Variability in the resistance of the Nigerian West African Dwarf goat to abbreviated escalating trickle and challenge infections with Haemonchus contortus

The West African Dwarf (WAD) goat is known to be relatively resistant to infection with Haemonchus contortus. An experiment was carried out to assess variability in resistance under escalating trickle and single pulse challenge infection protocols. Thirty two 7-8 month old goats were subjected to trickle infection beginning with a thrice weekly dose of 100 L3 and increasing to 500 L3 by week 5, the last dose being administered on D30. Infection was abbreviated with fenbendazole on D32 and the animals challenged with 4000 L3 (=404-741L3/kg body weight) on D46 and necropsied on D81. The goats exhibited marked individual variability in faecal egg counts (FEC) and worm burdens (Wb). For convenience, we arranged the goats into FEC classes 1, 2 and 3, corresponding to peak counts of <1000, 1000-2500 and >2500 epg and Wb classes 1, 2 and 3, with counts of <50, 50-100 and >100 worms, respectively, broadly reflecting relatively resistant, intermediate and poorly resistant response phenotypes. Wb classes broadly reflected the FEC phenotypes and although there were no significant differences between the Wb of the FEC classes at necropsy, significant differences in FEC were detected when analysis was carried out only on Wb classes. There was also a significant positive correlation between FEC class on each of the days of challenge (D67 to D80) and worm burden at necropsy. In general, the goats showed strong resistance/tolerance to challenge, with more than 80% having Wb not exceeding 5% of the challenge dose, and only in class three goats were weight loss and anaemia detected in some animals. PCV and weight changes also closely reflected the FEC and Wb classes, with a significant negative correlation between each and Wb. Infection level did not significantly affect the sizes of either male or female worms and no significant difference was detected between the levels of circulating eosinophils in any FEC or Wb class. Overall, the data are consistent with the existence of a range of response phenotypes, which reflect variations in resistance of WAD goats to H. contortus infection, the genetic basis of which will be explored in future experiments.     

Purebreeding of Red Maasai and crossbreeding with Dorper sheep in different environments in Kenya

The aim of this article was to study opportunities for improvement of the indigenous and threatened Red Maasai sheep (RM) in Kenya, by comparing purebreeding with crossbreeding with Dorper sheep (D) as a terminal breed, in two different environments (Env. A and a harsher Env. B), assuming different levels of genotype‐by‐environment interaction (G × E). Breeding goals differed between environments and breeds. Four scenarios of nucleus breeding schemes were stochastically simulated, with the nucleus in Env. A. Overall, results showed an increase in carcass weight produced per ewe by more than 10% over 15 years. Genetic gain in carcass weight was 0.17 genetic SD/year (0.2 kg/year) across scenarios for RM in the less harsh Env. A. For survival and milk yield, the gain was lower (0.04–0.05 genetic SD/year). With stronger G × E, the gain in the commercial tier for RM in the harsher Env. B became increasingly lower. Selection of females also within the commercial tier gave slightly higher genetic gain. The scenario with purebreeding of RM and a subnucleus in Env. B gave the highest total income and quantity of meat. However, quantity of meat in Env. A increased slightly from having crossbreeding with D, whereas that in Env. B decreased. A simple and well‐designed nucleus breeding programme would increase the genetic potential of RM. Crossbreeding of RM with D is not recommended for harsh environmental conditions due to the large breed differences expected in that environment.     

Serum antibody response of Castellana sheep to Haemonchus contortus infection and challenge: relationship to abomasal worm burdens

Primary and secondary serum antibody responses to Haemonchus contortus were studied in Castellana sheep. Ten-month-old sheep were infected (200 L3/kg live weight (lw)) and challenged (400 L3/kg lw) or uninfected and equally challenged with H. contortus. Primary infections induced a partially protective response upon challenge, characterized by higher serum protein levels, longer prepatent periods, lower fecal egg counts, and significant reduction in the establishment rate of the parasite and abomasal adult and L4 worm burdens. The resistant status of the infected and challenged sheep was not clearly related either to the serum specific antibody levels (IgG: IgG1, IgG2; IgM; IgA) estimated by ELISA or to immunodetection patterns in the Western blots.     

