Haemonchus Contortus Infection Research Articles

Goats are more susceptible to Haemonchus contortus infection than sheep under similar experimental settings

Due to differences in their feeding behavior, sheep and goats are often assumed to respond differently to helminth infections. The present study compared Haemonchus contortus infection profile between sheep and goats under the same experimental setting. Experimental infection was conducted using a randomized block design in four groups of intact sheep (InfSH and ConSH) and goats (InfG, and ConG). Groups InfSH and InfG (N = 7 each) received 10,000L3 of H. contortus whereas the control groups ConSH and ConG (N = 7 each) remained uninfected. Faecal egg counts and PCV were measured from Day 0 to day 56 post infection (PI). On day 56 PI, animals were humanely slaughtered and abomasal contents were recovered to measure worm burden, worm length and in utero egg count. The findings show that: (1) Haemonchus infected animals showed an increase in FEC starting from day 21 PI, (2) progressive reduction in PCV was registered from day 7 PI and continued to the end of the experiment while this remained at pre-infection levels in control groups, (3) FEC was much higher (P < 0.001) and PCV was significantly lower (P < 0.05) in infected goats than in infected sheep, (4) at necropsy, total worm burden with worm establishment rates of 63% and 28.87% were registered respectively for infected goats and sheep with significant difference (P < 0.05), (5) Female worms were significantly longer (P < 0.05) in InfG (22.8(± 1.2) compared to InfSH (20.5 ± 0.67 mm) while (6) Mean worm fecundity was 974.8 ± 239.4 and 1162.5 ± 89.4 respectively for groups InfSH and InfG with no significant difference (P = 0.07), and (7) Parasite traits such as worm burden, FEC and female worm length were well correlated in sheep whereas few such patterns were observed in goats. In conclusion, under the same experimental infection, Arsi-Bale goats are more susceptible to H. contortus infection than Arsi-Bale sheep and hence deserve special attention when they are forced to live on grazing rather than browsing.

Immunohistochemistry analyses of the abomasal mucosa show differences in cellular-mediated immune responses to Haemonchus contortus infection in resistant and susceptible young lambs

Haemonchus contortus is known for its high pathogenicity in sheep, and the uncontrolled use of anthelmintics resulted in the emergence of multiple drug-resistant populations. Breeding sheep for gastrointestinal nematode resistance is a sustainable alternative to reduce dependence of anthelmintic drugs, and differences in the degree of resistance between breeds have been reported. Here we compare two sheep breeds (Santa Ines and Ile de France), concerning the differences in innate and adaptive immune response involved in the resistance against H. contortus infection. Immunohistochemical analyses of the abomasum were conducted in naïve Santa Ines (n = 14) and Ile de France (n = 12) lambs randomized into four groups: infected Santa Ines (n = 8), non-infected control Santa Ines (n = 6), infected Ile de France (n = 8), and non-infected control Ile de France (n = 4). The infected lambs were initially infected with H. contortus infective larvae at 14 days of age, and multiple infections were conducted every second day until they reached 66 days of age. There was a significant effect (P < 0.001) of the infection with increase in numbers of CD3+ T; CD79α+ B; GATA3+ Th2/ILC2; POU2F3+ tuft cells; FOXP3+ T reg; and IgE + cells in the fundus of the abomasal mucosa in both Santa Ines and Ile de France lambs. Nevertheless, the infected Santa Ines lambs presented the highest averages for CD79α+ B; GATA3+ Th2/ILC; IgE + cells; and POU2F3+ tuft cells and there was a significant association of the breed and infection status with regards to POU2F3+ tuft cells, with the highest mean in the infected Santa Ines group. The infected Santa Ines group had three lambs with high degree of resistance and five lambs that showed a moderate infection. Our results suggest a mechanism of synergistic coordination between different immune-cell types in promoting resistance of suckling lambs under H. contortus infection.

Molecular signatures of Haemonchus contortus infection in sheep: A comparative serum proteomic study on susceptible and resistant sheep breeds

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.

