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PubMed2005

Haemonchus contortus egg excretion and female length reduction in sheep previously infected with Oestrus ovis (Diptera: Oestridae) larvae.

Mixed parasitic infection of animals is a common phenomenon in nature. The existence of one species often positively or negatively influences the survival of the other. Our experimental study was started with the objectives to demonstrate the interaction of Haemonchus contortus and Oestrus ovis in relation to cellular and humoral immune responses in sheep. Twenty-two sheep of Tarasconnais breed (France) were divided into four groups (O, OH, H and C) of five or six animals. Group O and OH received 5 weekly consecutive inoculations with O. ovis L1 larvae (total = 82 L1) in the first phase of the experiment between days 0 and 28. On the second phase, groups OH and H received 5000 L3 of H. contortus on day 48 while group C served as our control throughout the experimental period. Parasitological, haematological, serological and histopathological examinations were made according to standard procedures and all animals were slaughtered at day 95. There was no significant variation in the number and degree of development of O. ovis larvae between the two infected groups. Furthermore, in tissues examined in the upper respiratory tract (nasal septum, turbinate, ethmoide and sinus), group O and OH has responded similarly on the basis of cellular inflammatory responses (blood and tissue eosinophils, mast cells and globule leucocytes (GL)) and serum antibody responses against the nasal bots. This may indicate that the presence of H. contortus in the abomasa of group OH had no marked influence over the development of O. ovis larvae in the upper respiratory tract. On the other hand, we have observed a significantly lower H. contortus female worm length, fecal egg count (FEC) and in utero egg count in animals harbouring the nasal bot (OH) than in the mono-infected group (H). This was significantly associated with higher blood eosinophilia, higher packed cell volume (PCV) and increased number of tissue eosinophils and globule leucocytes. We conclude that, the establishment of O. ovis larvae in the upper respiratory tract has initiated higher inflammatory cellular activity in group OH there by influencing the development and fecundity of H. contortus in the abomasum.

Animals↗

PubMed2005

Effect of ivermectin on feeding by Haemonchus contortus in vivo.

While several in vitro studies have shown that the anthelmintic ivermectin inhibits feeding by parasites, the relevance of this putative site of action in vivo has not been demonstrated. For this study, techniques to measure feeding by Haemonchus contortus in vivo relied on the blood feeding characteristics of the worm, and utilised tritiated inulin administered to sheep intravenously and subsequently measured in worms recovered from abomasa. Nematodes recovered from sheep treated with ivermectin 4 h prior to the [3H]inulin administration showed equivalent feeding levels (over a 1 h period) to those recovered from sheep not treated with ivermectin. In addition, there was no difference in the radioactivity in nematodes of an ivermectin-susceptible and an ivermectin-resistant isolate recovered from individual sheep with concurrent infections after a dose with ivermectin. Ivermectin, therefore, had no effect on feeding by H. contortus in vivo under these experimental conditions. The results are discussed in relation to the dynamics of the expulsion of H. contortus from sheep following ivermectin treatment.

Abomasum↗

PubMed2005

Multiple anthelmintic resistance in Haemonchus contortus isolated from South African Boer goats in Switzerland.

A suspected case of multiple anthelmintic resistance on a farm in the canton of Zurich, Switzerland, into which South African Boer goats had previously been imported, was confirmed in a controlled test. Twenty sheep were allocated into one control group and three treatment groups to determine the efficacy of mebendazole, ivermectin and moxidectin applying the faecal egg count reduction test (FECRT). The sheep were slaughtered 1 week later and post-mortem worm counts were performed. Benzimidazole and ivermectin resistance were found in Haemonchus contortus with an efficacy of 55 and 61%, respectively. Moxidectin appeared to be effective when the data was analysed according to the recommended analytical techniques with an efficacy of 96%. This is the first description of resistance of gastrointestinal nematodes to one of the macrocyclic lactones in small ruminants in Switzerland. The results are discussed in relation to the importance of controlling livestock before importation.

Abomasum↗

PubMed2005

Rapid selection for ivermectin resistance in Haemonchus contortus.

