Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 2.594
Filter
1.
Vet Parasitol Reg Stud Reports ; 52: 101047, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38880566

ABSTRACT

The variability in the expression of different P-glycoprotein (P-gp) genes in parasitic nematodes of ruminants such as Haemonchus contortus (Hco-pgp) may be caused by different factors including nematode biology, geographical region and anthelmintic pressure. This study analysed the relative expression level of 10 P-gp genes in two H. contortus (Hco-pgp) field isolates from Yucatan, Mexico: 1) PARAISO (IVM-resistant) and 2) FMVZ-UADY (IVM-susceptible). These isolates were compared with a susceptible reference isolate from Puebla, Mexico, namely "CENID-SAI". In all cases H. contortus adult males were used. The Hco-pgp genes (1, 2, 3, 4, 9, 10, 11, 12, 14 and 16) were analysed for each isolate using the RT-qPCR technique. The Hco-pgp expressions were pairwise compared using the 2-ΔΔCt method and a t-test. The PARAISO isolate showed upregulation compared to the CENID-SAI isolate for Hco-pgp 1, 3, 9, 10 and 16 (P < 0.05), and the PARAISO isolate showed upregulation vs. FMVZ-UADY isolate for Hco-pgp 2 and 9 (P < 0.05), displaying 6.58- and 5.93-fold differences (P < 0.05), respectively. In contrast, similar Hco-pgp gene expression levels were recorded for FMVZ-UADY and CENID-SAI isolates except for Hco-pgp1 (P <0.1), which presented a significant upregulation (6.08-fold). The relative expression of Hco-pgp allowed confirming the IVM-resistant status of the PARAISO isolate and the IVM-susceptible status of the FMVZ-UADY isolate when compared to the CENID-SAI reference isolate. Therefore, understanding the association between the Hco-pgp genes expression of H. contortus and its IVM resistance status could help identifying the genes that could be used as molecular markers in the diagnosis of IVM resistance. However, it is important to consider the geographic origin of the nematode isolate and the deworming history at the farm of origin.


Subject(s)
ATP Binding Cassette Transporter, Subfamily B, Member 1 , Drug Resistance , Haemonchiasis , Haemonchus , Ivermectin , Animals , Haemonchus/drug effects , Haemonchus/genetics , Ivermectin/pharmacology , Mexico , Male , Drug Resistance/genetics , ATP Binding Cassette Transporter, Subfamily B, Member 1/genetics , Haemonchiasis/veterinary , Haemonchiasis/parasitology , Phenotype , Anthelmintics/pharmacology , Gene Expression , Sheep Diseases/parasitology , Sheep
2.
PLoS One ; 19(6): e0301554, 2024.
Article in English | MEDLINE | ID: mdl-38861496

ABSTRACT

In Sudan, resistance to benzimidazoles has been reported recently in cattle and goats from South Darfur. Herein, ivermectin efficacy against gastrointestinal nematodes (GINs) was evaluated in sheep and goats in three study areas in South Darfur. The faecal egg count reduction test (FECRT) was used to evaluate the efficacy of ivermectin in sheep and goats naturally infected with GINs in the region of Bulbul (goats: n = 106), Kass (goats: n = 40) and Nyala (Domaia (sheep: n = 47, goats: n = 77) and the University farm (goats: n = 52)), using different treatment plans, and the efficacy was evaluated 12 days after treatment. Ivermectin efficacy was also evaluated in goats experimentally infected using local Haemonchus contortus isolates from Kass and Nyala. Nematodes surviving ivermectin treatment in goats in Bulbul and Nyala were harvested and larvae used to infect worm-free male sheep (n = 6, ≤6 months old). Infected sheep were dosed subcutaneously with ivermectin every eight days with increasing doses from 0.2 mg/kg to 1.6 mg/kg bodyweight (bw). Reduced ivermectin efficacy was identified in sheep and goats in the four study locations. Using a paired statistic, the efficacy of a therapeutic dose in sheep was 75.6% (90% upper credible limit (UCrL): 77.5%), while twice the recommended dose led to a reduction of 92.6% (90% UCrL: 93.3%). In goats, the FECRs of a therapeutic dose were 72.9-95.3% (90% UCrL range: 73.6-95.7%) in Bulbul, Nyala Domaia, Nyala University farm and Kass. Twice the dose recommended for goats in Bulbul revealed a 90% UCrL of 87.6%. All post-treatment faecal cultures contained only Haemonchus spp. larvae. The experimental infection trials in sheep and goats supported our findings from field trials and calculated upper 90% CrL of below 98.9%. For the first time highly ivermectin resistant H. contortus populations have been identified in sheep and goats in Sudan, and resistance was experimentally confirmed.