Divergent humoral responses between males and females against 24 kDa excretory-secretory protein of Haemonchus contortus and influence of ovine β-globin polymorphism

Lambs harboring the Hb-AA β-globin haplotype present improved cell-mediated responses and increased resistance against Haemonchus contortus infection. The aim of the present study was to compare the effect of sex and β-globin haplotypes on specific humoral responses and phenotypes of resistance during H. contortus infection in Morada Nova sheep. As expected, females displayed stronger resistance during the first and second experimental challenges. Differential systemic humoral immune responses were observed comparing sex groups, in which higher levels of specific antibodies targeting 24 kDa excretory-secretory (ES24) protein of H. contortus of IgG and IgM antibodies were respectively observed as predominant isotypes in males and females. The IgM levels were significantly correlated with phenotypes of resistance, evaluated by packed cell volume and fecal egg counts. To our knowledge this is the first study reporting divergent humoral responses profiles to H. contortus infection between male and female sheep. The impact of β-globin haplotypes was less pronounced in females compared to males. Notably, only males showed significant weight differences across haplotypes, with Hb-AA lambs being the heaviest. Additionally, Hb-AA males had significantly higher PCV (indicating better red blood cell health) and lower FEC (indicating lower parasite burden). These findings suggest a more pronounced effect of β-globin polymorphisms on H. contortus infection in males, potentially due to their generally weaker resistance compared to females. This study highlights the importance of sex and β-globin haplotypes in shaping immune responses to H. contortus infection. Specifically, IgM antibodies targeting the ES24 protein appear to play a crucial role in host-parasite interactions and may hold promise for therapeutic development.

Single-Cell Transcriptomic Profiling Unveils Dynamic Immune Cell Responses during Haemonchus contortus Infection.

Haemonchus contortus is a parasite widely distributed in tropical, subtropical, and warm temperate regions, causing significant economic losses in the livestock industry worldwide. However, little is known about the genetics of H. contortus resistance in livestock. In this study, we monitor the dynamic immune cell responses in diverse peripheral blood mononuclear cells (PBMCs) during H. contortus infection in goats through single-cell RNA sequencing (scRNA-Seq) analysis. A total of four Boer goats, two goats with oral infection with the L3 larvae of H. contortus and two healthy goats as controls, were used in the animal test. The infection model in goats was established and validated by the fecal egg count (FEC) test and qPCR analysis of the gene expression of IL-5 and IL-6. Using scRNA-Seq, we identified seven cell types, including T cells, monocytes, natural killer cells, B cells, and dendritic cells with distinct gene expression signatures. After identifying cell subpopulations of differentially expressed genes (DEGs) in the case and control groups, we observed the upregulation of multiple inflammation-associated genes, including NFKBIA and NFKBID. Kyoto Encyclopedia of the Genome (KEGG) enrichment analysis revealed significant enrichment of NOD-like receptor pathways and Th1/Th2 cell differentiation signaling pathways in CD4 T cells DEGs. Furthermore, the analysis of ligand-receptor interaction networks showed a more active state of cellular communication in the PBMCs from the case group, and the inflammatory response associated MIF-(CD74 + CXCR4) ligand receptor complex was significantly more activated in the case group, suggesting a potential inflammatory response. Our study preliminarily revealed transcriptomic profiling characterizing the cell type specific mechanisms in host PBMCs at the single-cell level during H. contortus infection.

Molecular Characterization of Abomasal-Related Bacteria in Sheep with Haemonchus contortus Infection

Molecular Characterization of Abomasal-Related Bacteria in Sheep with Haemonchus contortus Infection

Abomasal RNA-seq reveals a strong local cellular response in suckling lambs with resistance against Haemonchus contortus