Lambs infected with two isolates, one British and one American, of Haemonchus contortus were treated with increasing doses of ivermectin. Eggs from the highest dose that had not eliminated the infection were cultured and larvae used to infect another lamb. After three generations the H. contortus was resistant to 0.2 mg/kg ivermectin. The results stress the ease with which ivermectin resistance can be selected if high selection pressure is applied.

Animals↗

PubMed2005

The predatory capability of Arthrobotrys cladodes var. macroides in the control of Haemonchus contortus infective larvae.

One hundred compost samples were examined for the presence of nematophagous fungi on the sheep farms of Mazanderan, province, Iran. Arthrobotrys cladodes var. macroides (IRAN 677C=CBS 143565) was isolated from 3% of the samples examined. Nematophagous activity of this fungus which was shown for the first time in this study, revealed the addition of 1000, 8000, 20000 and 100000 conidia per gram of feces of sheep reduced significantly (P<0.001) the number of Haemonchus contortus infective larvae in the feces by 41.71%, 63, 27%, 73.49% and 94.96%, respectively. These results show that A. cladodes var. macroides is a promising candidate for biological control of H. contortus.

Animals↗

PubMed2005

Influence of dietary protein supply on resistance to experimental infections with Haemonchus contortus in Ile de France and Santa Ines lambs.

The effect of Haemonchus contortus infection in sheep fed with a moderate and high protein content diet was evaluated in two breeds of sheep. Forty-eight Ile de France and Santa Ines lambs were maintained indoors since birth, in worm-free conditions. The lambs were allocated after weaning in four groups of six animals per breed, which were either infected or remain uninfected and given access to either a moderately or highly metabolizable protein diet. The moderately and highly metabolizable protein diets were calculated to supply 75 and 129 g metabolizable protein per kg of dry matter (MP/kg DM), respectively. The infection consisted of a trickle infection with 300 infective larvae, three times a week, for 12 weeks. Significant differences were observed for mast cell, globule leukocyte and eosinophil counts in the abomasal mucosa of the infected groups compared to the control of both breeds (P<0.05), regardless of the diet supplied. Significantly higher IgA anti-L5 antibody was detected in the infected Santa Ines groups than in the infected Ile de France groups (P<0.05). Increased metabolizable protein supply resulted in larger body weight gain and higher packed cell volumes for both breeds (P<0.05). Both breeds showed an increased ability to withstand the pathophysiological effects of H. contortus infection when given access to the highly metabolizable protein diet. However, increased metabolizable protein supply resulted in reduced worm burdens in Santa Ines lambs but not in the Ile de France lambs (P<0.05). The present results show that the increase in protein content in growing lamb diets may benefit resistance and resilience to gastrointestinal parasites but that these benefits may vary among breeds.

Abomasum↗

PubMed2005

Haemonchus contortus and Trichostrongylus colubriformis in pen-trials with Javanese thin tail sheep and Kacang cross Etawah goats.

Weight gain costs due to infection were higher in sheep than goats, 28 and 17.5%, respectively, for Trichostrongylus colubriformis and 48.7 and 32.2%, respectively, for Haemonchus contortus. The extent of bodyweight cost attributed to anorexia in sheep infected with H. contortus was higher (13.5 g/day) than in sheep infected with T. colubriformis (2.3 g/day). On the other hand, bodyweight cost due to the other pathogenic effects in sheep infected with T. colubriformis were higher (35.6 g/day) compared to sheep infected with H. contortus (10.9 g/day). A strong relationship between faecal egg count and worm count (r=0.79, P=0.006) was shown only in sheep infected with T. colubriformis. About half of the infected sheep and goats had low or zero faecal egg counts throughout the study. In about 40% the egg count rose initially but became low by weeks 10-16, whereas in about 10% counts increased progressively throughout the period of observation and these animals also had the highest numbers of worms at slaughter. Packed cell volume was reduced in sheep and goats infected with H. contortus but serum protein and haemoglobin levels were unaffected. Sheep infected with T. colubriformis had a higher level of eosinophilia after 8 weeks (18.4%) than sheep infected with H. contortus (11.4%), whereas this pattern was reversed in goats and levels were also lower (4.1 and 8.9%, respectively). There was no apparent relationship between eosinophilia and resistance to infection with H. contortus or T. colubriformis.

