Evaluation of the anthelmintic activity of Khaya senegalensis extract against gastrointestinal nematodes of sheep: in vitro and in vivo studies

The anthelmintic effect of Khaya senegalensis is described. In vitro and in vivo studies were conducted to determine possible direct anthelmintic effects of ethanolic and aqueous extracts of K. senegalensis towards different ovine gastrointestinal nematode. A larval development assay was used to investigate in vitro, the effect of aqueous and ethanolic extracts towards larvae of strongyles. The LC50 values of the effects of both the aqueous and ethanolic extracts were calculated. Another study was conducted in vivo to evaluate the therapeutic efficacy of the extracts administered orally at a dose rate of 125, 250 and 500 mg/kg of sheep harbouring naturally acquired infection of gastrointestinal nematodes. The presence of K. senegalensis extracts in the cultures decreased the viability of larvae. The LC50 of the aqueous extract (0.69 mg/ml) is not significantly different (P >0.05, t-test) from the ethanolic extract (0.51 mg/ml). The activity of the extract is concentration dependent in vivo. Sheep drenched with 500 mg/kg K. senegalensis ethanolic extract had a mean faecal egg count (FEC) reduction of 88.82%. The extract of K. senegalensis could find application in anthelmintic therapy in veterinary practice.     

Ovicidal and larvicidal activity of the crude extracts from Phytolacca icosandra against Haemonchus contortus

The development of anthelmintic resistance has impacted on the success of conventional anthelmintics (AH) for the control of gastrointestinal nematodes in grazing/browsing sheep and goats. Medicinal plants from the traditional herbolary in Mexico may provide new candidates that can be explored as alternative sources of AHs for ruminants. This study evaluated the leaf extracts derived from Phytolacca icosandra against infective L(3) larvae and eggs from Haemonchus contortus collected from sheep. Three extracts of different polarities were obtained from the leaf plants using ethanol, n-hexane and dichloromethane as the solvents. The effectiveness of the in vitro AH activity of the plant extracts was evaluated using larval migration inhibition (LMI) and egg hatch (EHA) assays. For the LMI assays, the ethanolic extract of P. icosandra showed 55.4% inhibition of larval migration at 2mg/mL (p<0.05). The dichloromethane extract of P. icosandra showed 67.1% inhibition of migration at 3mg/mL (p<0.05) and a dose-dependent response with an LD(50) of 0.90 mg/mL. The n-hexane extract failed to show inhibition of larval migration at any concentration explored. In the EHA for the ethanol extract, the lowest concentration tested (0.15 mg/mL) resulted in inhibition of egg hatching greater than 72.6%. Therefore, the LD(50) could not be calculated for this extract. The LD(50) of the dichloromethane extract of P. icosandra was 0. 28 mg/mL. An egg hatch inhibition greater than 90% was observed with both the ethanolic and dichloromethane extracts when using a concentration of 0.90 mg/mL or higher. The n-hexane extract failed to show egg hatch inhibition at any concentration tested. The AH activity reported for P. icosandra could be attributable to the flavonoids, steroids, terpenoids, coumarins and/or saponins that were present in the ethanolic and dichloromethane extracts. A combination of more than one component may also explain the observed AH activity against the H. contortus life stages that were evaluated. In conclusion, the ethanolic and dichloromethane extracts of P. icosandra showed clear in vitro AH activity against the H. contortus eggs and the L(3) larvae. However, the hexanic extract of the plant leaves failed to show any in vitro AH activity.     

Resistance to thiabendazole, fenbendazole and levamisole in Haemonchus and Trichostrongylus species in sheep on a Kenyan farm

Resistance to thiabendazole (TBZ), fenbendazole (FBZ) and levamisole (LVM) in naturally acquired gastrointestinal nematode parasites in sheep was investigated on a farm where anthelmintic resistance was suspected. This was measured by both the in vitro egg hatch assay, and reductions in faecal egg and worm counts in treated animals. In the egg hatch assay, nematode eggs were incubated in various concentrations of either TBZ or LVM. The level of resistance was expressed as the drug concentration inhibiting 50% of the eggs from hatching (LC50). The nematode population had LC50 values of 0.26 microgram ml-1 TBZ and 3.12 micrograms ml-1 LVM. In the faecal egg and worm count reduction test, naturally infected sheep were treated with either TBZ (88 mg kg-1), FBZ (10 mg kg-1) or LVM (15 mg kg-1). Faecal egg and total worm counts from these sheep were then compared with counts from untreated sheep. TBZ, FBZ and LVM failed to reduce the faecal egg counts and total worm counts by more than 90%. Based on the identification of larvae from faecal cultures, the most predominant nematode species in the resistant population were Haemonchus (62%) and Trichostrongylus (28%). TBZ reduced faecal egg counts for both species by less than 90%. FBZ and LVM also reduced Haemonchus spp. eggs by less than 90%. Other nematode species numbers did not satisfy criteria for the determination of efficacy.     

