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Genetic variability and differentiation of natural populations in three Japanese lung flukes, Paragonimus ohirai, Paragonimus iloktsuenensis and Paragonimus sadoensis (Digenea: Troglotrematidae).

The genetic differences among 3 closely related species, Paragonimus ohirai, Paragonimus iloktsuenensis and Paragonimus sadoensis, were assessed by electrophoretic analysis of allozymes. Fifteen enzymes were analyzed using starch gel electrophoresis. Seven of 18 loci were polymorphic. The most variable population (POs) was polymorphic at 6 of the loci (P = 0.333) with a heterozygosity H of 0.054. Genetic distances between P. ohirai and P. iloktsuenensis, between P. ohirai and P. sadoensis, and between P. iloktsuenensis and P. sadoensis were, on the average, 0.0375, 0.0809 and 0.0567, respectively; values which are well within the usual range for conspecific populations. An especially low estimate of genetic distance was obtained between sympatric populations of P. ohirai and P. iloktsuenensis (0.00059) in Sendai, indicating that gene flow occurred between the 2 species.

Animals↗

Human lung fluke Paragonimus heterotremus: differential diagnosis between Paragonimus heterotremus and Paragonimus westermani infections by EITB.

The specificity of three major polypeptides (35, 33 and 32.5 kD) from Paragonimus heterotremus antigens prepared from ether-extracted adult worms was tested against sera from heterologous infections as well as against P. westermani-infected sera. Only the 35 kD polypeptide was not present, its antigenic determinant being bound to the antibodies from all P. westermani-infected cases. Its cross-reactivity against various sera from heterologous helminthiases and other lung infections showed that it is not bound to these antigenic polypeptides. These major bands cannot be detected by Concanavalin A detector. Our research encourages the pattern (35, 33 and 32.5 kD) of immunoblot reactions for the diagnosis of P. heterotremus infections; the 35 kD antigen is specific for corresponding species and able to differentiate infections between both species of Paragonimus.

Animals↗

The antigens of Paragonimus westermani, Schistosoma mansoni, and Fasciola hepatica adult worms. Evidence for the presence of cross-reactive antigens and for cross-protection to Schistosoma mansoni infection using antigens of Paragonimus westermani.

The presence of cross-reacting antigens between Paragonimus westermani, Schistosoma mansoni, and Fasciola hepatica adult worms was demonstrated by Ouchterlony immunodiffusion and enzyme-linked immunosorbent assay (ELISA). A serum bank was developed against the three trematode genera to serve as probes to determine the presence of cross-reacting antibodies to P. westermani worm extracts. In this manner, it was possible to demonstrate that antigens common to F. hepatica and S. mansoni tegument were also present in P. westermani worm extracts. Likewise, it was possible to demonstrate that the F. hepatica antigens which bind to Concanavalin A, as well as the subfraction which in isoelectric focusing has a pI of 4.2, were also found in P. westermani worms. Also, a monospecific polyclonal serum to a Fasciola/Schistosoma cross-reacting antigen and the anti-P. westermani serum both reacted in Ouchterlony immunodiffusion with the P. westermani antigenic extract, each producing a line which linked with each other indicating common antigenic determinants and suggesting a common antigen among the digenetic trematodes. Finally, the P. westermani antigenic extracts induced in mice the production of antibodies which reacted with S. mansoni adult worm antigens by ELISA. As all of the Fasciola and Schistosoma sera were prepared against antigenic preparations which induced in mice protection to challenge infection with S. mansoni, this suggested that the P. westermani worms also contain protective antigens against S. mansoni. Immunity to Schistosoma mansoni infection was induced in mice by vaccination with Paragonimus westermani whole worm extracts (PwWWE). Immunized mice showed as high as a 67% worm burden reduction over controls. High doses of PwWWE did not confer protection to S. mansoni infection. Thus, in this study, immunity in heterologous systems was demonstrated and the existence of a common protective antigen shared by the digenetic trematodes was suggested.

