Resistance In Gastrointestinal Nematodes Research Articles

Resistance of gastrointestinal nematodes to the most commonly used anthelmintics in sheep, cattle and horses in Spain

The objective of this study was to evaluate the status of anthelmintic resistance (AR) in ruminants and horses in Spain. The efficacy of commonly used macrocyclic lactones (MLs) – ivermectin (IVM) and moxidectin (MOX) – was measured in sheep, cattle and horses. In addition, albendazole (ABZ) and levamisole (LEV) were evaluated in sheep and oxibendazole (OXI) and pyrantel (PYR) in horses. Efficacy was evaluated based on the difference between the arithmetic mean pre- and post-treatment faecal egg count (in cattle and horses), or compared to an untreated control group (in sheep). AR was present when the percentage reduction in egg count was <95% and the lower 95% confidence interval (CI) was <90%; if only one of these two criteria was met, the finding was recorded as suspected AR (SAR). In horses, AR–PYR and OXI was considered when the percentage reduction in egg count was ≤90% and the lower 95% CI was ≤80%. For each animal species, at least 10 study sites were selected.AR to at least one of the drugs was detected in all 10 sheep flocks; the main parasite identified after treatment was Teladorsagia circumcincta. Moreover, in 5 flocks multidrug resistance was identified, on 4 farms to drugs from different families, on one farm to both MOX and IVM and on another farm to all drugs tested. In cattle, the efficacy of both MOX and IVM was 100% on 4 and 3 farms, respectively, and therefore 60% of these farms were considered to have AR or SAR to both MLs. The most frequent parasite identified after treatment was Trichostrongylus spp., although Ostertagia ostertagi was also identified after treatment on one farm. In contrast to ruminants, the 4 drugs evaluated in horses were highly efficacious against strongyles, with efficacies for the MLs and OXI between 95 and 100% and between 94 and 100% for PYR, although 3 herds were SAR against PYR.In conclusion, AR to at least one of the commonly used drugs was identified on all sheep flocks investigated in the northwest of Spain. The occurrence of AR to MLs in cattle was higher than expected but consistent with what was observed in sheep. In horses, all currently used drugs were confirmed as effective against strongyles.

Anthelmintic resistance in gastrointestinal nematodes from grazing beef cattle in Campeche State, Mexico.

Production of beef cattle is one of the most important economic activities in Mexico. However, anthelmintic resistance (AR) has affected animal productivity. The aim of this study was to determine the presence of AR in gastrointestinal nematodes (GIN) of beef cattle in Candelaria Municipality of Campeche State, Mexico. Sixty-five-month-old beef calves were selected for the faecal egg count reduction test (FECRT) and the inhibition of egg hatch (IEH) assay. These parameters were determined using albendazole (benzimidazole, BZ), ivermectin (IVM, Macrocyclic lactone, ML) and levamisole (LEV, imidazothiazole, IMZ). Allele-specific PCR (AS-PCR) confirmed polymorphisms at codon 200 of isotype 1 of the β-tubulin gene of Haemonchus placei. The results showed 32% IVM toxicity by FECRT, indicating problems of AR in the GIN population. In contrast, BZ and LEV showed 95 and 100% toxicity, respectively, against GIN from infected beef calves. The infective larvae (L3) of Cooperia, Haemonchus and Oesophagostomum were identified before anthelmintic treatment, and Cooperia L3 larvae were identified after treatment with IVM. The IEH assays had lethal dose 50 (LD50) of 187nM to BZ, confirming the ovicidal effect of BZ. In contrast, the LD50 for LEV and IVM were 3.3 and 0.4mM, respectively. The results obtained by AS-PCR confirmed two DNA fragments of 250 and 550bp, corresponding to the resistant and susceptible alleles in the H. placei population. The nematode Cooperia showed AR against IVM, while the toxicity effect of BZ against GIN with both FECRT and IEH was confirmed.

