Comparative Analysis of Humoral Immune Response and Cognate Antigen Detection in Experimentally Infected Sprague Dawley Rats with Brucella abortus Biotype 1.

Background: This study investigated the IgG-specific humoral immune responses against specific antigen-like whole-cell antigen (WCA), outer membrane protein (OMP), periplasmic protein (PP), and cytoplasmic protein (CP) during the acute and subacute stages of Brucella abortus biotype 1 infection in Sprague Dawley (SD) rats. Materials and Methods: The intraperitoneal method was used to experimentally infect forty-four 6- to 8-week-old SD rats with 1 × 109 colony-forming units (CFUs) of B. abortus biotype 1. Following inoculation, the rat was serially sampled for serum at 0, 3, 7, 14, 21, 28, 35, 42, 60, 90, and 120 days. The IgG-specific immune responses and recognition of immunodominant antigens in WCA, OMP, PP, and CP of B. abortus were assessed by indirect enzyme-linked immunosorbent assay (IELISA) and western blot (WB) assay using infected rat sera. Results: The IgG antibody response was detectable at 3 days after infection. The peak serum IgG antibody titers were recorded against CP and PP at 28 days after infection. The highest serum IgG antibody titers were recorded at 42 days after infection against WCA and 90 days after infection only against OMP. WB assay revealed a wide array of protein bands between molecular weight of 13 and 95 kDa for WCA, 13 and 95 kDa for OMP, 15 and 65 kDa for PP, and 12 and 85 kDa for CP. Proteins bands of 10, 13, 20, 24, 46, and 76 kDa for WCA; 28, 35, 39, 85, and 95 for OMP; 20, 30, 40, 43, 46, and 65 kDa for PP, and 12, 23, 68, and 85 for CP were intensely recognized. Conclusion: Data of this study indicated that WCA, CP, and PP of B. abortus could be useful for diagnosis of acute and subacute brucellosis in SD rat model. OMP of B. abortus could be useful for differential diagnosis of subacute brucellosis.

In Vitro and In Vivo IFN-γ and IL-10 Measurement in Experimental Brucella abortus Biotype 1 Infection in Sprague-Dawley Rats.

The immune response to Brucella abortus mainly depends on antigen-specific T cell activation, CD4+ and CD8+ T cells, and Brucella-specific humoral response. Protective immune response against Brucella infection has not been performed in the Sprague-Dawley (SD) rat model. We measured bacterial kinetics in addition to in vivo and in vitro interferon gamma (IFN-γ) and interleukin-10 (IL-10) production against crude Brucella protein in the SD rats at different days of postinfection with B. abortus biotype 1 by indirect enzyme-linked immunosorbent assay. Forty SD rats were inoculated intraperitoneally with 0.1 mL sterile injectable pyrogen-free solution containing 1 × 1010 colony-forming units/mL of B. abortus biotype 1 obtained from cattle in Korea. Four rats were used as uninfected control. Serum IFN-γ level at 3 and 7 days postinfection were significantly higher (p > 0.001) compared with the IL-10 level. On the contrary, serum IL-10 levels were observed significantly higher at 21 and 28 days postinfection compared with the serum IFN-γ levels (p < 0.001). The production of IFN-γ by spleen cells was significantly higher at 7 and 14 days postinfection compared with IL-10 (p < 0.001). On the contrary, IL-10 productions were found to be significantly higher at 21, 28, 35, and 42 days postinfection compared with IFN-γ (p < 0.001). The presence of B. abortus in blood was marked till 5 weeks of infection, throughout the experiment in case of spleen, and no bacteria were isolated from the kidney and liver at 6 weeks postinfection. The in vivo and in vitro IFN-γ and IL-10 measurement in our study reported that B. abortus infection in rats primarily educe T helper (Th)1-dominant immune response in acute infection accompanied by Th2-dominant immune response in chronic infection.

The Profile of Immunoglobulin A and Immunoglobulin G Subclasses in Sprague Dawley Rats Experimentally Infected with Brucella abortus Biotype 1.

