is directed almost exclusively at the parasitic stage in the host using conventional anthelmintic therapy. However, to complete their life cycle, parasites have to develop through a series of free-living stages on pasture. It is within this environment that there is a vast range of cohabiting micro-organisms, some of which are known to infect or prey on nematodes. These include bacteria, protozoa, viruses and fungi. To date, most interest has been focused on those organisms producing chemical toxins which have been developed as anthelmintics, e.g. Streptomyces/avermectins. Yet the potential of predacious and nematoxic fungi and bacteria for the biological control of nematode parasites of livestock remains largely untapped. These organisms may well offer a cheaper and more sustainable approach to reducing the impact of parasitism in livestock.     

Biological control of nematode parasites of small ruminants in Malaysia using the nematophagous fungus Duddingtonia flagrans

Control of nematode parasites of small ruminants in a wet, tropical environment using the nematophagous fungus, Duddingtonia flagrans, was assessed in this study. Two methods of fungal delivery were tested, namely as a daily feed supplement, or incorporated into feed blocks. Initially, pen trials were conducted with individually penned groups of sheep and goats at dose rates of 125,000 spores and 250,000 spores/kg live weight per day. At the lower dose rate this reduction was between 80 and 90% compared with the pre-treatment levels. At the higher dose rate, there was virtually complete suppression (>99% reduction) of larval recovery. Trials using the fungal feed blocks, showed that when animals were individually penned, they consumed only small amounts of the block (particularly goats), hence little effect on larval recovery in faecal cultures was observed. Grouping animals according to species and dose rate induced satisfactory block consumption and subsequent high levels of larval reduction in faecal cultures. These larval reductions were mirrored by the presence of fungus in faecal cultures. This work was followed by a small paddock trial, whereby three groups of sheep were fed either a feed supplement without fungal spores, supplement with spores, or offered fungal blocks. The dose rate of spores in the latter two groups was 500,000 spores/kg live weight per day. Egg counts were significantly reduced in the two fungal groups, compared with the control group and the latter required two salvage anthelmintic treatments to prevent mortality due to haemonchosis. Pasture larval numbers on the two fungal group plots were also much lower than on the control plot.     

Drug resistance makes new control measures of stomach parasites in small ruminants necessary

Control of parasitic gastroenteritis in small ruminants is threatened by the worldwide growing problem of anthelmintic resistance. Therefore, alternative approaches for worm control are imperative. Of utmost importance is to slow down selection pressure for anthelmintic resistance by using alternative control measures combined with minimal anthelmintic usage. New control options for the Netherlands are discussed.     

Molecular mechanisms for anthelmintic resistance in strongyle nematode parasites of veterinary importance

Veterinarians rely on a relatively limited spectrum of anthelmintic agents to control nematode parasites in domestic animals. Unfortunately, anthelmintic resistance has been an emerging problem in veterinary medicine. In particular, resistance has emerged among the strongyles, a group of gastrointestinal nematodes that infect a variety of hosts that range from large herbivores to small companion animals. Over the last several decades, a great deal of research effort has been directed toward developing an understanding of the mechanisms conferring resistance against the three major groups of anthelmintics: macrocyclic lactones, benzimidazoles, and nicotinic agonists. Our understanding of anthelmintic resistance has been largely formed by determining the mechanism of action for each drug class and then evaluating drug‐resistant nematode isolates for mutations or differences in expression of target genes. More recently, drug efflux pumps have been recognized for their potential contribution to anthelmintic resistance. In this mini‐review, we summarize the evidence for mechanisms of resistance in strongyle nematodes.     

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.     

Genetic variation among calves in resistance to nematode parasites

Dutch Friesian male calves, which received a dose of 100 000 Cooperia spp. larvae at an age of 3 months, displayed a great variability in reaction, measured by number and length of worms, egg output, serum antibodies and liveweight gain. In an experiment with half-sib groups from randomly chosen sires, only the antibody response was significantly different between sire-groups. In a further experiment with half-sibs from two selected sires, egg output, worm numbers, worm length and antibody response differed. A third trial with half-sib groups of the same two sires was carried out under natural conditions of grazing. It is concluded that, within this breed, genetical variation exists in resistance to Cooperia spp.     

Anthelmintic resistance in nematode parasites of goats

Cases of anthelmintic resistance on 3 goat farms in Gippsland were investigated. On the first farm Haemonchus contortus, Trichostrongylus colubriformis and Ostertagia (Teladorsagia) circumcincta were found to be resistant to fenbendazole, but the first 2 species were fully susceptible to levamisole. On the second farm a population of T. colubriformis, resistant to concurrent full doses of levamisole and a benzimidazole, was found to retain this resistance when transferred to sheep. On the third farm, heavy mortality due to Ostertagia and Trichostrongylus spp burdens was arrested only by the use of concurrent full doses of levamisole and oxfendazole; no single anthelmintic was found to be effective. A disturbing rise in the incidence of clinical helminthiosis, often accompanied by anthelmintic resistance, occurred on goat farms in Gippsland early in 1984.     

