Towards finding effective indicators (diarrhoea and anaemia scores and weight gains) for the implementation of targeted selective treatment against the gastro-intestinal nematodes in lambs in a steppic environment

The effective application of targeted selective treatment (TST) against gastro-intestinal nematodes depends on the accurate identification of those animals in need of anthelmintic treatment. Finding cost and labour effective measures to identify such animals is paramount. This study tested the efficacy of three indicators on lambs in a farm in Eastern Algeria: anaemia score (FAMACHA(?)), diarrhoea score (DISCO) and weight gain. These were contrasted against traditional parasitological infection measures (nematode eggs per gram of faeces: EPG). Thirty lambs were used in the study; every second month they were sampled (FAMACHA(?), faecal samples for EPG and DISCO, weight gains) and two of the original 30 lambs were necropsied for adult worm counts. The main gastro-intestinal nematodes were Teladorsagia circumcincta, Marshallagia marshalli, Nematodirus helvetianus, Trichostrongylus vitrinus, and Haemonchus contortus. DISCO proved to be the most effective indicator correctly identifying 80% of the sheep in need of treatment. This was followed by FAMACHA(?) with a 50% accuracy level and finally weight gains, which were not found to be a useful indicator at all.     

Predicting resistance of sheep to Haemonchus contortus infections

In vitro cultures of lymphocytes from worm-free sheep responded to larval and adult H. contortus antigens by undergoing blast transformation. The level of response varied considerably between animals but was heritable and positively correlated with resistance to subsequent primary, secondary and trickle infection by the parasite, but not with age of the sheep older than 5 months. Lymphocyte responses to phytohaemagglutinin and H. contortus antigens were not present at birth in lambs but developed during the first 5 months in the absence of infection. The proportion of the infecting dose of larvae which was inhibited at the early fourth stage correlated with age of the sheep and with the increase in lymphocyte response to H. contortus antigens during primary infections with H. contortus. Sheep which were most susceptible to infection had the lowest lymphocyte responses to H. contortus antigens but had the highest rate of weight gain during infection. It is suggested that man has selected the most productive animals at the expense of resistance to H. contortus. The definition of genetically controlled markers on lymphocytes is seen as a means by which disease resistant strains of animals may be selected.     

Pathogenicity of different strains of Histophilus somni in the experimental induction of ovine epididymitis

The purpose of this study was to determine any differences in pathogenicity when sheep are experimentally infected with different Histophilus somni isolates: a) 2336 bovine origin strain; b) an isolate from ram orchitis and epididymitis; c) an isolate from the brain of a sheep with neurological signs; d) an isolate from the vagina of a clinically healthy ewe. A total of 20 rams divided in groups of 5 animals each were inoculated in the epididymis with 1 × 10⁷ CFU/mL of H. somni; a negative control group of 5 rams was used. All groups inoculated with H. somni showed some epididymitis, but the most pathology was caused by the epididymitis isolate, followed by the vaginal isolate. It was demonstrated that there is a difference in experimental infection capacity among isolates from different origins, as epididymitis occurred and the bacteria was recovered only from groups inoculated with isolates originating from epididymitis and vaginal exudate.     

The effect of phenothiazine treatment on arrested Haemonchus contortus larvae in sheep

An experimental infection of freshly cultured Haemonchus contortus larvae was given to housed, non-pregnant sheep in October. Phenothiazine was administered to all animals shortly after patency, but the egg output remained negative until mid-April indicating that removal of adult worms from the abomasum did not stimulate the resumption of development of arrested larvae. Worm burdens at post mortem suggest that adults were not being expelled to any great degree during the winter from the host, but this information is clouded by the difference in numbers of both adult worms and arrested larvae in sheep of different haemoglobin type. The results suggest that haemoglobin type may also be a factor in arrest of larvae as it is in the resistance of sheep to adult H. contortus.     

