Parasitic Nematode Of Small Ruminants Research Articles

Non-coding RNA in the gut of the blood-feeding parasitic worm, Haemonchus contortus

The intestine of Haemonchus contortus is an essential tissue that has been indicated to be a major target for the prevention of haemonchosis caused by this parasitic nematode of small ruminants. Biological peculiarities of the intestine warrant in-depth exploitation, which can be leveraged for future disease control efforts. Here, we determined the intestinal ncRNA (lncRNA, circRNA and miRNA) atlas using whole-transcriptome sequencing and bioinformatics approaches. In total, 4846 novel lncRNA, 982 circRNA, 96 miRNA (65 known and 31 novel) and 8821 mRNA were identified from the H. contortus intestine. The features of lncRNA, circRNA and miRNA were fully characterized. Comparison of miRNA from the intestines and extracellular vesicles supported the speculation that the miRNA from the latter were of intestinal origin in H. contortus. Further function analysis suggests that the cis-lncRNA targeted genes were involved in protein binding, intracellular anatomical structure, organelle and cellular process, whereas the circRNA parental genes were mainly enriched in molecular function categories, such as ribonucleotide binding, nucleotide binding, ATP binding and carbohydrate derivative binding. The miRNA target genes were related to the cellular process, cellular response to stimulus, cellular protein modification process and signal transduction. Moreover, competing endogenous RNA network analysis revealed that the majority of lncRNA, circRNA and mRNA only have one or two binding sites with specific miRNA. Lastly, randomly selected circRNA, lncRNA and miRNA were verified successfully using RT-PCR. Collectively, these data provide the most comprehensive compilation of intestinal transcripts and their functions, and it will be helpful to decipher the biological and molecular complexity of the intestine and lay the foundation for further functional research.

Assembly and Analysis of Haemonchus contortus Transcriptome as a Tool for the Knowledge of Ivermectin Resistance Mechanisms.

Haemonchus contortus (Hc) is an important parasitic nematode of small ruminants. In this study we assembled the transcriptome of Hc as a model to contribute to the knowledge about the profile of the differential gene expression between two Mexican Hc strains under different anthelmintic resistance statuses, one susceptible and the other resistant to ivermectin (IVMs and IVMr, respectively), in order to improve and/or to have new strategies of control and diagnosis. The transcript sequence reads were assembled and annotated. Overall, ~127 Mbp were assembled and distributed into 77,422 transcript sequences, and 4394 transcripts of the de novo transcriptome were matched base on at least one of the following criteria: (1) Phylum Nemathelminthes and Platyhelminthes, important for animal health care, and (2) ≥55% of sequence identity with other organisms. The gene ontology (GO) enrichment analysis (GOEA) was performed to study the level of gene regulation to IVMr and IVMs strains using Log Fold Change (LFC) filtering values ≥ 1 and ≥ 2. The upregulated-displayed genes obtained via GOEA were: 1993 (for LFC ≥ 1) and 1241 (for LFC ≥ 2) in IVMr and 1929 (for LFC ≥ 1) and 835 (for LFC ≥ 2) in IVMs. The enriched GO terms upregulated per category identified the intracellular structure, intracellular membrane-bounded organelle and integral component of the cell membrane as some principal cellular components. Meanwhile, efflux transmembrane transporter activity, ABC-type xenobiotic transporter activity and ATPase-coupled transmembrane transporter activity were associated with molecular function. Responses to nematicide activity, pharyngeal pumping and positive regulation of synaptic assembly were classified as biological processes that might be involved in events related to the anthelmintic resistance (AR) and nematode biology. The filtering analysis of both LFC values showed similar genes related to AR. This study deepens our knowledge about the mechanisms behind the processes of H. contortus in order to help in tool production and to facilitate the reduction of AR and promote the development of other control strategies, such as anthelmintic drug targets and vaccines.

Comparative Metabolome Analyses of Ivermectin-Resistant and -Susceptible Strains of Haemonchus contortus.

