scholarly journals Comparative Analysis on Transcriptomics of Ivermectin Resistant and Sensitive Strains of Haemonchus Contortus

2022 ◽  
Author(s):  
Waresi Tuersong ◽  
Caixian Zhou ◽  
Simin WU ◽  
Peixi Qin ◽  
Chunqun Wang ◽  
...  
Keyword(s):  
Resistant Strain ◽  
Haemonchus Contortus ◽  
Small Ruminants ◽  
Reduction Process ◽  
Parasitic Nematodes ◽  
Collagen Formation ◽  
Oxidation Reduction ◽  
Blood Sucking ◽  
Gated Channel ◽  
Transcriptional Changes

Abstract Background: Ivermectin (IVM) is one of the most important and widely used anthelmintics in veterinary medicine. However, its efficacy is increasingly compromised by widespread resistance, and the exact mechanism of IVM resistance remains unclear for most parasitic nematodes including Haemonchus contortus, a blood-sucking parasitic nematode of small ruminants.Methods: In this study, we isolated and assessed an IVM resistant strain from Zhaosu, Xinjiang, China. Subsequently, the comparative analyses on transcriptomics of IVM susceptible and resistant H. contortus adult worms were carried out using RNA sequencing and bioinformatics.Results: In total, 543 and 359 differentially expressed genes (DEGs) were identified in male and female adult worms of the resistant strain compared with the susceptible strain, respectively. The DEGs encode molecules involved in receptor activities, transport, detoxification, lipid metabolism and cuticle collagen formation. In addition, Gene Ontology (GO) analysis revealed that transcriptional changes were dominant in genes associated with ligand-gated channel activity, oxidation-reduction process, lipid metabolic process, and structural constituent of cuticle. The results support previous proposal that the IVM resistant mechanism of H. contortus involved in both neuromuscular and non-neuromuscular pathways. Finally, the quantitative RT-PCR results confirmed that the transcriptional profiles of selected DEGs (male: 8 genes, female: 10 genes) were consistent with those obtained by the RNA-Seq.Conclusions: The findings from this work provided valuable information for further studies on the IVM resistance in H. contortus.

scholarly journals A novel BR-SMAD is required for larval development in barber’s pole worm Haemonchus contortus

2021 ◽  
Vol 8 (2) ◽  
pp. 57-64
Author(s):  
Fangfang Li ◽  
Peixi Qin ◽  
Lisha Ye ◽  
Nishith Gupta ◽  
Min Hu
Keyword(s):  
Haemonchus Contortus ◽  
Developmental Stages ◽  
Small Ruminants ◽  
Bmp Signaling ◽  
Bimolecular Fluorescence Complementation ◽  
Blood Sucking ◽  
Morphogenetic Protein ◽  
Smad Proteins ◽  
Family Protein ◽  
Parasitic Worms

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.

scholarly journals Characterization of a Rhodanese Homologue from Haemonchus Contortus and its Immune-Modulatory Effects on Goat Immune Cells in Vitro

2020 ◽  
Author(s):  
Yujian Wang ◽  
Muhammad Ehsan ◽  
Jianmei Huang ◽  
Kalibixiati Aimulajiang ◽  
RuoFeng Yan ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Mononuclear Cells ◽  
Small Ruminants ◽  
Parasitic Nematodes ◽  
Dependent Manner ◽  
Peripheral Blood Mononuclear ◽  
Nematode Parasites ◽  
Wide Range

Abstract Background: Suppression and modulation of the immune response of the host by nematode parasites have been reported widely. Rhodaneses or thiosulfate: cyanide sulfurtransferases are present in a wide range of organisms, such as archea, bacteria, fungi, plants and animals. Previously, it was reported that a rhodanese homology could bind by goat peripheral blood mononuclear cells (PBMCs) in vivo.Results: In the present study, we cloned and produced recombinant rhodanese protein originated from Haemonchus contortus (rHCRD), which was one of the parasitic nematodes of small ruminants. The effect of this protein on modulating the immunity of goat PBMC and monocyte was studied in the current work. The predominant localization of the natural HCRD protein was verified as the bowel wall and body surface of worms, according to the immunohistochemical tests. It was proved in this study that the serum produced by artificially infecting goats with H. contortus successfully recognized rHCRD which conjugated goat PBMCs. The rHCRD was co-incubated with goat PBMCs to observe the immunomodulatory effect on proliferation, apoptosis and secretion of cytokines exerted by HCRD. The results showed that the interaction of rHCRD suppressed proliferation of goat PBMCs stimulated by ConA but did not induce the apoptosis of goat PBMCs. After rHCRD exposure, the production of TNF-α and IFN-γ were significantly decreased, however, it significantly increased the secretion of IL-10 and TGF-β1 in goat PBMCs. Phagocytotic assay by FITC-dextran internalization showed that rHCRD inhibited the phagocytosis of goat monocytes. Moreover, rHCRD could down-regulate the expression of MHC-II on goat monocytes in a dose-dependent manner. Conclusions: These discoveries proposed a possible target as immunomodulator, which was potentially beneficial to illuminate the interaction between parasites and hosts in the molecular level and hunt for innovative protein species as candidate targets of drug and vaccine.