This study measured total serum immunoglobulin A (IgA), immunoglobulin G (IgG)1, IgG2a response against whole cell antigen (WCA), outer membrane protein (OMP), periplasmic protein (PP), cytoplasmic protein (CP), and crude Brucella protein (CBP) of Brucella abortus in experimental brucellosis induced with B. abortus biotype 1 in Sprague Dawley (SD) rats during a 17-week infection period. Six- to 8-week-old SD rats (n = 44) were experimentally infected with 1 × 109 colony forming unit of B. abortus biotype 1 through the intraperitoneal route. Serial serum samples were collected from the rat at 0, 3, 7, 14, 21, 28, 35, 42, 60, 90, and 120 days after inoculation. The sera were tested by enzyme linked immunosorbent assay. We have noticed a very low level and short persistence of IgA antibody in our experiment. The low level and short persistence of IgA antibody suggest that this antibody isotype might not be protective against brucellosis in rats. Both Th1 and Th2 specific immune responses were recorded in our study with the production of IgG1 and IgG2a antibody isotopes, respectively. We noticed significant dominant IgG2a antibody responses over IgG1 responses throughout the experiment (p < 0.001) against WCA and OMP. The mixed Th1 and Th2 dominant immune responses mediated by IgG2a and IgG1 antibody isotypes were observed against CP, PP, and CBP. Data of our study suggest that IgG2a dominant responses in the early stages of disease play the main role in conferring protection against brucellosis and with the progress of disease IgG1 dominant responses were elicited.

Male rats transmit Brucella abortus biotype 1 through sexual intercourse.

The aim of this study was to evaluate transmission of Brucella abortus biotype 1 via sexual intercourse in rats. Male and female virgin Sprague-Dawley (SD) rats were experimentally infected intraperitoneally with 1×10(9)colony forming units (CFU) of B. abortus biotype 1, a Korean bovine isolate. At 14 days after infection, infected male rats (n=10) were housed with uninfected female rats (n=10) and infected female rats (n=10) were housed with uninfected male rats (n=10) for a period of one month. During this period all uninfected female rats became pregnant and 6 of 10 infected female rats became pregnant. Serum from two out of 10 female uninfected rats had positive reactions in the Rose Bengal Plate Agglutination Test (RBPAT), Tube Agglutination Test (TAT) or the Enzyme-Linked Immunosorbent Assay (ELISA); whereas none of the uninfected male rat had positive reactions in these tests. Using bacteriological culture and AMOS-PCR assay, B. abortus biotype 1 was isolated and identified from two uninfected female rats and all of the uninfected male rats were found negative for B. abortus biotype 1. It was concluded that transmission of B. abortus biotype 1 from infected male to uninfected female rats resulted from sexual intercourse.

Rats born to Brucella abortus infected mothers become latent carriers of Brucella.

INTRODUCTION: Rats are known to be infected with Brucella. Vertical transmission of brucellosis was recorded in rats. The study was performed to judge whether rats born from Brucella abortus infected mothers can act as latent carriers of Brucella infection. METHODOLOGY: Female Sprague Dawley (SD) rats were experimentally infected with B. abortus biotype 1 and subsequently bred 10 days post infection (PI). Serum samples of rats (n = 48) born from infected dams were tested using the Rose Bengal plate test (RBPT), tube agglutination test (TAT), and enzyme-linked immunosorbent assay (ELISA) at one, two and three months of age. Tissue samples were plated onto Brucella agar and blood agar media and incubated at 37 °C with 5% CO2 for five to seven days for isolation of bacteria. RESULTS: B. abortus was isolated from 18 out of 48 rats born to infected dams, and the isolates were confirmed as B. abortus by AMOS (B. abortus, melitensis, ovis and suis) PCR assay with the production of a 498 bp PCR amplicon. Serum samples of rats (n = 48) born from infected dams were tested negative using the RBPT, TAT and ELISA at all time points. CONCLUSION: We conclude from the study that rats born to infected dams may become latent carriers of Brucella infection potentially providing a reservoir for future transmission.

Immunoglobulin profiles in acute Brucellosis experimentally induced by Brucella canis in BALB/c mice.

