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Due to the negative impact of Haemonchus contortus in the tropics and subtropics, the detection of serum protein profiles that occur in infected sheep is of high relevance for targeted selective treatment strategies (TST). Herein, we integrated proteomics with phenotypic traits to elucidate physiological mechanisms associated to H. contortus infection in susceptible (Dorper - D) and resistant (Santa Inês - S) sheep breeds. Naïve female lambs were infected with H. contortus third-stage larvae on day zero (D0), and samples were collected weekly, for 28 days. Feces were used for individual fecal egg counts (FEC) blood for packed cell volume (PCV) and serum for specific antibody quantification through ELISA. Sera was collected on D0 (-) and D21 (+), and analyzed using a LC-MS/MS based proteomics approach. FEC, PCV, and anti-H. contortus antibody levels confirmed the absence of infection on D0. On D28 there was a significant difference between the two breeds for logFEC means (D = 3774 and S = 3141, p=0.033) and PCV means (D = 16.3 % and S = 24.3 %, p=0.038). From a total of 754 proteins identified, 68 differentially abundant proteins (DAPs) were noted. Phosphopyruvate hydratase (ENO3) was a DAP in all comparisons, while S+ vs D+ and S- vs D- shared the highest number of DAPs (8). Each of the four experimental groups clustered separately in a principal component analysis (PCA) of protein profile. Among the DAPs, proteins associated with the innate and adaptive immune system were detected when comparing S- vs D- and S+ vs D+. In D-, some proteins were linked to stress response to handling, sampling and heat. Focusing on the consequences of infection in each breed, in the D+ vs D- comparison, upregulated proteins were associated with inflammation control and immune response, where downregulated proteins pointed to a negative impact of infection on tissue anabolism, compromising muscle growth and fat deposition. In the S+ vs S- comparison, upregulated proteins were related to immune response, while the downregulated proteins were possibly linked to muscular development and growth, impaired by infection. Collectively, it can be concluded that ENO3 regulation emerges as a potential factor underlying the differential immune response observed between Santa Inês and Dorper sheep infected with H. contortus. In turn, detected acute phase proteins (APPs) reinforce their relation with infection, inflammation and stress conditions, whereas THEMIS-like may contribute to the immune system in Dorper. GSDMD, Guanylate-binding protein and ACAN warrant further investigation as possible biomarkers for TST strategy development., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ana Carolina S. Chagas reports financial support was provided by State of Sao Paulo Research Foundation. Andre M. Almeida reports financial support was provided by Foundation for Science and Technology. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Haemonchus contortus is a nematode parasite that causes anaemia and affects the health of sheep. The mean corpuscular haemoglobin concentration (MCHC) is an excellent indicator to detect anaemia that could help to characterize resistant or susceptible lambs to gastrointestinal nematodes. The aim of this study was to evaluate the predictive value of MCHC in detecting changes in red blood cells and their relation to anaemia in lambs re-infected with H. contortus. An analysis of information was performed using 24 Pelibuey lambs previously infected in grazing, dewormed and experimentally re-infected with H. contortus. At the first haematological sampling (admission) the lambs were classified based on MCHC quartiles (Q). Subsequently, the lambs were housed for 56 days. Blood samples were taken every seven days to determine the haematological parameters using an impedance haematological instrument. Confidence limits were constructed with the records of the lambs that recovered their haematological parameters. Each quartile was analysed as a treatment in a repeated measures design over time. To know the optimal combination of sensitivity and specificity of MCHC to detect anaemia a curve of receiver operating characteristic (ROC) curve and the cut-off values were evaluated. In quartile 4 (Q4), lambs showed the highest faecal egg count (FEC, 764 eggs/g of faeces), mean corpuscular haemoglobin (17.0 pg) and MCHC (54.6 g/dL). This group also presented the lowest RBC values (5.8 × 10 6 /mL), haematocrit (HCT, 18.3%), total plasma protein (5.7 g/dL), and HGB (9.7 g/dL). The optimal point of MCHC with ROC curve was 42.4 (sensitivity 88.2% and specificity 86.5%); the area under the curve was 0.91 (CI 95%, 0.86-0.96). These results are related to the haematological effects caused by H. contortus in susceptible lambs. In conclusion, the highest FEC and lower HCT in Q4 are important elements of the haematological damage caused by H. contortus and could identify susceptible lambs., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

In this study we evaluated the efficacy of trichlorfon against Haemonchus contortus, monitoring its influence on blood parameters and plasma enzymes of lambs with haemonchosis. A lamb group was orally treated with trichlorfon at 100 mg kg -1 while the other group was untreated. Split-plot design analysis was performed with the lamb groups defined as plots while the subplots were the four periods (weeks) of collection. The trichlorfon treatment promoted a significant and effective reduction of fecal egg counts after one week, with efficacies > 99%. After 21 days of treatment, detected blood parameters and serum levels of plasma enzymes were normal. Additionally, serum albumin and urea concentrations increased to normal values, which were not observed in untreated lambs. The treatment with this organophosphate, using a correct oral administration, may represent an effective therapeutic alternative for sheep infected with multi resistant strain of H. contortus., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The relationship between packed cell volume (PCV) and fecal egg count (FEC) in different breeds of meat goats and hair sheep infected with gastrointestinal nematodes, including Haemonchus contortus, was characterized. Growing males from eight commercial and two research farms (one Kiko, Spanish, Dorper, and St. Croix; three Boer; four Katahdin) in the southcentral United States were evaluated in a central performance test with ad libitum intake of a 50% concentrate pelleted diet. There were 84 Boer, 55 Kiko, and 57 Spanish goats and 52 Dorper, 129 Katahdin, and 49 St. Croix sheep. During adaptation, animals were dewormed then dosed with 10,000 infective H. contortus larvae. PCV and FEC were determined before deworming (i.e., natural infection potentially with multiple internal parasites) and 21, 28, 35, 42, and 49 days after artificial infection. Effects of species, breed, and year were analyzed with mixed effects models including day of sampling post dosing as a repeated measure and FEC and FEC × breed as covariates. Moreover, differences in correlation coefficients between PCV and logarithmic FEC (lnFEC) among species, breed, year, and day of sampling were evaluated. Breed affected (P ≤ 0.001) PCV in goats (24.8, 27.2, and 26.0% for Boer, Kiko, and Spanish, respectively; SEM = 0.42) and sheep (29.8, 26.7, and 31.0% for Dorper, Katahdin, and St. Croix, respectively; SEM = 0.28). There were effects of FEC × breed (P ≤ 0.029) on PCV for Boer, Kiko, Dorper, Katahdin, and St. Croix (-0.31, -0.33, -0.46, -0.46, and - 0.49% per 1000 eggs, respectively) but not for Spanish goats (P = 0.451). With all data, PCV and lnFEC with natural infection were highly correlated (P < 0.001) for Boer and Kiko goats and Dorper and Katahdin sheep (r = -0.59, -0.67, -0.77, and - 0.84, respectively) but not for Spanish goats or St. Croix sheep (P ≥ 0.323). Correlation coefficients for artificial infection with H. contortus were significant (P ≤ 0.002) except for Spanish goats, although values were lower (-0.40, -0.21, -0.23, -0.47, and - 0.28 for Boer, Kiko, Dorper, Katahdin, and St. Croix, respectively) compared with natural infection. In conclusion, PCV was not related to FEC in Spanish goats infected either naturally or artificially, and the nature of the relationship varied among breeds of goats and sheep. Based on the magnitude of the FEC × breed coefficient, sheep incurred a relatively greater reduction in PCV as FEC increased, and correlation coefficients indicate stronger relationships with natural than artificial infection., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The aim of this research was to evaluate the sensitivity and specificity of the FAMACHA © (F © ) system in Morada Nova ewes. The conjunctivae of 562 ewes were evaluated using the F © system. Packed cell volume (PCV) served as the gold standard for clinical F © evaluation. To calculate the sensitivity and specificity of the F © system, different criteria were adopted: animals classified as (I) F © 4 and 5 or (II) 3, 4, and 5 were considered to be anemic and animals classified as (I) F © 1, 2, and 3, or (II) 1 and 2 were considered to be non-anemic. Three standard values of PCV, namely, ≤ 19%, ≤ 18%, or ≤ 15%, were used to confirm anemia. The percentage of correct treatments was always high when the F © values 4 and 5 were used as criteria for positive tests. For all the PCV cut-off values, more animals were classified as false positives when evaluated using F © 3, 4, and 5 as criteria for a positive test and more true negative animals when evaluated using only F © 4 and 5 as criteria for a positive test. For both sets of criteria for the positive tests, few animals were classified as false negatives and true positives. Eliminating the classification of F © 3 as anemic decreased the sensitivity and increased the specificity for all the PCV cut-off values for the ewes. The F © system can be used as a reliable alternative to reduce selection pressure for anthelmintics in relation to routine non-selective blanket treatment for worm control in the Morada Nova ewes.