Infection of goats with Haemonchus contortus and Trichostrongylus colubriformis: histopathology and pH changes.

Two groups of goats were experimentally infected with Haemonchus contortus and Trichostrongylus colubriformis, and killed at various days after infection (DAI). The percentage of worms that established in the abomasum and the small intestine was low. At necropsy, abomasums from infected goats had thickened walls and oedematous folds. At 7 DAI there was an initial infiltration of eosinophils and some neutrophils which tend to increase with age of infection. The mean pH of the abomasum in goats infected with H. contortus was 5.43 (range 5.3-5.7), while that of the control goats was 3.30 (range 2.8-3.7).     

Partial protection of lambs against Haemonchus contortus by vaccination with a fractionated preparation of the parasite

Florida Native lambs, less than 6 months of age, were successfully vaccinated against Haemonchus contortus with a high mol. wt fraction (greater than 30,000 daltons) derived from a somatic extract of H. contortus larvae (SEL) and excretions and secretions (ES) of larvae isolated during in vitro development from the infective 3rd to 4th stage. A 59% reduction in adult worm numbers was obtained in vaccinates compared to naive lambs following challenge. The protection in vaccinated lambs was similar to that seen in lambs exposed to a primary infection of H. contortus larvae which had been cleared with anthelmintic prior to the challenge infection. The unfractionated SEL/ES preparation and a low mol. wt fraction gave no significant protection against challenge infection.     

The modulatory influence of Trypanosoma brucei on challenge infection with Haemonchus contortus in Nigerian West African Dwarf goats segregated into weak and strong responders to the nematode

Although Nigerian West African Dwarf (WAD) goats are relatively resistant to infection with Haemonchus contortus and are also trypanotolerant, natural outbreaks of both infections are known to occur. Despite their relative resistance to H. contortus WAD goats nevertheless show variability in response phenotype and it was of interest to examine the effect of this variability on the outcome of concurrent trypanosome infection. Trypanosoma brucei infections were established in goats that were initially classified as good or poor responders to H. contortus. Thirty-nine goats were exposed to an escalating infection with H. contortus, and on the basis of their mean faecal egg counts (FEC) were allocated to high FEC (poor responders, 18 goats with the highest FEC) or low FEC (good responders, 18 goats with the lowest FEC) classes. Nine uninfected naive control goats were included to provide reference baseline values. Retrospective analysis of parasitological and pathological parameters after allocation into high/low FEC classes showed that FECs differed significantly, in both classes packed cell volume (PCV) values fell relative to naive controls, neither class lost weight and both generated marked IgG responses. All goats received anthelmintic on day 61, half of each group was infected with 50 million trypanosomes and on day 67, excepting the controls, all goats were challenged with 3000 L3 of H. contortus. Trypanosome parasitaemia was generally low, and marginally, but not significantly, higher in the low compared with high FEC class, peaking 12-16 days after exposure in both groups and then falling to below microscopically detectable levels (although still detectable by sub-inoculation into mice) by week 3. At autopsy (days 109/110), worm burdens were significantly higher in the trypanosome-infected goats from the high FEC class, relative to all other groups. Trypanosome infected goats showed a tendency (although not significant) towards higher FEC and, irrespective of their FEC class, had lower PCV values although body weight did not vary significantly. All goats challenged with H. contortus had higher antibody levels than naive controls, but neither trypanosome infection nor FEC class affected the magnitude of responses. These results confirm that WAD goats comprise a range of response phenotypes to initial H. contortus infection and that trypanotolerance is a key trait of this breed. Although immunity to nematode infection develops even in poor responders, these animals harbour higher nematode burdens during concurrent infection with T. brucei.     