Resistance to anthelmintics such as ivermectin (IVM) is currently a major problem in the treatment of Haemonchus contortus, an important parasitic nematode of small ruminants. Although many advances have been made in understanding the IVM resistance mechanism, its exact mechanism remains unclear for H. contortus. Therefore, understanding the resistance mechanism becomes increasingly important for controlling haemonchosis. Recent research showed that the metabolic state of bacteria influences their susceptibility to antibiotics. However, little information is available on the roles of metabolites and metabolic pathways in IVM resistance of H. contortus. In this study, comparative analyses of the metabolomics of IVM-susceptible and -resistant adult H. contortus worms were carried out to explore the role of H. contortus metabolism in IVM resistance. In total, 705 metabolites belonging to 42 categories were detected, and 86 differential metabolites (17 upregulated and 69 downregulated) were identified in the IVM-resistant strain compared to the susceptible one. A KEGG pathway analysis showed that these 86 differential metabolites were enriched in 42 pathways that mainly included purine metabolism; the biosynthesis of amino acids; glycine, serine, and threonine metabolism; and cysteine and methionine metabolism. These results showed that amino acid metabolism may be mediated by the uptake of IVM and related with IVM resistance in H. contortus. This study contributes to our understanding of the mechanisms of IVM resistance and may provide effective approaches to manage infection by resistant strains of H. contortus.

Improved Traditional Drugs: State of knowledge for the sustainable management of gastrointestinal parasitic nematodes of small ruminants

This article aims to present the current state of the fight against gastrointestinal parasites in small ruminants from Improved Traditional Drugs (DTI). A host of scientific documents were consulted by means of search engines to gather the information useful for this synthesis. The failure of conventional treatments has led to the use of new methods to treat digestive pathologies due to internal parasitism in herds of sheep and goats. These include, among others, herbal medicine, homeopathy, aromatherapy, the mixed grazing system for small and large ruminants, ... All plant parts of plants can be used in the preparation of remedies. Several scientific studies show that plants are full of chemical compounds that cause anthelmintic effects on gastrointestinal parasites in small ruminants. Literature searches have revealed that studies on DTIs in the management of gastrointestinal nematodes in small ruminants are non-existent. In addition to the forms (decocted, macerated, infused, etc.) traditionally used by breeders, there are other forms that can be used to make DTIs from plant extracts. There are capsules, capsules, tablets etc. The placing on the market of DTIs requires official authorization based on the safety and reproducibility of the products. From the documents consulted, it appears that scientific research has not yet focused on the formulation of DTIs for the treatment of gastrointestinal parasitic nematodes in small ruminants. It will be useful to manufacture them to facilitate access to veterinary care and the sustainable management of animal health.
 Keywords: Improved Traditional Drugs; Gastrointestinal nematodes; Small ruminants, West Africa; Central Africa.

A novel BR-SMAD is required for larval development in barber's pole worm Haemonchus contortus.

SMAD proteins mediate TGF-β signaling and thereby regulate the metazoan development; however, they are poorly defined in Haemonchus contortus–a common blood-sucking parasitic nematode of small ruminants. Here, we characterized an R-SMAD family protein in H. contortus termed HcSMA2, which is closely related to Caenorhabditis elegans SMA2 (CeSMA2) involved in the bone morphogenetic protein (BMP) signaling. Hcsma2 is transcribed in all developmental stages of H. contortus but highly induced in the adult male worms. The RNA interference with Hcsma2 retarded the transition of infective L3 into L4 larvae. Besides, the bimolecular fluorescence complementation revealed the interaction of HcSMA2 with a TGF-β-activated-R-SMAD (HcDAF8). Together these results show a BMP-like receptor-regulated SMAD in H. contortus that is required for larval differentiation and underscore an adaptive functional repurposing of BMP-signaling in parasitic worms.