Prevalence of <Emphasis Type="Italic">Haemonchus contortus</Emphasis> infection in sheep slaughtered at Jimma town municipal abattoir,Ethiopia

Haemonchus contortus is a parasite of major economic importance of most sheep-rearing areas of the world. A cross-sectional study was conducted from November 2015 to March 2016 with objectives of determining the prevalence and associated risk factors of Haemonchus contortus infection in sheep slaughtered at Jimma town municipal abattoir, Ethiopia. Of the total 7000 sheep slaughtered during the study time period, 384 sheep were randomly selected and examined for parasites. The overall prevalence of Haemonchus contortus infection was 264 (68.75%). Infection rate of poor body condition animals was significantly (P?=?0.000) higher than good body conditioned animals. The mean packed cell volume (PCV) values (%) of parasitemic and aparasitemic sheep were 23.73?±?3.7 SD and 37.66?±?4.736 SD, respectively. The prevalence in adult sheep (>?1 year) was insignificantly (P?=?0.653) higher than that of young sheep (≤?1 year). The highest prevalence was recorded in sheep that originated from Seka district (73.3%) and the lowest in Kersa district (63.4%) with non-significant variation (P?=?0.691). The highest monthly mean worm burdens and prevalence of Haemonchus contortus infection were recorded in November and lowest in March. The current study revealed that Haemonchus contortus infection is an important and common parasitic disease and requires special attention to its control.     

Use of pathophysiological indicators for individual decision of anthelmintic treatment of ewes against gastro-intestinal nematodes in Morocco

The targeted selective treatments (TST) aim at reducing the number of anthelmintic treatments but also to maintain productivity of animals. The aim of this work was to assess the validity of pathophysiological indicators for detecting individually ewes in need for treatments in two regions of Morocco with different management and climatic environment (Chaouia plain-seven farms, and Middle-Atlas-three farms). Although resistance to benzimidazoles was already present the same drug was used for TST. The indicators tested were: FAMACHA(?) (associated with anaemia), DISCO (diarrhoea score), and BODCON (body condition score). Only FAMACHA(?) and DISCO indicators were well correlated to the EPGs. DISCO only did permit a substantial reduction (up to 85%) of the number of treatment and EPG (nematode eggs per gramme) remained low on average (less than 160).     

Anthelmintic resistance survey in New Zealand

The prevalence and level of thiabendazole resistance of sheep nematodes in the North Island of New Zealand was investigated by means of an in vitro egg-hatch technique; samples from sheep on 52 properties in six country areas were examined. Resistance of Haemonchus contortus and Trichostrongylus spp. was identified on the basis of egg hatch in 0.1 ppm thiabendazole. Evidence of resistance was obtained on 11 (21.2%) of the properties. LD 50's and resistance ratios were calculated for the resistant nematode populations found.     

Efficacy of albendazole against nematode parasites isolated from a goat farm in Ethiopia: relationship between dose and efficacy in goats

A suspected case of albendazole resistance in a goat farm of Hawassa University was examined using faecal egg count reduction test (FECRT), controlled anthelmintic efficacy test and egg hatch assay (EHA) to verify the development of resistance and/or the need for higher doses of the drug in goats than in sheep. The experiment was conducted in 12 sheep (2 groups: treatment versus control) and 24 goats (4 groups: 3 treatments versus control, n = 6; per group) following artificial infection with infective larvae of Haemonchus contortus and Oesophagostomum columbianum. The first group of sheep and goats were treated orally with albendazole at the dose rate of 3.8 mg/kg body weight (i.e. manufacturer's recommended dose for sheep) while the second group of sheep and the fourth group of goats were left untreated. The second and the third group of goats were treated with albendazole at 5.7 and 7.6 mg/kg respectively. The FECRT showed an efficacy of albendazole in goats to be 65.5, 81.4 and 84.1% at the dose rate of 3.8, 5.7 and 7.6 mg/kg body weight respectively while in sheep it was 62% at the dose rate of 3.8 mg/kg. Increasing the dose to 1.5 the sheep recommended dose induced minor improvement of efficacy in goats; however the efficacy was almost the same at 1.5 and twice the dose recommended for sheep. Worm counts at day 15 post-treatment revealed that H. contortus has developed resistance to albendazole. EHA results also supported these findings. On the other hand, O. columbianum was 100% susceptible at all dose levels tested.     

