Human sparganosis is a zoonotic disease caused by infection with larval forms (procercoid/plerocercoid) of Spirometra spp. The purpose of this study was to identify Spirometra spp. of infected snakes using a multiplex PCR assay and phylogenetic analysis of mitochondrial DNA sequence data from the spargana of terrestrial snakes obtained from Korea and China. A total of 283 snakes were obtained that included 4 species of Colubridae comprising Rhabdophis tigrinus tigrinus (n=150), Dinodon rufozonatum rufozonatum (n=64), Elaphe davidi (n=2), and Elaphe schrenkii (n=7), and 1 species of Viperidae, Agkistrodon saxatilis (n=60). The snakes were collected from the provinces of Chungbuk, Chungnam, and Gyeongbuk in Korea (n=161), and from China (n=122). The overall infection rate with spargana was 83% (235/283). The highest was recorded for D. rufozonatum rufozonatum (100%), followed by A. saxatilis (85%) and R. tigrinus tigrinus (80%), with a negative result for E. davidi (0%) and E. schrenkii (0%). The sequence identities between the spargana from snakes (n=50) and Spirometra erinaceieuropaei (KJ599680) or S. decipiens (KJ599679) control specimens were 90.8% and 99.2%, respectively. Pairwise genetic distances between spargana (n=50) and S. decipiens ranged from 0.0080 to 0.0107, while those between spargana and S. erinaceieuropaei ranged from 0.1070 to 0.1096. In this study, all of the 904 spargana analyzed were identified as S. decipiens either by a multiplex PCR assay (n=854) or mitochondrial cox1 sequence analysis (n=50).
Agkistrodon ; China* ; Chungcheongbuk-do ; Chungcheongnam-do ; Colubridae ; DNA, Mitochondrial ; Gyeongsangbuk-do ; Humans ; Korea* ; Multiplex Polymerase Chain Reaction ; Sequence Analysis ; Snakes* ; Sparganosis ; Sparganum ; Spirometra* ; Viperidae ; Zoonoses

Cryptosporidium is an important pathogen causing gastrointestinal disease in snakes and is distributed worldwide. The main objectives of this study were to detect and identify Cryptosporidium species in captive snakes from exotic pet shops and snake farms in Thailand. In total, 165 fecal samples were examined from 8 snake species, boa constrictor (Boa constrictor constrictor), corn snake (Elaphe guttata), ball python (Python regius), milk snake (Lampropeltis triangulum), king snake (Lampropeltis getula), rock python (Python sebae), rainbow boa (Epicrates cenchria), and carpet python (Morelia spilota). Cryptosporidium oocysts were examined using the dimethyl sulfoxide (DMSO)-modified acid-fast staining and a molecular method based on nested-PCR, PCR-RFLP analysis, and sequencing amplification of the SSU rRNA gene. DMSO-modified acid-fast staining revealed the presence of Cryptosporidium oocysts in 12 out of 165 (7.3%) samples, whereas PCR produced positive results in 40 (24.2%) samples. Molecular characterization indicated the presence of Cryptosporidium parvum (mouse genotype) as the most common species in 24 samples (60%) from 5 species of snake followed by Cryptosporidium serpentis in 9 samples (22.5%) from 2 species of snake and Cryptosporidium muris in 3 samples (7.5%) from P. regius.
Agriculture ; Animals, Exotic ; Boidae ; Colubridae ; Cryptosporidium parvum ; Cryptosporidium* ; Dimethyl Sulfoxide ; Floors and Floorcoverings ; Gastrointestinal Diseases ; Genes, rRNA ; Methods ; Milk ; Oocysts ; Polymerase Chain Reaction ; Snakes* ; Thailand* ; Zea mays

A total of 205 larval gnathostomes were collected from 18 (22.5%) of 80 red banded odd-tooth snakes, Dinodon rufozonatum rufozonatum, which had been smuggled from China and confiscated at Customs in Busan, Republic of Korea. In order to identify the species, some of the larvae were observed by a light microscope and a scanning electron microscope (SEM). The larvae were 2.18 x 0.29 mm in average size, and had a pair of lips at the anterior end, a muscular esophagus, 2 pairs of cervical sacs, and brownish intestines. The head bulb was characteristically equipped with 4 rows of hooklets; the average number of hooklets in each respective row was 38.6, 40.5, 41.5, and 43.7. In SEM views, the mouth evidenced a pair of lateral lips of equal size in a half-moon shape. Each lip featured a couple of labial papillae and a small amphid located between the 2 papillae. The hooklets on the head bulb had single-pointed, posteriorly-curved tips. The cuticular spines were larger and more densely distributed on the anterior part of the body, and decreased gradually in size and number toward the posterior body. On the basis of these morphological characteristics, the larvae were identified as the third stage larvae of Gnathostoma hispidum.
Animals ; China ; Colubridae/*parasitology ; Gnathostoma/*isolation & purification/pathogenicity/*ultrastructure ; Larva/ultrastructure ; Microscopy, Electron, Scanning/veterinary ; Muscles/parasitology ; Species Specificity ; Spirurida Infections/parasitology/*veterinary

OBJECTIVETo observe three species of Kalicephalus found in three species of snake (Ophiophagus hannah, Ptyas mucosus, and Naja Sputatrix) during research on Capture Snake for Trading in Java and Snake Biodiversity in Kalimantan Islands.

METHODSSpecimens for light microscopy examination were fixed with warm 70% alcohol, cleared and mounted in lactophenol for wet mounting. Drawings were made with the aid of a drawing tube attached to a Nikon compound microscope. Measurements were given in micrometers (µ) as the average of findings, followed by the range in parentheses, unless otherwise stated.