Modulatory functions of recombinant electron transfer flavoprotein α subunit protein from Haemonchus contortus on goat immune cells in vitro

Suppression and modulation of the host immune response to parasitic nematodes have been extensively studied. In the present study, we cloned and produced recombinant electron transfer flavoprotein α subunit (ETFα) protein from Haemonchus contortus (rHCETFα), a parasitic nematode of small ruminants, and studied the effect of this protein on modulating the immune response of goat peripheral blood mononuclear cells (PBMCs) and monocytes. Immunohistochemical tests verified that the HCETFα protein was localized mainly in the intestinal wall and on the body surface of worms. Immunoblot analysis revealed that rHCETFα was recognized by the serum of goats artificially infected with H. contortus. Immunofluorescence analysis indicated that rHCETFα bound to the surface of PBMCs. rHCETFα was co-incubated with goat PBMCs to observe the immunomodulatory effects exerted by HCETFα on proliferation, apoptosis, cytokine secretion and nitric oxide (NO) production. The results showed that rHCETFα suppressed the proliferation of goat PBMCs stimulated by concanavalin A and induced apoptosis in goat PBMCs. After rHCETFα exposure, IL-2, IL-4, IL-17A and TNF-α expression was markedly reduced, whereas secretion of TGF-β1 was significantly elevated, in goat PBMCs. Moreover, rHCETFα up-regulated NO production in a dose-dependent manner. FITC-dextran internalization assays showed that rHCETFα inhibited phagocytosis of goat monocytes. These results elucidate the interaction between parasites and hosts at the molecular level, suggest a possible immunomodulatory target and contribute to the search for innovative proteins that may be candidate targets for drugs and vaccines.

Characterization of a rhodanese homologue from Haemonchus contortus and its immune-modulatory effects on goat immune cells in vitro

BackgroundModulation of the host immune response by nematode parasites has been widely reported. Rhodaneses (thiosulfate: cyanide sulfurtransferases) are present in a wide range of organisms, such as archaea, bacteria, fungi, plants and animals. Previously, it was reported that a rhodanese homologue could be bound by goat peripheral blood mononuclear cells (PBMCs) in vivo.MethodsIn the present study, we cloned and produced a recombinant rhodanese protein originating from Haemonchus contortus (rHCRD), a parasitic nematode of small ruminants. rHCRD was co-incubated with goat PBMCs to assess its immunomodulatory effects on proliferation, apoptosis and cytokine secretion.ResultsWe verified that the natural HCRD protein localized predominantly to the bowel wall and body surface of the parasite. We further demonstrated that serum produced by goats artificially infected with H. contortus successfully recognized rHCRD, which bound to goat PBMCs. rHCRD suppressed proliferation of goat PBMCs stimulated by concanavalin A but did not induce apoptosis in goat PBMCs. The production of TNF-α and IFN-γ decreased significantly, whereas secretion of IL-10 and TGF-β1 increased, in goat PBMCs after exposure to rHCRD. rHCRD also inhibited phagocytosis by goat monocytes. Moreover, rHCRD downregulated the expression of major histocompatibility complex (MHC)-II on goat monocytes in a dose-dependent manner, but did not alter MHC-I expression.ConclusionsThese results propose a possible immunomodulatory target that may help illuminate the interactions between parasites and their hosts at the molecular level and reveal innovative protein species as candidate drug and vaccine targets.

Characterization of a phosphotyrosyl phosphatase activator homologue of the parasitic nematode Haemonchus contortus and its immunomodulatory effect on goat peripheral blood mononuclear cells in vitro

Suppression and modulation of the host immune response to parasitic nematodes have been extensively studied. In the present study, we cloned and produced recombinant phosphotyrosyl phosphatase activator protein from Haemonchus contortus (rHCPTPA), a parasitic nematode of small ruminants, and studied the effect of this protein on modulating the immune response of goat peripheral blood mononuclear cells. Enzymatic assays revealed that rHCPTPA enhanced the p-nitrophenylphosphate phosphatase activity of bovine PP2A1. Immunohistochemical tests verified that the HCPTPA protein was localised mainly in the bowel wall and on the body surface of worms. It was also shown that serum produced by goats artificially infected with H. contortus successfully recognised rHCPTPA, which conjugated with goat peripheral blood mononuclear cells. The rHCPTPA was then co-incubated with goat peripheral blood mononuclear cells to assess its immunomodulatory effects on proliferation, apoptosis, cytokine secretion, migration and nitric oxide production. Our results showed that rHCPTPA suppressed the proliferation of goat peripheral blood mononuclear cells stimulated by concanavalin A and induced apoptosis in goat peripheral blood mononuclear cells. After rHCPTPA exposure, IFN-γ and IL-2 expression was markedly reduced, whereas secretion of IL-10 and IL-4 was significantly elevated, in goat peripheral blood mononuclear cells. Moreover, rHCPTPA down-regulated nitric oxide production and migration of goat peripheral blood mononuclear cells in a dose-dependent manner. These results illuminate the interaction between parasites and hosts at the molecular level, suggest a possible immunomodulatory target and contribute to the search for innovative proteins that might be candidate targets for drugs and vaccines.