Doramectin resistance in Haemonchus contortus on an alpaca farm in Belgium

Parasitism by gastrointestinal nematodes is a health concern in New World Camelids (NWC) worldwide, and anthelmintic treatment is often needed for parasite control. Although anthelmintic resistance has been reported in ruminants worldwide, data in NWC are only scarce. In the present study, a case of suspected doramectin resistance in alpacas was examined. A field efficacy study was conducted for the evaluation of two different dosages of doramectin using a faecal egg count reduction test. A group of 8 alpacas was treated with a subcutaneous injection of doramectin at 0.2mg/kg bodyweight. Individual faecal samples were collected before treatment and 7 days after treatment. The faecal egg counts indicated a treatment efficacy of only 68%. To determine whether the treatment failure was caused by true anthelmintic resistance or suboptimal dosage in this animal species, a group of 4 alpacas was subsequently treated at 0.3mg/kg bodyweight. Faecal egg counts 7 days post treatment were reduced by only 41%, indicating that the treatment failure was more likely to be caused by the presence of resistant parasites on this farm. Coprocultures of faecal samples collected after treatment indicated the presence of 98.5% Haemonchus contortus and a small percentage of Cooperia oncophora (<1.5%). A controlled efficacy trial in sheep, for which the optimal dosage of doramectin is known, was conducted to ensure that this truly was a case of resistant parasites. Infective larvae collected from the faeces of these alpacas were used to infect eight nematode-free lambs. These lambs were assigned to one of two groups based on faecal egg counts post infection. One group was treated with doramectin injectable at 0.2mg/kg bodyweight, the other group served as a non treated control group. Pharmacokinetics indicated that the doramectin treatment was adequate, yet an efficacy of only 16% was determined on day 7 after treatment. Identification of the larvae after treatment revealed 100% H. contortus. On day 7 after treatment, H. contortus worm counts were only reduced by 8% in the treated lambs. The results of the present study report for the first time a case of doramectin resistance in alpacas, mainly in H. contortus.     

Anthelmintic activity of Trianthema portulacastrum L. and Musa paradisiaca L. against gastrointestinal nematodes of sheep

Evaluation of anthelmintic effects of Trianthema (T.) portulacastrum L. (Aizoaceae) whole plant and Musa (M.) paradisiaca L. (Musaceae) leaves against prevalent gastrointestinal worms of sheep was done that may justify their traditional use in veterinary clinical medicine. In vitro anthelmintic activity of the crude aqueous methanolic extract (CAME) of both the plants was determined using mature female Haemonchus (H.) contortus and their eggs in adult motility assay (AMA) and egg hatch test (EHT), respectively. In vivo anthelmintic activity of crude powder (CP) and CAME in increasing doses (1.0-8.0 g kg(-1)) was determined in sheep naturally infected with mixed species of nematodes using fecal egg count reduction test (FECRT) and larval counts. The study design also included untreated as well as treated controls. Fecal egg count reduction and larval counts from coprocultures were performed pre- and post-treatments to assess the anthelmintic activity of the plants. CAME of T. portulacastrum and M. paradisiaca showed a strong in vitro anthelmintic activity and pronounced inhibitory effects on H. contortus egg hatching as observed through AMA and EHT, respectively. Both plants exhibited dose and time dependent anthelmintic effects on live worms as well as egg hatching. M. paradisiaca (LC(50)=2.13 μg mL(-1)) was found to be more potent than T. portulacastrum (LC(50)=2.41 μg mL(-1)) in EHT. However, in vivo, maximum reduction in eggs per gram (EPG) of faeces was recorded as 85.6% and 80.7% with CAME of T. portulacastrum and M. paradisiaca at 8.0 g kg(-1) on 15th day post-treatment, respectively as compared to that of Levamisole (7.5 mg kg(-1)) that caused 97.0% reduction in EPG. All the species of gastrointestinal nematodes (GINs), i.e. Haemonchus contortus, Trichostronglyus spp., Oesophagostomum columbianum and Trichuris ovis which were prevalent, found susceptible (P<0.01) to the different doses of CP and CAME of both plants. The data showed that both T. portulacastrum and M. paradisiaca possess strong anthelmintic activity in vitro and in vivo, thus, justifying their use in the traditional medicine system of Pakistan.     