Subject(s)
Drug Resistance , Goat Diseases , Goats , Ivermectin , Nematode Infections , Sheep Diseases , Animals , Goats/parasitology , Ivermectin/pharmacology , Ivermectin/therapeutic use , Sheep/parasitology , Sheep Diseases/drug therapy , Sheep Diseases/parasitology , Goat Diseases/drug therapy , Goat Diseases/parasitology , Sudan , Nematode Infections/drug therapy , Nematode Infections/veterinary , Nematode Infections/parasitology , Feces/parasitology , Male , Parasite Egg Count/veterinary , Nematoda/drug effects , Anthelmintics/therapeutic use , Anthelmintics/pharmacology , Haemonchus/drug effects
3.
PLoS One ; 19(6): e0305155, 2024.
Article in English | MEDLINE | ID: mdl-38865346

ABSTRACT

Haemonchus contortus is a blood-feeding gastrointestinal parasite that impacts grazing sheep, causing economic losses in animal production. Due to its anthelmintic resistance, alternative antiparasitic treatments like plant-based anthelmintics are necessary to explore. Artemisia cina (Asteraceae) is a plant whose n-hexane extract and ethyl acetate extract exhibit anthelmintic activity against H. contortus, the n-hexane more active. To discover additional bioactive metabolites, a chemical analysis was performed on ethyl acetate extract, which presented an LC90 of 3.30 mg/mL and allowed the isolation of 11-[(1R,5S,7R,8R,10S,)-1,8-dihydroxy-5,10-dimethyl-4-oxodecahydroazulen-7-yl] acrylic acid. This new sesquiterpene was identified through one and two-dimensional NMR. The compound was named cinic acid and displayed an LC50 of 0.13 (0.11-0.14) mg/mL and LC90 of 0.40 (0.37-0.44) mg/mL, which, compared with ethyl acetate extract larvicidal activity, was 256-fold more active at LC50 and 15.71-fold at LC90. In this study, a new sesquiterpene with larvicidal activity against H. contortus L3 infective larvae was isolated from the ethyl acetate extract of Artemisia cina.


Subject(s)
Anthelmintics , Artemisia , Haemonchus , Larva , Plant Extracts , Sesquiterpenes , Artemisia/chemistry , Haemonchus/drug effects , Animals , Anthelmintics/pharmacology , Anthelmintics/isolation & purification , Anthelmintics/chemistry , Larva/drug effects , Plant Extracts/pharmacology , Plant Extracts/chemistry , Sesquiterpenes/pharmacology , Sesquiterpenes/chemistry , Sesquiterpenes/isolation & purification , Sheep , Magnetic Resonance Spectroscopy
4.
Parasitol Res ; 123(5): 201, 2024 May 03.
Article in English | MEDLINE | ID: mdl-38698272

ABSTRACT

Gastrointestinal nematodes (GINs) are a common threat faced by pastoral livestock. Since their major introduction to the UK in the early 1990s, South American camelids have been cograzed with sheep, horses, and other livestock, allowing exposure to a range of GIN species. However, there have been no molecular-based studies to investigate the GIN populations present in these camelids. In the current study, we sampled nine alpaca herds from northern England and southern Scotland and used high-throughput metabarcoded sequencing to describe their GIN species composition. A total of 71 amplicon sequence variants (ASVs) were identified representing eight known GIN species. Haemonchus contortus was the most prevalent species found in almost all herds in significant proportions. The identification of H. contortus in other livestock species is unusual in the northern UK, implying that alpacas may be suitable hosts and potential reservoirs for infection in other hosts. In addition, the camelid-adapted GIN species Camelostrongylus mentulatus was identified predominantly in herds with higher faecal egg counts. These findings highlight the value of applying advanced molecular methods, such as nemabiome metabarcoding to describe the dynamics of gastrointestinal nematode infections in novel situations. The results provide a strong base for further studies involving cograzing animals to confirm the potential role of alpacas in transmitting GIN species between hosts.


Subject(s)
Camelids, New World , Haemonchiasis , Haemonchus , Animals , Camelids, New World/parasitology , Haemonchus/genetics , Haemonchus/classification , Haemonchus/isolation & purification , Prevalence , Haemonchiasis/veterinary , Haemonchiasis/parasitology , Haemonchiasis/epidemiology , DNA Barcoding, Taxonomic , United Kingdom/epidemiology , Strongylida Infections/veterinary , Strongylida Infections/parasitology , Strongylida Infections/epidemiology , Feces/parasitology , England/epidemiology , Scotland/epidemiology
5.
Vet Parasitol ; 328: 110191, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38723410

ABSTRACT

Small ruminants (sheep and goats) constantly suffer from endoparasitoses caused by gastrointestinal nematodes. Among these, the species Haemonchus contortus (Rudolphi, 1803) is considered to be the one of greatest importance within sheep farming. This nematode is difficult to control due to its resistance to most commercial anthelmintics. The aim of the present study was to assess the potential of macrochelid mites as macrobiological agents for controlling endoparasitoses of sheep caused by the nematode, H. contortus. For this, novel in vitro methodology was used, in which assessments were made not only of the predatory ability but also the population growth of mite species (Macrocheles merdarius, Macrocheles robustulus and Holostaspella bifoliata) when offered larvae of the nematode, H. contortus. The predatory ability of the mites, M. merdarius and H. bifoliata were efficient regarding their predatory ability against H. contortus nematode larvae. The mite, M. merdarius exhibited the highest predation rate with mean distribution values for the treated group of 18656 ± 10091 and for the control group of 1178 ± 712 (P < 0.0001). The species, H. bifoliata presented the highest population growth rate, with a percentage acarid recovery rate of 263% in relation to the number added initially. The data from this in vitro predation experiment suggest that, M. merdarius and H. bifoliata showed promise as macrobiological agents for controlling gastrointestinal endoparasitoses of sheep caused by the nematode, H. contortus given that both species reduced the population of this helminth by more 70% and the number of mites recovered was three times greater than the number added.