Santa Ines (SI) and Ile de France (IF) sheep are known to be resistant and susceptible to Haemonchus contortus infection, respectively. Several studies have shown some genes as potential biological markers for sheep resistance against gastrointestinal nematodes using molecular tools, including transcriptomic analysis. In this study, we sequenced the polyadenylated RNA of the abomasal tissue of SI and IF suckling lambs to identify mucosa-specific transcript alterations between breeds artificially infected with H. contortus. Naïve SI (n = 4) and IF (n = 4) lambs were artificially infected every other day, over a period of 52 days, from 14 to 66 days old, with a total of 5,400 H. contortus infective larvae. Fundic abomasal tissue samples were collected at 68 days old, and submitted to high-throughput RNA sequencing (RNA-seq). Differential expression analysis (P value < 0.001 and False Discovery Rate (FDR) < 0.05) between SI and IF samples identified 292 genes, most of which showed greater expression in SI lambs. To help annotate and assign possible function to differentially expressed genes (DEGs), we used previously available single-cell RNA-seq (scRNA-seq) data from ovine abomasal mucosa to putatively identify cell types and possible mechanisms involved in resistance to H. contortus. In particular, genes associated with endothelial and tuft cells showed the greatest increases in expression in SI relative to IF lambs. SI lambs had higher percentages of tuft cells than IF lambs in the fundic abomasal mucosa. Although we found innate immunity (cell-mediated in mucosa) acting as a protagonist in impairing H. contortus infection, a stronger acquired immune response was being modulated at an earlier stage by SI lambs. We suggest that the complex connection between innate and adaptive immunity is via cellular antigen processing and presentation (APP). Based on comparison with scRNA-seq data, SI lambs showed a robust APP mechanism characterized mainly by greater T cell APP, macrophage differentiation, and cytokine signalling. We identified potential mechanisms and markers to advance knowledge for selection of H. contortus resistance at a very early age, in SI as well as in other commercial sheep breeds.

Metagenome reveals caprine abomasal microbiota diversity at early and late stages of Haemonchus contortus infection

Haemonchus contortus is one of the most detrimental gastrointestinal nematode parasites for small ruminants, especially in tropics and subtropics. Gastrointestinal nematode and microbiota share the same microhabitat; thus they interact with each other and their host. Metagenomics tools provide a promising way to examine the alterations in the gastric microbial composition induces by gastrointestinal parasites. In this study, we used metagenomics tools to characterize the impact of H. contortus infection on the caprine abomasal microbiota at early and late stage of infection and compared it with non-infected control. Our results showed that H. contortus infection caused a significant increase in abomasal pH at early (7 days post-infection) and late stage of infection (56 days post-infection). The analysis of alpha and beta diversity showed that the microbiota diversity both in number and in proportion was significantly affected at early and late stage of infection. All microbiota classes are impacted by H. contortus infection but Clostridia and Bacteroidia are more concerned. In infected animals, the genera Prevotella decreased at 7 and 56 days post-infection. Here we showed that the abomasal microbiota was significantly affected early after H. contortus infection, and these changes persist at late stage of the infection.

Physiological and Coprological Changes Associated with Single and conjunct experimental infections of trypanosoma brucei and haemonchus contortus in west African dwarf goats

An experimental study on the interaction between Trypanosoma brucei and Haemonchus contortus infections was conducted on 36 male West African Dwarf Goats (WADG) of 8–9 months old. Coprological and physiological observations were made on these animals in different groups of 6 groups of 6 animals each. In this study, experimental single infections of either T. brucei or H. contortus and conjunct infection of both parasites produced an acute infection in WADG with initial parasitaemia occurring 7- 14 days post-infection and initial detection of egg in faeces occurring 14 days post-infection. The pre-patent period of infection of T. brucei and H. contortus were influenced by conjunct infections resulting in the early detection of nematode egg in the faeces and trypanosome in the blood of the infected conjunct groups. A high level of parasitaemia was observed more in group 3 compared to other groups. The faecal egg count and worm burden were more in groups 3, 4, and 5 than group 2. Judging from the degree of clinical manifestation, animals in the conjunct group appeared to be more severely affected. Haematological parameter changes were particularly more marked in conjunct infected groups (3 and 5). These observations may not be unconnected with the stress of the conjunct infections and immunosuppressive effects of trypanosomosis. This was very evident in the significant sustained decrease in the levels of packed cell volume, mean haemoglobin concentration and total red blood cell counts following the onset of parasitaemia and patency in the infected goats. The above observation confirmed the long-held view that both parasites precipitate anaemia and immunosuppression in infected animals.&#x0D; &#x0D; &#x0D; &#x0D; &#x0D; Une étude expérimentale sur l'interaction entre les infections à Trypanosoma brucei et à Haemonchus contortus a été menée sur 36 West African Dwarf Goat (WADG) mâles âgées de 8 à 9 mois. Des observations coprologiques et physiologiques ont été faites sur ces animaux dans différents groupes de 6 groupes de 6 animaux chacun. Dans cette étude, des infections expérimentales uniques de T. brucei ou de H. contortus et une infection conjointe des deux parasites ont produit une infection aiguë dans le WADG avec une parasitémie initiale survenant 7 à 14 jours après l'infection et une détection initiale d'œufs dans les fèces survenant 14 jours après. -infection. La période d'infection pré-patente de T. brucei et H. contortus était influencée par des infections conjointes résultant en la détection précoced'œufs de nématodes dans les fèces et de trypanosomes dans le sang des groupes conjointsm infectés. Un niveau élevé de parasitémie a été observé davantage dans le groupe 3 par rapport aux autres groupes. Le nombre d'œufs fécaux et la charge de vers étaient plus élevés dans les groupes 3, 4 et 5 que dans le groupe 2. À en juger par le degré de manifestation clinique, les animaux du groupe conjoint semblaient être plus gravement touchés. Les modifications des paramètres hématologiques étaient particulièrement plus marquées dans les groupes infectés conjointement (3 et 5). Ces observations ne sont peut-être pas sans lien avec le stress des infections conjointes et les effets immunosuppresseurs de la trypanosomose. Cela était très évident dans la diminution significative et soutenue des niveaux d'hématocrite, de la concentration moyenne d'hémoglobine et du nombre total de globules rouges après l'apparition de la parasitémie et de la perméabilité chez les chèvres infectées. L'observation ci-dessus a confirmé l'opinion de longue date selon laquelle les deux parasites précipitent l'anémie et l'immunosuppression chez les animaux infectés.