Effects of concurrent copper deficiency and gastro-intestinal nematodiasis on circulating copper and protein levels, liver copper and bodyweight in sheep

Sheep were rendered hypocupraemic using parenteral ammonium tetrathiomolybdate (ATM). Fifteen thousand third stage larvae of Trichostrongylus axei and T. colubriformis in the ratio 1:1 were administered three times per week for six weeks, starting four weeks after cessation of ATM treatment. The changes in liver and plasma copper (Cu), caeruloplasmin activity, serum proteins, faecal nematode egg counts and total nematode counts were measured in the sheep for 10 weeks after infection. Decreases in liver Cu, plasma Cu and caeruloplasmin activities were detected soon after infection. There was a significant (P less than 0.05) interaction of the effects of Cu deficiency and nematode infection on these changes. Hypoproteinaemia, attributed to serum albumin loss, was demonstrated seven weeks after infection, but this was not associated with the interaction of Cu deficiency and nematode infection. No changes in serum globulins were detected. Although the results support the contention that gastro-intestinal nematodiasis can significantly exacerbate an existing Cu deficiency in sheep, there was no evidence that hypocuprosis would predispose sheep to higher nematode burdens.     

Efficacy of the amino-acetonitrile derivative,monepantel, against experimental and natural adult stage gastro-intestinal nematode infections in sheep

Multiple drug resistance by nematodes, against anthelmintics has become an important economic problem in sheep farming worldwide. Here we describe the efficacy of monepantel, a developmental molecule from the recently discovered anthelmintic class, the amino-acetonitrile derivatives (AADs). Efficacy was tested against adult stage gastro-intestinal nematodes (GINs) in experimentally and naturally infected sheep at a dose of 2.5 mg/kg body weight when administered as an oral solution. Some of the isolates used in experimental infection studies were known to be resistant to the benzimidazoles or levamisole anthelmintics; strains resistant to the macrocyclic lactones were not available for these tests. Worm count-based efficacies of >98% were determined in these studies. As an exception, Oesophagostomum venulosum was only reduced by 88% in one study, albeit with a low worm burden in the untreated controls (geometric mean 15.4 worms). Similar efficacies for monepantel were also confirmed in naturally infected sheep. While the efficacy against most species was >99%, the least susceptible species was identified as Nematodirus spathiger, and although efficacy was 92.4% in one study it was generally >99%. Several animals were infected with Trichuris ovis, which was not eliminated after the treatment. Monepantel demonstrated high activity against a broad range of the important GINs of sheep, which makes this molecule an interesting candidate for use in this species, particularly in regions with problems of anthelmintic resistance. Monepantel was well tolerated by the treated sheep, with no treatment related adverse events documented.     

Control of gastro-intestinal parasitism in sheep with ivermectin delivered via an intraruminal controlled-release capsule

The efficacy of ivermectin delivered by an intraruminal controlled-release capsule against gastro-intestinal nematodes of sheep was evaluated under controlled conditions. In seven Australian studies involving 170 Merino or Merino x Border Leicester sheep, intraruminal capsules developed for 20-40 kg or 40-80 kg sheep, and delivering 0.8 or 1.6 mg of ivermectin/day respectively for 100 days (minimum dose 20 microg/kg/day), were evaluated. Studies were designed to test the therapeutic efficacy against naturally acquired and induced infections treated at the adult and fourth larval stage, and the prophylactic efficacy against naturally acquired and induced infections with third stage infective larvae. The predominant pathogenic nematodes of sheep were represented. Two studies included known benzimidazole- and levamisole-resistant nematode strains. Sheep were necropsied for total nematode counts 21-8.5 days after treatment. The efficacy of the ivermectin controlled-release capsule was generally >99% against all nematode species tested, including those confirmed to be benzimidazole- and levamisole-resistant. High therapeutic activity was demonstrated against existing adult and fourth larval stage nematode infections at the time of treatment, and high prophylactic efficacy was shown against incoming third stage larvae of all species and strains tested.     

A survey of seasonal changes in nematode faecal egg count levels of sheep and goats in Senegal

A survey of the gastro-intestinal nematodes in domestic sheep and goats of the Sahelian zone of Senegal was undertaken. Their prevalence and the egg production were estimated by faecal egg count in 2234 sheep and 577 goats slaughtered at the Dakar slaughterhouse during a 12 month period. The highest prevalence (%) and highest mean egg counts (EPG) were recorded at the end of the wet season. Amongst the species of nematodes encountered there are two of potential importance: Haemonchus contortus and Oesophagostomum columbianum. It is suggested that the most effective control can be achieved by a critical treatment at the end of dry season (May–June) when the EPG starts to increase, and when hypobiosis is supposed to be terminating. This would insure the greatest proportion of the existing worm population to be exposed to the anthelmintic.     