Background: Pattern recognition receptors (PRRs) can recognize pathogen-associated molecular patterns and activate downstream signalling pathways, resulting in modulation of host immunity against pathogens. Here, we investigated whether PRR-mediated recognition is involved in host immune responses to the blood-feeding nematode Haemonchus contortus., Methods: During blood-feeding, H. contortus secretes immune-modulating antigens into host blood. Therefore, we stimulated sheep peripheral blood mononuclear cells (PBMCs) with H. contortus soluble extract (HcAg) and performed transcriptional profiling., Results: HcAg upregulated two genetically linked CLRs (CLEC2L and KLRG2), two NLRs attenuating inflammation (NLRP12 and NLRC3) and one G protein-coupled receptor with potent anti-inflammatory effects (HCAR2). Furthermore, several Th2-related transcription factors (ATF3, IRF4, BCL3 and NFATC) were also upregulated, which may confer anti-inflammatory type 2 immune responses to HcAg., Conclusions: Together, our preliminary studies provide new insights into how the host innate immune system controls type 2 immunity to H. contortus. Further work will be needed to identify H. contortus products recognized by the host innate immune system and determine the Th2 polarization ability of these putative PRR ligands.

The objectives of this study were to measure Haemonchus contortus larval aggregation by complement/antibody complexes, determine effect of breed resistance and infection status and determine the effect of larval maturation on larval aggregation in vitro. Larval binding assays were performed on H. contortus L3, exsheathed L3 and L4 incubated with serum from either parasite naïve or H. contortus primed St. Croix (resistant) and Suffolk (susceptible) lambs. No differences in L3 aggregation were observed between serum from either breed or infection status. Exsheathed L3 (60%) and L4 (42%) aggregation by primed Suffolk serum was significantly reduced compared with L3 (80%, P<.001). Removal of either complement or antibody effectively eliminated L3 aggregation (P<.001). Combination of antibody-depleted and complement-inactivated serum restored L3 aggregation to levels consistent with unprocessed serum, supporting a role for antibody and complement in aggregation (P<.001). Use of fluorescence-labelled anti-sheep IgG antibody allowed documentation of IgG bound to serum complexes within L3 masses and was present only in larvae incubated with normal serum, and complement- and antibody-depleted serum combination. These data indicate that complement/antibody complexes inhibit larval motility through larval aggregation which may be critical in early larval clearance of H. contortus., (© 2017 John Wiley & Sons Ltd.)

The present study had the primary objective of evaluating clinical, hematological and biochemical parameters, as well as observing anatomical and histopathological characteristics of abomasums, from calves prime-infected with Haemonchus contortus or H. placei. Ten male Holstein newborns were subdivided in three groups (GI placebo; GII infected with H. contortus; GIII inoculated with H. placei). Eye mucosa staining was evaluated. Hematological and biochemical tests were performed on animals. The euthanasia of all ten experimental calves was performed on the 42nd day post-inoculation. Fragments were collected from each of all 10 abomasums for histopathological analysis. Discrete submandibular edema was diagnosed in animals from both infected groups (H. contortus or H. placei). However, there were no significant changes (P > 0.05) in the color of the ocular mucosa of calves from all three experimental groups across the entire experimental period. Hematological and biochemical changes diagnosed on animals could not be linked to infections by species of Haemonchus spp. Regarding histopathological exams, it was possible to diagnose hypertrophy, hyperplasia, binucleated cells, inflammatory infiltrate, multifocal hemorrhage and edema in abomasums from calves of both groups infected with H. placei and H. contortus. It can, thus, be concluded that not only are calves susceptible to infections by both Haemonchus species, but they can also present clinical changes and similar anatomic histopathological lesions independent of being infected by Haemonchus placei or Haemonchus contortus. These results reflect a negative effect on helminth control by mixed grazing between sheep and cattle, especially when using calves., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The limitations associated with the use of anthelmintic drugs in the control of gastrotintestinal nematodosis, such as the emergence of anthelmintic resistance, have stimulated the study of the immunological control of many parasites. In the case of Haemonchus contortus, several vaccination trials using native and recombinant antigens have been conducted. A group of antigens with demonstrated immunoprotective value are cathepsin B - like proteolytic enzymes of the cysteine proteinase type. These enzymes, which have been observed in both excretory-secretory products and somatic extracts of H. contortus, may vary among different geographic isolates and on strains isolated from different hosts, or even from the same host, as has been demonstrated in some comparative studies of genetic variability. In the present study, we evaluated the genetic variability of the worms that fully developed their endogenous cycle in immunised sheep and goat in order to identify the alleles of most immunoprotective value. To address these objectives, groups of sheep and goats were immunised with PBS soluble fractions enriched for cysteine proteinases from adult worms of H. contortus from either a strain of H. contortus isolated from goats of Gran Canaria Island (SP) or a strain isolated from sheep of North America (NA). The results confirmed the immunoprophylactic value of this type of enzyme against haemonchosis in both sheep and goats in association with increased levels of specific IgG. The genetic analysis demonstrated that the immunisation had a genetic selection on proteinase-encoding genes. In all the immunised animals, allelic frequencies were statistically different from those observed in non-immunised control animals in the four analysed genes. The reduction in the allelic frequencies suggests that parasites expressing these proteases are selectively targeted by the vaccine, and hence they should be considered in any subunit vaccine approach to control haemonchosis in small ruminants., (Copyright © 2015 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.)

The aim of this study was to evaluate the nitric oxide (NO) levels, and oxidative and antioxidant markers of lambs experimentally and naturally infected by Haemonchus contortus, and its relation to lesions in the abomasum. For experimental study, a total of 14 healthy lambs were divided into two groups with seven animals each. Group A represented the uninfected animals (control), and Group B was formed by infected animals with 15,000 larvae of H. contortus. Blood was collected on days 15, 45, and 75 post-infection (PI) to obtain serum for biochemical analysis: lactate dehydrogenase (LDH), nitrite/nitrate (NOx), advanced oxidation protein products (AOPP), and ferric reducing ability of plasma (FRAP). Parasitological stool examination (eggs per gram of feces--EPG) was performed on days 15, 45, and 75 PI to verify the evolution of the infection. On day 15 PI EPG was negative, but on days 45 and 75 PI the EPG was positive for animals from Group B. In the three periods evaluated it was observed an increase of LDH levels in serum of lambs infected with gastrointestinal nematodes, but on the other hand NOx levels were reduced on the same periods in infected animals. The AOPP and FRAP levels did not differ between groups on days 15 and 45 PI, but increased significantly on day 75 PI in infected lambs. The same variables were studied in 10 lambs naturally infected with helminths, where more than 97% corresponded to H. contortus (hematocrit and EPG values were 18.8 ± 2.5% and 7120 ± 2940, respectively). Similar to the experimental study, the levels of NOx reduced, and the levels of LDH, FRAP, and AOPP increased in serum of this animal associated inflammatory infiltrate in the mucosa of the abomasum. Therefore, during the infection by H. contortus it was observed alterations in oxidative markers, indicators of cell lesion confirmed by histological examination of the abomasum, and consequently there were changes in antioxidant levels, with the purpose of cell protection. We also conclude that helminth infection interferes with the nitric oxide metabolism., (Copyright © 2014 Elsevier GmbH. All rights reserved.)

