Innate Immune Responses Associated with Resistance against Haemonchus contortus in Morada Nova Sheep.

The immune response against Haemonchus contortus infections is primarily associated with the Th2 profile. However, the exact mechanisms associated with increased sheep resistance against this parasite remains poorly elucidated. The present study is aimed at evaluating mediators from the innate immune response in lambs of the Morada Nova Brazilian breed with contrasting H. contortus resistance phenotypes. Briefly, 287 lambs were characterized through fecal egg counts (FEC) and packed cell volume (PCV) after two independent experimental parasitic challenges with 4,000 H. contortus L3. 20 extreme resistance phenotypes (10 most resistant and 10 most susceptible) were selected, subjected to a third artificial infection with 4,000 L3, and euthanized 7 days later. Tissue samples were collected from abomasal fundic and pyloric mucosa and abomasal lymph nodes. Blood samples were collected at days 0 and 7 of the third parasitic challenge. RNA was extracted from tissue and blood samples for relative quantification of innate immune-related genes by RT-qPCR. For the abomasal fundic mucosa, increased TNFα and IL1ß expression levels (P < 0.05) were found in the susceptible animals, while resistant animals had IL33 superiorly expressed (P < 0.05). Higher levels (P < 0.05) of TLR2 and CFI were found in the abomasal pyloric mucosa of resistant animals. TNFα was at higher levels (P < 0.05) in the blood of susceptible lambs, at day 0 of the third artificial infection. The exacerbated proinflammatory response observed in susceptible animals, at both local and systemic levels, may be a consequence of high H. contortus parasitism. This hypothesis is corroborated by the higher blood levels of TNFα before the onset of infection, which probably remained elevated from the previous parasitic challenges. On the other hand, resistant lambs had an enhanced response mediated by TLR recognition and complement activation. Nevertheless, this is the first study to directly associate sheep parasitic resistance with IL33, an innate trigger of the Th2-polarized response.

Microbial community and ovine host response varies with early and late stages of Haemonchus contortus infection.

The interactions between gastric microbiota, ovine host, and Haemonchus contortus portray the ovine gastric environment as a complex ecosystem, where all factors play a pertinent role in fine-tuning each other and in haemeostasis. We delineated the impact of early and late Haemonchus infection on abomasal and ruminal microbial community, as well as the ovine host. Twelve, parasite-naive lambs were divided into four groups, 7 days post-infection (dpi) and time-matched uninfected-control groups; 50 dpi and time-matched uninfected control groups were used for the experiment. Six sheep were inoculated with 5000 H. contortus infective larvae and followed for 7 or 50 days with their corresponding uninfected-control ones. Ovine abomasal tissues were collected for histological analysis and gastric fluids were collected for PH value measurements, microbial community isolation and Illumina MiSeq platform and bioinformatic analysis. Our results showed that Haemonchus infection increased the abomasal gastric pH (P = 0.05) and resulted in necrotizing and inflammatory changes that were more severe during acute infection. Furthermore, infection increased the abomasal bacterial load and decreased the ruminal microbiome. A 7-day infection of sheep with H. contortus significantly altered approximately 98% and 94% of genera in the abomasal and ruminal bacterial profile, respectively (P = 0.04-0.05). However, the approximate altered genera 50 days after infection in the ovine abomasal and ruminal microbiome were about 62% and 69%, correspondingly (P = 0.04-0.05) with increase in some bacteria and decrease in others. Overall, these results indicate that Haemonchus infection plays a crucial role in shaping stomach microbial community composition, and diversity.

Experimental infection of calves with Haemonchus placei or Haemonchus contortus: Assessment of clinical, hematological and biochemical parameters and histopathological characteristics of abomasums.

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.

Pathology of Haemonchus contortus in New World camelids in the southeastern United States: a retrospective review.