In vitro analysis of the anthelmintic activity of forage chicory (Cichorium intybus L.) sesquiterpene lactones against a predominantly Haemonchus contortus egg population

The anthelmintic activity of chicory (Cichorium intybus L.) herbage has been attributed to sesquiterpene lactones. Chicory leaves contain significant amounts of lactucin (LAC), 8-deoxylactucin (DOL), and lactucopicrin (LPIC), but the proportions of these three sesquiterpene lactones vary among forage chicory cultivars. To determine whether the individual compounds differ in anthelmintic activity, we prepared sesquiterpene lactone-enriched extracts from leaves of two forage chicory cultivars, 'Grasslands Puna' (Puna) and 'Forage Feast', and tested their effects on the hatching of a predominantly Haemonchus contortus egg population. The dominant constituents in the Puna and Forage Feast extracts were DOL and LAC, respectively; LPIC concentrations in the two extracts were similar. Extracts from both cultivars inhibited egg hatching at all concentrations tested (P<0.001), but there were significant differences in egg responses to the two extracts (P<0.001). With Puna, egg hatching decreased sharply in a linear fashion when the combined LAC, DOL, and LPIC concentrations ranged from 0 to 5.0mg/ml. A biphasic effect on egg hatching occurred with the Forage Feast extract. The fraction of eggs that hatched decreased gradually to 65% as the sesquiterpene lactone concentrations increased from 0 to 6.7 mg/ml. Treatment with higher concentrations resulted in a sharp decline in egg hatchability. Concentrations of sesquiterpene lactones required for 50% lethality were determined by probit dose-effect analysis to be 2.6 mg/ml (95% confidence interval: 2.4-2.8 mg/ml) for the Puna extract and 6.4 mg/ml (95% confidence interval: 5.9-7.2mg/ml) for the Forage Feast extract (P<0.0001). These concentrations provided 1.3 and 1.5mg/ml of DOL and 0.8 and 3.9 mg/ml of LAC for Puna and Forage Feast extracts, respectively. Results suggest that LAC has minimal effect on egg hatching and that DOL or other constituent(s) in the extracts is inhibitory. Quantitative analyses of free sesquiterpene lactones in chicory leaf extracts suggest that Puna may be a better cultivar than Forage Feast for use in bioactive pastures for gastrointestinal parasite control in small ruminants.     

Humoral responses in rabbits immunized with two fractions of Haemonchus contortus: the antigen specificity of such antibodies.

Humoral responses were examined in rabbits immunized with either 28-40 kDa (Fraction 1) or a 19-24 kDa (Fraction 2) antigenic fraction from soluble antigens (Sol L3 Ag) from infective larvae (L3) of Haemonchus contortus. These fractions were eluted from electrophoretically separated Sol L3 Ag. Immunoblots revealed antibodies to Fraction 1 (fr. 1) or Fraction 2 (fr. 2) polypeptides as well as to several other molecular weight polypeptides of the Sol L3 Ag. The latter antibodies were shown by absorption studies not to be Sol L3 Ag cross-reactive anti-bacterial rabbit antibodies. When Sol L3 Ag was affinity-purified using monoclonal antibody to phosphorylcholine (PC) and the resulting fractions were further analysed by immunoblotting using rabbit anti fr. 1 or anti fr. 2 antiserum, the PC antigen was found to be shared between fr. 1 and other polypeptides of Sol L3 Ag. Using the rabbit antibody fractions eluted from nitrocellulose membranes containing fr. 1 or 2 polypeptides, it was found that these fractions contained antibody that bound mainly to fr. 1 and only to fr. 2 polypeptides of Sol L3 Ag. It is concluded that, from the present immune rabbit sera, antibodies specific for either fr. 1 or fr. 2 may be isolated and then used to purify small amounts of the corresponding antigens.     