Ingestion of host immunoglobulin by three non-blood-feeding nematode parasites of ruminants

Host immunoglobulin was detected in homogenates of adult worms or fourth stage larvae of Haemonchus contortus, Ostertagia ostertagi, O circumcincta and Dictyocaulus viviparus, but not in preparasitic third stage larvae of H contortus. All the worms had been washed in detergent before being homogenised to remove any immunoglobulin adhering to the cuticle. This procedure, the concentrations of immunoglobulin detected and observations on sections of the worms stained with fluorescent antisheep immunoglobulin, all suggested that the immunoglobulin had been ingested, even by the non-blood-feeding species. This finding suggests that the Ostertagia species and D viviparus may be susceptible to vaccination by the gut antigen approach, a method with considerable promise for blood-feeding parasites like H contortus and Boophilus microplus.     

An egg-hatch assay for resistance to levamisole in trichostrongyloid nematode parasites

An in vitro technique is described for detecting resistance of nematodes to the anthelmintic levamisole hydrochloride. Samples of eggs are developed under controlled temperature conditions until just prior to the commencement of hatching. They are then exposed to different concentrations of the drug and, when hatching is almost complete, the test samples are killed and preserved. The proportion of unhatched eggs at each drug concentration can then be counted at leisure. Provided a suitable range of drug concentrations is chosen for each test isolate, this assay provides results which may be satisfactorily fitted to a log-concentration-probit regression model. Comparisons with in vivo anthelmintic assays have shown that the technique provides an accurate reflection of the resistance status of parasite populations.     

The contribution of molecular biology to the development of vaccines against nematode and trematode parasites of domestic ruminants.

Rapid developments in molecular biology have had an enormous impact on the prospects for the development of vaccines to control the major nematode and trematode infestations of livestock. Vaccine candidates are purified using conventional protein chemistry techniques but the limitations imposed by the scarcity of parasite material provide an insurmountable barrier for commercial vaccine production by this means. The ability to purify mRNA from different parasite life-cycle stages and to prepare cDNA expression libraries from it has proven central to the identification of immunogenic parasite proteins. Potentially, protective parasite antigens can now be produced in recombinant form in a variety of vectors and this represents a key breakthrough on the road to commercial vaccine production. The contribution of molecular biology to this process is discussed using several examples, particularly in vaccine development against the pathogenic abomasal nematode of sheep and goats, Haemonchus contortus, and the liver fluke of sheep and cattle, Fasciola hepatica. The difficulties of producing recombinant proteins in the correct form, with appropriate post-translational modification and conformation, are discussed as well as emerging means of antigen delivery including DNA vaccination. The opportunities offered by genome and expressed sequence tag analyses programmes for antigen targeting are discussed in association with developing microarray and proteomics technologies which offer the prospect of large scale, rapid antigen screening and identification.     

Genetic resistance to gastrointestinal nematode parasites in Rh?n sheep following natural infection.

Genetic parameters of resistance to gastrointestinal nematodes were estimated in Rh?n sheep, a local sheep breed in Germany. In the first year, a total of 131 lambs (born between January and February 1998) from five rams were evaluated. Faecal egg counts (FEC) (n=1528) were measured in all lambs every 2 weeks until 4 months of age and every 4 weeks until 12 months of age in female lambs. In the second year, a total of 129 lambs (born between January and February 1999) from the same rams were evaluated. FEC (n=645) were measured in the lambs every 4 weeks until 5 months of age. Blood was taken in both years at the time of faecal sampling from the jugular vein and haematocrit was measured. Heritabilities for log(10)(FEC+25) were between 0.11 and 0.44. Heritabilities for the corresponding haematocrits were between 0.19 and 0.26 depending on the number of samples taken in account. A significant negative correlation was estimated for the parameters of FEC and daily weight gain (r=-0.57). The results confirm the feasibility of genetic selection in Rh?n sheep for resistance to nematode parasites in an environment where Trichostrongylus spp./Teladorsagia spp., Haemonchus contortus and Cooperia spp. are the dominant species. Breeding decisions can be based on one or two samples depending on the mean values and ranges of FEC and haematocrit, respectively.     

Molecular approaches for inferring evolutionary relationships among protistan parasites.