Classification Performance of Machine Learning Methods for Identifying Resistance, Resilience, and Susceptibility to Haemonchus contortus Infections in Sheep.

This study investigated the feasibility of using easy-to-measure phenotypic traits to predict sheep resistant, resilient, and susceptible to gastrointestinal nematodes, compared the classification performance of multinomial logistic regression (MLR), linear discriminant analysis (LDA), random forest (RF), and artificial neural network (ANN) methods, and evaluated the applicability of the best classification model on each farm. The database comprised 3654 records of 1250 Santa Inês sheep from 6 farms. The animals were classified into resistant (2605 records), resilient (939 records), and susceptible (110 records) according to fecal egg count and packed cell volume. A random oversampling method was performed to balance the dataset. The classification methods were fitted using the information of age class, the month of record, farm, sex, Famacha© degree, body weight, and body condition score as predictors, and the resistance, resilience, and susceptibility to gastrointestinal nematodes as the target classes to be predicted considering data from all farms randomly. An additional leave-one-farm-out cross-validation technique was used to assess prediction quality across farms. The MLR and LDA models presented good performances in predicting susceptible and resistant animals. The results suggest that the use of readily available records and easily measurable traits may provide useful information for supporting management decisions at the farm level.

Haemato-biochemical responses, worm burdens and Haemonchus contortus ultrastructural changes in lambs fed diets containing graded inclusion levels of sericea lespedeza

Context Globally, sheep productivity is constrained by scarcity of high-quality feed and haemonchosis. These challenges may simultaneously be addressed by valorisation of nutraceutical forage legumes such as sericea lespedeza (SL) into sheep diets. Aim The current study determined the effects of feeding diets containing increasing inclusion levels of SL (0%, 6.25%, 12.5%, 18.75% and 25%) as replacement for lucerne on haematological, biochemical, and parasitological parameters in Haemonchus contortus-infected and non-infected lambs. Methods Bodyweights, body condition scores (BCS), FAMACHA© scores, faecal worm egg counts (FWEC), and haematological and biochemical parameters were recorded from 80 Dohne Merino intact male lambs assigned to diet-by-infection status-based treatments in a completely randomised design, with a 5 × 2 factorial arrangement of treatments comprising of eight lambs per treatment during a 42-day feeding trial. Adult worms were collected, enumerated, and their ultra-structure was evaluated using scanning electron microscopy after slaughter. Results Diet, and diet × infection status had no effects on red blood-cell parameters of the lambs (P &gt; 0.05). Eosinophils were affected by diet × infection status, being highest in non-infected lambs fed the control diet and lowest in infected lambs fed the 25% SL diet (P ≤ 0.05). Serum albumin, calcium and body condition scores declined (P ≤ 0.05) with increasing inclusion levels of SL in the diet. There was no influence of diet on lamb bodyweights, FAMACHA© scores, FWEC, and worm burdens. Dietary increments of SL disrupted the structural integrity of the head (χ2 = 28.43; ϕc = 0.493; P ≤ 0.05) of adult worms. Conclusions Sericea lespedeza may replace lucerne up to 25% in lamb feedlot diets, with minor negative effects on haematological and biochemical parameters, but offers little benefits for modulating Haemonchus contortus infection. Implications Beneficiation of biologically active alternative feed resources such as sericea lespedeza could help farmers adapt better to market changes through the development of enhanced livestock feed and pharmaceutical products, which could help alleviate losses associated with impoverished nutrition and gastrointestinal health of lambs.