Extending from PARs in Caenorhabditis elegans to homologues in Haemonchus contortus and other parasitic nematodes

Parasitology ◽  
2006 ◽  
Vol 134 (4) ◽  
pp. 461-482 ◽  
Author(s):  
S. NIKOLAOU ◽  
R. B. GASSER
Keyword(s):  
Caenorhabditis Elegans ◽  
Haemonchus Contortus ◽  
Asymmetric Cell Division ◽  
Small Ruminants ◽  
Future Research ◽  
Parasitic Nematodes ◽  
Par Proteins ◽  
Surface Receptors ◽  
Cytoskeletal Dynamics ◽  
Free Living Nematode

Signal transduction molecules play key roles in the regulation of developmental processes, such as morphogenesis, organogenesis and cell differentiation in all organisms. They are organized into ‘pathways’ that represent a coordinated network of cell-surface receptors and intracellular molecules, being involved in sensing environmental stimuli and transducing signals to regulate or modulate cellular processes, such as gene expression and cytoskeletal dynamics. A particularly important group of molecules implicated in the regulation of the cytoskeleton for the establishment and maintenance of cell polarity is the PAR proteins (derived from partition defective in asymmetric cell division). The present article reviews salient aspects of PAR proteins involved in the early embryonic development and morphogenesis of the free-living nematode Caenorhabditis elegans and some other organisms, with an emphasis on the molecule PAR-1. Recent advances in the knowledge and understanding of PAR-1 homologues from the economically important parasitic nematode, Haemonchus contortus, of small ruminants is summarized and discussed in the context of exploring avenues for future research in this area for parasitic nematodes.

Characterization of heat shock protein 70 gene fromHaemonchus contortusand its expression and promoter analysis inCaenorhabditis elegans

Parasitology ◽  
2013 ◽  
Vol 140 (6) ◽  
pp. 683-694 ◽  
Author(s):  
HONGLI ZHANG ◽  
QIANJIN ZHOU ◽  
YI YANG ◽  
XUEQIU CHEN ◽  
BAOLONG YAN ◽  
...  
Keyword(s):  
Heat Shock ◽  
Haemonchus Contortus ◽  
Expression Patterns ◽  
Small Ruminants ◽  
Green Fluorescence Protein ◽  
Bioinformatic Analysis ◽  
Parasitic Nematodes ◽  
Hsp70 Mrna ◽  
C Elegans ◽  
Larval Stages

SUMMARYHaemonchus contortusinfections in small ruminants are of major economic importance worldwide. Heat shock proteins (HSPs) are a family of molecular chaperones that play important roles in the process of invasion and survival of nematodes. Although HSP70 has been identified in several parasitic nematodes, little is known of its distribution and function inHaemonchus contortus. The aims of this study were to characterize HSP70 fromHaemonchus contortus(designed as Hc-hsp70), express Hc-hsp70 and analyse the promoter activity inCaenorhabditis elegans. Bioinformatic analysis revealed that the open reading frame of the Hc-hsp70 cDNA encodes a 646-amino acid peptide, which is highly conserved in comparison to HSP70 in other nematodes. Phylogenetic analysis indicated thatH. contortusis closely related toCaenorhabditis. The 5′-flanking region promoted green fluorescence protein (GFP) expression in the intestine in all larval stages and adult with 2 expression patterns inC. elegans. Expression of Hc-hsp70 mRNA transcripts inC. elegansincreased following 2, 4, 6 h of heat shock and peaked at 4 h. However, its expression induced down-regulation ofhsp-1ofC. elegans. These results suggest that theH. contortushsp70 might have a similar function to that ofC. elegans hsp-1.

scholarly journals Genome-Wide Identification of CircRNAs of Infective Larvae and Adult Worms of Parasitic Nematode, Haemonchus contortus

