Infection rate of Schistosoma japonicum in the snail Oncomelania hupensis quadrasi in endemic villages in the Philippines: Need for snail surveillance technique.

Schistosomiasis japonica is one of seven NTDs endemic in the Philippines that continues to threaten public health in the country. The causative agent, the blood fluke Schistosoma japonicum, uses an amphibious snail Oncomelania hupensis quadrasi which can harbor larval stages that multiply asexually, eventually producing the infective cercariae which are shed into the water. Contamination of freshwater bodies inhabited by the snail intermediate host occurs through release of human and animal feces containing S. japonicum eggs. Miracidia hatching from these eggs subsequently infect the snails that inhabit these water bodies. The degree of fecal contamination can vary across snail sites and influences snail infection rates in these sites. In this study, conventional malacological surveys using intensive manual search for snails were conducted from 2015 to 2016 in seven selected endemic provinces, namely Leyte and Bohol in the Visayas and Surigao del Norte, Agusan del Sur, Bukidnon, Lanao del Norte and Compostela Valley in Mindanao. A total of 6,279 O. hupensis quadrasi snails were collected from 38 snail sites. The municipality of Trento in Agusan del Sur recorded the highest number of snail sites (7) that yielded O. hupensis quadrasi snails while only one snail site was found positive for O. hupensis quadrasi snails in Kapatagan in Lanao del Norte and Talibon in Bohol. Alegria in Surigao del Norte yielded the highest number of snail sites (5) that were found to harbor snails positive for S. japonicum infection. The snail infection rates in this municipality ranged from 0.43% to 14.71%. None of the snails collected from Talibon in Bohol was infected. Bohol is the only province among the 28 schistosomiasis-endemic provinces which has reached near elimination status. Snail infection rates were found to vary considerably across snail sites, which could be due to the degree of fecal contamination of the snail sites and their connectivity to water that can serve as contamination source.

Infection rate of Schistosoma japonicum in the snail Oncomelania hupensis quadrasi in endemic villages in the Philippines: Need for snail surveillance technique

@#Schistosomiasis japonica is one of seven NTDs endemic in the Philippines that continues to threaten public health in the country. The causative agent, the blood fluke Schistosoma japonicum, uses an amphibious snail Oncomelania hupensis quadrasi which can harbor larval stages that multiply asexually, eventually producing the infective cercariae which are shed into the water. Contamination of freshwater bodies inhabited by the snail intermediate host occurs through release of human and animal feces containing S. japonicum eggs. Miracidia hatching from these eggs subsequently infect the snails that inhabit these water bodies. The degree of fecal contamination can vary across snail sites and influences snail infection rates in these sites. In this study, conventional malacological surveys using intensive manual search for snails were conducted from 2015 to 2016 in seven selected endemic provinces, namely Leyte and Bohol in the Visayas and Surigao del Norte, Agusan del Sur, Bukidnon, Lanao del Norte and Compostela Valley in Mindanao. A total of 6,279 O. hupensis quadrasi snails were collected from 38 snail sites. The municipality of Trento in Agusan del Sur recorded the highest number of snail sites (7) that yielded O. hupensis quadrasi snails while only one snail site was found positive for O. hupensis quadrasi snails in Kapatagan in Lanao del Norte and Talibon in Bohol. Alegria in Surigao del Norte yielded the highest number of snail sites (5) that were found to harbor snails positive for S. japonicum infection. The snail infection rates in this municipality ranged from 0.43% to 14.71%. None of the snails collected from Talibon in Bohol was infected. Bohol is the only province among the 28 schistosomiasis-endemic provinces which has reached near elimination status. Snail infection rates were found to vary considerably across snail sites, which could be due to the degree of fecal contamination of the snail sites and their connectivity to water that can serve as contamination source.

Discovery of Liopiophila varipes and Protopiophila contecta (Diptera: Piophilidae) from human cadavers.