Genome-wide scan of gastrointestinal nematode resistance in closed Angus population selected for minimized influence of MHC.

Genetic markers associated with parasite indicator traits are ideal targets for study of marker assisted selection aimed at controlling infections that reduce herd use of anthelminthics. For this study, we collected gastrointestinal (GI) nematode fecal egg count (FEC) data from post-weaning animals of an Angus resource population challenged to a 26 week natural exposure on pasture. In all, data from 487 animals was collected over a 16 year period between 1992 and 2007, most of which were selected for a specific DRB1 allele to reduce the influence of potential allelic variant effects of the MHC locus. A genome-wide association study (GWAS) based on BovineSNP50 genotypes revealed six genomic regions located on bovine Chromosomes 3, 5, 8, 15 and 27; which were significantly associated (-log10 p=4.3) with Box-Cox transformed mean FEC (BC-MFEC). DAVID analysis of the genes within the significant genomic regions suggested a correlation between our results and annotation for genes involved in inflammatory response to infection. Furthermore, ROH and selection signature analyses provided strong evidence that the genomic regions associated BC-MFEC have not been affected by local autozygosity or recent experimental selection. These findings provide useful information for parasite resistance prediction for young grazing cattle and suggest new candidate gene targets for development of disease-modifying therapies or future studies of host response to GI parasite infection.

Emergence of multiple resistances against gastrointestinal nematodes of Mehsana-cross goats in a semi-organized farm of semi-arid region of India

Faecal samples from 140 adult goats were collected and positive samples were examined quantitatively; goats having at least 500 eggs/g of faeces were selected and randomly divided into four groups. Faecal egg count reduction test was used to detect the efficacy of albendazole (ALB), levamisole (LEV) and ivermectin (IVM) in goats naturally infected with gastrointestinal nematodes in a semi-organized farm located in semi-arid region of Gujarat, India. ALB, LEV and IVM reduced faecal egg counts on 14th day post-treatment varied from 53% to 76%, revealing presence of resistance of worms to all these drugs. Finding IVM resistance in gastrointestinal nematodes is found to be alarming. Pre-treatment faecal culture of all the groups collectively revealed Haemonchus spp. as the predominant parasite again post-treatment faecal cultures further emphasize the pre-dominance of H. contortus in ALB- and LEV-treated groups. However, faecal cultures from IVM-treated group revealed predominance of Trichostrongylus species which were scarce in pre-treatment group. Study further reveals that only ALB was found to be effective against Oesophagostomum spp., and most of the gastrointestinal nematodes were found to have some degree of resistance against rest of the other drugs used in goats.

Widespread anthelmintic resistance in European farmed ruminants: a systematic review

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La résistance aux antiparasitaires: risques, prévention

Resistance against pesticides : threats, prevention. Chemoresistance of parasites has been reported worldwide in multiple groups of species (fungi, protozoa, helminths and arthropods) in regard to several classes of drugs. Anthelmintics resistance of gastrointestinal nematodes of small ruminants is a major constraint on production and induces severe economic losses ; idem with cyathostominæ of horses and very limited choice of anthelminthic drugs, with coccidia in poultry breeding and ticks in cattle. Because of the high number of immunocompromised patients (HIV+, low-birth-weight neonates, patients with organ transplantation or chemotherapy), this chemoresistance has a very disquieting dimension for the public health (zoonotic arthropod-borne diseases). The veterinary practitionner has the responsability of the choice of the optimal drug and the prescription in the interest of the best animal productions and the public health

Haemonchotolerance in West African Dwarf goats: contribution to sustainable, anthelmintics-free helminth control in traditionally managed Nigerian dwarf goats.

