The Cathepsin L-like cysteine proteinase genes (cpls) are multifunction genes related to the parasitic abilities of plant parasitic nematodes. A new cathepsin L-like cysteine proteinase gene (Dd-cpl-1) (GenBank Accession GQ 180107) was cloned from Ditylenchus destructor by RT-PCR and RACE. The cDNA sequence consisted of a 1 131 bp open reading frame (ORF) encoding 376 amino acid residues that were franked by a 29 bp 5'-untranslated region (UTR) and a 159 bp 3'-UTR. Genomic sequence analysis showed that Dd-cpl-1 contained 7 introns, obeyed the GT/AG rule in the splice-site junctions. Homology analysis showed that the identity was 77% between Dd-cpl-1 deduced protein Dd-CPL-1 and cathepsin L-like cysteine proteinase of Bursaphelenchus xylophilus. Multi-sequence alignment indicated that there were the catalytic triad (Cys183, His322 and Asn343) and two motifs ERFNIN motif and GNFD motif in deduced protein Dd-CPL-1. Cysteine proteinases phylogenetic analysis showed that Dd-cpl-1 belonged to the sub-clade of cathepsin L-like cysteine proteinases.
Amino Acid Sequence ; Animals ; Cathepsin L ; genetics ; Cloning, Molecular ; Cysteine Proteases ; genetics ; Genes, Helminth ; genetics ; Molecular Sequence Data ; Nematoda ; enzymology ; genetics ; Phylogeny ; Sequence Alignment ; Sequence Analysis, Protein ; Sequence Homology, Amino Acid ; Solanum tuberosum ; parasitology

A cDNA, named Dd-ace-2, encoding an acetylcholinesterase (AChE, EC3.1.1.7), was isolated from sweet-potato-stem nematode, Ditylenchus destructor. The nucleotide and amino acid sequences among different nematode species were compared and analyzed with DNAMAN5.0, MEGA3.0 softwares. The results showed that the complete nucleotide sequence of Dd-ace-2 gene of Ditylenchus destructor contains 2425 base pairs from which deduced 734 amino acids (GenBank accession No. EF583058). The homology rates of amino acid sequences of Dd-ace-2 gene between Ditylenchus destructor and Meloidogyne incognita, Caenorhabditis elegans, Dictyocaulus viviparous were 48.0%, 42.7%, 42.1% respectively. The mature acetylcholinesterase sequences of Ditylenchus destructor may encode by the first 701 residues of deduced 734 amino acids.The conserved motifs involved in the catalytic triad, the choline binding site and 10 aromatic residues lining the catalytic gorge were present in the Dd-ace-2 deduced protein. Phylogenetic analysis based on AChEs of other nematodes and species showed that the deduced AChE formed the same cluster with ACE-2s.
Acetylcholinesterase ; genetics ; Amino Acid Sequence ; Animals ; Cloning, Molecular ; DNA, Complementary ; genetics ; Genes, Helminth ; genetics ; Ipomoea batatas ; parasitology ; Molecular Sequence Data ; Nematoda ; enzymology ; genetics ; Plant Stems ; parasitology ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, Protein

Bursaphelenchus rainulfi isolated from dead pine trees in Zhejiang, China, is described and illustrated. It also provided some molecular characters of the Chinese population, including the PCR-RFLP and sequences of ITS region and D2-D3 expansion region of the large subunit (LSU) rRNA gene. Both the morphological characters and ITS-RFLP patterns match with the original description. The phylogenetic trees based on the 13 sequences of D2-D3 expansion region of the LSU rRNA gene and ITS region of Bursaphelenchus species were constructed, respectively, with the results showing the similar clades. The phylogenetic relationship based on the molecular data is similar to that with morphological characters. This is the first report of the species on pine wood in eastern China.
Animals ; Biological Evolution ; China ; DNA, Helminth ; genetics ; Nematoda ; anatomy & histology ; genetics ; Phylogeny ; Pinus ; parasitology ; Species Specificity