This study evaluated profiles of immunoglobulin (Ig; IgA, IgG, IgG1, and IgG2a) response in experimental brucellosis induced with Brucella canis in BALB/c mice during an 8-week infection period. Six- to 8-week-old BALB/c mice (n = 36) were experimentally infected with 1 × 10(9) CFU of B. canis via the intraperitoneal route. Serial serum samples were collected from the mice at 0, 3, 7, 14, 21, 28, 35, 42, 49, and 56 days after inoculation. The sera were tested by the rapid slide agglutination test (RSAT) and 2-mercaptoethanol-RSAT and indirect enzyme-linked immunosorbent assay. Sera tested positive for B. canis by the RSAT and 2-mercaptoethanol-RSAT beginning from 7 days after inoculation until the end of the experiment. The IgA response was detected at 14 days after infection and reached peak levels at 21 days after infection. The IgG antibody responses were detected at 7 days after infection and reached the peak value at 35 days after infection. Data of our study demonstrated IgG2a-dominant responses over IgG1 during the course of infection (p > 0.05).

Characteristics of the immune response during acute brucellosis in Sprague-Dawley rats.

BACKGROUND: Brucella is a facultative, intracellular pathogen that causes severe disease in animals and humans. Immunity against Brucella involves both humoral and cellular responses. To investigate the characteristics of immune response in acute brucellosis in Sprague-Dawley (SD) rats, IgG and its subclass specific immunoglobulins' (IgG1 and IgG2a) response in sera against B. abortus biotype 1 infection were studied. METHODOLOGY: Thirty-six rats were inoculated intraperitoneally with 0.1 ml apyrogenic saline containing 1x10(10) colony forming unit (CFU) of B. abortus biotype 1 Korean bovine isolate. Four rats were used as uninfected controls. The sera were collected from infected rats at 3, 7, 14, 21, 28, 35, 42, 49, and 56 days post infection (DPI) and screened for Brucella specific antibody response by the rose bengal plate test (RBPT). IgG and its subclass specific immunoglobulins' (IgG1 and IgG2a) response in the sera were measured by a lipopolysaccharide (LPS) based indirect enzyme-linked immunosorbent assay (IELISA). RESULTS: Brucella specific IgG, IgG1 and IgG2a responses in the sera of infected rats were detected from 3 DPI by IELISA. IgG and IgG1 concentrations in sera reached the peak level at 35 DPI, and then the concentrations gradually declined to the end of the experiment. IgG2a concentrations in the sera remained almost constant from 7 DPI until the end of this study. CONCLUSION: In acute brucellosis, IgG2a response (indicative of a Th1 response) was found to be significantly dominant over IgG1 response (indicative of Th2 response) (P < 0.001).

Efficacy of strain RB51 vaccine in protecting infection and vertical transmission against Brucella abortus in Sprague-Dawley rats.

Immunizing animals in the wild against Brucella (B.) abortus is essential to control bovine brucellosis because cattle can get the disease through close contact with infected wildlife. The aim of this experiment was to evaluate the effectiveness of the B. abortus strain RB51 vaccine in protecting infection as well as vertical transmission in Sprague-Dawley (SD) rats against B. abortus biotype 1. Virgin female SD rats (n = 48) two months of age were divided into two groups: one group (n = 24) received RB51 vaccine intraperitoneally with 3 x 10(10) colony forming units (CFU) and the other group (n = 24) was used as non-vaccinated control. Non-vaccinated and RB51-vaccinated rats were challenged with 1.5 x 10(9) CFU of virulent B. abortus biotype 1 six weeks after vaccination. Three weeks after challenge, all rats were bred. Verification of RB51-vaccine induced protection in SD rats was determined by bacteriological, serological and molecular screening of maternal and fetal tissues at necropsy. The RB51 vaccine elicited 81.25% protection in SD rats against infection with B. abortus biotype 1. Offspring from rats vaccinated with RB51 had a decreased (p < 0.05) prevalence of vertical transmission of B. abortus biotype 1 compared to the offspring from non-vaccinated rats (20.23% and 87.50%, respectively). This is the first report of RB51 vaccination efficacy against the vertical transmission of B. abortus in the SD rat model.

Effects of Brucella abortus biotype 1 infection on the reproductive performance of Sprague-Dawley rats.