West African Dwarf (WAD) goats are extremely important in the rural village economy of West Africa, but still little is known about their biology, ecology and capacity to cope with gastrointestinal nematode (GIN) infections. Here, we summarise the history of this breed and explain its economic importance in rural West Africa. We review recent work showing that Nigerian WAD goats are highly trypanotolerant and resist infections with Haemonchus contortus more effectively than other breeds of domestic goat (haemonchotolerance). We believe that haemonchotolerance is largely responsible for the generally low level GIN infections and absence of clinical haemonchosis in WADs under field conditions, and has contributed to the relatively successful and sustainable, anthelmintics-free, small-scale system of goat husbandry in Nigeria’s humid zone, and is immunologically based and genetically controlled. If haemonchotolerance can be shown to be genetically controlled, it should be possible to exploit the underlying genes to improve GIN resistance among productive fibre and milk producing breeds of goats, most of which are highly susceptible to nematode infections. Genetic resistance to GIN and trypanosome infections would obviate the need for expensive chemotherapy, mostly unaffordable to small-holder farmers in Africa, and a significant cost of goat husbandry in more developed countries. Either introgression of resistance alleles into susceptible breeds by conventional breeding, or transgenesis could be used to develop novel parasite-resistant, but highly productive breeds, or to improve the resistance of existing breeds, benefitting the local West African rural economy as well as global caprine livestock agriculture.

Prevalence and severity of anthelmintic resistance in ovine gastrointestinal nematodes in Australia (2009-2012).

This study aimed to provide an indication of the prevalence and severity of anthelmintic resistance (AR) in the Australian sheep industry by compiling the results of faecal worm egg count reduction tests (FECRTs). Government and private parasitology laboratories, pharmaceutical companies and veterinarians known to have conducted FECRTs were asked to provide results that conformed to Australian and New Zealand standard diagnostic procedures. Data were available from a total of 390 tests, with larval differentiation conducted in 222 cases. Pooled results from all states for the macrocyclic lactone (ML) class showed a lower prevalence of AR against combined species for moxidectin (54%) compared with abamectin (77%) and ivermectin (87%). Analysis by state revealed higher levels of ML-resistant Teladorsagia sp. in Tasmania and Western Australia than in other states and ML-resistant Haemonchus sp. was more frequently detected in New South Wales. This compilation of results of FECRTs conducted by Australian parasitology laboratories in 2009-12 showed widespread AR of the common sheep nematodes (Teladorsagia, Trichostrongylus and Haemonchus) to all broad-spectrum anthelmintics, with the exception of monepantel, whether used singly or in combination.

Anthelmintic resistance in ovine gastrointestinal nematodes in inland southern Queensland.

To establish the prevalence of anthelmintic resistance in ovine gastrointestinal nematodes in southern Queensland. An observational parasitological study using the faecal egg count reduction test. Sheep farms (n = 20) enrolled in this study met the twin criteria of using worm testing for drench decisions and having concerns about anthelmintic efficacy. On each farm, 105 sheep were randomly allocated to one of six treatment groups or an untreated control group. Faecal samples were collected on day 0 and days 10-14 for worm egg counts and larval differentiation. Single- and multi-combination anthelmintics, persistent and non-persistent, oral liquid or capsule, pour-on and injectable formulations were tested. Monepantel was not tested. Farmers also responded to a questionnaire on drenching practices. Haemonchus contortus was the predominant species. Efficacy <95% was recorded on 85% of farms for one or more anthelmintics and on 10% of farms for six anthelmintics. No resistance was identified on three farms. The 4-way combination product was efficacious (n = 4 farms). Napthalophos resistance was detected on one farm only. Resistance to levamisole (42% of farms), moxidectin injection (50% of farms) and the closantel/abamectin combination (67% of farms) was identified. Moxidectin oral was efficacious against Trichostrongylus colubriformis, which was predominant on only one farm. Of the farms tested, 55% ran meat breeds, 60% dosed more than the recommended dose rate and 70% always, mostly or when possible practised a 'drench and move' strategy. This level of anthelmintic resistance in southern Queensland will severely compromise worm control and force increased use of monepantel.