放牧绵羊主要寄生虫病药物防治技术试验与示范效果观察

为评价伊维菌素注射剂的驱虫效果与对放牧绵羊线虫病及外寄生虫病的防治示范效果,选择1.5岁感染线虫和部分外寄生虫的绵羊150只,设伊维菌素注射剂0.1,0.2和0.3mg/kg体重剂量组和伊维菌素片剂对照组,进行驱虫效果评价;在冬季应用伊维菌素注射剂按0.2mg/kg体重剂量对放牧绵羊进行规模防治技术示范,检查防治效果和考核防治效益。结果:药效试验中伊维菌素注射剂0.2mg/kg对绵羊消化道线虫虫卵转阴率和减少率分别为93.3%,99.3%,对原圆科线虫幼虫转阴率和减少率分别为90.0%和96.1%;0.3mg/kg剂量对消化道线虫虫卵及原圆科线虫幼虫转阴率和减少率均为100.0%;0.1mg/kg剂量对消化道线虫虫卵转阴率和减少率分别为76.7%,88.6%,原圆科线虫幼虫转阴率和减少率分别为66.7%和86.1%。防治示范群绵羊消化道线虫虫卵转阴率为93.3%,虫卵减少率为96.8%;对原圆科线虫幼虫转阴率和减少率分别为90.0%和96.2%。同期检查未防治对照组虫卵EPG和幼虫数略有增加。技术示范群比未示范群每只成年羊平均多增重5.47kg、幼年羊成活率平均提高2.1个百分点。结果表明该防治技术对放牧绵羊主要寄生虫病高效安全,效益显著。     

Duration of Anthelmintic Effect of Three Formulations of Ivermectin (Oral,Injectable and Pour-on) Against Multiple Anthelmintic-Resistant <Emphasis Type="Italic">Haemonchus contortus</Emphasis> in Sheep

We report the results of investigations that were conducted in a sheep flock in Uttaranchal, India where repeated failure of anthelmintic medication was noted. The study revealed that Haemonchus contortus in sheep had developed resistance to benzimidazoles (fenbendazole, mebendazole and albendazole), imidazothiazole (levamisole) and salicylanide (rafoxanide), while it was fully susceptible to avermectins (ivermectin). Further, the suppression of nematode egg output in faeces of sheep naturally infected with multiple anthelmintic-resistant H. contortus following treatment with ivermectin tablet (0.4 mg/kg body weight (bw), orally), ivermectin injection (1% w/v, 0.2 mg/kg bw, subcutaneously) and ivermectin pour-on (0.5 w/v, 0.5 mg/kg bw) was also studied over a period of 10 weeks post treatment. It was noted that ivermectin tablet after initial clearance of infection (faecal egg count reduction 100%), could not prevent establishment of new patent natural infection for even a single day, while ivermectin pour-on and injection prevented the establishment of new infection for 7 and 14 days post treatment, respectively. Maximum protection period (duration for which mean faecal egg count of sheep reaches 500 eggs per gram of faeces or more) of 68 days was recorded in sheep treated with injectable ivermectin, followed by pour-on (60 days) and oral (53 days) preparations.     