Most small ruminant farms in tropical climates are plagued by Haemonchus contortus, a hematophagous, abomasal parasite. Heavy burdens of this parasite can cause anemia, hypoproteinemia, weight loss, and mortality in susceptible animals. Haemonchus contortus is becoming a major health concern in New World camelids as well, namely llamas (Llama glama) and alpacas (Vicugna pacos), yet little research has been conducted regarding its prevalence or pathology in these species. Herein, we present a retrospective review of llamas and alpacas that were admitted to The University of Georgia Veterinary Teaching Hospital and Athens Diagnostic Laboratory between the years 2002 and 2013. Antemortem fecal egg count (FEC) estimates performed on 30 alpacas were negatively correlated with hematocrit, hemoglobin, and red blood cell count. Total protein was not significantly correlated with FEC. On postmortem examination, 55 of 198 camelids, including 2 from the aforementioned antemortem review, were infected with H. contortus, with llamas (42.6%) having a significantly higher infection rate than alpacas (22.2%). In 15.7% of the total cases, the parasite was the major cause of death. Common gross lesions included peritoneal, thoracic, and pericardial effusions, visceral pallor, subcutaneous edema, and serous atrophy of fat. Histologic lesions included centrilobular hepatic necrosis, hepatic atrophy, lymphoplasmacytic inflammation of the mucosa of the third gastric compartment (C3), extramedullary hematopoiesis in both the liver and spleen, and the presence of nematodes in C3. Our study emphasizes the importance of H. contortus diagnosis and herd monitoring in New World camelids, particularly llamas.

The effect of helminth infection on the microbial composition and structure of the caprine abomasal microbiome.

Haemonchus contortus is arguably the most injurious helminth parasite for small ruminants. We characterized the impact of H. contortus infection on the caprine abomasal microbiome. Fourteen parasite naive goats were inoculated with 5,000 H. contortus infective larvae and followed for 50 days. Six age-matched naïve goats served as uninfected controls. Reduced bodyweight gain and a significant increase in the abosamal pH was observed in infected goats compared to uninfected controls. Infection also increased the bacterial load while reducing the abundance of the Archaea in the abomasum but did not appear to affect microbial diversity. Nevertheless, the infection altered the abundance of approximately 19% of the 432 species-level operational taxonomic units (OTU) detected per sample. A total of 30 taxa displayed a significantly different abundance between control and infected goats. Furthermore, the infection resulted in a distinct difference in the microbiome structure. As many as 8 KEGG pathways were predicted to be significantly affected by infection. In addition, H. contortus-induced changes in butyrate producing bacteria could regulate mucosal inflammation and tissue repair. Our results provided insight into physiological consequences of helminth infection in small ruminants and could facilitate the development of novel control strategies to improve animal and human health.

Relationship between oxidative stress and pathological findings in abomasum of infected lambs by Haemonchus contortus.

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.

Anthelmintic activity of Securidaca longepedunculata (Family: Polygalaceae) root extract in mice, in vitro and in vivo.

OBJECTIVE: To elucidate the pharmacological bases of oral administration of Securidaca longepedunculata (S. longepedunculata) root extract as an anthelmintic in folkloric medicine. METHODS: Albino mice were infected with infective third (L3) larval stage of Heligmosomoides polygyrus (H. polygyrus) by esophageal intubation. Following establishment of the adult worms in the intestine, the mice were treated with 0-2 000 mg/kg body weight (bw) of methanolic root extract of S. longepedunculata and 100 mg/kg bw of pyrantel embonate, the reference drug in vivo. Bioactivity and larvicidal effects of the extract were tested by exposing brine shrimps (Artemia salina) to 0.00-1.00 mg/mL and the L3 stage of Heligmosomoidescontortus (H. contortus) and H. polygyrus to 0.00-2.50 mg/mL of the extract in vitro. RESULTS: The percentage yield of the extract was 7.13% w/w dry matter. The brine shrimps toxicity bioassay resulted in an LC50 of 74.18 µg/mL. The extract had a significant, dose-dependent larvicidal effect on the L3 stage of H. contortus and H. polygyrus with the terminal effect of 75% and 70% at the highest exposure concentrations, respectively. The extract however, did not affect the number of worm eggs per gram (epg) of fecal materials (P<0.05) and total worm burden (twb) of adult H. polygyrus in infected mice. Treatment with pyrantel embonate significant reduced both the fecal egg count and twb to 0 compared to the untreated control (P<0.05). CONCLUSIONS: These results indicate that S. longepedunculata root extract contains potent bioactive compounds and has larvicidal effect on L3 stage of H. contortus and H. polygyrus, substantiating its use as anthelmintic in alternative medicine.

Changes in the body condition scores of Nigerian West African Dwarf sheep experimentally infected with mixed infections of Haemonchus contortus and Trichostrongylus colubriformis.