Subject(s)
Haemonchiasis , Mites , Pest Control, Biological , Sheep Diseases , Haemonchus , Haemonchiasis/prevention & control , Mites/physiology , Larva , Predatory Behavior , Pest Control, Biological/standards , Population Growth , Female , Animals , Sheep , Sheep Diseases/parasitology , Sheep Diseases/prevention & control , Feces/parasitology , Species Specificity , In Vitro Techniques
6.
Exp Parasitol ; 262: 108778, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38735517

ABSTRACT

Sheep haemonchosis is a disease that causes serious losses in livestock production, particularly with the increase of cases of anthelmintic resistance around the world. This justifies the urgent need of alternative solutions. The aim of this study was to determine the chemical profile, in vitro, and, in vivo, anthelmintic properties of Thymus capitatus essential oil. To evaluate the, in vitro, anthelmintic activity of the T. capitatus EO on Haemonchus contortus, two tests were used: egg hatch assay (EHA) and adult worm motility (AWM) assay. The nematicidal effect of this oil was evaluated, in vivo, in mice infected artificially with Heligmosomoides polygyrus using faecal egg count reduction (FECR) and total worm count reduction (TWCR). Chromatographic characterization of T.capitatus composition using gas chromatography coupled to mass spectrometry (GC-MS) demonstrated the presence of carvacrol (81.16%), as the major constituents. The IC50 values obtained was 1.9 mg/mL in the EHT. In the AWM assay; T. capitatus essential oil achieved 70.8% inhibition at 1 mg/mL after 8 h incubation. The in vivo, evaluation on H. polygyrus revealed a significant nematicidal effect 7 days post-treatment by inducing 49.5% FECR and 64.5% TWCR, using the highest dose (1600 mg/kg). The results of present study, demonstrate that T.capitatus EO possess a significant anthelmintic properties. Furthermore, it could be an alternative source of anthelmintic agents against gastrointestinal infections caused by H. contortus.


Subject(s)
Anthelmintics , Feces , Flowers , Gas Chromatography-Mass Spectrometry , Haemonchiasis , Haemonchus , Nematospiroides dubius , Oils, Volatile , Parasite Egg Count , Strongylida Infections , Thymus Plant , Animals , Haemonchus/drug effects , Oils, Volatile/pharmacology , Oils, Volatile/chemistry , Oils, Volatile/isolation & purification , Mice , Nematospiroides dubius/drug effects , Thymus Plant/chemistry , Haemonchiasis/veterinary , Haemonchiasis/drug therapy , Haemonchiasis/parasitology , Strongylida Infections/drug therapy , Strongylida Infections/veterinary , Strongylida Infections/parasitology , Anthelmintics/pharmacology , Anthelmintics/isolation & purification , Anthelmintics/therapeutic use , Anthelmintics/chemistry , Feces/parasitology , Parasite Egg Count/veterinary , Flowers/chemistry , Female , Sheep , Inhibitory Concentration 50 , Monoterpenes/pharmacology , Monoterpenes/isolation & purification , Monoterpenes/chemistry , Male , Sheep Diseases/parasitology , Sheep Diseases/drug therapy , Cymenes
7.
Exp Parasitol ; 262: 108777, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38735520

ABSTRACT

This study describes the in vitro anthelmintic effect of a hydroalcoholic extract (HA-E) and its fractions from Cyrtocarpa procera fruits against Haemonchus contortus eggs and infective larvae. The HA-E was subjected to bipartition using ethyl acetate, which resulted in an aqueous fraction (Aq-F) and an organic fraction (EtOAc-F). The HA-E and both fractions were tested using the egg hatching inhibition assay (EHIA) and the larval mortality test (LMT). Fractionation of the EtOAc-F was achieved using different chromatographic processes, i.e., open glass column and HPLC analysis. Fractionation of the EtOAc-F gave 18 subfractions (C1R1-C1R18), and those that showed the highest yields (C1R15, C1R16, C1R17 and C1R18) were subjected to anthelmintic assays. The HA-E and the EtOAc-F displayed 100% egg hatching inhibition at 3 and 1 mg/mL, respectively, whereas Aq-F exhibited 92.57% EHI at 3 mg/mL. All subfractions tested showed ovicidal effect. Regarding the larval mortality test, HA-E and EtOAc-F exhibited a larvicidal effect higher than 50% at 50 and 30 mg/mL, respectively. The subfractions that showed the highest larval mortality against H. contortus were C1R15 and C1R17, with larval mortalities of 53.57% and 60.23% at 10 mg/mL, respectively. Chemical analysis of these bioactive subfractions (C1R15 and C1R17) revealed the presence of gallic acid, protocatechuic acid, and ellagic acid. This study shows evidence about the ovicidal and larvicidal properties of C. procera fruits that could make these plant products to be considered as a natural potential anthelmintic agents for controlling haemonchosis in goats and sheep.