In vitro and in vivo anthelmintic efficacy of condensed tannins extracted from the seeds of alfalfa (Medicago sativa L.) against Haemonchus contortus infection

This study was designed to examine in vitro and in vivo anthelmintic efficacy of condensed tannins (CT) extracted from seeds of Medicago sativa on Haemonchus contortus in sheep. CT's in vitro anthelmintic effect was assessed at a 300 μg/ml concentration compared with albendazole (reference drug) at 10 μg/ml. The results showed that CT had a nematocidal effect on H. contortus, and the cuticle of the adult worm appeared to be its initial target. For the in vivo experiment, nine 3-month-old helminths-free lambs were distributed into three groups. Group 1 (n=3) was challenged only as infected untreated controls; Group 2 (n=3) was treated with condensed tannin, and Group 3 (n=3) was treated with albendazole. Fecal and blood samples were collected every 3 days until the end of the experiment; for fecal egg count (FEC) and anti- H. contortus IgG titers determination, respectively. The lambs treated with the CT in G2 exhibited a pronounced decrease of mean FEC with great FECR% detected from the first-week post-treatment (PT) until the end of the experiment compared with G1 animals. The antibody levels gradually increased in G2 following the 2nd dose of CT treatment compared to other groups. A brilliant consistent relation between the elevation of IgG response and reduction of FEC was observed following the second booster dosing of the CT in G2. In conclusion, the CT evoked strongly in vitro and in vivo anthelmintic activity against H. contortus and could be used as a natural alternative treatment of high potency against haemonchosis in sheep.

H11-induced immunoprotection is predominantly linked to N-glycan moieties during Haemonchus contortus infection.

Nematodes are one of the largest groups of animals on the planet. Many of them are major pathogens of humans, animals and plants, and cause destructive diseases and socioeconomic losses worldwide. Despite their adverse impacts on human health and agriculture, nematodes can be challenging to control, because anthelmintic treatments do not prevent re-infection, and excessive treatment has led to widespread drug resistance in nematode populations. Indeed, many nematode species of livestock animals have become resistant to almost all classes of anthelmintics used. Most efforts to develop commercial anti-nematode vaccines (native or recombinant) for use in animals and humans have not succeeded, although one effective (dead) vaccine (Barbervax) has been developed to protect animals against one of the most pathogenic parasites of livestock animals - Haemonchus contortus (the barber's pole worm). This vaccine contains native molecules, called H11 and H-Gal-GP, derived from the intestine of this blood-feeding worm. In its native form, H11 alone consistently induces high levels (75-95%) of immunoprotection in animals against disease (haemonchosis), but recombinant forms thereof do not. Here, to test the hypothesis that post-translational modification (glycosylation) of H11 plays a crucial role in achieving such high immunoprotection, we explored the N-glycoproteome and N-glycome of H11 using the high-resolution mass spectrometry and assessed the roles of N-glycosylation in protective immunity against H. contortus. Our results showed conclusively that N-glycan moieties on H11 are the dominant immunogens, which induce high IgG serum antibody levels in immunised animals, and that anti-H11 IgG antibodies can confer specific, passive immunity in naïve animals. This work provides the first detailed account of the relevance and role of protein glycosylation in protective immunity against a parasitic nematode, with important implications for the design of vaccines against metazoan parasites.

Copy number variant-based genome wide association study reveals immune-related genes associated with parasite resistance in a heritage sheep breed from the United States.