Anthelmintic resistance in sheep gastrointestinal nematodes in Slovakia detected by in-vitro methods.

BackgroundThe intensive use of anthelmintics for the control of helminthic infections has resulted in the development of anthelmintic resistance, which has become a major practical problem in many countries. A variety of tests are available to monitor anthelmintic resistance but most of them are expensive, laborious and time consuming and therefore unpractical for large field surveys. The main aim of this survey was thus to detect the occurrence of benzimidazole (BZ) and macrocyclic lactone resistance on sheep farms in Slovakia by using novel and modified in vitro methods that are inexpensive, easy to use and quick and therefore practical for large surveys.ResultsBZ-resistant gastrointestinal nematodes were found on all 27 farms. Two farms (7.4%) had high levels of resistance (>40% of hatching), and 22 farms had low levels (<20% of hatching) of resistant nematodes. IVM-resistant populations were found on 14 of 49 sheep farms. The prevalence of BZ and IVM resistance has slightly increased on Slovak sheep farms during the last two decades.ConclusionsBoth the BZ and IVM surveys indicated that resistance against anthelmintics was present on Slovak sheep farms. Resistance against the BZ class of anthelmintics had been stable for two decades, but a slight increase on IVM resistance was confirmed. Farmers must thus observe the preventive measures to avoid a faster onset of IVM resistance, otherwise the presence of resistant parasites and ineffective treatment may harm the economy of their farms.

Anthelmintic resistance in gastrointestinal nematodes in goats and evaluation of FAMACHA diagnostic marker in Uganda

Gastrointestinal nematodes (GIN) are a challenge to goat production globally causing reduced growth, morbidity and mortality. We report here results of the first nation-wide anthelmintic resistance (AR) study and validation of assessment of clinical anaemia with FAMACHA eye scores in goats in Uganda. From August to December 2012 the efficacy of albendazole (7.5mg/kg), levamisole (10.5mg/kg) and ivermectin (0.3mg/kg) against strongyle nematodes was tested on 33 goat farms in Soroti, Gulu, Mpigi, Mbarara and Ssembabule districts of Uganda. Altogether 497 goats were subjected to a total of 45 different faecal egg count reduction tests (FECRT), each involving 5–20 goats. On one farm all substances were tested. Faecal and blood samples were collected and FAMACHA eye scores evaluated on the day of treatment and 15 days later. A questionnaire survey was conducted on frequency, type and dose of anthelmintics used, farm size and grazing management system. Examination of infective third stage larvae (L3) from pooled faecal cultures demonstrated Haemonchus to be the predominant genus (>75%). Resistance to at least one anthelmintic group was detected on 61% of the 33 farms and in 49% of the 45 test groups. Prevalence of resistance to ivermectin, levamisole and albendazole was respectively 58%, 52% and 38%. Correlation between pre-treatment packed cell volume determinations and FAMACHA scores (r498=−0.89) was significant. Paddock grazing system (Odds ratio 4.9, 95% CI 1.4–17.3) and large farm size of >40 goats (odds ratio 4.4, 95% CI 1.2–16.1) were significant predictors of AR. In all districts, resistance to all three anthelmintics was higher on large-scale goat farms practising mostly paddock grazing. Interestingly, resistance to albendazole, the most commonly used anthelmintic in Uganda, was lower than that to ivermectin and levamisole. We recommend adaptation of FAMACHA to goats to help restrict anthelmintic treatment to heavily infected individuals. This will limit selection pressure and hence delay development of anthelmintic resistance.

Gastrointestinal nematodes of dairy goats, anthelmintic resistance and practices of parasite control in Northern Italy.