Animals↗

PubMed2005

Synergism of rotenone by piperonyl butoxide in Haemonchus contortus and Trichostrongylus colubriformis in vitro: potential for drug-synergism through inhibition of nematode oxidative detoxification pathways.

The anthelmintic properties of rotenone and its activity in combination with the cytochrome P450 inhibitor piperonyl butoxide, were examined in in vitro assays with adults and larvae of Haemonchus contortus and larvae of Trichostrongylus colubriformis. Rotenone was toxic to larvae of both species, with LC(50) values in larval development assays of 0.54 and 0.64 microg/ml for H. contortus and T. colubriformis, respectively. The compound also caused complete cessation of movement in adult H. contortus after 72 h at a concentration of 20 microg/ml. Toxicity of rotenone towards the larvae of both species was increased in the presence of piperonyl butoxide (synergism ratios of 3-4-fold at the LC(50)) and the activity against adult H. contortus was also significantly enhanced following pre-treatment with piperonyl butoxide. This significant synergism suggests that these nematode species are able to utilize a cytochrome P450 enzyme system to detoxify rotenone and indicates that a role may exist for cytochrome P450 inhibitors to act as synergists for other anthelmintics which are susceptible to oxidative metabolism within the nematode.

Animals↗

PubMed2006

Towards the eradication of Haemonchus contortus from sheep flocks in Sweden.

Previous studies have shown that between-year transmission of Haemonchus contortus in Swedish sheep flocks is almost entirely as over-wintered populations within housed animals, and not on pasture. This suggests that eradication of this parasite is a realistic possibility. Thus, two sheep farms with a history of H. contortus infection on the Swedish island of Oland were selected for study. During the winter housing period of 2003/2004 all ruminants (sheep and cattle) on both farms were treated with ivermectin. Monitoring by faecal egg counts and infective larval differentials of ewes and lambs for the subsequent two grazing seasons, together with total abomasal worm counts of 10 lambs from each farm at the end of the first grazing year, showed that this objective was achieved.

Animals↗

PubMed2006

Effect of CD4+ T lymphocyte depletion on resistance of Gulf Coast Native lambs to Haemonchus contortus infection.

It has been reported that CD4(+) T lymphocytes are important in acquired immunity to gastrointestinal nematode infection. Whether these lymphocytes are also involved in the immune response of naturally resistant Gulf Coast Native (GCN) sheep to Haemonchus contortus infection remains to be defined. The objective of this study was to determine the role of CD4(+) T lymphocytes in this resistance. Ten GCN lambs were randomly assigned to a control (n=5) or a treatment (n=5) group. The treatment consisted of a series of IV injections with mouse anti-ovine CD4(+) T lymphocyte monoclonal antibodies for a period of 3 weeks. After the second treatment, all lambs were experimentally infected with 10,000 H. contortus infective larvae by oral inoculation. All lambs were monitored for fecal egg counts, blood packed cell volumes, white blood cell differential counts and serum antibody responses on a weekly basis. Fluorescence-activated cell sorter (FACS) analysis was done biweekly to enumerate CD4(+) T lymphocytes in peripheral blood. Necropsies were performed at the end of the study and 10% of the contents of the gastrointestinal tract were preserved for nematode enumeration and identification. Also at necropsy, mesenteric lymph nodes were extracted and FACS analysis was run on lymphoid cells. Mean fecal egg counts on day 21 and 28 post-infection and nematode counts at necropsy of the treated group were significantly (p<0.05) higher than that of the control group. Percent CD4(+) T lymphocytes in peripheral blood was significantly (p<0.05) lower in the treatment group than in the control group from day 9 to the end of the study. No differences were found in blood packed cell volumes, white blood cell differential counts, antibody titer or lymph node CD4(+) lymphocytes between groups. Lambs depleted of their CD4(+) T lymphocytes were more susceptible to H. contortus infection than undepleted lambs. The results of this study suggest that CD4(+) T lymphocytes are associated with the natural resistance of GCN sheep to H. contortus infection.