Multiple species of nematodes resistant to ivermectin and a benzimidazole-levamisole combination on a sheep farm in New Zealand

AIM: To confirm the presence of multiple anthelmintic resistance on a sheep farm in New Zealand. METHODS: Three groups of 10 weaned Romney-cross lambs were treated either with an oral dose of ivermectin (0.2 mg/kg), or a benzimidazole/levamisole (BZ/LEV) combination (4.75 albendazole and 7.5 mg/kg levamisole), or were left untreated. Ten days later, animals were necropsied, and adult worms recovered and identified from the abomasa and small intestines. Pre- and post-treatment faecal nematode egg counts (FEC) were recorded, and larval cultures were performed. RESULTS: In a faecal egg count reduction test (FECRT), adjusted to reflect pre- and post-treatment larval culture results, ivermectin resistance was detected in Teladorsagia (Ostertagia), Trichostrongylus and Haemonchus spp, while BZ/LEV combination- resistant Teladorsagia and Trichostrongylus spp were also present. Adult worm counts confirmed these results, and identified the species involved as Teladorsagia circumcincta, Trichostrongylus colubriformis and H. contortus. CONCLUSION: Multiple, multi-generic anthelmintic resistance was confirmed on a sheep property in New Zealand. This included the first confirmed case of ivermectin resistance in T. colubriformis from sheep in New Zealand.     

Teladorsagia circumcincta resistance to moxidectin and multiple anthelmintic groups in ewes following use of the persistent drug before lambing

This paper describes the nematode control strategy adopted by a Scottish hill sheep farmer. It provides an example of the limited use of anthelmintic drugs, targeted towards control of the periparturient rise in faecal nematode egg output in ewes and exploitation of grazing management, resulting in limited exposure of naive sheep to infective larvae on pasture. Resistance to benzimidazole, imidazothiazole and macrocyclic lactone anthelmintics, and to a combination of a macrocyclic lactone and imidazothiazole anthelmintic drugs was diagnosed. The targeted use of a persistent anthelmintic drug to control a periparturient rise in faecal nematode egg output in ewes may be unnecessary and selects strongly for resistance when the reservoir of anthelmintic-susceptible nematodes in refugia is small at the time of treatment. However, the use of a persistent anthelmintic drug in a selective proportion of ewes can be important and probably does not select strongly for resistance when the reservoir of anthelmintic-susceptible nematodes in refugia is large at the time of treatment. The former circumstances might arise on many Scottish hill sheep farms, whereas the latter may occur on upland and lowground farms, depending on previous grazing management, anthelmintic use and winter weather conditions. These factors must be taken into account when preparing sustainable health plans for nematode parasite control in individual sheep flocks.     

Use of a long acting injectable formulation of moxidectin to control the periparturient rise in faecal Teladorsagia circumcincta egg output of ewes

A field study was conducted in a sheep flock in the south east of Scotland with a history of ivermectin resistance in Teladorsagia circumcincta. The objective of the study was to compare the effects of single anthelmintic treatments in ewes before turn-out onto pasture that was contaminated with a moderate level of overwintered, ivermectin resistant, T. circumcincta infective larvae. The ewes were treated according to label directions with either a long acting injectable formulation of moxidectin (1mg/kg; affording up to 14weeks persistent action against macrocyclic lactone (ML)-susceptible T. circumcincta) or an oral formulation of moxidectin (0.2mg/kg; affording up to 5weeks persistent action against ML-susceptible T. circumcincta). The lambs were enrolled in the normal management of the farm, and received a total of three oral ivermectin treatments during the 16week study. The efficacy of both treatment strategies in controlling the periparturient rise in faecal nematode worm egg counts and subsequent pasture contamination was assessed from the faecal worm egg counts of the ewes and their lambs between lambing and weaning. Ewes that were treated with the oral formulation of moxidectin shed approximately 3.5 times more T. circumcincta eggs between lambing and weaning than ewes that were treated with the long acting formulation of moxidectin. This difference was reflected in the faecal worm egg counts of the lambs that were grazed alongside the different treatment groups of ewes. The results of the current study demonstrate persistent efficacy of the long acting formulation of moxidectin against an ivermectin resistant T. circumcincta population. The decreased pasture contamination after treatment could lead to improved lamb growth and a need for fewer anthelmintic treatments, thus potentially reducing one possible selection pressure for anthelmintic resistance. However, treatment with the long acting formulation of moxidectin would give rise to fewer susceptible nematodes being present in refugia, which could increase another possible selection pressure for anthelmintic resistance, depending on the subsequent grazing management of that pasture. The rationale for use of a persistent anthelmintic drug to control the periparturient rise in faecal ML-resistant T. circumcincta egg output of the ewes is discussed and potential differences in selection for macrocyclic lactone anthelmintic resistance using the different formulations of moxidectin are acknowledged.