Animals↗

[Study on isoenzymes of Paragonimus iloktsuenensis and Paragonimus westermani by isoelectric focusing].

The isoenzyme patterns of Paragonimus iloktsuenesis and P. westermani were studied by isoelectric focusing on polyacrylamide gels for PGM, MDH, HK, ME, PO and ACP. Interspecific differences were apparent as shown by the isoenzyme profiles of these enzymes examined. MDH was polymorphic with 2 recognizable phenotypes, designated common type and variant type respectively, in P. i. and with 3 phenotypes, designated MDH1, MDH2 and MDH1-2 respectively, in P. w. The result showed that the two species could be distinguished one from another by such species-specific band patterns of isoenzymes. This suggests that P. w. and P. i. have undergone genetically divergent development and P. i. can be regarded as a valid species different from P. w. In the light of MDH polymorphism, some implications of isoenzyme analysis on taxonomy of the genus Paragonimus were discussed.

Animals↗

Evidence for metacercarial polymorphism in lung flukes, Paragonimus ohirai and Paragonimus iloktsuenensis.

Cross breeding experiments between 2 species of lung fluke, Paragonimus ohirai and Paragonimus iloktsuenensis, were carried out using metacercarial characteristics as distinguishing markers. All metacercariae of F1 obtained were identical to those of P. ohirai. In the F2 and BF1 of F1 X P. iloktsuenensis, both the P. ohirai and P. iloktsuenensis types of metacercariae appeared. In the BF1 of F1 X P. ohirai, however, only the P. ohirai type metacercariae were produced. No intermediate type between the 2 species appeared. The results obtained demonstrate that the differences in the metacercariae, which were previously regarded as the most important characteristic for specific discrimination between these 2 flukes, are only a hereditary phenomenon within a single species. The metacercarial form, number of cyst layers, and body size seem to be controlled by a couple of alleles or very closely linked genes following simple Mendelian inheritance. Furthermore, we confirmed that reproduction in the lung flukes depends on cross-fertilization.

Alleles↗

Studies On The Lungfluke, Paragonimus Iloktsuenensis: IV. A Mathematical Analysis On Metacercarial Density Of Paragonimus Iloktsuenensis In Crab Host, Sesarma Dehaani.

Mathematical models such as the negative binomial, Poisson and Polya-Eggenberger distributions were applied to the observed data of the number of metacercariae of Paragonimus iloktsuenensis in crab hosts, Sesarma dehaani which were collected in Hadong, South Kyong-sang Do, Korea. From the above analysis, it was found that the pattern of density of metacercariae of this lungfluke among the crab hosts was well fitted to the negative binomial distribution, rather than to the Poisson or Polya-Eggenberger distribution.

Journal Article↗

Paragonimus heterotremus and other Paragonimus spp. in Thailand: pathogenesis, clinic and treatment.

Six species of Paragonimus have been reported in Thailand: P. siamensis in cat, bandicoot and rat; P. bangkokensis in mongoose; P. harinasutai in cat and dog (experiment); P. macrochis in bandicoot and rat; P. westermani in tiger and P. heterotremus in cat, dog and man. It is interesting to note that in 1965 two immature P. heterotremus worms were recovered for the first time in man, namely in subcutaneous swellings in a boy; in 1981 nine mature P. heterotremus worms were expectorated after praziquantel treatment. P. heterotremus has been postulated to be the main cause of human paragonimiasis in Thailand. The clinical manifestation of paragonimiasis heterotremus is similar to paragonimiasis westermani. In the 1960's and 1970's bithionol was used to treat paragonimiasis, the cure rate was only 50-60%, and side effects including urticaria, rash, abdominal pain, nausea, vomiting, diarrhoea and dizziness were common. In the past 4 years, niclofolan and praziquantel (2-cyclohexyl-carbonyl-1,2,3,6,7,11b-hexahydro - 4H - pyrazino [2,1-a]isoquinolin-4-one, EMBAY 8440, Biltricide) have been used. A single dose of 2 mg/kg body weight of niclofolan yielded 100% cure rate. Praziquantel at dosages of 3 X 25 mg/kg body weight daily for one day and two days gave 80% and 100% cure rates, respectively. The eggs disappeared in 2-3 weeks with improvement of symptoms and signs, but radiologically lesions took a few months or more to clear, depending on size and severity. Side effects in the niclofolan group were higher; in the praziquantel group side effects were minimal and no toxic effects were detected.

