Multiple DNases were identified from Haemonchus contortus intestine based on previous studies. The DNases detected at 34, 36 and 38.5 kDa had diverse characteristics. Some of them had characteristics similar to those of mammalians and others had unusual characteristics. This study was carried out to fractionate worm intestinal DNases from other proteins using phenyl Sepharose chromatographic methods. All DNases detected from Haemonchus contortus intestine were fractionated in the flowthrough of phenyl Sepharose, indicating the worm DNases are hydrophilic. The DNases were enriched five-fold in the flowthrough fraction while additional steps are required for isolation of the worm DNases. Thus, fractionation with phenyl Sepharose could be used as a good initial step to enrich and separate DNases from other proteins.
Chromatography ; Deoxyribonucleases ; Haemonchus ; Intestines ; Proteins ; Sepharose

Multiple DNases were identified from Haemonchus contortus intestine based on previous studies. The DNases detected at 34, 36 and 38.5 kDa had diverse characteristics. Some of them had characteristics similar to those of mammalians and others had unusual characteristics. This study was carried out to fractionate worm intestinal DNases from other proteins using phenyl Sepharose chromatographic methods. All DNases detected from Haemonchus contortus intestine were fractionated in the flowthrough of phenyl Sepharose, indicating the worm DNases are hydrophilic. The DNases were enriched five-fold in the flowthrough fraction while additional steps are required for isolation of the worm DNases. Thus, fractionation with phenyl Sepharose could be used as a good initial step to enrich and separate DNases from other proteins.
Chromatography ; Deoxyribonucleases ; Haemonchus ; Intestines ; Proteins ; Sepharose

The large stomach worm, Haemonchus contortus is an important pathogen of goats (Capra hircus) and sheep (Ovis aries). This paper describes characteristics of surface cuticular ridges (synlophe) of H. contortus adults from the two hosts. There were more ridges in H. contortus from goats compared to that from sheep. Total body length, vulvar morphology, spicule length and cervical papillae had been considered as markers of physical adaptation and were studied and described.
Sheep ; Haemonchus contortus ; Length ; upper case aitch ; Malaysia

There is no doubt that on a global basis, Haemonchus contortus is by far the most important parasite of small ruminants (sheep and goats). This is particularly more so now, with the development of high levels of resistance to both the broad and narrow spectrum anthelmintic drugs in H. contortus throughout the world. Epidemiological studies describe the lower environmental limits for haemonchosis to occur in sheep, as being a mean monthly temperature of 18C and approximately 50mm rainfall. Thus it has been generally recognised that H. contortus is a problem parasite restricted to the warm, wet countries where sheep and goats are raised. However, recent evidence shows that this parasite is apparently common even in northern Europe. Thus the need for sustainable control strategies for H. contortus is becoming much more pressing. This report highlights two examples of sustainable and highly efficient control programmes for H. contortus, that can be implemented in regions at the extremes of its geographic range (Malaysia and Sweden), where the authors have had direct involvement.
Parasites ; upper case aitch ; control ; Haemonchus contortus ; Sheep

Calreticulin (CALR), a multifunctional protein thoroughly researched in mammals, comprises N-, P-, and C-domain and has roles in calcium homeostasis, chaperoning, clearance of apoptotic cells, cell adhesion, and also angiogenesis. In this study, the spatial and temporal expression patterns of the Opisthorchis viverrini CALR gene were analyzed, and calcium-binding and chaperoning properties of recombinant O. viverrini CALR (OvCALR) investigated. OvCALR mRNA was detected from the newly excysted juvenile to the mature parasite by RT-PCR while specific antibodies showed a wide distribution of the protein. OvCALR was localized in tegumental cell bodies, testes, ovary, eggs, Mehlis’ gland, prostate gland, and vitelline cells of the mature parasite. Recombinant OvCALR showed an in vitro suppressive effect on the thermal aggregation of citrate synthase. The recombinant OvCALR C-domain showed a mobility shift in native gel electrophoresis in the presence of calcium. The results imply that OvCALR has comparable function to the mammalian homolog as a calcium-binding molecular chaperone. Inferred from the observed strong immunostaining of the reproductive tissues, OvCALR should be important for reproduction and might be an interesting target to disrupt parasite fecundity. Transacetylase activity of OvCALR as reported for calreticulin of Haemonchus contortus could not be observed.
Antibodies ; Calcium ; Calreticulin* ; Cell Adhesion ; Cell Body ; Citrate (si)-Synthase ; Eggs ; Electrophoresis ; Female ; Fertility ; Haemonchus ; Homeostasis ; In Vitro Techniques ; Mammals ; Molecular Chaperones ; Opisthorchis* ; Ovary ; Ovum ; Parasites ; Prostate ; Reproduction ; RNA, Messenger ; Testis ; Vitellins