Subject(s)
Anthelmintics , Fruit , Haemonchus , Larva , Ovum , Plant Extracts , Animals , Haemonchus/drug effects , Plant Extracts/pharmacology , Plant Extracts/chemistry , Larva/drug effects , Fruit/chemistry , Anthelmintics/pharmacology , Anthelmintics/isolation & purification , Anthelmintics/chemistry , Ovum/drug effects , Chromatography, High Pressure Liquid , Sheep , Haemonchiasis/parasitology , Haemonchiasis/veterinary , Sheep Diseases/parasitology
8.
Vet Parasitol ; 329: 110195, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38754156

ABSTRACT

The nematode Haemonchus contortus is, as a parasite, responsible for most mortality of small ruminants, causing significant economic losses. Numerous plant-derived compounds have exhibited promising anthelmintic activities against this nematode. Notably, the Annona genus stands out for demonstrated anthelmintic effects by extracts from several of its species against different nematodes. This study aimed to assess the effect of an Annona tomentosa fraction, rich in alkaloids, on H. contortus. This fraction, named Alk.F, is derived from the methanolic extract of the plant's stem bark. Chemical characterization of Alk.F was performed by liquid chromatography coupled with mass spectrometry. Among the nine predominant peaks obtained, seven alkaloids were identified: reticuline, reticuline N-oxide, reticuline N-oxide isomer, cyclanoline, asimilobine, tetrahydropalmatine and anonaine. Alk.F inhibited the larval development of H. contortus with an IC50 of 0.026 mg/mL, inhibited larval exsheathment with an IC50 of 0.38 mg/mL, and displayed low hemolytic activity towards sheep erythrocytes. Furthermore, atomic force microscopy revealed that Alk.F altered adhesive forces and the height profile on the surface of H. contortus larvae. In conclusion, A. tomentosa alkaloids alter the cuticle structure of H. contortus, inhibiting larval development and exsheathment, thus offering possibilities for contributing to the development of new anthelmintic drugs.


Subject(s)
Alkaloids , Annona , Anthelmintics , Haemonchus , Plant Extracts , Animals , Haemonchus/drug effects , Annona/chemistry , Plant Extracts/pharmacology , Plant Extracts/chemistry , Anthelmintics/pharmacology , Anthelmintics/chemistry , Alkaloids/pharmacology , Alkaloids/chemistry , Larva/drug effects , Plant Bark/chemistry , Sheep , Haemonchiasis/veterinary , Haemonchiasis/drug therapy , Haemonchiasis/parasitology
9.
Infect Genet Evol ; 122: 105609, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38806077

ABSTRACT

Nuclear hormone receptors (NHRs) are emerging target candidates against nematode infection and resistance. However, there is a lack of comprehensive information on NHR-coding genes in parasitic nematodes. In this study, we curated the nhr gene family for 60 major parasitic nematodes from humans and animals. Compared with the free-living model organism Caenorhabditis elegans, a remarkable contraction of the nhr family was revealed in parasitic species, with genetic diversification and conservation unveiled among nematode Clades I (10-13), III (16-42), IV (33-35) and V (25-64). Using an in vitro biosystem, we demonstrated that 40 nhr genes in a blood-feeding nematode Haemonchus contortus (clade V; barber's pole worm) were responsive to host serum and one nhr gene (i.e., nhr-64) was consistently stimulated by anthelmintics (i.e., ivermectin, thiabendazole and levamisole); Using a high-throughput RNA interference platform, we knocked down 43 nhr genes of H. contortus and identified at least two genes that are required for the viability (i.e., nhr-105) and development (i.e., nhr-17) of the infective larvae of this parasitic nematode in vitro. Harnessing this preliminary functional atlas of nhr genes for H. contortus will prime the biological studies of this gene family in nematode genetics, infection, and anthelmintic metabolism within host animals, as well as the promising discovery of novel intervention targets.


Subject(s)
Haemonchus , Larva , RNA Interference , Receptors, Cytoplasmic and Nuclear , Animals , Larva/genetics , Haemonchus/genetics , Receptors, Cytoplasmic and Nuclear/genetics , Multigene Family , Phylogeny , Anthelmintics/pharmacology , Genome, Helminth , Helminth Proteins/genetics , Helminth Proteins/metabolism , Humans
10.
FEMS Microbiol Ecol ; 100(6)2024 May 14.
Article in English | MEDLINE | ID: mdl-38821514

ABSTRACT

Complex cross-talk occurs between gastrointestinal nematodes and gut symbiotic microbiota, with consequences for animal metabolism. To investigate the connection between methane production and endoparasites, this study evaluated the effect of mixed infection with Haemonchus contortus and Trichostrongylus colubriformis on methanogenic and methanotrophic community in rumen microbiota of lambs using shotgun metagenomic and real-time quantitative PCR (qPCR). The rumen content was collected from six Santa Inês lambs, (7 months old) before and after 42 days infection by esophageal tube. The metagenomic analysis showed that the infection affected the microbial community structure leading to decreased abundance of methanotrophs bacteria, i.e. α-proteobacteria and ß-proteobacteria, anaerobic methanotrophic archaea (ANME), protozoa, sulfate-reducing bacteria, syntrophic bacteria with methanogens, geobacter, and genes related to pyruvate, fatty acid, nitrogen, and sulfur metabolisms, ribulose monophosphate cycle, and Entner-Doudoroff Pathway. Additionally, the abundance of methanogenic archaea and the mcrA gene did not change. The co-occurrence networks enabled us to identify the interactions between each taxon in microbial communities and to determine the reshaping of rumen microbiome associations by gastrointestinal nematode infection. Besides, the correlation between ANMEs was lower in the animal's postinfection. Our findings suggest that gastrointestinal parasites potentially lead to decreased methanotrophic metabolism-related microorganisms and genes.