Anthelmintics↗

An ecological study of Paragonimus in Malaysia.

An ecologic study on Paragonimus in Malaysia was attempted from May to September 1967. Seven streams located in various directions and distances from Kuala Lumpur were surveyed for the study of intermediate hosts, snail and crab. One Malayan village and one aborigine village where infected crabs were found, and two tuberculosis hospitals in K.L. were surveyed for the study of human population. Intradermal tests along with sputum or stool examination to detect human infection by Paragonimus were employed. Wild animals, only a few, were shot in the vicinity of the aborigine village and several domestic cats from the Malayan village were bought. These animals were autopsied and examined for adult Paragonimus. Among five species of crab collected from the study areas, only two species, Potamon jahorenes and Parathelphusa maculata were found to be infected with Paragonimus. P. maculata seemed to be better crab host for the Paragonimus because this species had higher infection rate and metacercarial density than the other in the very same area. Three out of seven streams had infected crabs and the infection rate as well as the infection intensity varied from one stream to another. Only avilable snail in the streams was identified as Brotia costula. The infection rate of the snail was very low, six snails out of 11,898, which is about the same rate reported from other countries. Infected snail, however, had thousands of rediae uncountable containing about twelve microcercocercariae in each redia, sufficient enough to maintain the life cycle of the parasite even with only a few infected snail, the amplifier. This is the first confirmed report on the snail host of Paragonimus from Malaysia where the existence of Paragonimus had been reported in 1923. The first trial to study human population by means of intradermal test, sputum and/or stool examinations in Malaysia showed no evidence of human infection of Paragonimus. The number of animals, wild and domestic, examined for natural infection was too small to draw any statement. These examined animals were all negative for adult Paragonimus. Even though more extensive studies on wild animals and human population may be necessary for the definite conclusion, the facts that infected crabs from jungle stream where human contacts are extreamely rare, and also highly infected crabs from the area where none of humans or domestic animals were infected, strongly suggest the life cycle of Paragonimus in this area may be maintained by wild animal hosts rather than by human host. The morphology of all stages of the parasite, the pattern of penetrating glands, flame cells and excretroy bladder of cercaria, lancet shaped single cuticular spines and 6 branched ovary of adult worm obtained from experimentally infected cat, and the shape of egg including all measurements agree well with the characteristics of Paragonimus westermani.

Journal Article↗

Prevalence of Paragonimus westermani in some Ulchin school children.

In order to estimate the prevalence of Paragonimus westermani among the school children in Ulchin county, Kyungpook Province, five primary schools were selected for sampling. The children were first examined using an intradermal test with Paragonimus antigen, Lot No. 890417, and then examined for Paragonimus eggs from sputa and stools collected from the intradermal-positive cases. In a total of 382 children examined, 53 cases or 13.9 percent revealed positive reactions for Paragonimus westermani. The sex specific rate of infection was a little higher in boys than in girls. The age specific rate of Paragonimus westermani infections varied from age to age. Paragonimus westermani was most prevalent in the 8-year-old group in both sexes and showed about the same proportion among boys and girls of each age group. A comparison of the reliability for the detection of Paragonimus eggs from intradermal positive cases in this study showed that the sputum test was more sensitive than the stool test. Summarizing the results, this study indicates that the prevalence for Paragonimus westermani is still high and the endemic foci of the fluke exists in Ulchin county, Kyungpook Province, Korea.

Animals↗