Characterization of the Complete Mitochondrial Genome of Ostertagia trifurcata of Small Ruminants and its Phylogenetic Associations for the Trichostrongyloidea Superfamily.

The complete mitochondrial (mt) genome of Ostertagia trifurcata, a parasitic nematode of small ruminants, has been sequenced and its phylogenetic relationship with selected members from the superfamily Trichostrongyloidea was investigated on the basis of deduced datasets of mt amino acid sequences. The entire mt genome of Ostertagia trifurcata is circular and 14,151 bp in length. It consists of a total of 36 genes comprising 12 genes coding for proteins (PCGs), 2 genes for ribosomal RNA (rRNA), 22 transfer RNA (tRNA) genes and 2 non-coding regions, since all genes are transcribed in the same direction. The phylogenetic analysis based on the concatenated datasets of predicted amino acid sequences of the 12 protein coding genes supported monophylies of the Haemonchidae, Dictyocaulidae and Molineidae families, but rejected monophylies of the Trichostrongylidae family. The complete characterization and provision of the mtDNA sequence of Ostertagia trifurcata provides novel genetic markers for molecular epidemiological investigations, systematics, diagnostics and population genetics of Ostertagia trifurcata and its correspondents.

Ovicidal and larvicidal activity of extracts from medicinal-plants against Haemonchus contortus

The use of medicinal plants (MP) containing bioactive compounds is an alternative strategy to control of parasitic nematode of small ruminants Haemonchus contortus at various stages of their life cycle. The aims of this study were to determine the in vitro anthelmintic activity of both aqueous and methanolic extracts from 13 medicinal plants typical for Central Europe, and to determine quantity of selected plant secondary metabolites (PSMs) in the methanolic extracts. In vitro egg hatch test and larval development tests were conducted to determine the possible anthelmintic effects of methanolic and aqueous extracts of the roots of Althaea officinalis L., Petasites hybridus L. and Inula helenium L.; flowers of Malva sylvestris L. and Chamomilla recutita L.; leaves of Plantago lanceolata L. and Rosmarinus officinalis L.; seeds of Foeniculum vulgare Mill. and stems of Solidago virgaurea L., Fumaria officinalis L., Hyssopus officinalis L., Melisa officinalis L. and Artemisia absinthium L. on eggs and larvae of H. contortus. Ultra-performance liquid chromatography and tandem mass spectroscopy was used for quantifying six PSMs: gallic acid (GA), rutin (RU), diosmin (DI), hesperidin (HE), quercetin (QU) and kaempferol (KA). RU content of the most effective methanolic extracts was in the order: M. sylvestris (9.33 mg/g DM) > A. absinthium (6.10 mg/g DM) > C. recutita (0.42 mg/g DM). The highest concentration of QU (44.8 mg/g DM) and KA (6.59 mg/g DM) were detected in stems of F. officinalis comparing to the other evaluated plants. The most significant (p < 0.05) anthelmintic effects exhibited methanolic extracts of A. absinthium in both in vitro tests (i.e., egg hatch test and larval development test). Additionally, only two methanolic extracts of C. recutita and M. sylvestris were comparable to activity of A. absinthium using the larval development test. Wider spectrum of aqueous extracts exhibited stronger ovicidal activity in comparison to methanolic extracts. The similar trend was observed in evaluating of larvicidal activity of aqueous and methanolic plant extracts.