Florida Native is a heritage sheep breed in the United States and expresses superior ability to regulate gastrointestinal nematodes. The objective of the present study was to investigate the importance of copy number variants (CNVs) on resistance to natural Haemonchus contortus infections. A total of 300 Florida Native sheep were evaluated. Phenotypic records included fecal egg count (FEC, eggs/gram), FAMACHA© score, percentage cell volume (PCV, %), body condition score (BCS) and average daily gain (ADG, kg). Sheep were genotyped using the GGP Ovine 50K single nucleotide polymorphism (SNP) chip. Log ratios from 45.2 k SNP markers spanning the entire genome were utilized for CNV detection. After quality control, 261 animals with CNVs and phenotypic records were used for the association testing. Association tests were carried out using correlation-trend test and principal component analysis correction to identify CNVs associated with FEC, FAMACHA©, PCV, BCS and ADG. Significant CNVs were detected when their adjusted p-value was <.05 after FDR correction. A total of 8124 CNVs were identified, which gave 246 non-overlapping CNVs. Fourteen CNVs were significantly associated with FEC and PCV. CNVs associated with FEC overlapped 14 Quantitative Trait Locus previously associated with H. contortus resistance. Our study demonstrated for the first time that CNVs could be potentially involved with parasite resistance in Florida Native sheep. Immune-related genes such as CCL1, CCL2, CCL8, CCL11, NOS2, TNF, CSF3 and STAT3 genes could play an important role for controlling H. contortus resistance. These genes could be potentially utilized as candidate markers for selection of parasite resistance in this breed.

Sensor-based detection of a Haemonchus contortus (Barber's pole worm) infection in sheep

Haemonchus contortus, or Barber's Pole worm, is considered one of the most pathogenic parasites of sheep due to the production, profitability and animal welfare implications associated with infections. However, detection of H. contortus infections by routine observation of sheep flocks is difficult, and producers are usually unaware of a problem until animals are severely infected. The aim of the current study was to explore the potential for accelerometer-based activity monitoring to detect H. contortus infections in sheep.A three-month clinical study was conducted in Central Queensland, Australia, whereby 45, 12-month-old merino wethers were randomly allocated to one of three treatment groups: control (n = 9), low H. contortus infection (n = 18) or high H. contortus infection (n = 18). Infection occurred on Day 0, and accelerometer sensors were attached to a halter and deployed on all animals from Day 21 to Day 70. Animals were sampled weekly (faecal and blood) and treated with an anthelmintic on Day 56. Following the clinical period, five study animals were observed for a total of five hours to compare direct behavioral observations against accelerometer data. This was then used to develop a machine learning (ML) model to classify the accelerometer data into active or inactive behavior, which was further summarized to determine the proportion of time spent active per day. The most valuable features in the selected ML model were movement variation (MV), maximum X-axis value (MaxX), standard deviation of the X-axis (SDX), maximum Y-axis value (MaxY) and standard deviation of the Z-axis (SDZ).Linear mixed-effects models identified a significant difference in all sheep activity pre- and post-anthelmintic drench on Day 56 (P < 0.001). Conversely, there was no significant difference between activity levels of infected animals compared to controls (P = 0.14). To further explore how the activity of sheep changed with clinical presentations of H. contortus infections, weekly faecal egg count (FEC) and blood packed cell volume (PCV) were compared to sheep activity using a Spearman's rank correlation test. There was a weak but significant negative correlation between FEC and sheep activity (P < 0.001) and a weak but significant positive correlation between PCV and sheep activity (P < 0.001).In conclusion, this study has shown that accelerometer-based activity monitoring may be able to identify changes in sheep activity associated with clinical presentations of H. contortus infections and between periods prior to and following anthelmintic treatment. However, the ability to detect activity differences between infected and control animals was less clear. Further research is needed to determine the impact of individual behavior variability and flock dynamics when using on-animal sensors for the detection of behavioral changes associated with disease.