Weekly changes in body condition scores and body weights were determined in West African Dwarf sheep following trickle exposure to weekly escalating mixed infections with Haemonchus contortus and Trichostrongylus colubriformis starting with single doses of 500 infective larvae (L(3)) at wk1, 1000 L(3) at wk2, 2000 L(3) at wk3 and 4000 L(3) at wk4. Thirty-three animals were randomly assigned to two groups namely, Group 1 (n=27) and Group 2 (n=6). Group 1 animals were used for experimental infections and Group 2 animals served as uninfected controls. Body condition scores (BCS), body weights (BW), faecal egg counts (FEC) per gram of faeces and worm burden (Wb) were determined using standard procedures. Lambs in group 1 with epg ≤ 1000 were classified as strong responders (n=16), those with epg between 1000 and 10,000 as intermediate (n=5) and lambs with epg>10,000 on any sampling day were classified as weak responders (n=6). Based on this classification, the mean epg of lambs classified as strong responders was significantly lower (P<0.001) than the epg of lambs classified as weak responders. The body condition scores and body weights of the strong responder lambs were comparable with those of the uninfected control and they were significantly higher than those of the weak and intermediate responders from wk5 till the end of the experiment. There was an inverse and significant relationship between the average BCS of days 56 and 59 and the average FEC of days 56 and 59 (r(s)=-572; P=0.017; n=27) as well as with Wb (r(s)=-531; P=0.028; n=27). We concluded that changes in the body scores are good indicators of the intensity of gastrointestinal nematode infection in the Nigerian WAD sheep.

Responses of the humid zone ecotype of the Nigerian West African Dwarf sheep to mixed infections with Haemonchus contortus and Trichostrongylus colubriformis.

The responses of the Nigerian West African Dwarf sheep to experimental infections with two of its most important gastrointestinal (GI) nematodes, namely, Haemonchus contortus and Trichostrongylus colubriformis were studied by means of two measures of parasitological response, namely, faecal egg count (FEC) and worm burden (Wb), and three measures of host pathology, namely, packed cell volume (PCV), body weight (Bwt) and body condition score (BCS). Following exposure to weekly escalating infections (60% H. contortus and 40% T. colubriformis) starting with single doses of 500 infective larvae (L(3)) at week 1, 1,000 L(3) at week 2, 2,000 L(3) at week 3 and 4,000 L(3) at week 4, two distinctive worm burden response phenotypes were readily recognisable, namely, low Wb (LWb) and high Wb (HWb) phenotypes. The percentage of inoculums (adults and immature stages) recovered at necropsy were 3.75% and 33.08% respectively for H. contortus and T. colubriformis. The results showed that the FECs of the lambs belonging to the LWb phenotype were significantly lower than the FECs of their HWb counterpart. Among the measures of host pathology tested, the LWb phenotype sheep had similar Bwt, PCV and BCS to the uninfected control sheep, whereas the HWb phenotype sheep had significantly lower values than their LWb counterparts and the control sheep towards the end of the experiment. There was a highly significant positive correlation between worm burden transformed as log(10) (Wb + 10) and the average of D56 and D59 FECs. A highly significant negative correlation also was obtained between log(10) (Wb + 10) and the PCV, Bwt and BCS. The low percentage of H. contortus inoculums recovered at necropsy suggested that the sheep may be resistant to their most important GI nematode, H. contortus but less so to T. colubriformis.

Comparison of parasitological and productive traits of Criollo lambs native to the central Mexican Plateau and Suffolk lambs experimentally infected with Haemonchus contortus.

The study compares the parasitological and productive traits of Criollo lambs native to the central Mexican Plateau (CNCMP) and Suffolk (SU) lambs experimentally infected with Haemonchus contortus. CNCMP lambs (n=20) and SU lambs (n=15) were infected with L3 of H. contortus while five lambs of each genotype were kept as controls. Fecal egg count (FEC), packed cell volume (PCV), blood eosinophil number (BEN), ocular mucous membrane color (as measured by the FAMACHA index), changes in body condition score (BCS) and cumulative live weight gain (CLWG) were measured weekly during a 20-week period. On week 20, all animals were euthanized and the number of adult worms (AW) in the abomasum was counted. Infected SU lambs had higher (p<0.05) FEC and AW mean values compared to CNCMP lambs, which had a higher mean BEN count (p<0.05). Infected lambs had lower PCV values than controls, regardless of genotype, and had a negative correlation (r=-0.84, p<0.05) with the FAMACHA index. BCS tended to decline for infected SU lambs and increased slightly for infected CNCMP lambs. CLWG differed in all groups (p<0.05); infected SU lambs gained 12.1+/-1.9kg, infected CNCMP lambs gained 18.8+/-0.7kg, control SU lambs gained 34.6+/-1.6kg, and control CNCMP lambs gained 26.9+/-0.8kg. In conclusion, CNCMP lambs had a smaller worm burden, a better ability to maintain their productive traits, and were less affected by infection with Haemonchus contortus.