Experiences with integrated concepts for the control of Haemonchus contortus in sheep and goats in the United States

The generally warm, moist environmental conditions in the southern United States (U.S.) are ideal for survival and growth of the egg and larval stages of Haemonchus contortus and other gastrointestinal nematodes (GIN) of sheep and goats. Consequently, infection with GIN is the greatest threat to economic small ruminant production in this region. With anthelmintic resistance now reaching epidemic proportions in small ruminants in the U.S., non-chemical control alternatives are critically needed. The Southern Consortium for Small Ruminant Parasite Control (SCSRPC) was formed in response to this crisis and over the last decade has successfully validated the use of several novel control technologies, including FAMACHA(?) for the implementation of targeted selective treatments (TST), copper oxide wire particles (COWP), nematode-trapping fungi, and grazing or feeding hay of the high-tannin perennial legume sericea lespedeza [Lespedeza cuneata (Dum.-Cours. G. Don)]. Producer attitudes toward GIN control in the U.S. have been shifting away from exclusive dependence upon anthelmintics toward more sustainable, integrated systems of parasite control. Some novel control technologies have been readily adopted by producers in combination with appropriate diagnostic tools, such as FAMACHA(?). Others techniques are still being developed, and will be available for producer use as they are validated. Although new drugs will likely be available to U.S. goat and sheep producers in the future, these will also be subject to development of anthelmintic resistance. Therefore, the adoption and implementation of sustainable GIN control principles will remain important. With emerging markets for grass-fed or organic livestock, there will continue to be a critical need for research and outreach on development and on-farm application of integrated GIN control systems for small ruminants in the U.S. for the foreseeable future.     

Pathological and immunohistochemical study of the abomasum and abomasal lymph nodes in goats experimentally infected with Haemonchus contortus

Histopathological changes and the distributions of T and B lymphocytes and IgG producing plasma cells were recorded in the abomasum and abomasal lymph nodes of goats 3, 7 and 21 weeks post-infection (wpi) after an experimental infection with H. contortus. The low rate of worm recovery by 3 wpi (5.6%) might have been due to larvae death as suggested by the presence of granulomas in the abomasal mucosa at 3 and 7 wpi, or simply due to a poor larval establishment. Marked increase in the secretion of mucus by mucous cells together with an abundant infiltration of eosinophils, mast cells, CD3+ T lymphocytes, CD79a+ B cells, IgG+ plasma cells and globule leukocytes were recorded in the abomasal mucosa, especially at 7 wpi. Except for the globule leukocytes, this reaction decreased substantially by week 21, suggesting this cell type may have been involved in rejection of adult nematodes. The abomasal lymph nodes showed marked hyperplasia, particularly of CD79a+ B cells and IgG+ plasma cells in all infected goats. These reactions may have been responsible for the reduction in the number of worms found in the abomasum between 3 and 7 wpi.     

Factors affecting the prevalence and severity of footrot in a Merino flock selected for resistance to Haemonchus contortus

SUMMARY An outbreak of virulent footrot occurred in three lines of sheep selected for different levels of resistance to Haemonchus contortus , providing an opportunity to examine the factors affecting footrot severity. The selection lines did not differ significantly in average footrot score, but various other factors were found to have an effect. In breeding ewes, the average score was 10.2 (maximum possible score 20) and the prevalence of footrot was 96%. There was a significant age effect, with average score increasing from 9.1 ± 0.8 in 2-year-old ewes, to 13.4 ± 0.9 in 7-year-old ewes. Ewes that lost their lambs tended to have higher scores than those that either failed to lamb or successfully reared their litter. Significant differences in footrot score were caused by the lambing paddock in which the ewes had been, during the period seven weeks to one week before infection. Among lambs, aged about 20 weeks, the average footrot score was lower (5.0), as was the prevalence (75%). The score increased by an average of 0.09 ± 0.03 per day of age and the prevalence increased by an average of 0.6%± 0.3% per day of age. Lambs born and reared as twins had significantly lower scores than singles, and progeny of maiden dams had lower scores than progeny of older ewes. There was no significant difference because of the sex of the lamb. The paternal half-sib estimate of heritability of footrot score was 0.20 ± 0.14 and of the genetic correlation between footrot score and worm egg count was essentially zero (0.02 ± 0.20), but these estimates were probably biased by the confounding of sires with lambing paddocks. Nevertheless, the results strongly suggest that there are no adverse genetic associations between resistance to footrot and resistance to internal parasites.     