The aim of this study was to assess the impact of Brucella abortus biotype 1 infection on the reproductive performance using Sprague-Dawley (SD) rat model. Virgin female SD rats (n = 24) were infected intraperitoneally with 0.1 mL of saline containing 1 x 10(11) Colony Forming Unit (CFU) of pathogenic B. abortus biotype 1 Korean bovine isolate. Control rats (n = 24) were inoculated with 0.1 mL of apyrogenic saline. Both inoculated and control rats were divided into six subgroups. Four rats in each subgroup were consistently bred at 3, 7, 14, 21, 28 and 60 days after infection. B. abortus infection induced 41.67% infertility in the infected rats. The mean number of offspring/litter was 8.71 +/- 2.01 for infected rats and 12.87 +/- 1.42 for control rats (p < 0.001). The mean weight of the viable offspring was 6.10 +/- 0.36 g for infected rats and 7.15 +/- 0.40 g for control rats (p < 0.001). The rate of stillbirth was 12.30% in the infected rats. B. abortus biotype 1 was isolated from the uteri of the infected rats. The data of this study indicate that B. abortus biotype 1 infections in SD rat model affect reproduction adversely by causing infertility, stillbirth and loss of number and weight of offspring.

Diagnostic efficacy of Brucella abortus strain RB51 in experimentally inoculated Sprague-Dawley rats using western blot assay.

BACKGROUND: To investigate the diagnosis and efficacy of Brucella abortus strain RB51 (SRB51) in experimentally inoculated Sprague-Dawley (SD) rat using western blot assay. METHODOLOGY: Female SD rats were orally administered with 1.0 x 10(7) colony forming unit (cfu) suspension of SRB51 and half of these SD rats were challenged at 4 weeks post inoculation with 1.0 x 10(9) cfu suspension of B. abortus biotype 1 isolated in South Korea. Sera of SD rats were monitored at regular intervals by western blot assay using whole cell antigen of B. abortus strain 1119-3 (S1119-3). The bacteriological examination of blood and clinical examination of the rats were also performed. RESULTS: There were several bands at 120, 70, 45, 30, 20 kDa and clear specific bands were found after vaccination (20, 70 kDa) and challenge (15, 20, 45, 70, 120 kDa). The highest immune response was observed in sera 4 weeks post SRB51 vaccination. SRB51 was recovered from the blood of all of SRB51 inoculated rats until one week post vaccination and there were no clinical signs in that inoculated rats. CONCLUSIONS: It is concluded that the SRB51 elicits antigen specific immunity in SD rats based on western blot assay.

Antimicrobial resistance of Escherichia coli O157 from cattle in Korea.

Of 45 Escherichia coli O157 isolates from cattle feces, which were collected between May 2000 and September 2003 in Korea, 32 were resistant to at least 1 antibiotic and 28 were resistant to 4 or more antibiotics, with 32, 30 and 30 of the isolates being resistant to streptomycin, tetracycline and sulfisoxazole, respectively. Two isolates were resistant to fluoroquinolones and to 10 or more of the 22 other antimicrobial agents that were tested. Thirteen antimicrobial resistant patterns were observed. The most frequent resistance type, which was found for 11 isolates, was streptomycin-tetracycline-kanamycin-ampicillin-piperacillin-cephalothin-sulfisoxazole-ticarcillin. Polymerase chain reaction (PCR) analysis of the isolates for E. coli O157 virulence markers revealed that 25 of the resistant E. coli O157 isolates tested positive for stx2 or both stx1 and stx2 genes. These findings suggest that many of the resistant E. coli O157 isolates might cause disease in humans.

Migration of Strongyloides venezuelensis in rats after oral inoculation of free-living infective larvae.

Strongyloides venezuelensis (SVZ) infection was chronologically monitored in 85 Sprague-Dawley rats (SDR), which were orally inoculated with approximately 1,000 infective larvae. In order to describe the characteristics of migrating larvae (MLS) in various visceral organs (the liver, lung, cardiac blood, and small intestine), 5 SDR were sacrificed at 20 min, 45 min, 1 hr, 2 hr, 3 hr, 4 hr, 8 hr, 12 hr, 16 hr, 48 hr, 72 hr, 96 hr, 120 hr, 144 hr, 168 hr and 192 hr post inoculation (PI). MLS were recovered from the liver and blood 20 and 45 min PI and measured 788 +/- 26 microm and 846 +/- 40 microm in length, respectively. MLS were first observed in the lung tissue 45 min PI and measured 925 +/- 38 microm on the average. In the trachea, MLS measuring 849 +/- 75 microm appeared 3 to 96 hrs PI. Adult worms (AWS) measuring 1,926 +/- 521 microm to 2,956 +/- 159 microm in length were observed in the small intestine from 120 hr PI. The worms appeared to mature more than 168 hr PI and attained the average maximum length of 2,420 +/- 532 microm. At 3 hr PI focal hyperemic and necrotic lesions were evidently observed in the liver and lung, together with eosinophilic infiltration in the stomach, liver, and lung. The parasites were histologically detectable in the lung tissues but were very difficult to find in the liver and the epithelial layer of small intestine. These data demonstrate that SVZ parasites take 20 min to reach the liver via the stomach and only three hours to reach the trachea through the same route. The development from eggs to adults takes 168 hr in the SDR model.