A TGF-β type I receptor-like molecule with a key functional role in Haemonchus contortus development

Here we investigated the gene of a transforming growth factor (TGF)-β type I receptor-like molecule in Haemonchus contortus, a highly pathogenic and economically important parasitic nematode of small ruminants. Designated Hc-tgfbr1, this gene is transcribed in all developmental stages of H. contortus, and the encoded protein has glycine-serine rich and kinase domains characteristic of a TGF-β family type I receptor. Expression of a GFP reporter driven by the putative Hc-tgfbr1 promoter localised to two intestinal rings, the anterior-most intestinal ring (int ring I) and the posterior-most intestinal ring (int ring IX) in Caenorhabditis elegans in vivo. Heterologous genetic complementation using a plasmid construct containing Hc-tgfbr1 genomic DNA failed to rescue the function of Ce-daf-1 (a known TGF-β type I receptor gene) in a daf-1-deficient mutant strain of C. elegans. In addition, a TGF-β type I receptor inhibitor, galunisertib, and double-stranded RNA interference (RNAi) were employed to assess the function of Hc-tgfbr1 in the transition from exsheathed L3 (xL3) to the L4 of H. contortus in vitro, revealing that both galunisertib and Hc-tgfbr1-specific double-stranded RNA could retard L4 development. Taken together, these results provide evidence that Hc-tgfbr1 is involved in developmental processes in H. contortus in the transition from the free-living to the parasitic stage.

Lungworm infection in small ruminants in Ethiopia: Systematic review and meta-analysis

Lungworms are important parasitic nematodes of small ruminants that colonize the lower respiratory tract and cause high morbidity and economic loss throughout the world. This systematic review was conducted with the aim of estimating the pooled prevalence of lungworm infection in small ruminants in Ethiopia. Besides, it examines the predictors thought to be responsible for heterogeneity noted between the reports. The search databases used were PubMed, CAB direct, AJOL and Web of science. Eligible studies were selected based on predefined inclusion and exclusion criteria. Thirty two studies published between 2000 and 2016 were found eligible and data were generated on 14,257 small ruminants (12,310 sheep and 1947 goats). Predictors including the study area, type of small ruminants affected, parasitic species involved, a diagnostic test used, study design, management systems, breed, agroecology, and year article published were used as milestone of data extraction. The statistical tool employed were meta-analysis, univariable and multivariable meta-regression and Egger's and begg's statistics for sensitivity analysis. Accordingly, the estimated pooled prevalence of lung worm infection in small ruminants was 40.8% (95% CI: 36.1, 45.6) with a very high heterogeneity between the study reports (I2 = 96.5%). However, diagnostic test used and administrative States were the only two predictors identified to explain 15.5% of the explainable heterogeneity (R2 = 0.1545, tau2 unexplained = 0.6032, tau2 total = 0.7135) noted between studies. The reviewed studies showed that three genera of lungworms, namely Dictyocaulus filaria, Mullerius capillaris and Protostrogylus rufescens were prevalent both in sheep and goats in Ethiopia. Mixed infection with two or three species was reported in 4.4 to 48.6% of the total infected animals. Indeed this review is informative and provides a better picture on lungworm infection in Ethiopia; however, it is not exhaustively detailed as there was lack of clear data on the role of some important factors like age, flock size, management system and seasonal variation on lungworm infection. Thus, thorough epidemiological studies including all seasons are required for formulation of meaning full control strategies.

In vitro and in vivo anthelmintic activity of extracts from Artemisia parviflora and A. sieversiana