Subject(s)
Gastrointestinal Microbiome , Methane , Rumen , Sheep Diseases , Animals , Rumen/microbiology , Rumen/parasitology , Sheep/microbiology , Methane/metabolism , Sheep Diseases/microbiology , Sheep Diseases/parasitology , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Archaea/genetics , Archaea/classification , Haemonchus/genetics , Trichostrongylus , Microbiota , Nematode Infections/microbiology , Nematode Infections/veterinary
11.
Parasitol Res ; 123(5): 226, 2024 May 30.
Article in English | MEDLINE | ID: mdl-38814484

ABSTRACT

In this study, 858 novel long non-coding RNAs (lncRNAs) were predicted as sensitive and resistant strains of Haemonchus contortus to ivermectin. These lncRNAs underwent bioinformatic analysis. In total, 205 lncRNAs significantly differed using log2 (difference multiplicity) > 1 or log2 (difference multiplicity) < - 1 and FDR < 0.05 as the threshold for significant difference analysis. We selected five lncRNAs based on significant differences in expression, cis-regulation, and their association with the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways. These expressions of lncRNAs, namely MSTRG.12610.1, MSTRG.8169.1, MSTRG.6355.1, MSTRG.980.1, and MSTRG.9045.1, were significantly downregulated. These findings were consistent with the results of transcriptomic sequencing. We further investigated the relative expression of target gene mRNAs and the regulation of mRNA and miRNA, starting with lncRNA cis-regulation of mRNA, and constructed a lncRNA-mRNA-miRNA network regulation. After a series of statistical analyses, we finally screened out UGT8, Unc-116, Fer-related kinase-1, GGPP synthase 1, and sart3, which may be involved in developing drug resistance under the regulation of their corresponding lncRNAs. The findings of this study provide a novel direction for future studies on drug resistance targets.


Subject(s)
Drug Resistance , Haemonchus , Ivermectin , RNA, Long Noncoding , Animals , Haemonchus/genetics , Haemonchus/drug effects , RNA, Long Noncoding/genetics , Ivermectin/pharmacology , Drug Resistance/genetics , Haemonchiasis/parasitology , Haemonchiasis/veterinary , Anthelmintics/pharmacology , MicroRNAs/genetics , Computational Biology , Gene Expression Profiling , Gene Expression Regulation/drug effects
12.
Parasitol Res ; 123(5): 227, 2024 May 30.
Article in English | MEDLINE | ID: mdl-38814495

ABSTRACT

The species Haemonchus contortus occurs in many regions worldwide, mainly parasitising small ruminants and economically impacting animal production. Climate change is considered a driving force for the risk of diseases caused by helminths and can also affect relationships between parasites and their hosts, with the potential to cause losses in both animal production and biodiversity in general. The aim of this study was to model the potential distribution of H. contortus in South America. We used MaxEnt to perform the analyses and describe the contribution of important bioclimatic variables involved in the species distribution. Our results show that H. contortus colonised most of the areas with habitats that suit the species' environmental requirements and that this parasite presents habitat suitability in a future scenario. Understanding the effects of climate change on the occurrence and distribution of parasite species is essential for monitoring these pathogens, in addition to predicting the areas that tend to present future parasite outbreaks and identify opportunities to mitigate the impacts of the emergence of diseases caused by these organisms.


Subject(s)
Haemonchiasis , Haemonchus , Animals , Haemonchus/classification , South America , Haemonchiasis/veterinary , Haemonchiasis/parasitology , Haemonchiasis/epidemiology , Climate Change , Ecosystem
13.
Cells ; 13(10)2024 May 15.
Article in English | MEDLINE | ID: mdl-38786064

ABSTRACT

BACKGROUND: 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. METHODS AND RESULTS: 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. CONCLUSIONS: Our study preliminarily revealed transcriptomic profiling characterizing the cell type specific mechanisms in host PBMCs at the single-cell level during H. contortus infection.


Subject(s)
Gene Expression Profiling , Goats , Haemonchiasis , Haemonchus , Single-Cell Analysis , Animals , Haemonchus/immunology , Haemonchiasis/veterinary , Haemonchiasis/immunology , Haemonchiasis/genetics , Haemonchiasis/parasitology , Transcriptome/genetics , Leukocytes, Mononuclear/metabolism , Leukocytes, Mononuclear/immunology , Goat Diseases/immunology , Goat Diseases/parasitology , Goat Diseases/genetics
14.
Vet Parasitol ; 328: 110177, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38583271

ABSTRACT

Infection by gastrointestinal nematodes (GIN), particularly Haemonchus contortus, can be detrimental to sheep health and performance. Genetic susceptibility to GIN varies between breeds, with those lacking high levels of natural resistance often requiring frequent anthelmintic treatment when facing parasitic challenge. Genetic technology can serve as a tool to decrease GIN susceptibility via selection for sheep with reduced fecal egg count (FEC) estimated breeding values (EBVs). However, the physiological changes that result from implementation of this strategy are not well described. Additionally, there is a need for comparison of animals from recent selective breeding against breeds with inherent GIN resistance. In this study we administered a challenge of H. contortus to Dorper x White Dorper (DWD; n = 92) lambs that have been genetically selected for either low (DWD-) or high (DWD+) FEC EBVs and Barbados Blackbelly x Mouflon (BBM; n = 19) lambs from a genetically resistant breed backgrounds. Lamb FEC, packed-cell volume (PCV) and serum IgG were measured at intermittent levels over 5 weeks. At day 21 and day 35, the selectively bred DWD- had a lower mean FEC compared to DWD+, but were higher than BBM. Reductions in both PCV and serum IgG from initial day 0 levels were observed in DWD lambs, but not in BBM. Furthermore, from a subset of lambs (n = 24) harvested at day 21, DWD- only tended (p = 0.056) to have lower mean worm counts than DWD+, with BBM having the lowest mean worm count. Differentially expressed genes (DEGs) identified via RNA-sequencing of abomasal tissue at day 21 indicate a more pronounced Th2 immune response and more rapid worm expulsion occurred in iBBM than iDWD- and iDWD+ lambs. However, gene expression in DWD- suggests an association between reduced FEC EBV and gastric acid secretion and the ability to limit worm fecundity. Ultimately, selection of Dorper sheep for low FEC EBV can reduce susceptibility to GIN, but it will likely require multiple generations with this trait as a breeding priority before presenting a similar resistance level to Caribbean breeds.