Within the diverse group of parasites broadly recognized as protists, there are limited morphological characters that can be used to distinguish species and even fewer characters that can infer evolutionary relationships among species. For this reason, molecular data are commonly used to infer relationships among species and strains. These studies most commonly rely on sequences associated with the ribosomal RNA genes but increasingly other nuclear, mitochondrial or plastid genes are contributing data. Molecular systematics has been invaluable in expanding the range of characters that are available for inferring relationships among protistan taxa. As an adjunct to morphological characters, sequence data allow us to better understand the evolutionary history of this group of parasites and thereby erect a well-supported taxonomic framework that reflects these historical relationships. Importantly, the predictive nature of such a framework can aid the search for therapeutic compounds (e.g. via shared biochemical pathways) and highlight organisms that should be tested for cross-reactivity in immunological or molecular diagnostic methods (e.g. use of the closest relatives to assess test specificity). For emergent and poorly known parasites, molecular characterization and placement within the broader phylogenetic framework can help predict likely life history traits, including possible or likely definitive hosts.     

Nematophagous fungi as a biological control agent for nematode parasites of small ruminants in Malaysia: a special emphasis on Duddingtonia flagrans

Approximately 2,800 fresh dung samples from animals, mainly ruminant livestock, were screened for the presence of nematophagous fungi in Malaysia. Arthrobotrys spp. was noted on numerous occasions, but only one isolate of Duddingtonia flagrans was made. For the purposes of producing sufficient quantities of this fungus for feeding trials in sheep, various, commonly available, cheap plant materials were tested as possible growth substrates. This showed that cereal grains (wheat, millet and rice) were the best media for fungal growth. Pen feeding trials were carried out using sheep, both naturally and experimentally infected with nematode parasites (predominantely Haemonchus contortus), to test the efficiency of D. flagrans when administered either in a grain supplement, or incorporated into a feed block. These showed that the fungus survived gut passage in sheep and that dose rates of approximately 1 x 10(6) D. flagrans spores / animal / day, reduced the percentage of infective larvae developing in faecal cultures by more than 90%. These results indicate that using D. flagrans as a biological control agent of nematode parasites, is a promising alternative to nematode parasite control of small ruminants in Malaysia, where anthelmintic resistance is now a major problem.     

Morphological identification of nematode larvae of small ruminants and cattle simplified

A simplified system has been developed to facilitate identification of nematode larvae of the common nematodes of cattle, sheep and goats. Firstly, in addition to the characteristics conventionally used for identification (such as the shape of the cranial extremity and numbers of intestinal cells), the lengths of the infective sheath tails of infective larvae of each genus/species are related to that of Trichostrongylus spp. instead of using measurements for differentiation. For instance, if the mean length of the sheath tail (the distance the sheath extends caudad beyond the caudal tip of the larva) of Trichostrongylus spp. is assumed to be "X", then that of Haemonchus contortus is 2-2.7"X", and that of Oesophagostomum spp. from sheep is 4-7"X", etc. Secondly, by estimating the proportion of the sheath tail of a larva comprised of a terminal thin whip-like filament, identification is aided, particularly in those L3 of species that resemble one another closely, such as Chabertia ovina and Oesophagostomum venulosum or Oesophagostomum columbianum. After some practice with the system it is usually necessary to measure only one or two sheath tails of L3 in a mixed population, whereupon the identity of most of the remaining L3 can be estimated in relation to those measured, without a need for further measurements. The keys were found to facilitate differential larval identification and are particularly useful for training.     

Anthelmintic resistance in nematode parasites of sheep in Denmark with special emphasis on levamisole resistance in Ostertagia circumcincta.

This study was undertaken to elucidate the presence of anthelmintic resistance in nematode parasites of sheep in Denmark. Twenty two flocks of sheep were selected for Faecal Egg Count Reduction (FECR) tests, based on a prior history of either the same anthelmintic, or anthelmintic class having been used 3 times or more over the previous 5 years. Evidence of anthelmintic resistance was detected in 7 flocks. Two flocks showed FECR of 88% and 94% after treatment with thiabendazole, FECR of another 2 were 90% and 94% following treatment with fenbendazole. Three flocks showed FECR of 73%, 89% and 94%, respectively following the use of levamisole. Ostertagia circumcincta was isolated from 1 of the latter flocks and subjected to an in vivo controlled slaughter assay. Following treatment with levamisole at the recommended dose rate of 7.5 mg/kg, FECR was 44.5% and worm counts were reduced by 67.7%. These results were further substantiated by an in vitro egg hatch paralysis assay and by measuring pepsinogen levels in treated and non-treated lambs. This is the first instance of anthelmintic resistance in sheep nematodes in Scandinavia.     

Mycoplasma infections in small ruminants.

Mycoplasmas have complex mechanisms of antigenic variation that allow them to evade the immune system. These organisms cause a variety of clinical syndromes that can have a significant economic effect on small ruminant production. The syndromes range from acute septicemia and death to chronic infection resulting in decreased production. Recent research findings have shed light on the means by which these organisms evade the host immune response and cause or contribute to the development of disease in the host. This article provides a review of the pathogenesis, clinical signs, and treatment options for common disease syndromes involving Mycoplasma spp. in small ruminants.