BackgroundGastrointestinal nematodes (GINs) are one of the main constraints to ruminant production worldwide. Anthelmintic resistance (AR) has been reported in goats throughout Europe, yet little is known about the AR status in Italy. The aims of the study were: i) determine the frequency of AR in GINs in goat flocks in Northern Italy, Italy, ii) survey goat farmers on the current practices of parasite control, iii) update the species composition of the gastrointestinal helminthofauna. Thirty three flocks were enrolled and 1288 individual fecal samples were collected. Based on the egg per gram (EPG), 15 flocks were selected to evaluate the presence of AR in GINs with the Fecal Egg Count Reduction Test (FECRT). A questionnaire surveyed 110 dairy goat farmers to acquire information about farm management and drenching practices against GINs. Further, the gastrointestinal tracts of 42 goats were analyzed.ResultsThe FECRs indicated that five of the 15 flocks had problems of AR, which was identified in all two of the anthelmintic classes tested. Resistance and suspected resistance was found in 40% of the flocks selected for AR testing that were treated with benzimidazoles while 20% of the flocks treated with eprinomectin had resistant GINs. Teladorsagia/Trichostrongylus L3 were isolated from the post-treatment coprocultures of all flocks with resistance but not from the flock with suspected oxfendazole resistance. Treatments against helminths were performed once annually in 73.63% of the flocks, but 20.00% of farmers declared not regularly treating their goats every year. Annual treatments usually occurred in autumn or winter at dose rate for sheep. Te. circumcincta, H. contortus, Tr. colubriformis, Skrjabinema caprae and Oesophagostomum venulosum were the most abundant and prevalent species of the gastrointestinal tract.ConclusionsStrategies to prevent the development of AR should be widely adopted in Northern Italy. Further, farmers and practitioners should be educated about the importance of using the correct dose rates in goats. In addition, the presence of highly pathogenic GINs coupled with high worm burden in all sector of gastrointestinal tract and the prevalence values further suggest that improved diagnosis and active surveillance of GINs infection is needed.

First report of cattle farms with gastrointestinal nematodes resistant to levamisole in Mexico

The objectives of the present study were: (1) to report the percentage of cattle farms with gastrointestinal nematodes (GINs) resistant to levamisole in Veracruz, Mexico, (2) to identify the genera of GINs involved in resistance, and (3) to identify factors associated with these resistances. The faecal egg count reduction test (McMaster technique) was used to detect the presence of resistant GINs. A questionnaire was given to owners to understand the history of anthelmintic use. The percentage of cattle farms with GINs resistant to levamisole was 36.4% (4/11). The percentage of faecal egg count reduction on resistant farms was 91%, 82%, 42% and 88%. A similar number of cattle farms (4/11) were identified as potentially having levamisole resistance. Only three farms had GIN populations susceptible to levamisole. Cooperia spp. was the genus most commonly found to be resistant, followed by Haemonchus spp., Ostertagia spp. and Oesophagostomum spp. No factors were identified that influenced the presence of GIN resistance. However, there were identified inappropriate anthelmintic practices in cattle farms that should be improved. None of the farmers weighed their animals in order to dose them correctly with anthelmintics. Six cattle farms (54.5%) applied anthelmintics to new arriving animals. This is the first report of levamisole resistant GINs in Mexico. Improving the use of anthelmintics and measures of quarantine for infected cattle will help control the spread of resistance.