Subject(s)
Feces , Haemonchiasis , Haemonchus , Parasite Egg Count , Sheep Diseases , Animals , Sheep , Sheep Diseases/parasitology , Sheep Diseases/immunology , Sheep Diseases/genetics , Haemonchiasis/veterinary , Haemonchiasis/parasitology , Haemonchiasis/immunology , Parasite Egg Count/veterinary , Feces/parasitology , Selective Breeding , Male , Female , Genetic Predisposition to Disease , Breeding
15.
Vet Parasitol ; 328: 110184, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38643645

ABSTRACT

This study applied the in vitro rumen exsheathment test (IVRET) to evaluate the exsheathment kinetics of Haemonchus contortus infective larvae (L3) incubated in ruminal liquor (RL) containing acetone:water extracts of Acacia pennatula (AP), Gymnopodium floribundum (GF), Havardia albicans (HA) or Lysiloma latisiliquum (LL). The role of polyphenols in the biological activity of the evaluated extracts was also determined. Larvae were incubated in RL either alone or added with a different plant extract (AP, GF, HA, or LL) at 1200 µg/mL. Polyethylene glycol (PEG) was added to block polyphenols in each treatment (RL+PEG, AP+PEG, GF+PEG, HA+PEG, and LL+PEG). After incubation times of 0, 1, 3, 6, 9, and 24 h, the exsheathment process was stopped to count the number of ensheathed and exsheathed L3. A Log-Logistic model was used to determine the L3 exsheathment kinetics in the different RL treatments. The inflection point of the respective kinetic curves, which indicates the time to reach 50 % exsheathed L3 (T50), was the only parameter that differed when comparing the exsheathment models (99 % probability of difference). The T50 values obtained for GF, HA, and LL treatments (T50 = 7.11 - 7.58 h) were higher in comparison to the T50 of RL (5.72 h) (≥ 70 % probability of difference). The L3 incubated in RL added with GF, HA, and LL extracts delayed their exsheathment at 3 and 6 h of incubation (28.71 - 48.06 % exsheathment reduction) compared to the RL treatment. The T50 value for AP, AP+PEG, GF+PEG, HA+PEG, and LL+PEG were similar to RL and RL+PEG (T50 = 5.34 - 6.97 h). In conclusion, the IVRET can be used to identify plants with the potential to delay the exsheathment of H. contortus L3 in the ruminal liquor. The acetone:water extracts of G. floribundum, H. albicans, and L. latisiliquum delayed the T50 of H. contortus exsheathment, which was evident at 3 and 6 h of incubation in ruminal liquor. The observed exsheathment delay was attributed to the polyphenol content of the extracts.


Subject(s)
Haemonchus , Larva , Plant Extracts , Rumen , Animals , Haemonchus/drug effects , Rumen/parasitology , Plant Extracts/pharmacology , Plant Extracts/chemistry , Larva/drug effects , Haemonchiasis/veterinary , Haemonchiasis/parasitology , Anthelmintics/pharmacology , Anthelmintics/chemistry
16.
Acta Parasitol ; 69(2): 1132-1140, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38568361

ABSTRACT

PURPOSE: We analysed the possible synergistic activity among active extracts from Artemisia cina and Tagetes lucida combinations on Haemonchus contortus, a nematode parasitising sheep. METHODS: The work was carried out in vitro on eggs and infective larvae (L3) of H. contortus. The results were analysed with SAS 9.1, applying the ANOVA and Tukey test, and the lethal concentration (LC) values LC50 and LC90 were determined with regression analysis, employing Proc Probit of SAS 9.1. Additionally, the lethal concentration (LC) was calculated with LC50 and LC90 to determine the synergistic effect. RESULTS: The results demonstrated a high efficacy of the two plants studied on both nematode eggs and L3 larvae as well as of their combinations. The highest egg hatching inhibition was obtained with a 50/50 combination, and the best larvae mortality was obtained with 25% A. cina and 75% T. lucida at 10 mg/mL. Additionally, this combination showed a synergistic effect. CONCLUSION: The two plant species studied here can be applied as natural anthelmintic alternatives due to their high bioactive effect and synergistic response.