A phage display single-chain variable fragment (scFv) library against Bursaphelenchus xylophilus cellulase (BXC) was constructed and used to screen the specific antibodies binding to BXC. The total RNA was extracted from fresh spleens of BALB/C mice immunized with BXC. Gene fragments encoding VH and VL were amplified by RT-PCR and assembled into a single chain by overlapping PCR with a linker DNA encoding the peptide (Gly4Ser)3. The recombinant fragments were cloned into the phagemids (pCANTABSE) and electroporated into E. coli TG1. The recombinant phagemids were rescued by reinfection of helper phage M13K07. The repertoire of the phage display antibody was about 5 x 10(4). The specific antibodies against BXC were obtained after five rounds of affinity selection. The positive phage clone was used to infect E. coli HB2151. SDS-PAGE and western blot analysis showed that the soluble scFv antibodies expressed bound specifically to BXC. The studies laid foundation for quarantine and pathological study of Bursaphelenchus xylophilu.
Animals ; Cellulase ; genetics ; immunology ; Cloning, Molecular ; Electroporation ; Escherichia coli ; genetics ; metabolism ; Female ; Helminth Proteins ; genetics ; immunology ; Immunoglobulin Variable Region ; genetics ; Mice ; Mice, Inbred BALB C ; Nematoda ; enzymology ; Peptide Library ; Pinus ; parasitology ; Plant Diseases ; parasitology ; Recombinant Proteins ; biosynthesis ; genetics

A survey of canine heartworm (Dirofilaria immitis) infections among German shepherds in five areas of South Korea was performed from October 1994 to August 1995 using a microfilarial test (modified Knott's test) and an antigen test (DiroCHEK , Synbiotics, USA). The infection rate of 127 German shepherds (71 males and 56 females) was 10.2% (13/127) by the microfilarial test, but was 28.3% (36/127) by the antigen test, revealing that 24 of the 36 antigen-positive dogs were microfilaria-negative in the peripheral blood. All dogs that were microfilaria-positive were also antigen-positive. Three of the microfilaria-negative and antigen-positive dogs contained 4 - 15 adult heartworms in the heart and pulmonary arteries upon necropsy. The infection rate among German shepherds was the highest in Hoengsong-gun (Kangwon-do, 84.4%), while those of Yechon-gun (Kyongsangbuk-do) and Chungwon-gun (Chungchongbuk-do) areas were 20.0% and 14.3%, respectively. None of the dogs in the Kimhae-shi (Kyongsangnam-do) and Kwangju areas was heartworm positive. The infection rates of heartworm in dogs at the age of 1-3, 4-6, and 7-11 years were 6.3%, 21.4%, and 56.4%, respectively. Based on the fact that the antigen test is more accurate than the microfilarial test, this study strongly indicates that the prevalence rate of canine heartworm in Korea may be higher than previously reported (3.1+=23.0%) which utilized microfilarial tests.
parasitology-helminth-nematoda ; Dirofilaria immitis ; dogs ; antigen ; microfilaria ; epidemiology

this report describes the structures of high-mannose-type N-linked oligosaccharides in glycoproteins synthesized by the microfilariae of Dirofioria immitis. Microfilariae of D. immitis were incubated in vitro in media contaning 2-[(3)H] mannose to allow metabolic radiolabeling of the oligosaccharide moieties of newly synthesized glycoproteins. Glycopeptides were prepared from the radiolabeled glycoproteins by digestion with pronase and fractionation by chromatography on concanavalin A-Sepharose. Thirty eight percent of 2-[(3)H] mannose incorporated into the microfilariae of D. immitis glycopeptides was recovered in high mannose-type asparagine-linked oligosaccharides whech were bound to the immobilized lectin. Upon treatment of 2-[(3)H] mannose labeled glycopeptides with endo-beta-N-acetylglucosaminidase H, the high mannose-type chains were released and their structures were determined by high performance liquid chromatography and exoglycosidase digestion. The major species of high mannose-type chains synthesized by microfilariae of D. immitis have the composition Man(5) GlcNAc(2), Man(6) ClcNAc(2), Man(8) GlcNA(2), and Man(8) GlcNAc(2). Structural analyses indicate that these oligosaccharides are similar to high mannose-type chains synthesized by vertebrates.
parasitology-helminth-nematoda ; Dirofilaria immitis ; mannose ; carbohydrates ; oligosaccharieds ; micrlfilariae ; metabolic labeling ; HPLC ; enzyme digestion ; biochemistry