The aim of this study was to evaluate the purine levels of lambs experimentally infected with Haemonchus contortus. A total of 12 healthy lambs were divided into two groups, composed of 6 animals each: Group A represented the healthy animals (uninfected), while in Group B the animals were infected with 15 000 larvae of H. contortus. Blood was drawn on days 15, 45 and 75 post-infection (PI) in order to perform the purine analysis (ATP, ADP, AMP, adenosine, inosine, hypoxanthine, xanthine and uric acid) by high pressure liquid chromatography (HPLC) in serum. On day 15 PI a significant (P<0·05) increase in the levels of ATP and inosine was observed in the infected animals, unlike the levels of ADP, adenosine, xanthine and uric acid which were reduced. On day 45 PI a significant (P<0·05) increase in the ATP and xanthine levels in infected animals was observed, contrasting with reduced levels of ADP and uric acid. Finally, on day 75 PI an increase occurred in the levels of ATP, adenosine and hypoxanthine in infected lambs, concomitant with a reduction in the levels of ADP and uric acid (P<0·05). These changes in purine levels may influence the inflammatory process and the pathological events.

The aim of this study was to evaluate the acetylcholinesterase (AChE) activity in lymphocytes of lambs experimentally infected by Haemonchus contortus. A total of 14 healthy lambs were used, divided into two groups of seven animals each. Group A (negative control) represented the uninfected animals, and Group B (positive control) was formed by animals infected with 15,000 larvae of H. contortus. Blood was drawn on the days 15, 45 and 75 post-infection (PI) in order to perform the white blood cells (WBC) count, as well as the evaluation of AChE activity in lymphocytes. Parasitological stool exam (eggs per gram of feces - EPG) was performed on the same days to follow up the evolution of the infection. On day 15 PI it was verified negative EPG; however, on days 45 and 75 PI it was observed positive EPG only in the animals of group B. In the three evaluated periods was observed a lower number of leukocytes, associated with decreased lymphocytes and neutrophils in lambs infected by this gastrointestinal nematodes. Lambs infected with H. contortus showed significant (P<0.01) lower AChE activity in lymphocytes compared uninfected. Statistically, there was a positive correlation (P<0.05) between AChE activity in lymphocytes and number of lymphocytes (r=0.69). The lymphocytes are cells with direct participation in the cholinergic system; therefore, based on these results, it can be concluded that the experimental infection with H. contortus influences the number of lymphocytes, and consequently the AChE activity in these cells., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The aim of the study was to compare the in vivo anthelmintic activity of sainfoin hay (Onobrychis viciifolia) and carob pod meal (Ceratonia siliqua) against gastrointestinal nematodes. Seven days before infection, 64 naive lambs were assigned to four different groups: Group S received sainfoin hay and group CAR was fed with carob pods. The remaining lambs received lucerne hay (Medicago sativa) and were assigned to positive (non-treated, NT) and negative (treated, T) control groups (treatment with albendazole). On day 0, lambs were artificially trickle infected for 6 weeks, with a mixture of infective larvae of Haemonchus contortus and Trichostrongylus colubriformis. Parasitological and pathophysiological parameters were measured repeatedly during the 2-month study. Compared to the NT group, decreases in egg excretion were observed in the CAR and S groups with significant differences only found for sainfoin (p < 0.05). At necropsy, group S showed decreases in the total worm numbers of both nematode species with significant differences for H. contortus. In contrast, no differences were noticed for the CAR group. Compared to the NT group, lower values for fecundity of female H. contortus were found in the S and CAR groups, however differences were non-significant. No differences in body weight gains were found between groups. Consistent results were found showing significantly higher packed cell volume (PCV) values in the T and S groups compared to NT and CAR groups. Overall, these results confirm a positive effect associated with the feeding of lambs with tanniniferous resources on host resilience (PCV values) and against gastrointestinal parasitic nematodes by affecting some biological traits of worm populations (e.g. eggs per gram of faeces and worm numbers). However, the anthelmintic effects differed between the two tannin-containing resources, which might be associated with the quantity and/or quality of secondary metabolites (condensed tannins and/or other polyphenols)., (© C. Arroyo-Lopez et al., published by EDP Sciences, 2014.)

The aim of this study was to evaluate adenosine deaminase (E-ADA) activity in sera of lambs experimentally infected with Haemonchus contortus. We used 12 lambs divided into 2 groups; Group A had 5 healthy, non-infected animals (control) and Group B had 7 healthy animals infected with H. contortus . Lambs were infected orally with 500 larvae (L3) per animal every 2 days, for a period of 20 days, and later the infection was confirmed by examination of feces (eggs per gram [EPG] via fecal egg count). Blood collection was performed at days 0, 20, 40, 60, and 80 post-infection (PI) for analysis of E-ADA activity. Animals in Group A showed negative EPG throughout the experiment unlike those from Group B that had elevated EPG counts. E-ADA activity was reduced in the serum of animals infected with H. contortus when compared to non-infected controls at days 20, 40, 60, and 80 PI. Therefore, it is concluded that infection with H. contortus influences the E-ADA activity in lambs.

The objective of this study was to compare the effects of the type of Haemonchus contortus experimental infection (trickle infection, TI versus single infection, SI) on feed intake, nutrients digestibility, parasitological and haematological measures, and plasma leptin in Creole kids. The animals were infected over 2 periods (challenge 1 and challenge 2) of 6 weeks each, corresponding respectively to the primary and the secondary infection. Periods prior infection (1 week each) were considered as controls. The primary infection was realized with 35 Creole kids (18.40±3.76 kg BW) housed in individual boxes and fed a hay-based diet. The secondary infection continued with 29 kids (21.90±3.40 kg BW) from the initial 35. A total of 6 kids and 8 kids were slaughtered for measuring nematode burden at the end of the primary and the secondary infection, respectively. Measurements of nutrients digestibility were made at 0, 3 and 5 weeks post-infection for both challenges. Faecal egg count (FEC), blood eosinophilia and packed cell volume (PCV) were monitored weekly. Feed intake (dry matter intake, DMI) and nutrients digestibility were negatively affected by H. contortus infection only during the primary infection. Plasma leptin changed significantly over time (P=0.0002) but was not affected by the infection type. Effect of infection type was observed only on crude protein digestibility during the primary infection, which was lower in the TI group (P<0.01). The overall level of blood eosinophilia was significantly higher in the TI group (P<0.0001) during both challenges. The overall FEC mean was significantly higher in the SI compared with the TI groups, during both challenges (P<0.02). These results were related to the mean female length significantly higher in the SI group compared with the TI group during challenge 1 (P=0.004), and the number of adult nematode significantly lower in the TI group compared with the SI group during the challenge 2 (P=0.05). The results showed that the response of Creole kids to H. contortus experimental infection was in part dependent on the type of experimental infection. Our data suggest that plasma leptin would not be involved in the response of Creole kids against H. contortus infection, as no relationship between its plasma level and the transient reduction in voluntary feed intake observed in both groups during the primary infection was observed., (Copyright © 2012 Elsevier B.V. All rights reserved.)