Eurasian otter (Lutra lutra), a definitive host for Dirofilaria immitis.

A mature male and a mature female Dirofilaria immitis were found in the right ventricle of the heart of a naturally infected 2-yr-old male Eurasian otter (Lutra lutra) that had died of severe lung congestion at a zoo in South Korea. Both developing embryos and microfilariae were present in the uterus of the female D. immitis. Although circulating microfilariae were not detected in blood or tissue, the Eurasian otter may serve as a definitive host for D. immitis.

Persistent infection with Strongyloides venezuelensis in the Mongolian gerbil (Meriones unguiculatus).

To examine the fate of Strongyloides venezuelensis. Mongolian gerbils (Meriones unguicalatus) were orally infected with 1,000 L3 larvae per animal. Altogether, 50 gerbils divided into 5 groups of 10 each were monitored for a period of 570 days to document the kinetics of faecal egg output, adults worm population, morphological development, fecundity, and hematological changes including peripheral blood eosinophilia. This study chronicled a life long parasitism of S. venezuelensis in the gerbil host, and showed that S. venezuelensis infection was quite stable throughout the course of infection and the worms maintained their normal development as evidenced by their body dimension. A progressive loss of body condition of the infected gerbils was observed as the level of infection advanced. However, no detectable pathological changes were observed in the gastrointestinal tract. The present findings indicate that an immunocompetent host, such as the Mongolian gerbil, can serve as a life long carrier model of S. venezuelensis if the worms are not expelled within 570 days after infection.

[Fractionation of antigen for ELISA of bovine fascioliasis]

In order to obtain the most specific and sensitive antigen from crude antigens of Fasciola hepatica for the immunodiagnosis of bovine fascioliasis by the enzyme linked immunosorbent assay(ELISA), phosphate buffered saline extract of F. hepatica was prepared. The crude extract was fractionated into 7 antigens using Sephadex G-100 column chromatography. Seven fractionated antigens were applied to ELISA, precipitation test and intradermal test, respectively. Results obtained are as follows: The specificity (95 per cent confidence interval in negative sera of bovine fascioliasis; Mean+2 x SD of absorbance ) of the first (MW>150,000) and the second antigens (MW 120,000) were 93.7 per cent, but those of others including crude antigen showed 100 per cen.t. The sensitivity (positive sera of bovine fascioliasis having higher values with compared to the criterion) of the first, the sixth (MW 16,000) and the seventh antigen (MW<5,000) were 91.6 per cent, 87.5 per cent and 0 per cent, respectively, but those of others showed all 100 per cent. The absorbance by ELISA using the fifth antigen (MW 26,000) was 8.43-folds higher in the positive sera than that in the negative sera. This could be used as one of the most specific antigens for the immunodiagnosis of bovine fascioliasis. In Ouchterlony test, precipitin lines were not found in the sera naturally infected with F. hepatica, but some were found in the sera of rabbits immunized with the crude antigens. The numbers of precipitin lines in the sera of rabbits were different in the different fractionated antigens. They were 6 in the crude, 2 in the second and the third antigens, 1 in the forth, the fifth and the sixth antigens and absent in the seventh antigen, respectively. The wheal size for the bovine infected with F. hepatica was 2.46+/-0.15 cm in the intradermal test antigen (saline extract of F. hepatica) supplied by the Veterinary Research Institute, Rural Development Administration, Korea. The wheal size of the first, the second and the third antigens were larger than that of intradermal test antigen, whereas those of the forth, the fifth, the sixth and the seventh antigens showed smaller than that of the intradermal test antigen. The results suggest that the fifth antigen may be specific antigen for the immunodiagnosis of bovine fascioliasis.