In order to know the species and frequency of human parasitic infection diagnosed by biopsy, 149 cases (0.18%) of parasitic infection were reviwed, which were selected from 80,947 biopsied materials submitted for routine histopathological examination during a period of 10 years from 1980 to 1989 at Department of Pathology, Chonnam National University Hospital. They consisted of 112 cases of cysticercosis, 17 paragonimiasis, 7 clonorchiasis, 4 amebiasis, 1 sparganosis, 1 enterbiasis, 1 aniskiasis, and 1 fascioliasis respectively. Based on morphological preservation of cysticercus, they could be divided into mild (20.2%), morderate (40.4%), and severe (39.4%) degeneration. Except 2 cases biopsied at the lungs, 15 cases of ectopic paragonimiasis were located at abdominal cavity (8 cases) and central nervous system (7 cases). One case of intrahepatic fascioliasis was observed. This is the 13th human fascioliasis reported in Korea. From the above results, the frequency of parasitic infections found in biopsied specimens was on the decrease as the cysticercosis and ectopic paragonimiasis.
parasitology-helminth-cestoda-nematoda-trematoda ; biopsy ; histopathology ; tissue parasite ; cysticercose ; ectopic ; Enterobius vermicularis ; Taenia solium ; Paragonimus westermani ; Clonorchis sinensis ; Entamoeba histolytica ; Anisakis ; Fasciola sp. ; paragonimiasis ; fascioliasis

This study was performed to observe the present status of intestinal parasitic infections in a remote island, Yondo, located in southern part of Jeonranam-do(province), Korea. In February and May 1988, total 1,011 individual stool samples were collected and examined for intestinal helminths and protozoa using formalin-ether centrifugal sedimentation technique. The results are summarized as follows: Of 1,011 ingabitants examined, 398(39.4%) were positive for intestinal parasites. Helminth positives were 372(36.8%), and protozoan cyst positives were 56(5.5%). Ten species of parasites were found. Trichuris trichiura revealed the highest infection rate of 27.5%, Ascaris lumbricoides 17.4%, Taenia sp. 5.8%, Entamoeba coli 3.3%, Giardia lamblia 1.5%, Endolimax nana 0.8%, Hymenolepis nana 0.4%, hookworm 0.2%, Trichostrongylus orientalis 0.2%, and Entamoeba histolytica 0.2%, respectively. The female group showed higher positive rate (44.0%) than males (34.7%). Also, higher positive rates were observed among adults as compared with the group younger than 10 years old. Average value of E.P.G. was 1,876(range 200-17,800) in A. lumbricoides positives, and 327(range 200-1,600) in T. trichiura positive cases. In helminth egg positive cases, single infection was 63.4%, double infection 34.7%, and triple infection 1.9%, respectively. Among protozoan cyst positives, single infection was 94.6%, and double infection was 5.4%. The present study revealed that the prevalence of intestinal parasites among inhabitants in Yondo island is still so high that special control measures should be performed.
parasitology-helminth-trematoda ; cestoda-nematoda-protozoa ; epidemiology ; Ascaris lumbricoides ; Taenia sp. ; Entamoeba coli ; Giardia lamblia ; Endolimax nana ; hookworm ; Trichostrongylus orientalis ; Entamoeba histolytica

Female and male nematodes isolated from an eye of a 36 year old Korean man in August 1987 were identified as Thelazia callipaeda. This is the 17th record of human thelaziasis in Korea. The internal structure of both female and male were described.
parasitology-helminth-nematoda ; thelaziasis ; Thelazia callipaeda ; eye ; case report

On October 26, 1987, the authors encountered a case of human thelaziasis involving both eyes. The case in Chollapuk-do, Korea, a farmer aged 23, complained of mild lacrimation for 3 weeks followed by 1 week duration of foreign body sensation in both eyes prior to examination of eyes. The conjunctivae of both eyes appeared slightly reddened. No other abnormalities were noticed. Total 6 worms, one male and two females from each eye, were removed, parasitologically studied, and identified as Thelazia callipaeda Railliet and Henry, 1910.
parasitology-helminth-nematoda ; thelaziasis ; Thelazia callipaeda ; eye ; case report