This study was carried out to evaluate the relationships of cellular changes in the abomasal mucosa and parasitological parameters, by comparing resistant and susceptible young Creole goats (kids) after experimental infection with Haemonchus contortus. The kids were infected over 2 periods (challenges 1 and 2) of 7 and 6 weeks, respectively. Fecal egg count (FEC), blood eosinophilia, packed cell volume (PCV), and body weight were weekly monitored. At the end of both challenges a subgroup of kids was slaughtered for nematode burden measurements and analysis of inflammatory cell infiltration in the abomasal mucosa. The average daily gain was higher in resistant kids after both challenges. Blood eosinophilia and FEC were higher in susceptible kids after both challenges. The number of immature worms and the means of female length were lower after challenge 2 whatever the genetic status. No differences were observed in the eosinophil and mononuclear cell infiltration between challenges 1 and 2 and resistant and susceptible kids. Globule leukocyte infiltration was found higher after the challenge 1 in resistant kids. This effect of the genetic status on globule leukocytes counts but not on the other inflammatory cell highlights the need for further study on the functional activity of these cell populations.

A preliminary analysis of the significance of genetic diversity in cysteine proteinase genes has been performed simultaneously in sheep and goats, with regard to the immunological control using these enzymes against haemonchosis. For this purpose, we have studied the cross-immunoprotective effect of cysteine protease-enriched protein fractions (CPFs) in adult worms of two Haemonchus contortus strains from North America and Spain that are adapted to sheep and goats, respectively. Previous genetic analysis of cysteine proteinase genes in both strains has shown that some of loci are polymorphic and these differences are translated into changes in the amino acid sequences. However, our results show that CPFs from H. contortus adult worms have a protective effect against the parasite in both sheep and goats. These results are similar regardless of whether they were obtained from sheep or goat-adapted H. contortus strains, which could be very important in case H. contortus CPFs were commercially used in different countries, as vaccines to prevent the negative effects of this parasite. Interestingly, this experimental inoculation of both species with a heterologous strain of H. contortus contributes to the idea shown in previous studies about how difficult is the interpretation and the comparison of vaccination where strains not adapted to a specific host are used. Therefore, the challenger of using heterologous strains could provide similar results to those observed in immunised animals. This study suggests the possibility of exploring the mechanisms involved in natural protection against non-adapted strains, in order to develop strategies to control haemonchosis., (Copyright © 2012 Elsevier B.V. All rights reserved.)

A pharmaco-parasitological assessment of four different albendazole (ABZ) formulations was carried out in lambs infected with multiple resistant gastrointestinal (GI) nematodes. The comparative drug systemic exposure profiles (ABZ sulphoxide plasma concentrations) and anthelmintic efficacies (clinical endpoint measured through the faecal nematode eggs reduction counts) were determined for a reference formulation (RF) and three different test (T1, T2, T3) generic ABZ preparations. Fifty (50) Corriedale lambs naturally infected with multiple resistant GI nematodes were allocated into five experimental groups (n = 10). Animals in each group received treatment with either the RF, one of the test ABZ formulations (5 mg/kg by the intraruminal route) or were kept as untreated control. Blood samples were collected over 48 h post-treatment. ABZ parent drug was not recovered in the bloodstream. The ABZ sulphoxide (ABZSO) and sulphone (ABZSO(2) ) metabolites were measured in plasma by ultraviolet high-performance liquid chromatography over 36-48 h post-treatment. A faecal nematode egg count reduction test (FECRT) was performed at day 10th post-treatment to lambs from all treated and untreated groups, which indicated the predominance of nematodes with high level of resistance to ABZ. Both ABZSO C(max) and AUC(0-LOQ) values obtained for the RF (pioneer product) were significantly higher (P < 0.05) than those obtained for the T1 and T3 preparations. Based on the currently available bioequivalence criteria, the test (generic) ABZ formulations under evaluation could not be considered equivalent to the RF regarding the rate (C(max) ) and extent (AUC(0-LOD) ) of drug absorption (indirectly estimated through the ABZSO metabolite). A large variation in nematode egg counts did not permit to obtain statistically significant differences among formulations. However, a favourable trend in the efficacy against the most resistant nematodes was observed for the formulations with the highest ABZSO systemic exposure., (© 2011 Blackwell Publishing Ltd.)

This experiment aimed to evaluate the effect of periparturient metabolizable protein (MP) nutrition on resistance to Haemonchus contortus in single rearing Ile de France and Santa Ines ewes. The restriction-fed iso-energetic diet was calculated to provide either 0.8 (low MP diet) or 1.3 (high MP diet) times MP, from three weeks before parturition until eight weeks into lactation. The ewes were experimentally infected with 1000 H. contortus infective larvae (L3) three times a week (Mondays, Wednesdays and Fridays), starting five weeks before the predicted date for parturition until a total of 15,000 L3 had been administered. While both breeds showed elevated fecal egg counts (FEC), these values were significantly lower for Santa Ines ewes than Ile de France ewes, but were independent of level of MP feeding. The latter also did not affect lamb weight gain and ewe body weight variation in each breed. Packed cell volume and total plasma protein for Santa Ines in all periods were significantly higher than those for Ile de France ewes (P<0.01) but were not affected by nutrition. In contrast, levels of serum IgG and IgA antibodies against somatic H. contortus infective larvae and adult antigens were similar between breeds but higher in animals that received high MP diets (P<0.05). The reduced body score of ewes at the beginning of the experiment probably influenced their high susceptibility to incoming larvae. Since, unexpectedly, MP scarcity was not achieved in this experiment, our data support the view that Santa Ines ewes are more resistant to H. contortus than Ile de France ewes., (Copyright © 2011. Published by Elsevier B.V.)

The objective of this study was to assess the correlation among traits associated with resistance or susceptibility to Haemonchus contortus infestation and to evaluate the participation of the ovine major histocompatibility complex (MHC) in Pelibuey sheep, a prevalent breed in tropical and sub-tropical regions in Mexico and elsewhere. Association among the fecal egg count (FEC), blood packed cell volume (PCV), antibody (AB) levels, serum proteins (SP) and blood eosinophil count (EOS) was assessed in 52 lambs experimentally infected with H. contortus, and the participation of the MHC was evaluated using polymorphisms in three microsatellites, located at the class I (OMHC1) and class II (OLADRB1, OLADRB2) regions of the MHC. Spearman correlation analysis among the traits showed a negative association (P<0.01) between FEC and PCV (-0.35), EOS (-0.50), SP (-0.30) and AB (-0.57), and a positive correlation of antibodies with EOS (0.50). The homozygotes for the OMHC1-188 and OLADRB2-282 alleles were associated with a reduction in FEC (-813 and -551, respectively). Conversely, the OMHC1-200 and OMHC1-206 alleles were associated with an increase in FEC (1704 and 1008, respectively). Furthermore, the OLADRB1-482 allele was associated with an increase of 163 EOS by allele copy, while the OMHC1-200 allele showed a reduction of 95 EOS in homozygotes. The associations among microsatellite MHC loci and the remaining variables were not significant. These results reinforce the evidence that MHC polymorphisms have an important role in parasite resistance or susceptibility in Pelibuey sheep and could be used as genetic markers to assist selection and improve parasite resistance to H. contortus., (Copyright © 2010 Elsevier B.V. All rights reserved.)