Summary In the northern areas of Pakistan, the use of Artemisia based therapeutics is a common practice. Plants of genus Artemisia are known to possess anthelmintic and therapeutic effect. Infections caused by gastrointestinal nematodes are major threat to livestock industry across the world resulting in loss of production and indirect economic losses due to high cost of anthelmintic drugs. Present study was carried out to evaluate in vitro and in vivo effect of Artemisia sieversiana and Artemisia parviflora on Haemonchus contortus, a parasitic nematode of small ruminants. Methanolic plant extract was tested against three different developmental stages using an egg hatch assay, infective larvae and adult worm motility assay. Different concentrations were used for the bioassays and post exposure mortality was recorded after 8 hr for adult worms and infective larvae, while egg inhibition percentage was observed after 27 hr. A highly significant ability to inhibit the egg hatching (100 %) was recorded for both plant extracts while, the highest activity for adult worm assay and larvicidal assay was 90 % for A. sieversiana. The highest activity for adult motility and larvicidal assay for A. parviflora was 89 % and 86.6 % respectively. For in vivo trials maximum parentage reduction was 77.0 % for A. sieversiana and 73.6 % for A. parviflora. It is concluded that selected plant extracts were effective in reducing worm burden in animals.

Gastrointestinal Nematode Parasites of Small Ruminants and Anthelmintics Efficacy Test in Sheep of Haramaya District, Eastern Ethiopia

The combination of cross-section and controlled field trail were carried out from December 2015 to June 2016 at Haramaya District, Eastern Hararhge, Ethiopia to evaluate the prevalence ofgastrointestinal nematodes and anthelmintic efficacy test in naturally infected small ruminants using faecal egg count reduction (FECR) test. From a total of 372 faecal samples examined using fecal flotation technique about 57% (212/372) samples were positive to gastrointestinal nematodes. Coprological investigation revealed that sheep and goats in district were infested with varieties of nematode parasites. The strongyles species were highly prevalent nematode parasites identified at about 22.3% (83/372) followed by trichuris 9.1% (34/372) and strongyloides 5.6% (21/372); whereas mixed infections of strongyles and strongyloides were recovered at lowest rate which accounted for about 5.1% (19/372). From considered risk factors age and kebele showed significant association (p&lt;0.05) with prevalence of nematodes parasites. The study revealed that 53.29% (97/182) of goats and 60.53% (115/190) sheep were found positive, while 58% (115/198) in female and 55.74% (97/174) in male and showed no significant difference (p&gt;0.05) between these factors. Mc Master egg counting in sheep harbored nematode parasites showed relatively similar percentage with no significant difference; but more of examined animals were classified in the range of moderate to heavy infections. The faecal egg count reduction test against albendazole, Tetraclozash and ivermectin indicated that 97.59%, 98.79% and 99.56% respectively which indicated an overall of very good state of efficacy against gastrointestinal nematodes of sheep. Therefore this study revealed that small ruminants of the study area harbored considerable level of gastrointestinal nematodes parasites which need further studies on species identification, larval ecology and strategic deworming with proper practice of available anthelmintics in small ruminants of the study area.

In vitro control of parasitic nematodes of small ruminants using some plant species containing flavonoids.

This study determined in vitro anthelmintic efficacy of three plant species: Trema orientalis, Urtica dioica and Zanthozylum capense on nematode larvae of small ruminants. Dried leaf samples (40g) were extracted in 70% ethanol, in portions of 10g and concentrated to 100ml. Half and one quarter of the original crude extract were both made to 100ml. Rectal faecal material from 10 Merino sheep and 25 Nguni goats was pooled within species and thoroughly hand-mixed. Dung samples, each of 5g were cultured for 12days at 27°C. On day 13, 4 plates were watered and 4 others treated with ethanol to correct for solvent effect on mortality. The design was 2 (animal species) × 3 (plant species) × 3 (extract concentrations). In each of three runs, three plates were treated with each crude extract in three incremental concentrations. Surviving L3 larvae were isolated, counted and mortalities became indices of anthelmintic efficacy. Data from nematode larval mortality were analysed to determine the effect of animal species, plant species, concentration and their interactions. Efficacy was affected by concentration (P = 0.0001), animal species (P = 0.0046), plant species (P = 0.0572), the interactions of animal species and concentration (P = 0.0010), plant species and concentration (P = 0.0123) and concentration × animal × plant species (P = 0.0435).