SHORT COMMUNICATION: Neutrophil oxidative metabolism and haemogram of sheep experimentally infected with Haemonchus contortus and supplemented with selenium and vitamin E

The objective of this study was to evaluate neutrophil oxidative metabolism and haemogram in sheep experimentally infected with Haemonchus contortus and supplemented with selenium and vitamin E. Twenty male Corriedale sheep were utilised and distributed into four experimental groups each with five animals infected with larvae: G1‐ supplemented with sodium selenite, 0.2 mg/kg body weight (bw) given intramuscularly (IM); G2‐ supplemented with sodium selenite and vitamin E, 0.2 mg/kg bw and 2000 IU per animal, respectively, both IM; G3‐ supplemented with vitamin E, 2000 IU per animal IM; G4‐ not supplemented. A haemogram and the number of parasite eggs were determined in samples of blood and faeces, respectively, on days zero (T0), 20 (T1), 40 (T2) and 60 (T3) and nitroblue tetrazolium (NBT) assays were performed on heparinized blood samples taken on days zero, 30 and 60. A lower total leucocyte count was detected in G1 in relation to G4 at T4. Lymphocytes were reduced in G1 in relation to G3 and G4 at T3. In both non‐stimulated (NBT‐NS) and stimulated (NBT‐S) dye reduction assays, there was reduced activity at 60 days, in relation to other times in the groups treated with selenium (G1 and G2). On the basis of results obtained, we conclude that supplementation with selenium provides better antioxidant protection to neutrophils.     

Changes in liveweight gain and blood constituents in experimental infection of goats with a goat-derived compared with a sheep-derived strain of Haemonchus contortus.

Two groups of goats were experimentally infected with two different strains of Haemonchus contortus and compared with the controls. Group A animals were infected with a goat-derived strain (GDS) while Group B animals were infected with a sheep-derived strain (SDS). Changes in the liveweights and some blood constitutents between the two infected groups were compared. The pathogenic effects of the GDS larvae were generally more serious when compared with those of the SDS larvae.     

Immune exclusion of Haemonchus contortus larvae in sheep: dose dependency, steroid sensitivity and persistence of the response

Naive and immune sheep, treated with anthelmintic, challenged directly into the abomasum and killed 48 hours later were used to examine the steroid sensitivity, persistence and dose dependency of immune mechanisms which result in the failure of incoming Haemonchus contortus larvae to establish (immune exclusion). Immune exclusion was almost totally abolished by corticosteroid treatment, the mean percentage of larvae establishing in the mucosa of immune sheep was 1.4 per cent of the challenge infection whereas in those treated with dexamethasone it was 24.4 per cent and in naive sheep 27.3 per cent. Immune animals challenged after seven and 42 days without antigenic experience excluded more than 90 per cent of the challenge larvae from their mucosae, whereas those challenged after 84 days without antigenic experience were as susceptible as naive animals. Immune exclusion was dose dependent, animals challenged with 10(6) and 10(5) larvae excluded 93 per cent and 82.5 per cent of the challenge dose whereas those challenged with 10(4) larvae failed to exclude larvae.     