This research was designed to determine if divergent selection for resistance to Haemonchus contortus had produced correlated changes in voluntary feed intake and diet selection. Voluntary feed intake, diet selection and production were determined in 54 Merino weaner rams from the CSIRO Haemonchus selection flock, increased resistance to Haemonchus (IRH), decreased resistance to Haemonchus (DRH) and random bred control (C) selection lines. Weaner rams were fed ad libitum either a high (9.2 MJ ME/kg DM, 90 g MP/kg DM) or moderate (6.3 MJ ME/kg DM, 30 g MP/kg DM) quality diet and given the choice between the two diets, when uninfected (NIL) or infected with H. contortus (INF). Symmetrical response to divergent selection for worm egg count (WEC) was not matched by a symmetrical change in feed intake and there was no difference in diet selection between selection lines. Feed intake, growth and wool production of DRH animals remained the same as that of IRH, yet DRH animals had five times greater WEC than IRH. This study begins to explain the mechanisms that allow resistant animals to effectively prevent establishment and/or development of H. contortus, by maintaining a greater immune response to infection through higher circulating eosinophils, plasma globulin and IgG(1) antibody titres. Susceptible animals have displayed resilience by improving feed conversion efficiency and increasing protein synthesis., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Test sensitivity and specificity for the FAMACHA(©) clinical test for anaemia due to haemonchosis have previously been shown to be adequate in differentiating between heavily/less heavily infected sheep, but these properties give no objective guidance for setting the optimum threshold at which anthelmintic treatment should occur. The aim of this work was to use Receiver Operating Characteristic (ROC) curves to evaluate the diagnostic accuracy of FAMACHA(©) testing by estimating the area under the ROC curve, and to use two-graph ROC curves to decrease subjectivity in selecting treatment thresholds on two farms with contrasting management. Test diagnostic accuracy, and thus discriminating power as determined by the area under the ROC curves, ranged from "moderate to good" on the first farm, and from "moderate to high" on the second farm for haematocrit (the Gold Standard for the test) cut-offs of ≤ 22% and ≤ 19% on both farms respectively. Accuracy of classification between haematocrit cut-offs was not significantly different within farms, but did differ significantly between farms, with test accuracy being highest on the second farm at both haematocrit cut-offs (p<0.05). The results also showed the suitability of the two-graph ROC curve approach for discriminating not only between different levels of accuracy of evaluators, but also to give an indication of the so-called ROC cut point (i.e. the desired threshold level) at which animals should be treated for a given level of risk of loss. The approach appears to have the potential not only to validate the diagnostic accuracy of the test across the complete testing range (i.e. all FAMACHA(©) categories from 1 to 5), but also to compensate for such inaccuracy by allowing objective adjustment of the threshold treatment level according to the output of the two-graph ROC method., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The benefit of long-term feeding of fresh or ensiled cassava foliage on gastrointestinal parasite in goats was evaluated. Eighteen male goats (15.15 +/- 2.83 kg and between 4-6 months) were randomly allocated into three treatments supplemented with 200 g of wheat bran head(-1) day(-1). All groups were fed ad-libitum on either grass (CO), fresh cassava (CaF) or ensiled cassava foliage (CaS). At the beginning of the trial, each goat was inoculated with 3000 L3 containing approximately 50% Haemonchus contortus. Individual LWt, FEC and PCV were measured at weekly intervals for 10 weeks. At the termination of the experiment all goats were slaughtered for worm recovery and enumeration. The goats in CaF and CaS had similar weight gains while those in CO lost weight (p < 0.05) through the trial. FEC in CaF and CaS were lower (p < 0.05) than CO during the patency of parasite infections, but there was no difference between CaF and CaS goats. PCV of all groups decreased from above 30% to around 25% at the end of the trial. The compositions of established worm burdens were mainly H. contortus (19-40%) and Trichostrongylus colubriformis (55-76%). TWB did not differ among the groups, however, CaS significantly reduced H. contortus burdens, as compared to CaF and CO (p < or = 0.005). Thus, ensiled cassava foliage reduced the H. contortus population while the fresh foliage only reduced worm fecundity.

The objective of this study was to evaluate the relationship of parasite-specific serum antibodies with the resistance status of Creole kids. The average breeding values on egg output predicted in a context of natural infection at 11 months of age were distant of 1.07 genetic standard deviation between resistant and susceptible animals. After drenching the animals were maintained worm-free during 1 month until experimental infection with 10,000 Haemonchus contortus infective larvae (L3). Enzyme-linked immunosorbent assay was carried out in serum samples to determine the level of IgG, IgA and IgE anti-H. contortus L3 crude extracts and adult excretion/secretion products (ESP). Parasitological and blood immunological parameters were measured on the 2 extreme groups. Despite the absence of any typical signs of haemonchosis, susceptible kids had more than 11 times higher faecal egg counts (FEC) at 35 days post-infection (d.p.i.) than resistant kids had. Levels of immunoglobulin against H. contortus L3 and ESP increased significantly after infection in both groups. However, no difference in the host immune response mediated by immunoglobulin against H. contortus was evidenced between groups. This finding suggests that, in goats previously infected by H. contortus, a degree of protection occurred and the phenotypic and genetic segregation in resistant and susceptible animals were not related to the humoral immune response. The correlation coefficients between FEC and IgE anti-ESP (r=0.593; P<0.05 was significant in both resistant and susceptible animals. Such correlation suggesting a hypersensitivity reaction dependent on worm prolificacy has never been described. This result needs further studies to understand the mechanisms underlying this observation.

The study was conducted to examine the occurrence and interaction between Oestrus ovis and Haemonchus contortus in experimentally infected Ethiopian Afar breed of goats. Twenty goats were divided into four groups (O, OH, H, and C) of five animals each. Each animal of groups O and OH received weekly infections for 5 weeks with 66 first instar larvae (L1) of O. ovis. Then animals of groups OH and H were infected with a single dose of 5000 third stage larvae (L3) of H. contortus. Goats of group C were kept free of any infection as non-infected control. Faecal egg count (FEC), blood cell count, total serum protein level and body weight were recorded weekly throughout the study period. At necropsy worm burden, female worm length, fecundity and larval burden of O. ovis in the nasal-sinus cavities of infected animals were assessed. The results showed that the presence of H. contortus in the abomasum of goats of group OH had no influence on the development of O. ovis. On the contrary, a significant reduction (P<0.05) in FEC, worm burden, fecundity and female worm length was revealed in group OH animals compared to the mono-infected animals (group H). This was associated with eosinophilia and reduced packed cell volume.

The gastrintestinal helmintiases occupy prominence place among factors that limit the farming goat in entire world, especially on tropical regions, where economical losses are more accentuated. The objective of this work was to evaluate FAMACHA© method utilization as an auxiliary strategy on the control of parasites of goats in semiarid paraibano. Moreover the clinical assistance, values of hematocrit and OPG were determinate to evaluate the relation between the exams. They were used 30 moxotó female goats, monthly evaluated for 90 days. The female goats that presented degree three of mucose in accordance to the FAMACHA© method do not need to be vermifuged in accordance with hematocrit and OPG values. The FAMACHA© method, although too indicated for use in goats, needs little adaptation to be best administered.

From May to July 2005 a study was conducted on the prevalence and effects of haemonchosis in goats farmed on communal grazing belonging to resource-poor farmers at two study sites, namely Mlungisi (26 degrees 53'E, 31 degrees 53'S) and Ezibeleni (26 degrees 58'E, 31 degrees 53'S), situated within the Chris Hani District Municipality in Eastern Cape Province of South Africa. The usefulness of the FAMACHA system for identifying goats needing treatment was confirmed. It allowed stock owners to identify badly affected goats accurately, as borne out by the agreement between FAMACHA scores and corresponding haematocrits.

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.

Gastrointestinal nematode control has been performed through use of anthelmintics. However, the development of resistant populations has required research into new alternatives. There are popular reports about anti-parasitic activity of Azadirachta indica in animals and plants. The aim of this study was to evaluate the anthelmintic activity of A. indica after feeding sheep with the dried leaves. In this experiment, 40 sheep were allotted into four treatment groups. Group I received a treatment of A. indica dry leaves mixed in a concentrate at a rate of 0.1 g/kg dose for 3 months. Group II was treated with double the dose of Group I. Group III was treated with closantel (Diantel) at the manufacturer-recommended dose once at the beginning of the study and Group IV was not treated. To compare treatment effects, the following parameters were evaluated: egg count per gram of feces (EPG), worm burden, weight gain and haematocrit. EPG and worm burden results were statistically evaluated using the Kruskal-Wallis test. Haematocrit and live weight gain were submitted to analysis of variance (ANOVA) and the means evaluated by Tukey's test with 95% probability. None of the evaluated parameters of the treatment groups were statistically different when compared to the control group, demonstrating that, with the protocol used, A. indica has no anthelmintic effect.

