Chronological observations of intestinal histopathology in rats (Rattus norvegicus) infected with Centrocestus caninus.

Intestinal pathological enzyme activity changes were studied chronologically in rats after Centrocestus caninus infection. A single inoculation of 300 metacercariae isolated from the gills of goldfish (Carassius auratus), was orally administered to male rats (n = 15). Uninfected animals were used as controls (n = 5). At days 3, 7, 14, 21, and 28 post-infection (PI), three infected rats, and one from each control group, were sacrificed. The duodenum, jejunum, and ileum were removed separately and fixed in 10% formalin and 10% cold formal calcium solution for histopathological and alkaline phosphatase activity investigations, respectively. The worms were found intruded into the intervillous space of the mucosa and the mucosa showed villous atrophy, crypt hyperplasia and stromal inflammation with inflammatory cell accumulations. Alkaline phosphatase (ALP) activity also showed retardation. However, it seemed that these phenomena would return to normal at the end of the experiment. It can be concluded, from our data, that C. caninus could cause mild histopathological alterations and reduce ALP activity in the small intestines.

A new record of Clinostomum philippinensis (Valasquez, 1959) in Trichogaster microlepis (Gunther, 1861) from Bung Borapet, Nakhon Sawan, Thailand.

Thirty-six freshwater fish from Bung Borapet, a large reservoir, were examined for helminths in July 2005. The prevalence of infection was 19.44 (7/36) and the intensity was 4.67 (168/36) per fish. Seven fish were infected with 4 species of helminths: 1 trematode, Clinostomum philippinensis (metacercariae); 1 cestode, Senga chiangmaiensis; 1 acanthocephalan, Pallisentis sp and 1 nematode, Camallanus anabantis. An encysted metacercaria of Clinostomum philippinensis was found in the gills of Trichogaster microlepis. The present study is a new report of Clinostomum metacercariae in the second intermediate host, T. microlepsis, in Thailand. Future work should investigate the epidemiology of the metacercarial stage in the intermediate host and the adult in the experimental definitive host.

Dna quantities and qualities from various stages of some trematodes using optical and HAT-RAPD methods.

The aim of this experiment was to minimize DNA quantity and quality for detection by optical (spectrophotometer at 260 nm and 280 nm) and HAT-RAPD methods. Total DNA from different stages, adult, metacercaria and eggs of 6 trematode species were isolated for analysis. In this experiment, the adult trematodes were classified into 3 groups by size: small, Haplorchis taichui and Stellantchasmus falcatus; medium, Opisthorchis viverrini and Ganeo tigrinus; and large, Paramphistomum epiclitum and Fischoederius elongatus. The adult minimal DNA quantities and qualities of all specimen samples detected by optical method were 97.22,72.28, 3,167.00, 1,490.62, 21,382.66, and 27,321.77 ng; eggs were 3.92, 3.57, 3.72, 6.23, 17.53, and 14.01 ng, respectively; and metacercarial stages 50.70 and 40.98 ng in H. taichui and S. falcatus. In addition, the HAT-RAPD technique was chosen to amplify the minimal DNA qualities and quantities of all trematode specimens. Total DNA was 1-1 x 10(-12) ng; DNA templates in each dilution were used for amplification by primer OPA-09. DNA concentrations ranging between 1 x 10(-8) and 1 x 10(-11) ng were amplified with high polymorphism. Our experiment concluded that only a single specimen of each egg, metacercaria, or adult stage could be amplified with distinct bands.

Mucosal mast cell responses in rats (Rattus norvegicus) experimentally infected with Centrocestus caninus.

Mucosal mast cell (MMC) responses and worm recovery rates in rats experimentally infected with Centrocestus caninus were investigated. Metacercariae of C. caninus, procured from goldfish, Carassius auratus, were orally administered to twenty-five male rats (300 metacercariae each rat). The infected rats were sacrificed on days 3, 7, 14, 21 and 28 post-infection (PI) along with the control rats. Worm recovery was performed from each part of small intestine. To investigate MMC, duodenal, jejunal and ileal paraffinized-tissue sections were processed and stained with 1% alcian blue and 0.5% safranin-O. The average worm recovery rates were 42.8, 37.7, 21.2, 12.5 and 3.7% on days 3, 7, 14, 21 and 28 PI, respectively. The majority of the worms (98.9%) were collected from the duodenum and jejunum. The MMC numbers in the infected rats were significantly higher than those of the controls (p<0.05). A peak level was observed on days 14 PI and the numbers gradually decreased thereafter. The results reveal that MMC plays an important role in the expulsion of C. caninus from the host intestine. A more precise description of the role the MMC plays in helminth expulsion is still needed to understand the mechanism of host defense against intestinal helminthic infection, along with other effector cells, such as goblet cells.