Differences in immune parameters are associated with resistance to Haemonchus contortus in Caribbean hair sheep.

Caribbean hair sheep are more resistant to gastrointestinal nematodes than conventional wool breeds, but mechanisms that confer resistance are not fully understood. This study compared immune effector cell populations and antibody concentrations in 12 hair and 12 wool lambs infected with the abomasal parasite Haemonchus contortus and sacrificed at 3 or 27 days post-infection (p.i.) and 14 uninfected animals of each breed. Faecal egg counts were over 2.5-fold higher (P = 0.12) and packed cell volumes approximately 8% lower (P < 0.10) in infected wool lambs. Abomasal lymph nodes were heavier in infected animals (P < 0.05) and infected hair sheep had larger lymph nodes than infected wool sheep (P < 0.05). Tissue eosinophil concentrations were likewise larger (P = 0.07) in hair compared with wool sheep at 3 days p.i. Circulating levels of IgE and IgA in uninfected lambs were higher in hair sheep (P < 0.05) and during infection, hair sheep had higher serum IgA than wool sheep at 3, 5, and 21 days p.i. (P < 0.05). Serum IgE in infected lambs did not differ between breeds, but concentrations of IgE in lymph nodes were higher (P < 0.01) at 27 days p.i. in infected hair sheep.

The effect of dietary protein on the productivity of West African Dwarf (WAD) goats infected with Haemonchus contortus.

The effects of increased dietary protein on the performance of West African Dwarf (WAD) goats infected with Haemonchus contortus were investigated. 28 pubertal 9-12-month-old female goats were divided into two equal groups A and B and fed on high and low protein diets, respectively, from day 1 of pregnancy (day of mating) to 6 weeks post-partum. Each animal was trickle infected with a total of 2400 infective larvae of H. contortus over 4 weeks starting from day 1 of pregnancy and the prepatent period recorded. Live weights and body condition scores were measured weekly and the changes determined by subtracting the initial value from each of the subsequent values. Birth and weaning weights of kids as well as stillbirths and foetal loses were also determined. High protein diet improved the ability of goats to resist worm establishment and patency, which was manifested as significant increase in the prepatent period in group A than in the low protein diet group B (p<0.001). Also high protein diet resulted in significantly higher increase in body weight during pregnancy (p<0.01). During lactation both groups rapidly lost weight although body weight increase relative to preinfection value remained significantly higher in group A than B (p=0.05). Between weeks 3 and 13 post infection, the body condition scores increased but were significantly higher in group A than in group B (p<0.001). From weeks 16 to 27, the body condition scores remained significantly higher in group B than group A although both experienced severe losses during lactation. Group A delivered significantly heavier kids than group B (p<0.001) and had no foetal losses as occurred in the latter. However, the level of supplementation had no influence on weaning weights as there was no significant difference in the weaning weights of kids of both group (p>0.05). It is concluded that lactation demand for dietary protein is higher than that for gestation since both body weights and body condition scores deteriorated in both group during lactation, and that improved dietary protein enhances resistance to parasite establishment (increased prepatent period) and resilience in terms of kidding performance, birth weight and survival of neonates.

Sequential microarray to identify timing of molecular responses to Haemonchus contortus infection in sheep.