The effects of cold storage of infective third-stage larvae (L3) of different isolates of the parasitic nematode Haemonchus contortus were studied with respect to infectivity, pre-patent period and propensity for larval arrestment. Two complementary experiments were conducted with 2 groups of lambs, each animal being inoculated with 2000 L3 of either Swedish or Kenyan origin. In a first experiment, L3s were cold treated at 5 degrees C for 9 months prior to infection, whereas in a second experiment larvae were newly hatched. Individual faecal egg counts (FECs), and worm burdens were determined for each experiment. The results showed that the greatest differences were associated with the pre-treatment of larvae. The pre-patent period and the FECs differed significantly between the experiments but not between the isolates used in each experiment. However, the extent of hypobiosis was significantly different between the two isolates when fresh larvae were used (36% Kenyan isolate and 70% Swedish). The storage of H. contortus at 5 degrees C had no apparent effect on the infectivity of L3s, as high establishment ranging from 43 to 74% were observed, irrespective of isolates used. This study showed that H. contortus exhibited similar phenotypic traits regardless of geographical origin. Thus, there was limited evidence for adaptations to temperate climatic conditions.

Gastrointestinal nematodes are of concern in sheep production because of production and economic losses. Control of these nematodes is primarily based on the use of anthelmintic treatment and pasture management. The almost exclusive use of anthelmintic treatment has resulted in development of anthelmintic resistance which has led to the need for other parasite control options to be explored. The blood sucking abomasal parasitic nematode Haemonchus contortus causes severe losses in small ruminant production in the warm, humid sub-tropic and tropics. This study evaluated the effectiveness of a nematode trapping fungus, Duddingtonia flagrans, in reducing availability of parasitic nematode larvae, specifically H. contortus, on pasture. Chlamydospores of D. flagrans were mixed with a supplement feed which was fed daily to a group of crossbred ewes for the duration of the summer grazing season. A control group was fed the same supplement feed without chlamydospores. A reduction in infective larval numbers was observed in fecal cultures of the fungus-fed group. Herbage samples from the pasture grazed by the fungus-fed group also showed a reduction in infective larvae. There were no significant (P > 0.05) differences in overall fecal egg count, packed cell volume or animal weight between fungus-fed and control groups. Tracer animals were placed on the study pastures at the end of the study to assess pasture infectivity. Although tracer animals were only two per group, those that grazed with the fungus-fed group had substantially reduced (96.8%) nematode burdens as compared to those from the control group pasture. Results demonstrated that the fungus did have activity against nematode larvae in the feces which reduced pasture infectivity and subsequently nematode burdens in tracer animals. This study showed that D. flagrans, fed daily to grazing ewes, was an effective biological control agent in reducing a predominantly H. contortus larval population on pasture.

Forty pure West African Dwarf (WAD) goats and 35 of its F1 crosses with the Sahelian breed were used in a multifactorial experimental design to evaluate the effects of an experimental Trypanosoma congolense infection and interactions with natural helminth infections and different levels of diet on health and productivity of these two breeds. Trypanosome infection caused a severe drop in packed cell volume (PCV), but this was not significantly affected by breed. Neither deworming nor diet had any effect on the course of anaemia after trypanosome infection. The mean score of parasitaemia tended to be higher in crossbreeds than in WAD goats although this was not significant (P>0.05). Similarly, the antibody response to trypanosome infection was not significantly different between breeds. Parasitaemia level was significantly influenced by the level of diet with the group under high supplementation having a higher mean parasitaemia score than the group under low supplementation. Weight loss due to trypanosome infection tended to be greater in crossbreeds than in WAD goats (P>0.05). During this study, there was no difference in mean helminth egg output between crossbred and WAD goats. However, between weeks 4 and 10 after trypanosome infection (corresponding to a period of heavy rainfall and highly infective pastures), the mean egg output was higher in the crossbreeds. The immunosuppressive effect of trypanosome infections was revealed by a lower antibody response to Haemonchus contortus in infected animals compared to the uninfected controls. Trypanosome infection tended to increase strongyle egg output. This study did no reveal any superior trypanotolerance of WAD goats compared to crossbreeds.

A longitudinal study was conducted on the differential faecal egg counts, haematocrits and body condition scores of sheep belonging to resource-poor farmers at Rust de Winter, Gauteng province, and Kraaipan, North West Province, South Africa. The animals were scored for level of anaemia using the FAMACHA method, an assay for the clinical evaluation of anaemia caused by Haemonchus spp. Periods of higher Haemonchus egg counts occurred from October to March for sheep at Rust de Winter and from September/October to February or April for sheep at Kraaipan. Lower haematocrit values were registered during these periods as was a higher incidence of anaemic conjunctival mucous membrane colour scores compared to the period April to September. No clear relationship between the faecal egg counts and the body condition scores was evident. Although wider application of the FAMACHA system in sheep raised by resource-poor farmers should be investigated, the present study indicates that this method may certainly prove to be a valuable worm control strategy for such livestock owners.

The diversity and predominance of nematode genera in goats of resource-poor farmers at Rust de Winter, Gauteng Province, Impendle, KwaZulu-Natal Province, and Kraaipan, North-West Province, South Africa, was determined by means of a longitudinal study of the nematode faecal egg counts (FECs) and differential third-stage nematode larvae. The animals were bled for haematocrit determination and scored for pallor of ocular mucous membranes using the FAMACHA( Copyright) method, an assay for clinical evaluation of anaemia caused by Haemonchus spp. Animals considered to be in danger of dying from anaemia caused by haemonchosis were selectively treated with an anthelmintic. Lower haematocrit values were registered during periods of heavier Haemonchus infection, which occurred from December/January to March for Rust de Winter; from December to March/April for Impendle; and from November/December to February or April for Kraaipan. There was agreement too between the lower haematocrits and paler mucous membranes scored according to the FAMACHA( Copyright) method. The use of this system may be recommended as part of an integrated approach to worm control in goats kept in the resource-poor areas studied.

One hundred faecal specimens and corresponding blood samples were evaluated for Haemonchus contortus infection in sheep by ELISA utilizing somatic, circulating and coproantigens. Results proved that coproantigen was more potent than the others in diagnosis of sheep haemonchosis. SDS-polyacrylamide gel electrophoresis demonstrated that the three antigens are structurally related. Immunoblot analysis revealed that somatic antigen contained 6 polypeptides of molecular weights 234, 205, 178, 83, 48 and 14 kDa, while, coproantigen showed 4 immunogenic polypeptides of 234, 205, 85 and 45 kDa. In circulating antigen only 2 polypeptides of molecular weights 165 and 99 kDa were immunoreactive. The potency of coproantigen may be attributed partially to one polypeptide of molecular weight 45 kDa. This low molecular weight polypeptide was only expressed in coproantigen as an immunogen, probably increases the immunogenicity, and may have potential diagnostic and protective values.

Trials were conducted on a farm in Mpumalanga Province in South Africa to test the possibility of grading the colour of the ocular mucous membranes of sheep as an indication of the extent to which the animals are affected by Haemonchus contortus infection. The range of observed colour shades were classified into five categories, from red, through red-pink, pink and pink-white to white. Over a period of 125 days routine drenching of a flock of 388 sheep on irrigated kikuyu (Pennisetum clandestinum) pasture was terminated. During this time the animals were examined at practically weekly intervals and haematocrit determinations done for all the sheep with pale conjunctivae. Only those sheep having a haematocrit of 15% or lower were treated. Compared to a previous drenching tempo of close to every 3 weeks during the Haemonchus season on the farm, drenching was reduced by approximately 90%, as 70% of the sheep did not require salvage drenching and only 10% of the flock had to be given more than one salvage treatment. At the time of the trial the five clinical classifications were not related to predetermined haematocrit categories. However, when compared to categories that were set in later trials, 94% of the clinical estimates in the present trial were either in the correct haematocrit category, or, if not, the sheep were probably not disadvantaged by the errors. In 2.6% of cases the incorrect estimate may have placed the sheep concerned in jeopardy, as the haematocrit values were so low that salvage drenching was required, while the sheep were not regarded as anaemic. Changes in the mean haematocrit values of drenched and undrenched sheep were mirrored reciprocally by the changes in clinical colour estimates. Lactating ewes were by far the most susceptible class of sheep, as only 44.6% of them were able to manage without drenching, compared to 83% of dry, and 70.6% of pregnant ewes. Correlations between the haematocrits and clinical estimates were highly significant, although the associations were not high enough to give reasonable surety that the haematocrit values of individual animals could be predicted with confidence from their clinical classifications. Exceptionally large numbers of worms were recovered from seven of the 14 sheep that were culled because of age at the end of the trial, but these were reflected neither in their faecal worm egg counts, nor, with one exception only, in clinical signs.