A study of ectoparasites of Canis lupus familiaris in Mueang district, Khon Kaen, Thailand.

We studied ectoparasites found on Canis lupus familiaris sampled in five areas in Mueang district, Khon Kaen Province, Thailand. The prevalence of fleas and ticks as well as their density were determined in 100 dogs that did not receive treatments. A total number of 458 ectoparasites was found corresponding to two species: 25.8% Ctenocephalides canis and 74.2% Rhipicephalus sanguineus. R. sanguineus was the most abundant species, and Ct. canis was the only flea species found. The stages of R. sanguineus were larvae (5.3%), nymphs (29.1%) and adults (39.1% in male and 26.5% in female). The stages of Ct. canis were larvae (41.5%) and adults (58.5%). Both species were commonly found on domestic dogs in all areas of the study. Ct. canis was not present on domestic dogs in one sub-district. The prevalence rates of tick-harboring domestic dogs was 80% (R. sanguineus), and flea-haboring domestic dogs was 26% (Ct. canis).

Some factors affecting Stellantchasmus falcatus metacercaria in laboratory.

Killing factors, various temperatures and solutions were studied in the laboratory on Stellantchasmus falcatus metacercaria in half-beaked fish (Dermogenus pusillus). Killing criteria followed the Movability Index from 1.000 within 24 hours. The metacercariae were collected from Chiang Mai moat. They were incubated in 0.85 % NaCl at -20 degrees C, room temperature, 4 degrees, 37 degrees, and 65 degrees C. The in vitro investigation showed that at -20 degrees C and 65 degrees C, the worms were killed within 18 and 2 hours, respectively, while other temperatures produced no effect. The solutions investigated were NaCl (10, 20, 30, and 40%), lemon juice (25, 50, 75, and 100%), acetic acid (5, 10, 20, and 30%), vinegar (1, 3, and 5%) and water as a control. The worms were killed in NaCl at 20, 30, and 40% within 12, 6, and 2 hours, respectively. Acetic acid at 5% and 10% killed the metacercaria within 12 and 6 hours while at 20% and 30%, within 2 hours. The killing effect of 3% vinegar was found within 18 hours and of 5% vinegar within 12 hours. Lemon juice showed no killing effect.

In vitro effects of aqueous extract from Artocarpus takoocha Roxb on tegumental surface of Haplorchis taichui.

The tegumental surface of the adult intestinal fluke, Haplorchis taichui, after in vitro incubation in Tyrode's solution containing 25, 50, 75 and 100% of aqueous extract of Artocarpus takoocha for 1, 6, 12 and 24 hours, was monitored by Scanning Electron Microscopy (SEM). The effect on worm motility was studied in contemporizing period. The adult worms stopped their motility immediately after being incubated with 25, 50, 75 and 100% of aqueous extract of A. takoocha. Observed under a light microscope, most worms showed bubbling from the oral sucker, ventral sucker and excretory pore. After exposure to the solution, SEM observation showed large blebs protruding from the oral sucker, ventral sucker and excretory pore. The surface damage was composed of numerous small blebs, followed later by the disruption of tegument. The severity of damage increased due to the increasing of aqueous extract of A. takoocha and incubation periods.

Survival of Haplorchis taichui metacercariae in Lab-Pla, Thai traditional food preparation.

Lab-Pla is a famous kind of Thai traditional food in northern and northeasthern Thailand. Consumption of uncooked Lab-Pla can lead to trematode infection because cyprinoid fish serve as the 2nd intermediate hosts of trematodes. The preparation of Lab-Pla can be made in different ways, depending on seasonings used in different places. The effect of the seasonings used in Lab-Pla on the survival of Haplorchis taichui metacercariae were, therefore, determined. The methodology was done by the following: 100 g of chopped Siamese mud carp fish (Henicorhynchus siamensis) were used for each experiment of seasonings used for completely mixed Lab-Pla. Mixed materials were blended and digested by 1% pepsin solution to obtain metacercariae and then checked for the movement under a stereomicroscope. The results showed that metacercariae remained active in 5 experimental groups: fermented fish treated, fish sauce treated, chili powder treated, ground roasted rice treated, and complete mixed treated indicated by average as follows: 27, 38.7, 33.3, 42.7 and 21%, respectively. In conclusion, fermented fish and completely mixed methods have more effectiveness in killing metacercariae in Lab-Pla preparation than other seasonings (p < 0.05).

