Genetic differences in the rat lungworm, Angiostrongylus cantonensis (Nematoda: Angiostrongylidae), in Thailand.

This study surveyed the genetic differences among Angiostrongylus cantonensis (A. cantonensis) using the mitochondrial cytochrome b (cytb) gene. Partial cytb sequences were determined for 91 worms from eight locations in Thailand. Using morphological techniques, the nematodes were found to be A. cantonensis. Phylogenetic analysis found two main clades, which were subdivided into four subclades (clusters). Haplotype network analysis showed that 11 distinct cytb haplotypes were also present in four groups of A. cantonensis. There was no observable relationship between the genetic differentiation of gene flow and geographical distance. This low genetic variation and geographical distribution of A. cantonensis in each location indicates a founder effect, which may have resulted from multiple independent origins, and suggests that haplotypes migrated from endemic areas via human-related activities.

Comparative morphometry, morphology of egg and adult surface topography under light and scanning electron microscopies, and metaphase karyotype among three size-races of Fasciola gigantica in Thailand.

Comparative morphometry of eggs and adults under light microscope, and morphology of adults under scanning electron microscope (SEM) were undertaken in the three size-races (< 25 mm, 25-35 mm, > 35 mm) of Fasciola gigantica (Thailand strain). Morphometric examination revealed intraspecific variation with respects to the dimensions of eggs and adults, whereas surface topography of the three size-race adults under SEM was morphologically similar. The observations on mitotic metaphase chromosomes of spermatogonial cells from testes of the three size-races revealed 2n=20 (diploid type), and no karyotypic difference was observed among them. The meiotic metaphase chromosome was 10 bivalents in primary spermatocyte in diplotene to diakinesis, and many mature spermatozoa were seen in the testicular preparations.

The karyotype of the lung fluke, Paragonimus siamensis.

The karyotype of chromosomes obtained from the germ cells of Paragonimus siamensis was analyzed by air-drying technique. The result revealed the diploid number of chromosomes to be 2n=22. The entire chromosomes consisted of one large-sized metacentric, four medium-sized subtelocentrics, three small-sized metacentrics or submetacentrics and three small-sized submeta-centrics or subtelecentrics. The relative arm lengths of the chromosomes were 20.13+/-1.21, 13.16+/-0.42, 11.07+/-0.48, 10.37+/-0.84, 9.29+/-0.52, 6.50+/-0.39, 6.42+/-0.34, 6.27+/-0.28, 6.04+/-0.58, 5.65+/-0.39 and 5.03+/-0.40% respectively. These data were comparable with those of other related species.

Comparative studies on protein, isoenzyme and DNA restriction endonuclease profiles of Thai Paragonimus metacercariae.

General proteins and 14 enzymes from metacercariae of Paragonimus heterotremus, P. siamensis and P. westermani were determined by vertical polyacrylamide gel electrophoresis. The isoenzyme profiles showed considerable interspecific polymorphism for general protein (PT), malate dehydrogenase (MDH), malic enzyme (ME) and tetrazolium oxidase (TO) while those of glucose phosphate isomerase (GPI) and phosphoglucomutase (PGM) showed similarity. The Pt-6 and To-I loci can be used as identification markers for these three species. The preliminary study of the molecular biology of Paragonimus heterotremus P. siamensis and P. westermani was based on analysis of metacercarial genomic DNA with restriction endonuclease Pst I. Agarose gel electrophoresis revealed restriction fragment length differences among the three species studied. The DNA restriction fragments were approximately 4-6 fragments, ranging from 5.35 to 14.67 kb. Among these. P westermani shared two homologous fragments with P. siamensis, ie, 5.35 and 7.22 kh, none with P. heterotremus, while P. heterotremus shared only one with P. siamensis, ie, 8.16 kb. Thus, the DNA restriction fragment length differences can be used to differentiate among these three species.

Angiostrongyliasis: analysis of antigens of Angiostrongylus costaricensis adult worms versus IgG from infected patients with Angiostrongylus cantonensis.

The possibility of cross-reactivity was previously investigated by indirect ELISA with sera from Angiostrongylus cantonensis infections, normal controls and A. costaricensis antigen. 5 microg/ml of crude antigen from both sexes of each species reacted with diluted serum samples (1:800) of each of 20 cases of angiostrongyliasis and normal controls, and further with anti-human IgG conjugate at 1:1,000. The mean absorbance values were evaluated as follows; normal controls showed a value of 0.033 using A. costaricensis antigen lower than (0.085) A. costaricensis antigen. Both mean values of angiostrongyliasis cases were rather close (0.491) using A. costaricensis antigen and the other antigen (0.518). The present study continued with a crude antigen of 13 A. costaricensis females and males. Serum samples were analyzed; 27 sera of angiostrongyliasis, 30 negative controls and 193 cases of other parasitic infections (91 cases of nematodiasis; 45 cases of cestodiasis; 47 cases of trematodiasis and 10 cases of HIV) and 7 cases of other brain infections. This antigen was evaluated for ELISA with a concentration of 5 microg/ml, serum dilution 1:400 and anti-human IgG conjugate at 1:2,000. The test gave sensitivity and specificity at cut-off value 0.261; 92.59% and 73% respectively. The antigen was cross-reactive with 30 cases from 9 out of 10 different kinds of nematodiasis (gnathostomiasis, strongyloidiasis, ascariasis, hookworm infections, trichinosis, toxocariasis, trichuriasis, onchocercosis and Wuchereria bancrofti infections. Five cases from 3 of 6 kinds of cestodiasis (neurocysticercosis, echinococcosis and Hymenolepis nana infections) and 18 cases of 4 out of 5 kinds of trematodiasis (Paragonimus heterotremus infections, opisthorchiasis, schistosomiasis and fascioliasis). One case of other brain infections was observed. The crude antigen of A. costaricensis showed a high percentage sensitivity with serum antibodies of angiostrongyliasis cases. Low specificity of the test was observed by reactions of those serum antibodies with various kinds of antigenic molecules. This study provides baseline data for further immunodiagnosis of human angiostrongyliasis.

Chromosomes and C-banding of Opisthorchis viverrini.

Chromosome of Opisthorchis viverrini was observed by air-drying and C-banding techniques. The chromosome number was 2n=12 and n=6 consisting of one large-sized metacentric, one medium-sized metacentric, one small-sized metacentric, one small-sized submetacentric or subtelecentric, one small-sized subtelocentric or acrocentric and one small-sized acrocentric. The relative lengths of the chromosomes were 32.02 +/- 2.52, 23.28 +/- 1.98, 15.24 +/- 3.40, 13.39 +/- 3.11, 10.18 +/- 1.56 and 5.82 +/- 0.59% respectively. After C-banding treatment, two of the small-sized chromosomes showed a remarkable constitutive heterochromatin.