The anthelmintic activity of Calotropis procera latex was investigated in sheep that had been infected with single oral doses of 12000 infective Haemonchus contortus larvae. Inappetence, dullness, erosive abomasitis, decreased haemoglobin concentration and increased eosinophils were the main features of haemonchosis in the sheep. In the sheep treated with single oral doses of 0.01 ml or 0.02 ml/kg body weight of C. procera latex, egg production was significantly reduced, but not completely suppressed, and fewer adult Haemonchus worms were found in the abomasum. Although the appetite improved, the haemoglobin concentration and serum copper, iron and zinc levels were still reduced after therapy with Calotropis latex. Calotropis latex showed a concentration-dependent larvicidal activity in vitro within 20 min of application.

One option for controlling haemonchosis in warm pastoral regions is improvement of resistance by selective breeding. Variation in acquired immunity to H. contortus and immunological correlates of infection were studied in West African Dwarf (WAD) goats. Following exposure to 5000 L3, 63 per cent of the inoculum established but 77 per cent of established worms were expelled by week 5. All infected animals were anaemic (day 14). When exposed to 2000L3, 36 per cent of the inoculum was still present (day 35) with no loss by day 49. Persisting primary infection worms survived a superimposed challenge (day 35), but their growth was slowed and resistance to challenge was significant. Most goats showed eosinophilia and parasite-specific IgG responses to primary infection, but only eosinophilia increased after challenge. No consistent associations were found between parasite burden and any immunological measures of infection, but parasite egg counts showed considerable variation. Overall, our results suggest that resistant genotypes exist among the WAD goat population., (Copyright 1999 W.B. Saunders Company Ltd.)

A study was made of the benefits of protein supplementation for parasitised and non-parasitised lambs. Sixty, 5-month-old crossbred wether lambs were placed in individual pens indoors for 9 weeks. Half of the animals were experimentally dosed with 1500 Haemonchus contortus larvae per head per week and were fed ad libitum and the other half were worm-free, pair-fed controls. Diets were formulated to be iso-energetic (9.0 MJ of calculated metabolisable energy per kg dry matter) with five levels of protein (10, 13, 16, 19 and 22% crude protein). These diets were based on oaten chaff, with barley, cotton-seed meal, urea and mineral mix (except for the 22% crude protein diet which did not contain barley). Dietary crude protein content increased live-weight gain, feed intake, rumen fluid ammonia-N, packed cell volume, eosinophil counts and antibody responses to H. contortus L3 antigen and decreased faecal worm egg counts significantly. Infection did not significantly affect packed cell volume of animals on diets with 16, 19 and 22% crude protein content. We conclude that extra dietary protein can prevent the adverse effects of H. contortus infection on animal production.

A total of 70 strongyle-free French Alpine dairy goats were exposed to a combination of sequential and challenge infections with Haemonchus contortus and Trichostrongylus colubriformis third-stage larvae. The sequential infection consisted of three inoculations at 50-day intervals, each infection being abbreviated by anthelmintic treatment at 40 days postinoculation. The challenge infection, composed of the same nematode strains, was undertaken 2 months later, when goats were at their 1st month of lactation. Fecal egg counts (FECs), packed cell volumes (PCVs), pepsinogen concentrations, inorganic phosphate concentrations, and peripheral eosinophil numbers were measured at 30-40 days after each inoculation. Goats were defined as being resistant or susceptible according to their level of nematode egg output following the first inoculation. Significant differences in FECs were recorded between the two groups throughout the further inoculations and the challenge infection. The reliability of FECs was supported by the high repeatability values found within and between infections. With regard to blood constituents, only PCVs related to H. contortus infection showed values that differed significantly between the two groups, resistant goats having higher PCVs after the first and the third inoculations than did susceptible animals. However, this difference was not detectable after the challenge infection. The milk production yield for the current lactation was significantly lower in the resistant goats. Moreover, resistant animals exhibited constantly greater body condition scores as compared with susceptible animals. These results indicate that the individual responsiveness of dairy goats to experimental nematode infection can be estimated on the basis of FECs and PCVs (for H. contortus) and is negatively related to the level of milk production of the animals.

Two cases of haemonchosis occurred in the Spanish ibex (Capra pyrenaica) population of the National Hunting Reserve of Tortosa and Beseit, northeastern Spain, in July 1992 and May 1993. The animals were cachectic and recumbent and had an acute hemorrhagic, macrocytic and hypochromic anemia; which was related to a high infection of 2,016 and 1,863 Haemonchus contortus, respectively, in the abomasum of the animals.

As part of a systematic examination of the protective epitopes on H11, groups of sheep were vaccinated with preparations of purified H11 used untreated (group A), or progressively denatured (linearized) by incubation with sodium dodecyl sulphate (SDS) (group B) or by boiling with SDS in the presence of dithiothreitol (group C). All the sheep developed antibodies which bound to the untreated H11. When challenged with 10,000 infective larvae of Haemonchus contortus the mean levels of protection relative to the mean values for adjuvant controls were 99.8%, 85% and 79% for faecal egg counts and 95%, 79% and 54% for worm burden at post-mortem for groups A, B and C respectively. The H11-specific antibodies inhibited the microsomal aminopeptidase activity of H11 in vitro up to 80%. The levels of inhibition by sera from individual animals correlated with levels of protection with r2, of 0.69-0.87.

The influence of gastrointestinal nematode infection on the kinetics of albendazole (ABZ) and its metabolites, albendazole sulphoxide (ABZSO) and sulphone (ABZSO2) in plasma and abomasal fluid was investigated in sheep. A micronised suspension of ABZ was administered intraruminally at 7.5 mg kg-1 to the following groups of sheep: (a) non-parasitised (control); (b) artificially infected with Haemonchus contortus; (c) naturally infected with Haemonchus contortus and other species of gastrointestinal nematodes. Plasma and abomasal fluid samples were obtained serially over 72 h post-treatment and they were analysed by HPLC for ABZ and its metabolites. The ABZ parent drug was not detected in plasma at any time post-treatment, however the metabolites ABZSO and ABZSO2 were recovered in the bloodstream. The active metabolite ABZSO was recovered in plasma between 0.5 and 48 (uninfected), 60 (H. contortus infected) or 72 h (naturally infected sheep) post-administration. The area under the plasma concentration vs time curve (AUC) values for ABZSO were higher in both artificially infected (64.0 micrograms h ml-1) and naturally infected (79.3 micrograms h ml-1) sheep as compared with non-infected animals (41.8 micrograms h ml-1). Peak plasma concentrations for ABZSO and ABZSO2 were higher in both artificially and naturally infected sheep than in non-parasitised animals. No changes in the half-lives and mean residence times for these metabolites were observed in infected sheep. ABZ and its metabolites were found in the abomasum between 0.5 and 48 (infected animals) or 72 h (uninfected) post-treatment. The availability (total AUCs) of ABZ and its metabolites in abomasal fluid were lower in H. contortus infected sheep than in the uninfected control animals. The increased abomasal pH induced by the presence of the H. contortus infection may reduce the plasma/abomasum pH gradient, which results in a decreased ionic-trapping of ABZ and its metabolites in the abomasum. Such a phenomenon correlates with: (a) the higher total AUC values obtained for ABZ metabolites in the bloodstream of the infected compared to the control sheep, (b) the lower concentration profiles of the ABZ parent drug and its metabolites found in the abomasal fluid of the infected animals.