Anthelmintics are currently the most common method of worm control. The emergence of worms with multiple-drug resistance and issues of residues in the food chain make alternative parasite control measures a priority. To develop improved and sustainable methods for controlling Haemonchus contortus such as genetic selection of resistant sheep, a better understanding of the host-parasite relationship is required. A trial was undertaken using sheep surgically implanted with abomasal fistulas to enable sequential biopsy of the abomasal mucosa during trickle infection with two strains of H. contortus. These were ivermectin-resistant CAVR and ivermectin-sensitive McMaster. From a gross parasitology perspective, this approach enabled the effect of developing immunity to be observed on both the establishment and maturation of two CAVR doses within and between groups. Since the only difference in parasite treatment between the groups was the staggering of the two CAVR doses, microarray results from biopsies taken on the same day in different groups were combined and compared between different biopsy dates to observe differential gene transcription over time. Differential gene transcription was detected by comparing transcription in our array data between different biopsy dates using a low P value screen (P<0.01) and by compiling a list of 82 immunoparasitology-related genes and examining transcription in this list with a higher P value screen (P<0.05). Our microarray data were validated in silico by comparison with intelectin 2, trefoil factor 3, calcium activated chloride channel and mucin 5 from other gene transcription studies and with phenotypic data such as the response by gammadelta T cells and immunoglobulins to H. contortus. The first four genes are involved in non-specific responses to infection and mucosal healing. These were upregulated at the early time points and intelectin 2 remained prominent throughout the trial. As the trial progressed, immunoglobulin genes became strongly upregulated. These included IgCgamma IgG2a heavy chain constant region, IGHE immunoglobulin heavy constant epsilon and IGHM immunoglobulin heavy constant mu.

Cellular phenotypes in the abomasal mucosa and abomasal lymph nodes of goats infected with Haemonchus contortus.

The distribution of T-cell subsets (CD2, CD4, CD8, and gammadelta) and B cells (IgM) was examined at 3, 6, 10 and 13 days post-infection (dpi) in the abomasal mucosa and abomasal lymph nodes of goats primarily infected with Haemonchus contortus. In the abomasal mucosa a mild (3 and 6 dpi) or marked (10 and 13 dpi) increase of T cells, particularly CD4+ and gammadelta+ lymphocytes, was observed, whereas the increase in CD8+ cells was less pronounced. B cells and IgG+ plasma cells also showed a marked increase in the abomasal mucosa at 10 and 13 dpi. The abomasal lymph nodes showed an increase in size, particularly at 10 and 13 dpi, and a decrease in the proportion of T cells, particularly CD8+ lymphocytes, due to the increased proportion of B cells. The proportion of CD4+ and gammadelta+ lymphocytes did not change significantly during the infection in the abomasal lymph nodes, but their absolute numbers were augmented as a result of the enlargement of the nodes. The results revealed a strong cellular and humoral immune response during the early post-infection stages. However, as indicated by the worm burdens, this rapid host response was unable to induce larval expulsion.

Experimental infection of Haemonchus contortus strains resistant and susceptible to benzimidazoles and the effect on mast cells distribution in the stomach of Mongolian gerbils (Meriones unguiculatus).

Establishment rate of Haemonchus contortus in non-suppressed and immunosuppressed gerbils within 14 days post-infection was compared after inoculation with 1,000 third-stage larvae (L3), exsheathed BZ-susceptible larvae. Based on significantly higher number of larvae in gerbils receiving low doses of immunosuppressant agent hydrocortisone, development of benzimidazole (BZ)-susceptible and BZ-resistant strain of nematode in the stomach was studied on days 4, 7, 10, and 14 p.i. Sections of stomach from both groups of animals were examined for overall histopathological response and dynamics of mucosal mast cells (MMC) and connective tissue mast cells (CTMC). In the immunosuppressed gerbils, H. contortus L3 stage larvae developed to the L4 stage on days 10 and 14 p.i., and their sex ratio was higher toward female worms. Significantly higher ratios of establishment rate were recorded for BZ-susceptible than BZ-resistant strain. Infection elicited strong inflammation mainly in the lamina propria mucosae, where MMC numbers peaked on day 7 p.i., being present in a significantly higher numbers in gerbils infected with BZ-susceptible strain. Infection with BZ-susceptible strain of nematode also resulted in a higher number of CTMC in comparison with the effect of BZ-resistant strain, which were observed in the tela submucosa only. Thus, H. contortus infection in gerbils seems to be a suitable model to study host-parasite interactions. Our results indicate that BZ-resistant strain of H. contortus have a decreased capacity to establish infection in direct relation with lower mucosal and connective tissue MCs counts in the stomach.

Excretory/secretory products of sheep abomasal nematode parasites cause vacuolation and increased neutral red uptake by HeLa cells.