Two benzimidazole resistance-associated SNPs in the isotype-1 β-tubulin gene predominate in Haemonchus contortus populations from eight regions in China

Haemonchus contortus is one of the most important parasitic nematodes of small ruminants around the world, particularly in tropical and subtropical regions. The control of haemonchosis relies mainly on anthelmintics, but the excessive and prolonged use of anthelmintics is causing serious drug resistance issues in many countries. As benzimidazole (BZ) anthelmintics have been broadly used in China, we hypothesized that resistance is widespread. Given the link between three known single nucleotide polymorphisms (SNPs, designated F167Y, E198A and F200Y) in the isotype-1 β-tubulin gene and BZ resistance, our goal here was to explore the presence of these mutations in H. contortus from small ruminants (sheep and goats) from eight provinces in China using PCR-coupled sequencing. In addition, the genetic diversity and genetic relationship of isotype-1 β-tubulin sequence haplotypes were also investigated. Among 192 H. contortus adult individuals representing the eight populations, we identified six distinct sequence types, five of which had SNP E198A (GCA) and/or F200Y (TAC). Sequence analysis showed that the frequencies of SNPs E198A and F200Y were 0–70% and 0–31%, respectively. SNP F167Y (TAC) was not detected in any population. In addition, high haplotype diversities (0.455–0.939) and nucleotide diversities (0.018–0.039) were calculated. A network analysis of the isotype-1 β-tubulin gene sequences showed that SNPs E198A and F200Y occurred in multiple distinct groupings, suggesting multiple independent origins of these SNPs. The findings of this first study of SNPs in the isotype-1 β-tubulin gene of H. contortus populations suggest that BZ resistance is prevalent in some regions of China, and that any control strategy might focus on monitoring BZ resistance in this country.

In vitro and in vivo anthelmintic activities of Iris kashmiriana Linn.

In search of a natural antiparasitic, in vitro and in vivo anthelmintic activities of methanol and aqueous extracts of Iris kashmiriana Linn. rhizome were tested against gastrointestinal nematodes of sheep. A worm motility inhibition assay was used for in vitro study and a faecal egg count reduction assay was used for in vivo study. Crude aqueous extracts of I. kashmiriana exhibited greater anthelmintic activity against Haemonchus contortus than crude methanolic extract (P<0.05). The aqueous extracts of I. kashmiriana resulted in a mean worm motility inhibition of 85.0%, while methanolic extracts resulted in a mean worm motility inhibition of 100.0%. The in vivo anthelmintic activity of aqueous and methanolic extracts of I. kashmiriana in sheep naturally infected with mixed gastrointestinal nematodes species demonstrated a maximum (70.27%) egg count reduction in sheep treated with aqueous extracts at 2gkg−1 body weight on day 15 after treatment, closely followed by methanolic extracts at 1gkg−1 body weight on day 15 after treatment (33.17% egg count reduction) while as lethal concentration (LC50) values of aqueous and methanolic extracts of I. kashmiriana on adult worms of gastrointestinal nematodes of sheep H. contortus are 18.50mg/ml and 16.66mg/ml respectively. Thus aqueous extracts exhibited greater anthelmintic activity under both in vitro and in vivo conditions; this could be due to the presence of water soluble active ingredients in I. kashmiriana. From the present study it can be suggested that I. kashmiriana rhizome exhibited significant anthelmintic activity against gastrointestinal nematodes of sheep and has the potential to contribute to the control of gastrointestinal nematode parasites of small ruminants.

A review of &lt;i&gt;Bridelia ferruginea, Combretum glutinosum&lt;/i&gt; and &lt;i&gt;Mitragina inermis&lt;/i&gt; plants used in zootherapeutic remedies in West Africa: historical origins, current uses and implications for conservation