[Experimental studies on the second intermediate hosts of Clonorchis sinensis IV. Observations on the fate of metacercariae of Clonorchis sinensis in the fish host, Cultriculus eigenmanni]

In order to judge appropriately the suitability as the second intermediate host of Clonorchis sinensis, the present experiments were proposed to survey the metacercaria of C. sinensis in the fish host, Cultriculus eigenmanni, in the field, and to observe, by experimental infection of the fish with Clonorchis cercaria, the penetrating ability of the cercaria, maturity, the process of degeneration and extinction, and infectivity of the metacercaria. The following is a brief summary of the leading facts gained through the experiments. All the ten fish (mean body weight 13.4 +/- 3.43g) which were caught in Ok-ku irrigation reservoir were infected with the metacercariae and the average number of the metacercariae found in the fish body/gram was 4.55. The fish was easily subject to invasion of the cercaria, and the examination of the cercariae after having the fish infected disclosed that 24 hours after the infection, all of the cercariae formed their cysts in muscle and the metacercariae kept growing, that 7 days later the metacercariae were found folding their bodies twice, and that in 15 days the characteristic structure of the metacercariae was complete and made a vigorous rotary movement intermittently. In the case of control fish, Pseudorasbora parva, both penetrating ability of the cercariae and the infective degree of the metacercariae to control host were stronger as compared with the fish host because that the control fish had higher suceptibility to C. sinensis. Afterwards, the metacercariae in the fish host came to a state of maturity and beyond this stage some metacercariae started the process of degeneration and extinction in as early as 30 days, and as more days elapsed, mortality of the metacercariae was not uniformly increased but difference of mortality appeared to be according to the individual of fish host. That is, in 390, 540 and 586 days, all of the metacercariae in the fish hosts became dead, meanwhile almost all of those found in the fish host of 596 days after the infection survived (mortality; 8.51 percent). Percentage of orally administered metacercariae developing to maturity in rabbit is 10.46 percent in the last case. As the days were progressed the shapes of the metacercariae in the fish host tended to change from ellipsoid to almost round shapes, the length became shorter while the width was wider. The results shown above revealed that C. eigenmanni was, though it could not be proper host than P. parva, suitable as the second intermediate host in addition to our previous observation results.

[Application of micro-ELISA in serodiagnosis of fascioliasis in cattle]

Fascioliasis in cattle is one of the most common and very serious trematode diseases in Korea. In the present study, the enzyme linked immunosorbent assay (ELISA) was applied in the diagnosis of fascioliasis using antigen of Fasciola hepatica, peroxidase of conjugate anti-cattle IgG and orthophenylenediamine as a substrate by micro-method technique of Voller et al. (1976b) and MacLaren (1978) with a slight modification. Results obtained from the present study are as follows: In assay for optimal dilution of stock antigen, the antigen (protein contents; 0.8 mg/ml) was diluted from 1/50 to 1/600 with carbonate buffer (pH 9.6), and then absorbance values were measured with 1/100 diluted sera. The regression equations between the OD values of ELISA and dilution of antigen were log Y=-0.181-0.00127X in infected sera, and log Y= -0.578- 0.000879X in normal sera. The significantly higher (p<0.05) OD value was observed in the former. In assay for optimal dilution of sera, the sera were diluted from 1/25 to 1/400 with in PBS/ Tween 20(pH 7.4), and absorbance values were measured with 1/200 diluted antigen. The regression equation between the OD values of ELISA and dilution of sera were log Y=-0.1540-0.0007238X in infected sera and log Y=-0.4834-0.00116X in normal sera. The former was higher than the latter (p<0.05). In the 27 cases of negative intradermal test, OD values of the ELISA are 0.447 +/- 0.144, the 95 percent confidence interval (Mean + 2 x SD) of the values was 0.735, and there was no case over the values. Therefore, the sensitivity of the antigen to diagnose fascioliasis was 100 percent in the negative case. The OD value 0.7 which is designed as a criterion (detection level of positive one) is useful for the performance of the ELISA in fascioliasis. According to the OD value of criterion in the regression equations, the optimal dilutions of stock antigen and serum were 1/250 and 1/100, respectively. In the 58 cases of fascioliasis from which the adult could be found in the bile ducts, the OD value was 0.846 +/- 0.224). The 75 percent (44 cattle) among them had higher value with compared to the criterion, and the 60 percent (20 cattle) of the cases of proliferative cholangitis of 33 cattle which had been infected previousely with Fasciola sp. is higher than the criterion. Prevalence of fascioliasis was 43.4 percent in the application of the ELISA to 272 cattle which were reared in Jeonbug district.