Abomasum↗

PubMed2006

A modified larval migration assay for detection of resistance to macrocyclic lactones in Haemonchus contortus, and drug screening with Trichostrongylidae parasites.

We have developed a modified migration assay system in 96-well plate format which is able to detect resistance to the macrocyclic lactone group of drugs in Haemonchus contortus. The assay involves exposure of infective stage larvae to drug for a 24 h period, then counting the numbers of larvae that are able to migrate through an agar and filter mesh system over a further 48 h. The agar barrier greatly increased the sensitivity of the assay for resistance detection compared to use of filter mesh alone. The assay was able to detect the presence of 10% resistant worms in an otherwise susceptible background. However, the assay was ineffective with Trichostrongylus colubriformis and Ostertagia circumcincta indicating that its usefulness for field monitoring will be restricted to situations where H. contortus is of most significance. A small-scale drug screening exercise showed that the assay identifies some anthelmintic activities distinct from those identified by larval development assays. The assay therefore also has a potential role in drug discovery programmes in screening for new anthelmintics.

Agar↗

PubMed2006

Identification of a 66 kDa Haemonchus contortus excretory/secretory antigen that inhibits host monocytes.

A 66 kDa adult Haemonchus contortus excretory/secretory (E/S) antigen was identified in Western blot by reaction with sera from the infected goats. The protein was purified from the adult worm extract and E/S products by anion exchange and ConA-Sepharose chromatography. The purified protein inhibited monocyte function in vitro as judged by decreased production of hydrogen peroxide and nitric oxide in the culture medium. The protein also caused proliferation of peripheral blood mononuclear cells. The absence of protein in the free living L3 larvae suggests that the expression of this protein coincides with the adaptation to the parasitic life. A correlation of antibody titre and worm burden was observed in the infected goats with higher antibody levels in high worm burdened animals. Anti-protein antibody caused loss of adult worm motility in vitro resulting in the death of the parasite. The fact that the protein is recognized by the host together with in vitro killing of adult parasites by antibodies makes this protein a promising candidate for vaccination trial.

Abomasum↗

PubMed2006

Detection of benzimidazole resistance in Haemonchus contortus using RFLP-PCR technique.

Benzimidazole (BZ) resistance in Haemonchus contortus is linked primarily with the mutation in the beta-tubulin isotype 1 gene that substitute phenylalanine (Phe) to tyrosine (Tyr) at 200 codon of the gene. In the present study, a new restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) technique has been developed for detection of BZ resistance in the beta-tubulin isotype 1 gene of H. contortus. The technique utilizes two primers viz. AvikaF and AvikaR to amplify the region containing mutation in the beta-tubulin gene followed by restriction digestion. After digestion, the 'rr' individuals (homozygous resistant) revealed 257 and 48 bp bands, the 'rS' individuals (heterozygous) showed 305, 257 and 48 bp bands, while 'SS' individuals (homozygous susceptible) revealed uncut 305 bp band. A total of 162 adult male H. contortus collected from Avikanagar, Jaipur and Bikaner regions (54 from each region) were genotyped for analyzing BZ resistance in the beta-tubulin gene. Out of which, 130 adults were 'rr' types, 20 'rS' types and 12 'SS' types. The results showed that genotypic frequencies of different genotypes (rr, rS and SS) were highly significant difference among the three regions (P<0.001). The 'rr' individuals were higher (98%) in Jaipur followed by Avikanagar (93%) and Bikaner (50%) regions. Overall, the prevalence of BZ resistant allele (r) was higher (86%) as compared to BZ susceptible allele (S) (14%). The technique was also found suitable for genotyping of larvae of H. contortus and yielded reproducible results. The study indicated that RFLP-PCR is an easy, reproducible and less expensive than allele specific PCR. This technique will be helpful in establishing the prevalence rate of BZ resistance in H. contortus and can also be utilized for existing worm control programme.

Amino Acid Substitution↗

PubMed2006

Evaluation of multiple low doses of copper oxide wire particles compared with levamisole for control of Haemonchus contortus in lambs.