High prevalence of Haplorchis taichui metacercariae in cyprinoid fish from Chiang Mai Province, Thailand.

This study aimed to investigate Haplorchis taichui metacercarial infection in fish collected from the Chom Thong and Mae Taeng districts, Chiang Mai Province during November 2001 to October 2002. A total 617 cyprinoid fish of 15 species were randomly collected and examined for H. taichui metacercariae. All the species of fish were found to be infected with H. taichui. The infection rates were 91.4% (266/290) and 83.8% (274/327), with mean intensities of 242.9 and 107.4 in the Chom Thong and Mae Taeng districts, respectively. The portion of the fish body with the highest metacercarial density was the muscles, and second, the head, in both districts. In addition, the fish had mixed-infection with other species of trematodes, namely: Centrocestus caninus, Haplorchoides sp, and Haplorchis pumilio.

Recovery and growth of Haplorchis taichui (Trematoda: Heterophyidae) in chicks.

An experimental study was performed to observe the recovery and growth of a minute intestinal fluke, Haplorchis taichui in chicks (Gallus gallus domesticus). Metacercariae of H. taichui were isolated from Jullien's mud carp, Henicorhynchus siamensis, which were collected in the Chiang Mai Province, Thailand. Two hundred metacercariae were orally force-fed to each chick. The intestine of the chicks were examined from day 1 to day 54 post-infection (PI). The incidence of infection was 84.9% (28/33) and the mean intensity was 19.9 (656/33), with the range 0-59. The worm recovery rate was the highest at day 11 PI (29.5%). On day 3 PI, mature adult worms were recovered and 1-200 eggs were observed in the uterus of the worms. The worms grew rapidly in the chicks and the genital organs were fully developed in 14 days. This parasite can survive in chicks up to day 48 PI. It is concluded that they are a suitable definitive host for infection with H. taichui.

Susceptibility of rodents to Stellantchasmus falcatus infection.

Two rodent hosts, rats (Rattus norvegicus) and mice (Mus musculus), were examined for their susceptibility to Stellantchasmus falcatus infection. The metacercariae were recovered from half-beaked fish, Dermogenus pusillus, collected from Hang Dong District, Chiang Mai Province. A single inoculation of three hundred metacercariae was orally conducted with male rats (n=15) and mice (n=15). Uninfected animals were used as controls (n=5). At days 3, 7, 14, 21 and 28, post-infection (PI), three rats and mice, also one from each control group, were sacrificed for adult worm recovery, and the blood was drawn by heart puncture and prepared for white blood cell and eosinophil counts. The results showed that adult worm recovery gradually decreased after day 21 PI in the rats, whereas, reduction in the mice was found after day 7 PI and the adult worms completely disappeared at day 28 PI. Hematologically, total white blood cell counts in two kinds of infected hosts were not statistically different from those of controls. However, eosinophil counts of infected rats slightly increased during days 7-14 PI, and started to decrease at day 21 PI. A similar finding was observed in the infected mice. From these results, it can be concluded that the rat is a more suitable host for S. falcatus than the mouse. To understand the mechanisms implicated in determining host susceptibility, intensive studies are required.

Investigation of Stellantchasmus falcatus metacercariae in half-beaked fish, Dermogenus pusillus from four districts of Chiang Mai Province, Thailand.

Half-beaked fish, Dermogenus pusillus, collected from Mueang, Hang Dong, Doi Saket and Saraphi Districts, Chiang Mai Province, were examined for their infection status with Stellantchasmus falcatus (Digenea: Heterophyidae) metacercariae. The infection rate of the fish was 100%. Fish in three of four districts were found only to have metacercariae of S. falcatus, whereas those in Saraphi District had mixed infections with metacercariae of Posthodiplostomum sp. The intensity of S. falcatus infection per fish varied; 652-1,342 (mean 999.5), 562-2,422 (1,323.1), 185-2,492 (502.6), and 22-550 (210.4) in Mueang, Hang Dong, Saraphi, and Doi Saket Districts, respectively. The body portions of the fish with the heaviest metacercarial infection were the muscles, in all districts. The present study confirms that half-beaked fish in Chiang Mai Province are heavily infected with S. falcatus metacercariae.