RESULTSKalicephalus (Costatus) indicus was found from 7 Ptyas mucosus, Kalicephalus bungari from 2 Naja sputatrix and 1 Kalicephalus (Costatus) indicus and Kalicephalus assimilis found from 1 Ophiophagus hannah. The morphology and measurement of three species of Kalicephalus found in this study were close to those described before.

CONCLUSIONSNew finding of host of Kalicephalus (Costatus) indicus and Kalicephalus bungari was a snake species of Naja sputatrix. New records of locality were Kalimantan island as the new locality of Kalicephalus assimilis, and Java island was new locality of Kalicephalus (Costatus) indicus.

Ancylostomatoidea ; anatomy & histology ; growth & development ; isolation & purification ; physiology ; Animal Structures ; anatomy & histology ; Animals ; Body Size ; Colubridae ; parasitology ; Elapidae ; parasitology ; Host Specificity ; Indonesia ; Intestines ; parasitology ; Male

The phylogenetic relationships of the 3 Neodiplostomum spp. (Digenea: Neodiplostomidae) occurring in Korea (N. seoulense, N. leei, and N. boryongense) were analyzed using the partial mitochondrial cytochrome c oxidase subunit 1 (CO1) gene. The adult flukes were recovered from Sprague-Dawley rats (N. seoulense) and newborn chicks (N. leei and N. boryongense) experimentally infected with the neodiplostomula from the grass snake, Rhabdophis tigrinus tigrinus. The genomic DNA was amplified using specific primers, and the sequence of CO1 was obtained. According to the results, the pairwise similarity was 96.1% between N. boryongense and N. seoulense, but was 95.0% between N. boryongense and N. leei and 94.2% between N. leei and N. seoulense. The results demonstrated a closer phylogenetic relationship between N. seoulense and N. boryongense. This high relationship of N. seoulense and N. boryongense may be related to their similar morphologic features including the limited distribution of vitellaria and the presence of a genital cone. N. leei is distinct on the other hand with an extensive distribution of vitellaria and the absence of a genital cone.
Animals ; Base Sequence ; Chickens ; Cluster Analysis ; Colubridae/*parasitology ; DNA, Helminth/chemistry/genetics ; Electron Transport Complex IV/*genetics ; Female ; Korea ; Molecular Sequence Data ; Phylogeny ; Rats, Sprague-Dawley ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid ; Trematoda/*classification/*genetics

OBJECTIVETo evaluate the effect of soluble total proteins of Zaocys dhumnades on the expressions of interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and IL-10 in human fibroblast-like synoviocytes (FLS) cultured in vitro.

METHODSPrimary cultured FLS isolated from the synovium of patients with rheumatoid arthritis (RA) were incubated in the presence of different concentrations (50, 150 and 450 microg/ml) of soluble total proteins of Zaocys dhumnades, with Tripterygium hypoglaucum Hutch (THH) and DMEM as the positive and negative controls, respectively. Enzyme-linked immunosorbent assay (ELISA) and RT-PCR were used to detect the expressions of IL-1beta, IL-10 and TNF-alpha in the FLS.

RESULTSThe protein and mRNA levels of IL-1beta and TNF-alpha in the supernatant of the FLS exposed to 150 and 450 microg/ml of the soluble total proteins of Zaocys dhumnades decreased, while IL-10 protein and mRNA increased significantly as compared with those in the negative control group (P<0.01).

CONCLUSIONThe soluble total proteins of Zaocys dhumnades produce therapeutic effect on RA possibly by inhibiting IL-1beta and TNF-alpha and promoting IL-10 expressions in the FLS.

Animals ; Arthritis, Rheumatoid ; Cells, Cultured ; Colubridae ; Humans ; Interleukin-10 ; secretion ; Interleukin-1beta ; secretion ; Materia Medica ; chemistry ; pharmacology ; Primary Cell Culture ; Proteins ; isolation & purification ; Synovial Membrane ; cytology ; metabolism ; pathology ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo study the effect of Xianlong granules (XLG) on immunological function in the rat of adjuvant arthritis (AA).

METHODRats were randomly divided into normal group, AA model group, prednisone group and low, middle and high dose XLG groups, 10 rats in each group. All rats were treated by intragastric administration from the 18 days after arthritis was induced by the complete Freud's adjuvant and the effect of XLG on toes swelling was observed. On the 30th days after modeling, proliferation of the splenic and thymic lymphocytes, and IgG secreted by splenocytes were detected respectively by MTT assay and ELISA.

RESULTCompared with the model group, both the high and middle dose XLG groups had significant therapeutic effects on toes dwelling in the rat of AA (P < 0.05 or P < 0.01); The low, middle and high dose XLG groups strengthened the PHAM-inhibited proliferation of splenic lymphocytes (P < 0.05), and inhibited the PHAM-augmented proliferation of thymic lymphocytes (P < 0.05); XLG did not significantly effect on IgG level secreted by splenocytes in rats of AA.

CONCLUSIONXLG can cure toes swelling in rats of AA, which is related with regulation of the abnormal immunlological function.

Animals ; Arthritis, Experimental ; immunology ; pathology ; prevention & control ; Cell Proliferation ; drug effects ; Colubridae ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Edema ; immunology ; pathology ; prevention & control ; Female ; Immunoglobulin G ; metabolism ; Lymphocytes ; drug effects ; pathology ; secretion ; Male ; Materia Medica ; isolation & purification ; pharmacology ; Medicine, Chinese Traditional ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Spleen ; pathology ; secretion ; Thymus Gland ; pathology ; Toes ; pathology