2021 ◽  
Vol 11 ◽  
Author(s):  
Caixian Zhou ◽  
Yao Zhang ◽  
Simin Wu ◽  
Zhiheng Wang ◽  
Waresi Tuersong ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Parasitic Nematode ◽  
Developmental Stages ◽  
Enrichment Analysis ◽  
Mapk Signaling ◽  
Differentially Expressed ◽  
Parasitic Nematodes ◽  
Transcriptional Level ◽  
Blood Sucking ◽  
Third Stage

CircRNAs, a novel class of ncRNA family, are endogenous transcriptional products involved in various biological and physiological processes in plants and animals. However, almost no information is available for circRNAs of parasitic helminths. In the present study, the circRNAs repertoire was comprehensively explored in Haemonchus contortus, a blood-sucking parasitic nematode of ruminants. In total, 20073 circRNAs were identified and annotated from three key developmental stages/genders of H. contortus including the free-living infective third-stage larvae (L3, 18883), parasitic adult female (Af, 3491), and male worms (Am, 2550) via deep-sequencing technology and bioinformatic analysis. Among these identified circRNAs, 71% were derived from exonic regions of protein-coding genes. The number of circRNAs transcribed from the X chromosome (4704) was higher than that from Chromosome I-V (3143, 3273, 3041, 3030, 2882). The amount of highly expressed circRNAs in third-stage larvae was significantly more abundant than that in adult stage. 15948 and 16847 circRNAs were differentially expressed between Af and L3s and between Am and L3, respectively. Among them, 13409 circRNAs existed in both comparisons. Furthermore, 1119 circRNAs were differentially expressed between Af_and_Am. GO enrichment analysis indicated that source genes of circRNAs differentially expressed between Am and L3 as well as between Af and L3 were significantly enriched in many biological processes, primarily including signaling, signal transduction and cell communication terms. KEGG analysis revealed that parental genes of differentially expressed circRNAs were mainly related to metabolism (pyruvate metabolism, glycerophospholipid metabolism, and carbon metabolism), MAPK signaling pathway, and phosphatidylinositol signaling system. Moreover, many circRNAs contained one or more miRNA potential binding sites, suggesting that they could regulate gene expression at the post-transcriptional level. Furthermore, the correctness of head-to-tail back splicing site and alternative circularization events were verified by Sanger sequencing using both divergent and convergent primers. Finally, the reliability of RNA-Seq data and the resistance of circRNAs to RNase R digestion were confirmed by quantitative RT-PCR. Taken together, our findings provide a foundation for elucidating the regulatory mechanisms of circRNAs in H. contortus, which will advance the understanding of circRNAs in parasitic nematodes.

scholarly journals Evaluation of resistance in a selected field strain of Haemonchus contortus to ivermectin and moxidectin using the Larval Migration on Agar Test

2013 ◽  
Vol 33 (2) ◽  
pp. 183-187 ◽  
Author(s):  
Fernanda S. Fortes ◽  
Fernando S. Kloster ◽  
Andressa S. Schafer ◽  
Daniele Bier ◽  
Andréia Buzatti ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Anthelmintic Resistance ◽  
Field Isolate ◽  
Small Ruminants ◽  
Field Strain ◽  
Parasitic Nematodes ◽  
Larval Migration ◽  
High Consistency ◽  
Anthelmintic Drugs

Haemonchus contortus is one of the most common and economically significant causes of disease in small ruminants worldwide, and the control programs of parasitic nematodes - including H. contortus - rely mostly on the use of anthelmintic drugs. The consequence of the use of this, as the sole sanitary strategy to avoid parasite infections, was the reduction of the efficacy of all chemotherapeutic products with a heavy selection for resistance. The widespread of anthelmintic resistance and the difficulty of its early diagnosis has been a major concern for the sustainable parasite management on farms. The objective of this research was to determine and compare the ivermectin (IVM) and moxidectin (MOX) effect in a selected field strain of H. contortus with a known resistance status, using the in vitro larval migration on agar test (LMAT). Third stage larvae of the selected isolate were obtained from faecal cultures of experimentally infected sheep and incubated in eleven increasing diluted concentrations of IVM and MOX (6, 12, 24, 48, 96, 192, 384, 768, 1536, 3072 and 6144µg/mL). The dose-response sigmoidal curves were obtained using the R² value of >0.90 and the lethal concentration (LC50) dose for the tested anthelmintic drugs using a four-parameter logistic model. The LC50 value for MOX was significantly lower than IVM (1.253µg/mL and 91.06µg/mL), identifying the H. contortus isolate as considerably less susceptible to IVM compared to MOX. Furthermore, the LMAT showed a high consistency (p<0.0001) and provided to be a useful diagnostic tool for monitoring the resistance status of IVM and MOX in H. contortus field isolate, as well as it may be used for official routine drug monitoring programs under the Ministry of Agriculture (MAPA) guidance.