Anthelmintic resistance in cattle nematodes in the US

The first documented case of macrocyclic lactone resistance in gastrointestinal (GI) nematodes of cattle was seen in the US approximately 10 years ago. Since that time the increase incidence of anthelmintic resistance has continued at an alarming rate. Currently parasites of the genera Cooperia and/or Haemonchus resistant to generic or brand-name macrocyclic lactones have be demonstrated in more than half of all operations examined. Both of these parasite genera are capable of causing economic losses by decreasing food intake and subsequently animal productivity. Currently, there are no easy and quick means to detect anthelmintic resistant GI nematodes. Definitive identification requires killing of cattle. The most commonly used field detection method is the fecal egg count reduction test (FECRT). This method can be adapted for use as a screening agent for Veterinarians and producers to identify less than desired clearance of the parasites after anthelmintic treatment. Further studies can then define the reasons for persistence of the egg counts. The appearance of anthelmintic resistance is largely due to the development of very effective nematode control programs that have significantly improved the productivity of the US cattle industry, but at the same time has placed a high level of selective pressure on the parasite genome. The challenges ahead include the development of programs that control the anthelmintic resistant nematodes but at the same time result in more sustainable parasite control. The goal is to maintain high levels of productivity but to exert less selective pressures on the parasites. One of the most effective means to slow the development of drug resistance is through the simultaneous use of multiple classes of anthelmintics, each of which has a different mode of action. Reduction of the selective pressure on the parasites can be attained through a more targeted approach to drug treatments where the producer's needs are met by selective treatment of different classes of animals and not by blanket treatment at a location. The implementation of such programs will vary by the sector of the industry and the individual site. In general, the feedlot will be the easiest sector for developing of programs, while stocker/backgrounder operations will provide the most challenging problems. A major question that must be addressed is whether it is important that parasite control programs to be sustainable over time, or if the pharmaceutical industry develop new control agents at a rate sufficient to render sustainability unimportant.

Frequency of eprinomectin resistance in gastrointestinal nematodes of goats in canton Berne, Switzerland

Eprinomectin (EPN) is a member of the avermectin class of compounds and the only anthelmintic registered for goats in Switzerland with a zero milk withdrawal period. The aim of the present study was to identify the actual efficacy of EPN in an area with a higher density of goat enterprises. Forty-three randomly chosen farms from canton Berne were investigated. At least eight goats were investigated on every farm. Conditions for inclusion in the study were the absence of anthelmintic treatment during the previous six weeks and a pooled faecal sample showing a mean faecal egg count (FEC) higher than 600epg faeces. Pre- and 14–16 days post-treatment samples were individually collected directly from the rectum. Animals were treated with the recommended dose of EPN (1mg/kg body weight) after taking the pre-treatment samples. Efficacy of EPN was tested with the faecal egg count reduction test (FECRT) and faecal cultures were performed on every farm from pooled faeces samples before and after treatment. Additionally the farmers completed a questionnaire. None of the gastrointestinal nematode populations of the 43 investigated farms were susceptible to EPN at the required level. The mean egg count reduction was 40%. None of the typical risk factors, such as production type, stocking rate, animal traffic and quarantine measures showed an association with the level of eprinomectin resistance. It can be concluded with 80% certainty that the prevalence of EPN resistance on goat farms is at least 95% in canton Berne.

Anthelmintic resistance and multidrug resistance in sheep gastro-intestinal nematodes in France, Greece and Italy

Anthelmintic resistance (AR) in ovine gastro-intestinal nematodes has been reported to affect the health and productivity of sheep globally. The objective of the present study was to evaluate the efficacy of commonly used oral drenches in sheep in France, Greece and Italy. In each country, 10 farms were selected. On each farm, 50 animals were blocked based on the pre-treatment faecal egg count (FEC). Within each block, animals were randomly allocated to one of 5 treatment groups. In addition to an untreated control group, there were 4 groups treated per oral route: moxidectin (MOX) and ivermectin (IVM), both at 0.2mg/kg bodyweight, levamisole (LEV; at 7.5mg/kg bodyweight) and a benzimidazole (BZ; at 3.75–5mg/kg bodyweight). In France, animals were not treated with LEV, but with netobimin (NET; at 7.5mg/kg bodyweight). The FEC was monitored using a modified McMaster technique. Two weeks after treatment, individual faecal samples were taken from all animals and efficacy was calculated as the difference between arithmetic mean FEC of the control group versus each respective treatment group. The results of the present study indicate the high efficacy of treatment with oral formulations of MOX (99–100%) and IVM (98–100%) on all farms, except on 1 farm in Greece. On this farm, multi drug resistance (MDR) was identified involving 4 anthelmintics (efficacy MOX: 91%; IVM: 0%; BZ: 58% and LEV: 87%). In Greece and Italy, AR against LEV and BZ was observed on some farms, with MDR involving both anthelmintics on 3 farms in Greece and on 2 farms in Italy. In France, AR against BZ and NET was observed on all 10 farms included. In all countries, Teladorsagia sp. was the most common nematode larva identified after treatment, followed by Haemonchus sp. and Trichostrongylus sp., with differences among farms and treatments. The current study confirms the high efficacy of oral treatments with MOX and IVM, even on farms with worm populations resistant to BZ, LEV or NET. This study also reports MDR against 4 anthelmintics on one farm in Greece.