High levels of anthelmintic resistance in gastrointestinal nematodes (GIN) of small ruminants have created the need for alternative approaches to parasite control. Copper oxide wire particles (COWP; 2g) have proven effective in decreasing GIN infection in lambs. However, the risk of copper toxicity has limited the usefulness of this approach. Recently, smaller doses (0.5 and 1g) have proven effective in GIN control, reducing the risk of toxicity. The objective of this study was to examine the effectiveness and risk of toxicity using multiple small doses of COWP for GIN control in lambs between weaning and market weight. Dorper crossbred ram lambs were orally administered levamisole (Levasol, 8.0mg/kg; n=8), 0.5g (n=9), or 1g COWP (n=9) at weaning (Day 0; 118+/-2 days of age; late May 2005) and again at 6-week intervals for a total of four treatments. A pooled fecal culture determined that Haemonchus contortus was the predominant gastrointestinal parasite at weaning. Lambs grazed bermudagrass pastures and were supplemented with up to 500g corn/soybean meal and free choice trace mineralized salt. Fecal egg counts (FEC), packed cell volume (PCV), and plasma aspartate aminotransferase (AST) activity were determined every 14 days and lambs weighed every 28 days. GIN infection reached a peak at Day 42 (high FEC, low PCV). COWP effectively reduced FEC on Days 0 and 42 compared with the previous week, but did not reduce FEC on Days 84 and 126 (treatment by time interaction, P<0.005). Plasma AST activity and weight gains were similar among treatment groups throughout the study period. Concentrations of copper in the liver on Day 155 were greater in COWP-treated lambs (P<0.001), but all concentrations were normal. Multiple doses of COWP were as effective as levamisole for control of H. contortus without risk of copper toxicity.

Animals↗

PubMed2006

Ovicidal and larvicidal activity of Melia azedarach extracts on Haemonchus contortus.

Haemonchus contortus is responsible for severe economic losses in sheep and goat breeding in the Northeast of Brazil. However, the effectiveness of control is compromised due to anthelmintic resistance and misuse. In the search for natural anthelmintics, Melia azedarach L., a plant indigenous to India but now distributed throughout Brazil, was selected due to the reported anthelmintic properties of its seeds. The aim of this study was to evaluate the anthelmintic activity of the seed and leaf extracts of the Brazilian adapted plant and investigate the type of organic chemical compounds present in the most active extracts. The ovicidal and larvicidal activity of M. azedarach extracts on H. contortus was evaluated through egg hatching and larval development tests. Hexane and ethanol extracts of seeds and chloroform and ethanol extracts of leaves of M. azedarach were used in the tests. To perform the larval development test, feces of an animal free from parasites were mixed with third instar H. contortus larvae and extracts in several concentrations. The coprocultures were incubated for 7 days at 30 degrees C, then the larvae were recovered and counted. LC50 was calculated by probits using the SPSS 8.0 program. The seed ethanol extract was the most active on eggs (LC50=0.36mgmL(-1)) and the leaf ethanol extract showed the best inhibition of larval development (LC50=9.18mgmL(-1)). Phytochemical analysis of the most active extracts revealed the presence of condensed tannins, triterpenes and alkaloids.

Animals↗

PubMed2006

Control of Haemonchus contortus in goats with a sustained-release multi-trace element/vitamin ruminal bolus containing copper.

The objective of this study was to determine the effectiveness of a sustained-release multi-trace element/vitamin ruminal bolus (TEB) containing copper administered to mature does for control of gastrointestinal nematodes (GIN) during summer months and during late gestation. Haemonchus contortus was the predominant nematode during these trials. In Experiment 1, yearling Spanish does were untreated or administered TEB (n=11/group) on Day 0 (August 2005) prior to breeding. In Experiment 2, Spanish and Boer does were untreated or administered TEB 6 weeks before kidding. Fecal egg counts (FEC) and blood packed cell volume (PCV) were determined weekly between Days 0 and 42. In both experiments, FEC were reduced within 7 days in TEB-treated compared with untreated does. PCV was similar between treatment groups (Experiment 1) or tended to be lower in the TEB group (Experiment 2). GIN control did not persist more than 28 days. These studies suggest that TEB may be an effective means of GIN control in mature goats, but additional control measures may be necessary.