Subject(s)
Anthelmintics , Artemisia , Drug Synergism , Haemonchus , Larva , Plant Extracts , Tagetes , Animals , Haemonchus/drug effects , Artemisia/chemistry , Plant Extracts/pharmacology , Plant Extracts/chemistry , Tagetes/chemistry , Larva/drug effects , Anthelmintics/pharmacology , Acetates/pharmacology , Sheep , Haemonchiasis/parasitology , Haemonchiasis/veterinary , Haemonchiasis/drug therapy , Sheep Diseases/parasitology , Sheep Diseases/drug therapy , Ovum/drug effects , Hexanes
17.
J Ethnopharmacol ; 328: 118132, 2024 Jun 28.
Article in English | MEDLINE | ID: mdl-38565411

ABSTRACT

ETHNOPHARMACOLOGICAL RELEVANCE: Infections caused by parasitic worms or helminth continue to pose a great burden on human and animal health, particularly in underdeveloped tropical and subtropical countries where they are endemic. Current anthelmintic drugs present serious limitations and the emergence of drug resistance has made it increasingly challenging to combat such infections (helminthiases). In Bangladesh, medicinal plants are often used by indigenous communities for the treatment of helminthiases. Knowledge on such plants along with screening for their anthelmintic activity has the potential to lead to the discovery of phytochemicals that could serve as novel molecular scaffolds for the development of new anthelminthic drugs. AIM OF THE STUDY: The purpose of this study was i) to conduct an ethnobotanical survey to gather data on Bangladeshi medicinal plants used in the treatment of helminthiases, ii) to test plants with the highest use values for their in vitro anthelmintic activity, and iii) to carry out in silico screening on phytochemicals present in the most active plant extract to investigate their ability to disrupt ß-tubulin function in helminths. METHODS: The ethnobotanical survey was conducted across three sub-districts of Bangladesh, namely Mathbaria, Phultala and Khan Jahan Ali. The in vitro screening for anthelmintic activity was performed in a motility test using adult Haemonchus contortus worms. Virtual screening using PyRx was performed on the phytochemicals reported from the most active plant, exploring their interactions with the colchicine binding site of the ß-tubulin protein target (PDB ID: 1SA0). RESULTS: The survey respondents reported a total of 32 plants for treating helminthiases. Based on their use values, the most popular choices were Ananas comosus (L.) Merr., Azadirachta indica A.Juss., Carica papaya L., Citrus maxima (Burm.) Merr., Curcuma longa L., Momordica charantia L., Nigella sativa L. and Syzygium cumini (L.) Skeels. In vitro anthelmintic testing revealed that A. indica leaves and bark had the highest activity with LC50 values of 16 mg/mL in both cases. Other plant extracts also exhibited good anthelmintic activity with LC50 values ranging from 16 to 52 mg/mL, while the value for albendazole (positive control) was 8.39 mg/mL. The limonoids nimbolide and 28-deoxonimbolide showed a binding affinity of -8.9 kcal/mol, and satisfied all drug-likeness parameters. The control ligand N-deacetyl-N-(2-mercaptoacetyl)colchicine had a binding affinity of -6.9 kcal/mol. CONCLUSION: Further in silico and in vitro studies are warranted on the identified limonoids to confirm the potential of these derivatives as novel drug templates for helminthiases. The current study supports the need for an ethnobotanical survey-based approach to discover novel drug templates for helminthiases.


Subject(s)
Anthelmintics , Haemonchus , Helminthiasis , Limonins , Plants, Medicinal , Adult , Animals , Humans , Plants, Medicinal/chemistry , Tubulin , Anthelmintics/pharmacology , Plant Extracts/pharmacology , Plant Extracts/chemistry , Phytochemicals/pharmacology , Colchicine
18.
Exp Parasitol ; 261: 108768, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38679124

ABSTRACT

This study describes the anthelmintic efficacy of an organic fraction (EtOAc-F) from Guazuma ulmifolia leaves and the evaluation of its reactive oxidative stress on Haemonchus contortus. The first step was to assess the anthelmintic effect of EtOAc-F at 0.0, 3.5, 7.0 and 14 mg kg of body weight (BW) in gerbil's (Meriones unguiculatus) artificially infected with H. contortus infective larvae (L3). The second step was to evaluate the preliminary toxicity after oral administration of the EtOAc-F in gerbils. Finally, the third step was to determine the relative expression of biomarkers such as glutathione (GPx), catalase (CAT), and superoxide dismutase (SOD) against H. contortus L3 post-exposition to EtOAc-F. Additionally, the less-polar compounds of EtOAc-F were identified by gas mass spectrophotometry (GC-MS). The highest anthelmintic efficacy (97.34%) of the organic fraction was found in the gerbils treated with the 14 mg/kg of BW. Histopathological analysis did not reveal changes in tissues. The relative expression reflects overexpression of GPx (p<0.05, fold change: 14.35) and over expression of SOD (p≤0.05, fold change: 0.18) in H. contortus L3 exposed to 97.44 mg/mL of EtOAc-F compared with negative control. The GC-MS analysis revealed the presence of 4-hydroxybenzaldehyde (1), leucoanthocyanidin derivative (2), coniferyl alcohol (3), ferulic acid methyl ester acetate (4), 2,3,4-trimethoxycinnamic acid (5) and epiyangambin (6) as major compounds. According to these results, the EtOAc-F from G. ulmifolia leaves exhibit anthelmintic effect and increased the stress biomarkers on H. contortus.