Antiparasitic Activity of Plants Extract against Gastrointestinal Nematodes and Rhipicephalus microplus

Parasites are the significant factors influencing the health and production status of animals all over the globe. The high cost of currently available drugs and the emergence of parasitic-resistant strains has highlighted the importance of alternate control strategies. One of the effective ways to combat parasitism is to develop plant-based medicines. The current study aimed to develop a formulation based on the leaves of three traditional plants, Calotropis procera, Syzygium cumini, and Ziziphus jujube. Crude methanolic extract of leaves was evaluated for its antiparasitic activity utilizing egg hatch test (against nematode eggs), adult motility assay (against adult nematodes), and modified larval immersion test (against ticks). Meanwhile, sheep acquiring natural infections of Haemonchus contortus were used to determine in vivo anthelmintic activity. Results from the egg hatch test indicated that 41.05% of the extract inhibited 50% hatching of eggs (LC50): much higher than oxfendazole (4.11%; control group). More interestingly, it was observed that the effects of plant extract on adult worms increased with time. LC50 of plant extract reduced from 176.03 to 4.74% after 12 hours of treatment of adult nematodes. A similar response was observed against ticks and reduced the exact dose required to induce LC50 after 24 hours of treatment (3641.11%) to 1576.55% after six days of treatment. On the other hand, the moderate activity of the extract was indicated in sheep acquiring natural infections of H. contortus. The results show that plants can induce antiparasitic activity; however, further research into plants' bioactive molecules is required to get a maximum reduction of parasites.

Early Transcriptome Differences Between Pre-Infected and Naïve Kid Goats Infected With Haemonchus contortus.

In small ruminant production, gastrointestinal nematode (GIN) infection is one of the major causes of economic losses. The aim of this study was to compare the abomasal mucosa transcriptome of naïve and pre-infected goats at early time points after Haemonchus contortus infection, in order to identify different pathways and upstream regulators involved in the host immune response. Naïve and pre-infected Creole kids were orally infected with 10,000 H. contortus infective larvae (L3), and abomasal mucosa was sampled at 0, 4, and 6 days post-infection (dpi). At 6 dpi, all the animals were slaughtered to perform parasite burden counts. The mean number of L4 recovered in naïve kids was more than twice as high as that recovered in the pre-infected ones (5,860 and 2,474 respectively, p < 0.001). RNA-seq analysis showed a number of differentially expressed genes (DEGs) very low for both naïve and pre-infected animals when comparing day 0 vs. day 4 post-infection. A total of 2,237 and 3,206 DEGs were identified comparing 0 vs. 6 dpi in naïve and pre-infected animals, respectively. Interestingly, only 18 DEGs were found for the comparison of pre-infected vs. naïve animals at 6 dpi. Ingenuity pathway analysis (IPA) showed that several immune responses were activated in pre-infected compared with naïve animals at 0 and 4 dpi such as Th2 and Th1 pathways, natural killer cell, B cell receptor, IL-2, and IL-15 signaling. On the other hand, both naïve and pre-infected animals showed activation for those pathways comparing 6 vs. 0 dpi, with no difference between them. A similar pattern was recorded for upstream regulator genes which were related to immunity like TNF, IL-1β, IL-2, IL-5, TGFβ1, IFNγ, TCR, IL-18, IL-6, and IL-4. Our results showed that at 0 and 4 dpi the immune response was activated toward Th1 and Th2 pathways in pre-infected kids compared to the naïve ones, however, the same immune response was developed in naïve kids as earlier as 6 dpi. We conclude that repeated H. contortus infection in kid goats induced a concomitant early activation of a Th1 and Th2 immune response resulting in the regulation of worm establishment.

Early-onset immune response to Haemonchus contortus infection in resistant Santa Ines suckling lambs compared with susceptible Ile de France