[Survey On Encysted Cercaria Of Trematodes From Fresh-Water Fishes In Tongjin Riverside Areas In Korea]

In an attempt to clarify the epidemiological feature of distomiasis in Tongjin riverside area, the prevalence of distomiasis in the residents and infection rates of the metacercariae in fresh-water fishes were investigated at the upper, middle and lower reaches of the river from January to April 1984. The results obtained were summarized as follows: Out of a total of 931 fresh-water fishes which composed of 33 different species, 611 fishes (65.6 %) of 31 species were found positive with digenetic trematode metacercariae of 16 different species, and there were some differences in infection rates of the metacercariae among the fishes in the 3 parts of the river; 53.8 percent in upper, 80.7 % in middle, and 61.0 % in lower reaches, respectively. Infection rates of the metacercariae of Exorchis oviformis, Metagonimus yokogawai, Echinochasmus japonicus, Metorchis orientalis and Clonorchis sinensis in the fishes were 48 %, 29 %, 11 %, 7.9 % and 6.3 %, respectively. The average number of the encysted larvae of Clonorchis found in fish body/gram showed 4.44 in Pseudorasbora parva, Gnathopogon coreanus (1.2), Microphysogobio yaluensis(0.76), Abbottina springeri(0.4), Acanthorhodeus asmussi (0.21) and Cultriculus eigenmanni (0.17), respectively. The average number of the metacercariae of Metagonimus found in fish body/gram disclosed 34.01 in Zacco platypus, Zacco temmincki (16.46), Carassius carassius (5.35), Moroco oxycephalus (1.54), Aphyocypris chinensis (1.5) and etc., respectively. Detection rates of the eggs of Clonorchis and Metagonimus among residents were 1.1 % and 0.8 %, respectively, out of a total 923 persons.

[Epidemiological Studies Of Clonorchis Sinensis In Mangyeong Riverside Areas In Korea]

In an attempt to clarify the epidemiological feature of C. sinensis in Mangyeong riverside area, the prevalence of clonorchiasis, infestation rate of the cercariae in Parafossarulus manchouricus, and detection rate of the metacercariae in fresh-water fishes were investigated from March 1 to September 30, 1983 at the upper, middle and lower reaches of the river. The results obtained were summarized as follows:Detection rate of C. sinensis egg among inhabitants was 8.2 % out of a total of 1,266 persons. but the differences in detection rates were not statistically signifcant among upper, middle and 1ower reaches. According to sex, the detection rates were 10.3 % in male and 6.1 % in female (p<0.05), but by age groups, increases of the rates were observed as increase in age (p<0.05). Out of a total of 380 fresh-water fishes of 32 different species, 93 fishes (25 %) of 12 species were found positive with Clonorchis metacercariae, and there were differences in infection rates of the metacercariae among the fishes in 3 parts of the river; 11 % in upper, 35 % in middle, and 34 % in lower reaches respectively. The metacercarial detection rates from various fishes were 97 % in Pseudorasbora parva, Cultriculus eigenmanni (85 %), Gnathopogon strigatus (67 %), Microphysogobio yaluensis (50 %), Gnathopogon coreanus (47 %), Pungtungia herzi(44 %), Abbottina rivularis (40 %), Moroco oxycephalus (33 %), Coreoleuciscus splendidus (32 %), Gnathopogon majimae (26 %), Rhodeus ocellatus (7 %), and Aphyocypris chinensis (3 %) respectively. Although very few P. manchouricus were collected at upper reach, 12 snails (0.7 %) among a total of 1,713 were found infected with Clonorchis cercariae. Also the cercariae of Echinochasmus japonicus (7.99 %), Lexogenes liberum (0.99 %), Cyathocotyle orientalis (0.75 %), Exorchis oviformis (0.23 %) and Asymphylodora japonica (0.05 %) were detected from the snails.