Because of the complexity of the cathepsin B-like (CBL) family, an information on the biological and biochemical characteristics of individual CBL genes is lacking. In this study, we investigated the degradative effects of the recombinant HC58 protein isolated from Haemonchus contortus parasites on protein substrates over a broad pH range in vitro. This protein, which hydrolyzed the synthetic peptide substrates Z-FR-AMC and Z-RR-AMC, had characteristics of the cysteine protease class of proteins. In the acidic pH range, the isolated protein actively degraded hemoglobin (Hb), the heavy chain of goat immunoglobulin G, and azocasein. By contrast, it degraded fibrinogen in the alkaline pH range. These activities were strongly inhibited in the presence of the cysteine protease inhibitor E-64. While the protein digested Hb, it did not induce the agglutination of erythrocytes from its natural host. These results suggest that the HC58 protein may play a role in the nutrition of this parasite.
Animals ; Caseins/metabolism ; Cathepsin B/antagonists&inhibitors/*genetics/isolation & purification/*metabolism ; Cysteine Proteinase Inhibitors/pharmacology ; DNA, Complementary/genetics ; Goat Diseases/*parasitology ; Goats ; Haemonchiasis/parasitology/*veterinary ; Haemonchus/*enzymology/genetics/isolation & purification ; Hemagglutination Tests/veterinary ; Hemoglobins/metabolism ; Hydrogen-Ion Concentration ; Immunoglobulin G/metabolism ; Leucine/analogs & derivatives/pharmacology ; RNA, Helminth/chemistry/genetics ; Recombinant Proteins/genetics/metabolism ; Reverse Transcriptase Polymerase Chain Reaction/veterinary

The complete coding sequence of Haemonchus (H.) contortus HC29 cDNA was generated by rapid amplification of cDNA ends in combination with PCR using primers targeting the 5'- and 3'-ends of the partial mRNA sequence. The cloned HC29 cDNA was shown to be 1,113 bp in size with an open reading frame of 507 bp, encoding a protein of 168 amino acid with a calculated molecular mass of 18.9 kDa. Amino acid sequence analysis revealed that the cloned HC29 cDNA contained the conserved catalytic triad and dimer interface of selenium-independent glutathione peroxidase (GPX). Alignment of the predicted amino acid sequences demonstrated that the protein shared 44.7~80.4% similarity with GPX homologues in the thioredoxin-like family. Phylogenetic analysis revealed close evolutionary proximity of the GPX sequence to the counterpart sequences. These results suggest that HC29 cDNA is a GPX, a member of the thioredoxin-like family. Alignment of the nucleic acid and amino acid sequences of HC29 with those of the reported selenium-independent GPX of H. contortus showed that HC29 contained different types of spliced leader sequences as well as dimer interface sites, although the active sites of both were identical. Enzymatic analysis of recombinant prokaryotic HC29 protein showed activity for the hydrolysis of H2O2. These findings indicate that HC29 is a selenium-independent GPX of H. contortus.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; DNA, Complementary/genetics/isolation & purification ; Glutathione Peroxidase/*genetics/*metabolism ; Goat Diseases/parasitology ; Goats ; Haemonchiasis/parasitology/prevention & control/*veterinary ; Haemonchus/*enzymology/*genetics ; Hydrogen Peroxide/metabolism ; Molecular Sequence Data ; Phylogeny ; RNA, Helminth/chemistry/genetics ; Random Amplified Polymorphic DNA Technique ; Rats ; Rats, Sprague-Dawley ; Sequence Alignment ; Sequence Analysis, DNA