Excretory/secretory (ES) products of Ostertagia (Teladorsagia) circumcincta and Haemonchus contortus have been implicated in the inhibition of gastric acid secretion and vacuolation, and the loss of parietal cells associated with abomasal parasitism. Vacuolation of epithelial (HeLa) cells caused by adult O. circumcincta or L3 O. circumcincta or H. contortus ES products have been examined by differential interference contrast microscopy and by the neutral red uptake assay. ES products caused visible vacuolation of HeLa cells, and this effect was enhanced by 8 mM NH4Cl. Some parasite ES products caused a marked detachment of cells from the coverslip. At lower concentrations of ES products, neutral red uptake was usually increased above the control, but at higher concentrations of ES products, uptake was often decreased, probably because of cell detachment. Although generally consistent with direct observations of HeLa cell vacuolation by parasite chemicals, neutral red uptake was not a satisfactory quantitative assay.

Abomasal lymph node responses to Haemonchus contortus intestinal antigens established in kid goats by infection or immunization with intestinal antigens.

Immune responses to Haemonchus contortus intestinal antigens were evaluated using abomasal lymph node (ALN) lymphocytes from kid goats protected against challenge infection by immunization with parasite intestinal antigen, and from kids that were challenged after immunization with ovalbumin. ALN lymphocytes from the intestinal antigen-immunized group produced significantly higher antibody levels against intestinal antigens than the ovalbumin group, supporting the theory that immunization contributed to that ALN response. In contrast, intestinal lysates and membrane enriched preparations from intestinal cells stimulated significant proliferation of ALN lymphocytes in both groups. The proliferation was antigen-dependent, since intestinal antigens failed to stimulate proliferation in ALN lymphocytes from unimmunized and uninfected kids. For both the intestinal antigen and ovalbumin immunized groups, CD4+ T lymphocytes predominated in ALN lymphocytes that were stimulated to proliferate by intestinal antigens. The results indicate that H. contortus infection alone can induce ALN lymphocyte responses to intestinal antigens. In contrast to ALN lymphocyte responses, serum antibody against intestinal antigens was generally low to undetectable in ovalbumin-immunized kids following infection. Abomasal mucus from an H. contortus infected lamb was probed with a monoclonal antibody that binds to a periodate sensitive determinant on numerous H. contortus intestinal membrane and secreted proteins. Numerous bands of reactivity were detected, indicating that multiple parasite intestinal antigens were released into abomasal mucus during infection. The results, challenge the general concept that H. contortus intestinal antigens are 'hidden' from the host immune system during an infection. On the contrary, parasite intestinal proteins may be relatively abundant antigens presented to the host during infection. In addition, ALN T lymphocytes appear to provide a more sensitive measure than serum antibody to detect presentation of these antigens to the host immune system.

Experimental haemonchosis in goats: effects of single and multiple infections in the host response.

Histopathological changes and the distribution of T lymphocytes (CD3), B cells (CD79alpha) and IgG secreting plasma cells were recorded in the abomasum and abomasal lymph nodes of goats during early and late post-infection stages with one to four doses of Haemonchus contortus L3. The infiltration of eosinophils, mast cells, CD3(+) T lymphocytes, CD79alpha(+)B cells and IgG(+) plasma cells in the abomasal mucosa increased dramatically from 10dpi onwards, whereas globule leukocytes were observed only during chronic infection. In late post-infection stages abomasal infiltration of globule leukocytes, CD3(+) T lymphocytes, CD79alpha(+)B cells and IgG(+) plasma cells was significantly higher (P<0.05) in reinfected (groups 6-8) than in primarily infected goats (group 5). In the abomasal lymph nodes, marked hyperplasia of lymphoid follicles and medullary cords, with increase of CD3(+) T lymphocytes, CD79alpha(+)B cells and IgG(+) plasma cells was recorded from 10dpi (group 3) onwards. Worm burdens and the severe abomasal response during the late post-infection stages suggests that a rapid expulsion of nematodes did not occur. The prolonged time required for generating globule leukocytes suggested that immune mechanisms dependent of this cell type are of crucial importance in the protective immunity against H. contortus in goats.

Incidence of Haemonchus spp. and effect on haematocrit and eye colour in goats farmed under resource-poor conditions in South Africa.

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.

Haemonchus spp. in sheep farmed under resource-poor conditions in South Africa--effect on haematocrit, conjunctival mucous membrane colour and body condition.

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.