The effect of risk factors of sheep flock management practices on the development of anthelmintic resistance in the Czech Republic

The in vivo faecal egg count reduction test was used for the evaluation of the benzimidazole and macrocylic lactone drug efficacies against gastrointestinal nematodes in 10 flocks of sheep. The same samples were tested concurrently in vitro for benzimidazole resistance using the egg hatch test, and for resistance to ivermectins using the microagar larval development test. The conformity of obtained results between in vivo and in vitro tests was recorded. When the selected methods were applied, anthelmintic resistance to benzimidazoles was detected at four (40%) farms while resistance to ivermectins was evident at one (10%) farm. At one farm (10%) ivermectin resistance was suspected. Moxidectin was effective at all surveyed farms. Teladorsagia was recognized as the only benzimidazole resistant genus in post-treatment coprocultures, whereas Haemonchus larvae were resistant to ivermectins. This represents the first recorded occurrence of ivermectin resistance in gastrointestinal nematodes of sheep in the Czech Republic. A linear mixed-effects model demonstrated that the majority of evaluated management practices have a significant effect on resistance to benzimidazoles. While application of preventive practices like quarantine and smart drenching maintains low levels of anthelmintic resistance, others like the dose-and-move strategy, administration of the same family of drugs over extended periods of time, and number of treatments per year are responsible for the increase of resistance to anthelmintics at evaluated farms. Only targeted selective treatments approaches had no effect on resistance status. This study indicates the importance of farm management practices in anthelmintic resistance development.

Evaluation of In Vitro Ovicidal Activity of Ethanolic Extracts of Prosopis juliflora (Sw.) DC (Fabaceae)

Gastrointestinal nematode infections in ruminants have direct effects such as mortality, weaknesses, feeding efficiency and decreased productivity. Animal deaths due to nematode infections are common in tropical regions where control programs based solely on the use of synthetic anthelmintics are no longer sustainable due to increased prevalence of gastrointestinal nematode resistance, slow development of new anthelmintics, high costs to poor farmers and concerns regarding residue in food and the environment. Alternative methods of control are thus required. Prosopis juliflora is a fast-growing, drought-resistant tree adapted to poor and saline soils in arid and semi-arid lands of Kenya, inhabited by nomadic pastoralists. Analysis of the EHA data using t- test revealed that there was no statistically significant difference in the activities of both LEE and REE (p>0.05). However, in comparison to ALB, the difference in activity was significantly different (p<0.05). The observed in vitro anthelmintic activity was attributed to saponins and condensed tannins and to alkaloids present in the plant. Phytochemical analysis confirmed presence of tannins, saponins and alkaloids, among other phytoconstituents. These phytochemicals are known to exhibit anthelmintic activity, therefore making the plant a potential candidate for drug development against gastrointestinal nematodes.