A total of 46 weaned kids of three genotypes aged about 4-5 months were used to evaluate the effects of trickle infection with a sheep strain of Haemonchus contortus. A completely randomized 3 x 2 factorial design was used. Factors were genotype (Thai native (TN), 75% TN x 25% Anglo-Nubian (AN) and 50% TN x 50% AN) and parasite (control and infected). The animals were infected with 750 infective larvae (L3) of H. contortus three times a week for 3 weeks, with a total of 6750 larvae. The experiment lasted 9 weeks. Each week animals were weighed, faecal egg counts done and blood examined for haematological and biochemical variables. Twenty-seven kids were slaughtered at the end of experiment for worm recovery. Weight gain of infected animals was lower than those of uninfected controls (P < 0.05). The genotype 50% TN x 50% AN had higher growth rate than TN and 75% TN x 25% AN genotypes (P < 0.05). Eggs per gram of faeces (EPG) were significantly higher in 50% TN x 50% AN kids than in TN (P < 0.0005) and 75% TN x 25% AN (P < 0.0001) kids. There was a large variation in the EPG of individual animals within a genotype. The percent establishment of L3 was 8.2% in TN, 16.97% in 50% TN x 50% AN and 17.91% in 75% TN x 25% AN kids. TN kids had worm counts lower than 50% TN x 50% AN (P < 0.05) and 75% TN x 25% AN (P = 0.07) kids. Infection had a significant effect on packed cell volume (PCV), haemoglobin, total protein and albumin. The decrease in the level of these blood parameters was less in TN kids than in 50% TN x 50% AN and 75% TN x 25% AN kids. There was no significant difference between genotypes in the values of total and differential leucocyte counts and mean corpuscular volume (MCV). It can be concluded that TN goats are more resistant to H. contortus than 50% TN x 50% AN goats. However, under the present experimental conditions, the liveweight gain of 50% TN x 50% AN was higher than the TN goats.

Six-month-old red Maasai lambs were more resistant than Dorper lambs to repeated infections at one to two week intervals with 1000 Haemonchus contortus infective larvae. Resistance after infection was assessed by means of faecal egg counts, packed cell volumes, eosinophil counts, total serum protein concentrations and mortality rates. The weight gains of the infected animals were only marginally lower than those of their uninfected controls, most probably because of their significantly higher feed consumption, and evidently the infected lambs were not utilising all of the extra feed for growth. This absence of anorexia in spite of the infection was probably due to the palatability of the high protein diet fed to the lambs.

Twenty helminth-naive Scottish Blackface lambs were given three infections with 10,000 infective larvae of Haemonchus contortus at 8 week intervals. An additional six lambs served as uninfected controls and eight lambs were infectivity controls. The lambs were 7 months old at the start of the infection. Four of the 20 lambs developed severe haemonchosis and were put down during the experiment. The remaining 16 lambs plus uninfected controls were necropsied 8 weeks after the third infection. The mean faecal egg count peaked 6-8 weeks after the first infection, gave a second smaller peak 6-8 weeks after the second infection but produced no peak after the third infection. Mean red blood cell counts fell rapidly during the first infection, then rose gradually during the second and third infections. The mean values suggested that two infections were sufficient to produce effective immunity in the sheep population but they masked considerable individual variation. Eleven animals appeared relatively resistant following the first infection, while two animals were relatively susceptible to even the third infection. The repeatability of mean faecal egg counts or mean red blood cell counts for each animal during the replicate infections were very high, because the rankings of the individual sheep remained remarkably stable. Faecal egg counts were very strongly correlated with red blood cell counts. Multiple regression analysis showed that four factors--faecal egg counts, red blood cell counts, weight and sex--accounted for essentially all of the observed variation in worm burdens among the lambs. Therefore, under these controlled experimental conditions, additional markers appear unnecessary for the detection of resistance status.

The influence of dietary protein supplementation on resistance to haemonchosis was examined in Hampshire down lambs fed either a basal diet or a diet supplemented with soyabean. At seven months of age the lambs were challenged with an initial loading dose of Haemonchus contortus, followed by a trickle infection three times a week. Blood and faecal samples were collected three times a week and bodyweights were recorded weekly. After 10 weeks the lambs were slaughtered and their worm burdens and carcase composition determined. Although their mean worm burdens were similar, the lambs given the basal diet had higher faecal egg counts, lower packed red cell volumes and lower concentrations of total plasma protein and plasma albumin than the lambs given the supplemented diet. The dietary supplementation also improved the carcase composition of the lambs.

In 8 healthy, noninfected lambs with an initial body weight (b. w.) of 24.79 +/- 1.16 kg as well as in 3 groups of lambs (n = 8 per group) of a similar body weight after an inoculation with 3rd stage larvae of Haemonchus contortus (2500) and Trichostrongylus colubriformis (10,000) analyses of the blood plasma were performed for 45 days thereafter. On the 37th day after infection ivermectin (0.2 mg/kg b. w.) was administered per os respectively subcutaneously to the lambs of 2 groups. After 7 and 8 days the content of ascorbic acid in the liver, the spleen and the adrenals as well as that of vitamin A in the liver was analysed. The behaviour of the values influenced by a moderate infection with trichostrongylid nematodes is discussed.

Some systemic responses to single-dose infection with 10,000 Haemonchus contortus infective larvae were examined in sheep already shown to have protective immunity against the parasite. The major haematological finding was a neutrophil leukocytosis that occurred after the infections became patent but not during the pre-patent period. There was no definitive eosinophilia and no discernible change in the erythrocyte parameters. Systemic hyperthermia was not conclusively evident during the pre-patent period. Enzyme-linked-immunosorbent assays (ELISA) were used to measure the secondary anti-helminth antibody response in serum during the pre-patent period when the establishment of patent infection is resisted. These ELISAs employed preparations from adult worms to represent the parasitic stages of the worm, preparations from infective larvae to represent the pre-parasitic stages of the worm, and exsheathing fluid, which is the soluble material obtained when H. contortus larvae undergo ecdysis and transform from the pre-parasitic to the parasitic phase. Antibody responses to the three preparations differed qualitatively, indicating the presence of three different but perhaps overlapping sets of antigens. The three peaks in antibody against exsheathing fluid may reflect the pulses of antigen delivered to sheep as the parasite undergoes its three moults within the host.

Weaned wether goats (n = 144) approximately 6 months of age were placed in a 2 x 3 factorial experiment to test the effects and interaction of two levels of nutrition (growth+maintenance, NUT1; and twice growth+maintenance, NUT2) and three levels of Haemonchus contortus burden (0, 500, and 2000 larvae administered every 2 weeks; W0, W500, and W2000, respectively) on packed cell volume, red blood cell count, total serum protein and leukocytes. The statistical analysis revealed clear and proportionate differences among levels of infection for all variables. A significant (P < 0.05) nutritional effect was also found associated with all the variables except leukocytes. Nutrition by worm load interactions were found for packed cell volume and leukocytes. The neutrophil/lymphocyte ratio was higher in the NUT1-infected animals, leading to the nutrition by worm load interaction for leukocytes. An analysis for the different leukocyte types revealed significant (P < 0.05) differences among infection levels for lymphocytes, while nutrition level was found to be a significant effect for basophil count and immature white cells. Several significant correlations were observed between pairs of variables. Faecal egg output could be predicted from actual worm count in three of the four necropsy periods. The clear differences observed for blood parameters were not present in production traits, suggesting that physiological thresholds may play an important role in framing the metabolic activity of biological organisms. Total serum protein was the best indicator of these effects on production parameters.