Context and objectives : West Africa has a very rich ethnozoological heritage due to its rich biodiversity, history and culture. Due to its location in a biodiversity hotspot, its agro-pastoralist traditions and its role during the maritime discoveries of the Renaissance, West Africa has a long list of animals and animal parts that have been used in its folk medicine. These uses can still be found in many historical documents and pharmacopeias. Bridelia ferruginea , Combretum glutinosum and Mitragyna inermis are tropical and subtropical medicinal plants widely used in general traditional African medicine and in West Africa in particularly, including Benin, Nigeria, Burkina Faso and Ivory Coast to treat many diseases such as bladder troubles, diabetes, dysentery, arterial hypertension, rheumatism pain. This study aims to do a literature review of these three tropical plants on their biological and pharmacological properties on veterinary medicine Methodology and Results : A thorough literature search was done and plants little studied Bridelia ferruginea, Mitragyna inermis and Combretum glutinosum were chosen in the search for their anthelmintic activity against gastrointestinal parasitic nematodes of small ruminants in Djallonke. Also to confirm their use in traditional veterinary medicine in West Africa. Conclusion and application of results : This literature review of their extracts properties showed that the plants possessed wide-reaching pharmacological actions, including anti-diabetic, cardiovascular disease antibacterial, anti-inflammatory, antispasmodic and antiplasmodial and justifies their use in traditional medicine for treating various diseases. This manuscript intends to be a starting point to these future investigations. Key words : Bridelia ferruginea, Combretum glutinosum, Mitragyna inermis, anthelmintic, Traditional folk medicine, Pharmacological action.

Variability of the egg hatch assay to survey benzimidazole resistance in nematodes of small ruminants under field conditions

The egg hatch assay (EHA) is one of the main in vitro methods for detection of benzimidazole resistance in nematode parasites of small ruminants. However, although the EHA has been standardised at the laboratory level, the diagnostic performance of this method has not been fully characterised for field screenings. In the present work, monthly variation of benzimidazole resistance estimated by EHA was surveyed over two years in three sheep flocks and in one goat and an additional sheep flock sharing the same pastures. Resistance was measured by calculating both the effective dose of thiabendazole (TBZ) that inhibited hatching of ≥50% of parasite eggs (ED50) and the proportion (Pdd) of eggs hatching at a discriminating dose of 0.1μg/ml TBZ. Pdd exhibited higher variability than ED50, in agreement with the higher sensitivity of Pdd to changes in resistance levels. Both resistance parameters, however, were highly correlated, and their variation was similarly related to the same factors. Resistance levels differed among sheep flocks, and the resistance level of the goat flock was higher than that measured for the sheep flock sharing the same pasture. Moreover, monthly variation of resistance in goats did not mirror that recorded in sheep. Resistance levels varied seasonally, with the highest values recorded in the spring, and they were inversely related to the number of days that samples were stored under anaerobic conditions. In addition, they were directly associated with the relative abundance of Teladorsagia spp. but inversely related to the relative abundance of Haemonchus spp. After controlling for the effects of these identified factors for variation, inter-monthly sampling variation due to unknown factors was the main source of variability, accounting for more than 60–70% of variance for both resistance parameters and yielding absolute estimation errors higher than 0.06 for ED50 or 0.2 for Pdd when resistance was estimated from a single sampling. Optimum sample size, estimated from variance components, suggested that at least 4–5 samplings would be needed to halve this absolute error, whereas additional samplings would slightly increase precision but at the cost of substantially increasing sampling effort. More research is needed to identify the main factors involved in this inter-sampling variation to standardise the implementation of EHA under field conditions.

In vitro effects of Musa x paradisiaca extracts on four developmental stages of Haemonchus contortus

This study was carried out to evaluate the in vitro effect of Musa x paradisiaca stem and leaf against the parasitic nematode of small ruminants Haemonchus contortus. Three extracts (aqueous, methanolic and/or dichloromethane) of Musa x paradisiaca stem and leaf were tested in vitro on four developmental stages of H. contortus using egg hatch assay (EHA), larval development assay (LDA), L3 migration inhibition assay (LMI) and adult worm motility assay (AWM). The highly significant (P<0.0001) ability to stop larval development (inhibition >67% for each extract) and the negative effect of the dichloromethane extract of leaf on adult worm motility (43% of inhibition of motility after 24h of incubation) compared to the negative controls, suggest anthelmintic properties of Musa x paradisiaca stem and leaf against H. contortus. The active principles responsible for the activity could be secondary metabolites such as terpenoid and flavonoid compounds present in the leaf and stem of the plant.