Evaluation of pharmacological interactions after administration of a levamisole, albendazole and ivermectin triple combination in lambs

The goals of the current trial were (a) to characterize the plasma disposition kinetics of levamisole (LEV), albendazole (ABZ) and ivermectin (IVM), each administered either alone (single active ingredient) or as a combined formulation to lambs; (b) to compare the clinical anthelmintic efficacy of the same drugs given either separately or co-administered to lambs infected with resistant nematodes. Fifty Corriedale lambs naturally infected with multiple resistant gastrointestinal nematodes were involved in the following experimental trials: (a) “Pharmacokinetic trial”: the animals were allocated into five groups (n=10 each) and intraruminally treated with either LEV (8mg/kg), ABZ (5mg/kg), IVM (0.2mg/kg), or with a LEV+ABZ+IVM combined formulation, where each active ingredient was administered at the same dose. Blood samples were collected over 15 days post-treatment and drug plasma concentrations measured by HPLC. (b) “Efficacy trial”: the same treated groups plus an untreated control group were used to assess the comparative anthelmintic efficacy by the faecal egg count reduction test (FECRT). Although the overall LEV disposition kinetics was unaffected, significantly lower (61%) ABZ-sulphoxide and higher (71%) IVM systemic availabilities were obtained after administration of the combined formulation in comparison to those obtained after treatment with each drug alone. A multiple drug resistance situation was observed for Haemonchus spp. The observed efficacies were 52% (LEV), 72% (ABZ), 80% (IVM) and 87% (triple combined formulation). The results reported here contribute to the pharmaco-therapeutic knowledge on drug combinations. This type of research is crucial before further development of combined anthelmintic preparations reaches the market to deal with resistant nematode control. The co-administration of LEV+ABZ+IVM did not result in a significant advantageous anthelmintic effect compared to the treatment with IVM alone. The simultaneous/combined administration of LEV, ABZ and IVM may account for a drug–drug pharmacological interaction in infected lambs. The pharmacokinetic interaction accounted for a reduced ABZ-sulphoxide and enhanced IVM systemic exposure following the combined treatment.

Methods to determine resistance to anthelmintics when continuing larval development occurs

The in vivo faecal egg count reduction test (FECRT) is the most commonly used test to detect anthelmintic resistance (AR) in gastrointestinal nematodes (GIN) of ruminants in pasture based systems. However, there are several variations on the method, some more appropriate than others in specific circumstances. While in some cases labour and time can be saved by just collecting post-drench faecal worm egg counts (FEC) of treatment groups with controls, or pre- and post-drench FEC of a treatment group with no controls, there are circumstances when pre- and post-drench FEC of an untreated control group as well as from the treatment groups are necessary. Computer simulation techniques were used to determine the most appropriate of several methods for calculating AR when there is continuing larval development during the testing period, as often occurs when anthelmintic treatments against genera of GIN with high biotic potential or high re-infection rates, such as Haemonchus contortus of sheep and Cooperia punctata of cattle, are less than 100% efficacious. Three field FECRT experimental designs were investigated: (I) post-drench FEC of treatment and controls groups, (II) pre- and post-drench FEC of a treatment group only and (III) pre- and post-drench FEC of treatment and control groups.To investigate the performance of methods of indicating AR for each of these designs, simulated animal FEC were generated from negative binominal distributions with subsequent sampling from the binomial distributions to account for drench effect, with varying parameters for worm burden, larval development and drench resistance. Calculations of percent reductions and confidence limits were based on those of the Standing Committee for Agriculture (SCA) guidelines. For the two field methods with pre-drench FEC, confidence limits were also determined from cumulative inverse Beta distributions of FEC, for eggs per gram (epg) and the number of eggs counted at detection levels of 50 and 25. Two rules for determining AR: (1) %reduction (%R)<95% and lower confidence limit <90%; and (2) upper confidence limit <95%, were also assessed. For each combination of worm burden, larval development and drench resistance parameters, 1000 simulations were run to determine the number of times the theoretical percent reduction fell within the estimated confidence limits and the number of times resistance would have been declared.When continuing larval development occurs during the testing period of the FECRT, the simulations showed AR should be calculated from pre- and post-drench worm egg counts of an untreated control group as well as from the treatment group. If the widely used resistance rule 1 is used to assess resistance, rule 2 should also be applied, especially when %R is in the range 90 to 95% and resistance is suspected.