Ostertagia ostertagi infection in the calf: effects of a trickle challenge on the hormonal control of digestive and metabolic function

Metabolic effects of a trickle challenge with the equivalent of 10,000 infective Ostertagia ostertagi larvae per day were investigated in 12 calves allocated to infected, pair-fed control or ad libitum-fed control groups. Changes in hormone levels reflecting abomasal, pituitary and pancreatic function were monitored using radioimmunoassay techniques previously validated for use in cattle. A range of metabolic profile parameters and blood metabolites was also measured. Feed intake of the infected calves began to decline as blood gastrin and pepsinogen levels reached a peak. The depression in appetite recorded in this group was responsible for significant increases in plasma urea and non-esterified fatty acid levels and associated with an increase in growth hormone/insulin ratio. No significant difference in glucagon levels was recorded between groups. A decline in blood albumin values was also shown in the infected group and associated with a drop in nitrogen digestibility. A significant depression in circulating calcium levels was related to either the hypoalbuminaemia or impaired mineral absorption in the intestine. A decrease in plasma cholesterol values in the infected group was associated with changes in digestive function.     

High-level ivermectin resistance in a field isolate of Haemonchus contortus associated with a low level of resistance in the larval stage: implications for resistance detection

The IVPro isolate of Haemonchus contortus was isolated in 1999 after significant numbers of the parasite survived an ivermectin capsule treatment of grazing sheep acquiring a natural infection at Prospect, NSW, Australia. The isolate shows high-level resistance to ivermectin (faecal egg count is unaffected by ivermectin oral treatment at 0.2mg kg(-1)). The larval LC(50), as assessed by larval development assays (LDAs), is only approximately two-fold higher than several susceptible isolates, making it difficult to detect the resistance using larval LC(50) as an indicator. However, the isolate shows the presence of a small proportion of the population (<20%) able to develop at significantly higher drug concentrations than the susceptible isolates. Hence, if the IVPro and susceptible isolates are compared at the LC(99) level, the IVPro isolate is readily identifiable as resistant. This degree of distinction at the LC(99) allows the IVPro isolate to be identified as resistant by simply observing the highest drug concentration in the development assay at which some larvae develop relative to the susceptible isolates. Assessing the development assay using this criterion allows a distinction between IVPro and the susceptible isolates equivalent to 10-fold differences in drug concentration, greatly increasing the likelihood of detecting the resistant isolate in routine resistance tests. This study highlights the need to examine this aspect of LDAs in order to detect the type of resistance displayed by IVPro H. contortus.     

Moisture requirements for the free-living development of Haemonchus contortus: quantitative and temporal effects under conditions of low evaporation

The key influence of moisture availability on development of the free-living stages of Haemonchus contortus is well-documented, although quantitative relationships between moisture variables and development are poorly defined. A factorial experiment (3x2x2) was conducted in programmable incubators to determine the effects of amount and distribution of simulated rainfall on H. contortus development at low evaporation rates (approximately 2 mm/day) under temperatures typical of summers on the Northern Tablelands of NSW, Australia. Sheep faeces containing H. contortus eggs were placed in experimental units containing sterile soil and had one of the three amounts (12, 24 and 32 mm) of simulated rainfall applied, in either a single event on the day after deposition (d 1) or three split events over 6 days (d 1, 3 and 6). Treatments were applied either in week 1 only, or in weeks 1 and 2. Recovery of infective larvae (L3) at 4, 7 and 14 days post-contamination increased with each incremental amount of simulated rainfall over the range of 12-32 mm and was significantly higher under the single "rain" event (2.8%), compared with the three smaller, split events (1.9%). The second application of simulated rainfall in week 2 had only a very small influence on L3 recovery, suggesting that the majority of development to L3 occurred in response to simulated rainfall events in the first 7 days. Both faecal moisture content and the cumulative ratio of precipitation and evaporation (P/E) were strongly and positively correlated with recovery of L3. Recovery of L3 from treatments, which received simulated rainfall in week 1 only was best described by P/E at d 5. Whether the relationships observed in this study hold under field conditions, where macroclimatic conditions such as evaporation rate are substantially more limiting to free-living development, is yet to be determined.