Subject(s)
Anthelmintics , Catalase , Gerbillinae , Glutathione , Haemonchiasis , Haemonchus , Oxidative Stress , Plant Extracts , Plant Leaves , Superoxide Dismutase , Animals , Haemonchus/drug effects , Plant Leaves/chemistry , Oxidative Stress/drug effects , Haemonchiasis/veterinary , Haemonchiasis/drug therapy , Haemonchiasis/parasitology , Anthelmintics/pharmacology , Anthelmintics/therapeutic use , Anthelmintics/chemistry , Plant Extracts/pharmacology , Plant Extracts/chemistry , Plant Extracts/therapeutic use , Superoxide Dismutase/metabolism , Catalase/metabolism , Catalase/analysis , Glutathione/metabolism , Glutathione/analysis , Gas Chromatography-Mass Spectrometry , Male , Parasite Egg Count/veterinary , Biomarkers , Glutathione Peroxidase/metabolism , Female
19.
Vet Parasitol ; 328: 110166, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38508098

ABSTRACT

The physiologic increase in some sex hormones has been associated with an increase in the parasite load caused by Haemonchus contortus in ewes, especially prolactin. In lambs that are especially susceptible to hemonchosis, the levels of sex hormones are low; in contrast, the levels of another pituitary hormone, growth hormone (GH), which is structurally very similar to prolactin, are high. In this study, the in vitro and in vivo effects of GH on H. contortus larvae development and establishment were evaluated. The addition of 20 ng/mL GH for 5 and 10 days to cultures of H. contortus larvae induced an enlargement (p<0.01) and an L3/L4 molting rate (p<0.03) greater than that of untreated larvae or those treated with other concentrations of the hormone. Flow cytometry showed that 3.8% of the largest and most complex cells of newly obtained larvae of H. contortus were positive for the GH receptor, and by immunofluorescence with confocal microscopy, it was observed that these receptors are located in the intestinal region larvae. In the in vivo assay, the administration of recombinant GH to gonadectomized lambs produced an increase in FEC (p<0.03), the number of female adult worms in the abomasum (p<0.05) and the levels of specific antibodies (p<0.04) in relation to the control lambs; however, it did not affect the fertility of H. contortus females. Although many factors affect the development and implantation of H. contortus in the abomasum of sheep, the results of this study strongly suggest that GH participates in the development and establishment of the parasite in sheep, mainly in young sheep.


Subject(s)
Growth Hormone , Haemonchiasis , Haemonchus , Larva , Sheep Diseases , Animals , Haemonchus/drug effects , Sheep , Haemonchiasis/veterinary , Haemonchiasis/parasitology , Sheep Diseases/parasitology , Growth Hormone/pharmacology , Larva/drug effects , Larva/growth & development , Female , Male , Receptors, Somatotropin/metabolism
20.
Vet Parasitol ; 328: 110163, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38513446

ABSTRACT

Gastrointestinal nematodes (GIN), especially Haemonchus contortus, represent a significant challenge for sheep production. Given the global concern about GIN anthelmintic resistance, alternative control methods able to reduce the dependence on these drugs are highly advisable. Since previous studies have shown that sheep carrying the Hb-A allele of ß-globin are more resistant to H. contortus, this study aimed to investigate the relationship between the different haplotypes (Hb-AA, Hb-AB and Hb-BB) and phenotypes in Santa Inês (SI), Texel (TX) and White Dorper (DO) breeds infected with H. contortus. Blood samples were collected from 180 ewes and 123 lambs of the three breeds for DNA extraction followed by qPCR using a hydrolysis probe to identify the ß-globin haplotypes. Phenotypic data, including fecal egg count (FEC), packed cell volume (PCV), FAMACHA score and body condition score for ewes and lambs, as well as weight gain for lambs, were collected. The genotypic frequencies of ß-globin for ewes and lambs were, respectively: 21.7% and 21.4% Hb-AA, 50% and 50% Hb-AB and 28.3% and 28.6% Hb-BB in SI; 0% and 0% Hb-AA, 18.6% and 9.4% Hb-AB and 81.4% and 90.6% Hb-BB in TX; and 0% and 0% Hb-AA, 13.1% and 0% Hb-AB and 86.9% and 100% Hb-BB in DO. In ewes, mean PCV differed (p<0.05) between the three haplotypes, with higher PCV in Hb-AA animals, followed by Hb-AB and Hb-BB. When considering each breed separately, SI Hb-AA ewes presented higher PCV (p<0.05), highlighting that even in a breed already considered resistant, animals with Hb-AA haplotype showed superior performance. Lambs with the Hb-AA haplotype exhibited a higher (p<0.05) mean PCV compared to those with Hb-AB and Hb-BB. The same pattern was found in SI when analyzing each breed separately. No significant association was found between ß-globin haplotypes and FEC, FAMACHA score, body condition score, or weight gain. Nevertheless, given that anemia is the major clinical sign of haemonchosis, our findings on PCV reinforce that sheep carrying the Hb-A allele of ß-globin are more tolerant to haemonchosis. This study may support the development of a valuable tool, targeting genetic selection for GIN control, reducing the dependence on anthelmintics and boosting sheep production worldwide.


Subject(s)
Haemonchiasis , Sheep Diseases , beta-Globins , Animals , Sheep , Sheep Diseases/parasitology , Sheep Diseases/genetics , beta-Globins/genetics , Haemonchiasis/veterinary , Haemonchiasis/parasitology , Female , Haplotypes , Polymorphism, Genetic , Haemonchus/genetics , Parasite Egg Count/veterinary , Feces/parasitology
SELECTION OF CITATIONS
SEARCH DETAIL
...