A study of some factors influencing the immunization of sheep against Haemonchus contortus using attenuated larvae

The respective roles of age of lamb, size of immunizing dose, previous exposure to infection and anthelmintic therapy on the immune response of Scottish Blackface sheep to gamma-irradiated H. contortus larvae were studied.As previously shown, young lambs immunised at 10 and 14 weeks of age failed to develop a significant resistance to subsequent challenge.In contrast, parasite-naive lambs immunised at 9 and 10 months of age with 10 000 H. contortus larvae irradiated at 60 kiloröntgens developed a highly significant immunity to subsequent challenge. This degree of immunity was not significantly altered by the administration of spaced doses of normal larvae from 10 weeks of age until the point of vaccination.Worm-free lambs vaccinated at 9 and 10 months of age failed to develop any immunity to subsequent challenge when the anthelmintic thiabendazole was administered 3 weeks after each immunizing infection.     

Identification and quantification of benzimidazole resistance polymorphisms in Haemonchus contortus isolated in Northeastern Brazil

Haemonchus contortus is the most prevalent nematode in Brazil. The objective of this study was to select 6 populations of H. contortus of known or suspected benzimidazole resistance status and characterize these using quantitative real-time polymerase chain reaction (qPCR) for single nucleotide polymorphisms (SNPs) F200Y, F167Y and E198A in the β-tubulin isotype 1 gene. qPCR was performed using DNA from a pool of 10 adult male H. contortus from a single animal per farm. Faecal egg count reduction test (FECRT) and egg hatch test (EHT) were used to determine the resistance status. Samples were obtained from 6 farms located in 5 counties in the Ceará State: Tauá, Boa Viagem, Quixadá, Santa Quitéria and Solonópole. The inbred-susceptible-Edinburgh (ISE) isolate was used as reference for comparative purposes in the qPCR. Benzimidazole resistance was detected by FECRT on all farms with efficacy values ranging from 0 to 51%. EC50 values as determined by EHT were all above 1.49 μg/ml. High frequencies of the resistant SNPs F200Y and F167Y alleles were detected but no resistance was detected at SNP E198A. Our results suggest that the SNPs F167Y and F200Y are both important for benzimidazole resistance in the studied populations.     

Epididymitis Caused by Brucella ovis in a Southern Ontario Sheep Flock

Epididymitis was diagnosed in three rams in a commercial sheep flock in southern Ontario. The affected rams had palpably enlarged epididymides and two rams had semen which contained inflammatory cells and was of poor quality. Serum compliment fixation titers for Brucella ovis were 1:20, 1:80 and 1:90. Five other rams in the flock were clinically normal and without titers. Two of the affected rams had lesions similar to those produced by experimental infection with B. ovis. The infection in the rams had no apparent affect on ewe performance. The source of the infection remains unknown, but the rams were purchased from a flock which had imported ewes from the western U.S.A.     

In vitro effects of Tabernaemontana citrifolia extracts on Haemonchus contortus

Tabernaemontana citrifolia (Apocynaceae) is traditionally used as an anthelmintic preparation for ruminants in Guadeloupe (French West Indies). This study was carried out to evaluate the in vitro effect of this plant against the parasitic nematode of small ruminants Haemonchus contortus. Three extracts (aqueous, methanolic and dichloromethane) of T. citrifolia fruit, leaf and root were tested on four developmental stages of the parasite, using egg hatch assay (EHA), larval development assay (LDA), L3 migration inhibition assay (LMI), and adult worm motility assay (AWM). Compared to the negative control, significant effects were observed for the different parts of T. citrifolia but with differences depending on the parasitic stage; efficacies on the larval development of H. contortus from 88.9% to 99.8% for fruit, from 72.1% to 83.8% for root and from 33.5% to 85% for leaf with dose-dependent effect for the methanolic extract. The root gave the best result on EHA (22.7% efficacy for dichloromethane extract) and AWM (56% efficacy, with dose-dependent effect for dichloromethane extract) and the leaf on LMI (49.4% efficacy). These results suggest that T. citrifolia possess anthelmintic activity against H. contortus. The active ingredients responsible for the activity could be the alkaloid compounds present in the plant parts of the plant.     