Animal Nutritional Physiological Phenomena↗

PubMed2006

Effect of nutritional status and fat reserves on the periparturient immune response to Haemonchus contortus infection in sheep.

A study was made of the effect of body fat stored by ewes in early pregnancy on the subsequent immune response to gastrointestinal parasites around parturition. Pregnant ewes were given access to a lucerne pelleted diet either ad libitum (H) or at approximately 0.70 of their maintenance requirements (L) from the 42nd day of pregnancy in order to achieve a clearly differentiated level of body reserves by the 90th day of pregnancy. Then, all animals were put on the same plane of nutrition till 3 weeks after parturition. All ewes in both groups received 7,000 infective larvae of Haemonchus contortus per week for 7 weeks prior to lambing. The dietary treatments led to large differences between both groups of ewes in back-fat thickness that had a high correlation with mean plasma concentrations of leptin. In spite of the fact that animals were on the same plane of nutrition at infection time, host resistance, as measured by faecal egg counts, was significantly affected by the nutritional treatment established during early pregnancy. This effect produced noticeable differences in worm size and in worm burden at lambing. The response was accompanied by a marked increase in circulating eosinophils in better-fed ewes than in those maintained on a restricted diet in early pregnancy. Serum pepsinogen concentration, however, was inversely affected by the nutritional treatment till lambing, showing a maximum difference as early as 2 weeks after infection. The results support the view that higher levels of nutrition during early pregnancy enhance the expression of immunity against gastrointestinal parasites around parturition. Furthermore, the differences in the immune response appeared associated with serum leptin levels suggesting that leptin may be a key link between nutritional status and the protective immune reactivity against GI nematode infection.

Adipose Tissue↗

PubMed2006

Accuracy of an anaemia scoring chart applied on goats in sub-humid Kenya and its potential for control of Haemonchus contortus infections.

We tested the practical application of an anaemia scoring chart (the FAMACHA chart) as a method for controlling Haemonchus contortus in goats kept under smallholder conditions in a sub-humid area of Central Kenya. The objectives were: (1) to test the accuracy of the FAMACHA chart in identifying anaemic goats (PCV< or =18); (2) to quantify the proportion of goats left untreated at farm level when using the chart. On each of two farms, Small East African goats of various ages were allocated to two treatment groups; a FAMACHA group (F1 (n=34) and F2 (n=31) on farms 1 and 2, respectively) and a control group (C1 (n=34) and C2 (n=30)). In F1 and F2 goats with a FAMACHA score of 3, 4 or 5 were treated with anthelmintic after scoring. In C1 and C2 goats were treated every 4 weeks from 15 February to 20 July. Every 2 weeks all goats were scored with the FAMACHA chart and weighed. Furthermore, faecal samples were collected for faecal egg counts (FEC) and blood samples were collected for packed cell volume (PCV) determination. H. contortus was found to be the predominant nematode on both farms. The mean FECs were higher on farm 1 compared to farm 2, while in contrast the mean PCV levels were lowest on farm 2. The latter was most likely due to the presence of Fasciola spp., flea and tick infections on farm 2. The accuracy of the chart was evaluated by using PCV as the gold standard for anaemia (PCV< or =18%). The mean percentage of false-negative scorings per sampling was 0.7% on farm 1 and 1.6% on farm 2, while the mean percentage of false-positive scorings was 9.7% and 21.4%, respectively. It is most likely that the accuracy of the chart was negatively affected by the concurrent parasite infections on farm 2. The mean proportion of untreated goats per sampling was 89% and 77% on farm 1 and farm 2. It was concluded that the FAMACHA chart can be a valuable tool for decision-making in control of H. contortus in goats kept under smallholder conditions, without morbidity or mortality unacceptable to the farmer. The application may further reduce the risk of development of anthelmintic resistance by increasing refugia. However, caution should be taken under conditions where other anaemia-causing parasites are present (e.g. Fasciola spp. and ecto-parasites), since this possibly decreases the accuracy of the FAMACHA chart.

Anemia↗