Effect of Jatropha curcas Latex on L3 Haemonchus contortus Larval Motility

2017 ◽  
Vol 9 (3) ◽  
Author(s):  
Noorzaid Muhamad ◽  
Syahirah Sazeli ◽  
Resni Mona ◽  
Jannathul Firdous
Keyword(s):  
Larvicidal Activity ◽  
Jatropha Curcas ◽  
Haemonchus Contortus ◽  
Anthelmintic Resistance ◽  
Small Ruminants ◽  
Gastrointestinal Nematodes ◽  
Dose Dependent ◽  
Plants Extract

The anthelmintic resistance has limited the control of gastrointestinal nematodes of small ruminants and thus has awakened interest in the study of plants extract as a source of anthelmintics. These experiments were carried out to evaluate the in vitro efficacy of Jatrophacurcas latex extract against Haemonchuscontortus larval motility. To evaluate the larvicidal activity, H.contortus L3 were incubated with the extracts with varying concentration of 5 mg/mL, 10 mg/mL, 15 mg/mL and 20 mg/mL at 27°C for 48, 72 and 96 hrs. The results were subjected to the Kruskal-Wallis test (P less than 0.05). The extracts showed dose-dependent larvicidal effects. These results suggest that J.curcas can be used to control gastrointestinal nematodes of small ruminants.

scholarly journals Susceptible trichostrongyloid species mask presence of benzimidazole-resistant Haemonchus contortus in cattle

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Khalid M. Mohammedsalih ◽  
Jürgen Krücken ◽  
Ahmed Bashar ◽  
Fathel-Rahman Juma ◽  
Abdalhakaim A. H. Abdalmalaik ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Epidemiological Data ◽  
Nematode Species ◽  
Parasitic Nematodes ◽  
Molecular Approaches ◽  
Tubulin Genes ◽  
Faecal Egg Count ◽  
Third Stage ◽  
Before And After ◽  
Β Tubulin

Abstract Background Benzimidazole (BZ) anthelmintics are widely used to control infections with parasitic nematodes, but BZ resistance is an emerging threat among several nematode species infecting humans and animals. In Sudan, BZ-resistant Haemonchus contortus populations were recently reported in goats in South Darfur State. The objective of this study was to collect data regarding the situation of BZ resistance in cattle parasitic nematodes in South Darfur using phenotypic and molecular approaches, besides providing some epidemiological data on nematodes in cattle. Methods The faecal egg count reduction test and the egg hatch test (EHT) were used to evaluate benzimidazole efficacy in cattle nematodes in five South Darfur study areas: Beleil, Kass, Nyala, Rehed Al-Birdi and Tulus. Genomic DNA was extracted from pools of third-stage larvae (L3) (n = 40) during trials, before and after treatment, and pools of adult male Haemonchus spp. (n = 18) from abattoirs. The polymorphisms F167Y, E198A and F200Y in isotype 1 β-tubulin genes of H. contortus and H. placei were analysed using Sanger and pyrosequencing. Results Prevalence of gastro-intestinal helminths in cattle was 71% (313/443). Reduced albendazole faecal egg count reduction efficacy was detected in three study areas: Nyala (93.7%), Rehed Al-Birdi (89.7%) and Tulus (88.2%). In the EHT, EC50 values of these study areas ranged between 0.032 and 0.037 µg/ml thiabendazole. Genus-specific PCRs detected the genera Haemonchus, Trichostrongylus and Cooperia in L3 samples collected after albendazole treatment. Sanger sequencing followed by pyrosequencing assays did not detect elevated frequencies of known BZ resistance-associated alleles in codon F167Y, E198A and F200Y in isotype 1 β-tubulin gene of H. placei (≤ 11.38%). However, polymorphisms were detected in H. contortus and in samples with mixed infections with H. contortus and H. placei at codon 198, including E198L (16/58), E198V (2/58) and potentially E198Stop (1/58). All pooled L3 samples post-albendazole treatment (n = 13) were identified as H. contortus with an E198L substitution at codon 198. Conclusions To the knowledge of the authors, this is the first report of reduced albendazole efficacy in cattle in Sudan and is the first study describing an E198L substitution in phenotypically BZ-resistant nematodes collected from cattle.

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