Santa Ines, an indigenous Brazilian hair sheep, in comparison with European sheep breeds (e.g., Ile de France), show greater resistance against gastrointestinal nematode (GIN) infections, mainly to Haemonchus contortus. Here we bring novel findings that address some gaps regarding the resistance traits involved in the development of the immunity of young lambs in the first few weeks of life to H. contortus infection. This study aimed to compare parasitological, humoral, and local effector cell-mediated responses, also the histopathological differences in the abomasal mucosa of Santa Ines (SI) and Ile de France (IF) suckling lambs serially infected with H. contortus. Parasitological variables, local and circulating humoral immunity, and local cellular response were evaluated in naïve Santa Ines (n = 14) and Ile de France (n = 12) lambs, randomized into four groups: infected SI (n = 8), non-infected control SI (n = 6), infected IF (n = 8) and non-infected control IF (n = 4). Lambs from infected groups were first infected at 14 days old, and multiple infections were conducted every second day, until the age of 66 days old (52 days post first infection). In comparison with infected Ile de France, infected Santa Ines lambs had lower mean eggs per gram of faeces, lower total H. contortus worm burden, lower females’ length, greater abomasal lymph node weight, greater mucosal thickness in the fundus, and also higher counts of eosinophils in the fundus, and mast cells and globule leukocytes in both fundic and pyloric mucosa of the abomasum. Intrabreed differences were observed into the infected Santa Ines group, with three of the eight lambs classified as highly resistant for displaying H. contortus burden ranging only from 1 to 42 worms. Overall, Santa Ines suckling lambs showed great resistance against H. contortus infection in comparison with Ile de France lambs, being able to mount a robust innate immune response at an early age, and before weaning.

A Review: Haemonchus contortus Infection in Pasture-Based Sheep Production Systems, with a Focus on the Pathogenesis of Anaemia and Changes in Haematological Parameters.

Simple SummaryInfection with Haemonchus contortus parasites (haemonchosis) is an important cause of anaemia in sheep. Haemonchosis is a global problem, although sheep that are kept in warm, high rainfall environments are at the greatest risk of infection due to the favourable conditions for H. contortus survival. Following ingestion, the parasites develop in the abomasum of sheep. Various factors such as age, breed, health, nutritional status, and larval challenge influence the severity of clinical disease. Hyperacute, acute, and chronic haemonchosis are reviewed, focusing on the pathophysiology of haemonchosis, associated clinical signs, and haematological and biochemical findings.Haemonchosis is an important cause of anaemia in sheep worldwide, particularly those that are kept in pasture-based systems in warm, high rainfall environments. Potential outcomes vary based on the severity of infection and the sheep’s immune response, however, in some sheep infection can lead to death. The consequences of Haemonchus contortus infection mean that it has been well-studied in a range of different farming systems. However, to our knowledge, there has not been a recent review focused on the pathophysiology of anaemia caused by haemonchosis. Thus, this review provides an in-depth discussion of the literature related to the pathophysiology of haemonchosis and associated clinical signs for hyperacute, acute, and chronic haemonchosis. Additionally, haematological and biochemical findings are presented, and various diagnostic methods are assessed.

Immunization With Recombinant Haemonchus contortus Y75B8A.8 Partially Protects Local Crossbred Female Goats From Haemonchus contortus Infection.

Haemonchus contortus Y75B8A.8 (Hc8) derived from H. contortus excretory–secretory (ES) products was identified as a functional inhibitor of goat interleukin 2 (IL-2). It may act as a vaccine candidate for the development of therapeutic strategies against H. contortus infection. In this research, recombinant Hc8 (rHc8) and goat anti-rHc8 polyclonal antibodies were employed to evaluate the protective capacities of Hc8 antigen against H. contortus infections via active and passive immunization trials, respectively. In both trials, local crossbred female goats aged 9–12 months old were randomly divided into three groups, five in each group, respectively. Parasitological examinations, including fecal egg counts (FEC), cumulative FEC (cFEC), and worm burdens, were performed. In addition, antibody levels in mucosal homogenate (MH) samples and hematological and immunological parameters were detected. In the passive trial, goats were intravenously immunized with 5 mg total IgG containing anti-rHc8 goat polyclonal antibodies. After twice immunization, compared with the challenged control group, cFEC was reduced by 39%. In addition, there was a 46% reduction of worm burdens compared with the challenged controls. In the active immunization trials, 500 μg of recombinant Hc8 protein was given subcutaneously twice to 9–12-month-old local crossbred female goats with a 2-week interval, resulting in the generation of high levels of antigen-specific circulating antibodies. Besides, cFEC and abomasal worm burden were reduced by 70 and 55%, respectively, compared with the challenged control group. In addition, immunized goats had higher mucosal homogenate IgA and hemoglobin levels than the challenged controls in both passive and active immunization trials. These preliminary results demonstrated the immunoprophylactic effects of Hc8 antigen and will inform new studies on ES proteins in developing subunit recombinant vaccines against H. contortus.