Genetic parameter estimates for gastrointestinal nematode parasite resistance and maternal efficiency indicator traits in Santa Inês breed

Infection with Haemonchus contortus is the most economically important gastrointestinal nematode parasitosis and the most important cause of mortality in sheep production. The aim of this study was to estimate variance components of gastrointestinal parasite resistance traits, maternal efficiency (ME) and ewe adult weight (AW) in Santa Inês breed in tropical conditions. The phenotypic records were collected from 700 animals, belonging to four flocks located south‐east and north‐east in Brazil. The evaluated traits were as follows: degree of anaemia assessed using the FAMACHA chart (FMC), haematocrit (HCT), white blood cell (WBC), red blood cell (RBC), haemoglobin (HGB), platelets (PLT), faecal egg count (EPG_log), ME, metabolic maternal efficiency (MME), AW and metabolic ewe adult weight (MAW). From the 700 animals, 576 (82% of the evaluated population) were genotyped with the Ovine SNP12k BeadChip (Illumina, Inc.). Markers with unknown genomic position, located on sex chromosomes, monomorphic, with minor allele frequency <0.05, call rate <90% and with excess heterozygosity were excluded. The variance components were estimated using a single‐trait animal model with ssGBLUP procedure. The correlation between the parasite's resistance indicators and the ME suggested that selecting animals with both higher adult weight and ME will also favour the selection of animals with better resistance to gastrointestinal nematodes parasites, specially H. contortus. Therefore, since there are few or no studies with Santa Inês breed in this area, it is important to study those traits to better manage selection programs.     

Synergistic action of mebendazole and levamisole in the treatment of a benzimidazole-resistant Haemonchus contortus in sheep

One-year old worm free, merino wethers, were each infected with 5000 H. contortus larvae of a strain resistant to mebendazole at a rate of 52 mg/kg body weight of sheep. After 21 days, they were assigned to two trials. The preliminary trial showed that mebendazole and levamisole acted synergistically on the H. contortus infection. In the second trial, sheep were treated with 0.35 mg/kg levamisole (one seventh the minimum effective dose against susceptible worms) or 40 mg/kg mebendazole (40 times the minimum effective dose against susceptible worms). In each case the anthelmintics did not reduce worm burdens, although mebendazole depressed egg production. However, when mebendazole and levamisole, at the above dose rates, were administered simultaneously, total worm counts in sheep were reduced by almost 60%. Similar results were obtained when the levamisole was administered 8 h or 14 h after mebendazole treatment. The implications of these observations for the treatment of benzimidazole-resistant haemonchiasis in sheep are discussed.     

Functional investigation of a QTL affecting resistance to Haemonchus contortus in sheep

This study reports a functional characterization of a limited segment (QTL) of sheep chromosome 12 associated with resistance to the abomasal nematode Haemonchus contortus. The first objective was to validate the identified QTL through the comparison of genetically susceptible (N) and resistant (R) sheep produced from Martinik × Romane back-cross sheep. The R and N genotype groups were then experimentally infected with 10 000 H. contortus larvae and measured for FEC (every three days from 18 to 30 days post-challenge), haematocrit, worm burden and fertility. Significant differences in FEC and haematocrit drop were found between R and N sheep. In addition, the female worms recovered from R sheep were less fecund. The second step of the characterization was to investigate functional mechanisms associated with the QTL, thanks to a gene expression analysis performed on the abomasal mucosa and the abomasal lymph node. The gene expression level of a candidate gene lying within the QTL region (PAPP-A2) was measured. In addition, putative interactions between the chromosome segment under study and the top ten differentially expressed genes between resistant MBB and susceptible RMN sheep highlighted in a previous microarray experiment were investigated. We found an induction of Th-2 related cytokine genes expression in the abomasal mucosa of R sheep. Down-regulation of the PAPP-A2 gene expression was observed between naïve and challenged sheep although no differential expression was recorded between challenged R and N sheep. The genotyping of this limited region should contribute to the ability to predict the intrinsic resistance level of sheep.