Here, we present two cases in which larvae of the family Piophilidae were detected in human cadavers. Both cases were found in Tochigi Prefecture, which is located in the middle of Honshu Island, Japan. Case 1: A corpse was found hanging in the sun lounge of a house. Dipteran larvae were collected from inside the spinal canal, despite no visible breach on the skin. The adults derived from these larvae were identified as Piophila casei (Linnaeus, 1758) and Liopiophila varipes (Meigen, 1830). Case 2: Skeletal human remains were found in a mountainous forest. Dipteran larvae were detected in the bone marrow cavity of a tibial section during autopsy. One adult fly derived from the larvae was identified as Protopiophila contecta (Walker, 1860). This is the first report of the identification of L. varipes and P. contecta in human cadavers.

Characteristics of granuloma formation and liver fibrosis in murine schistosomiasis mekongi: a morphological comparison between Schistosoma mekongi and S. japonicum infection.

A histopathological study was performed to clarify the characteristics of granuloma formation and liver fibrosis in Schistosoma mekongi infection in comparison with S. japonicum infection. Mice were exposed to S. mekongi (Laotian strain) and S. japonicum (Japanese strain) cercariae, and were dissected at 6, 8, 12, 16, and 20 weeks post-exposure. In the liver, granulomas in S. mekongi infection were cellular, initially organized with foam cells, and continuously appeared in the intralobular area, while granulomas in S. japonicum infection were fibrous and did not continuously appear in the intralobular area. Portal fibrosis was not seen in S. mekongi infection, but was commonly seen in S. japonicum infection in the later weeks. Granulomas in the small intestine were seen mainly in the submucosa with foam cells in S. mekongi infection and without foam cells in S. japonicum infection. The lung granulomas contained mainly histiocytes in both S. mekongi and S. japonicum infection. The absence of portal fibrosis in S. mekongi infection allows schistosome eggs to infiltrate into the intralobular area continuously, which can be what lies behind the ultrasonographic differences; the echogenic network pattern as was seen in S. japonicum infection, has not been noted in S. mekongi infection.

Establishment of a GIS monitoring system for schistosomiasis japonica in Kofu, Japan.

Although the incidence of human infection with Schistosoma japonicum in Japan fell to zero in 1977, the threat of the possible re-emergence of the disease caused by this trematode still exists. Surveillance of the parasite's intermediate host, Oncomelania nosophora, in Kofu basin therefore began in 1996. A simple, new method for monitoring O. nosophora in an at-risk area in Kofu, which is based on a geographical information system (GIS), was established. At each monitoring site (of which there were 120 from 1996 until 2000, and 60 from 2001 until 2003), the O. nosophora in two quadrats, each measuring 25 x 25 cm, were collected. During the study, the exact location of each site was determined using a hand-held global-positioning system (GPS). This allowed all the sites to be digitally mapped, so that anyone with a hand-held GPS could and can reach each site. The snail and location data were processed using commercial GPS/GIS software packages and used to create a risk map for schistosomiasis re-emergence. Although all snails collected between 1996 and 2003 were uninfected, the proportion of investigated sites in which O. nosophora was detected increased from 36.7% in 1996 to 56.7% in 2003. The mean number of O. nosophora collected per snail-positive site fluctuated widely, between 8.2 and 57.4, in each calendar year. Over the study period there appeared to be a shift southwards in the areas with high densities of O. nosophora. The present results indicate that it is possible to utilize a GIS-based method for the long-term monitoring of the possible re-emergence of schistosomiasis japonica in Japan.

Comparative studies on susceptibilities of two different Japanese isolates of Oncomelania nosophora to three strains of Schistosoma japonicum originating from Japan, China, and the Philippines.

Oncomelania nosophora (Gastropoda: Pomatiopsidae) is the intermediate host of Schistosoma japonicum in Japan. Although most of the snails were eliminated during the 20th century, they are still found in two areas in Japan. One area is in the Kofu Basin, including Nirasaki City, in Yamanashi Prefecture. The other is the Obitsu River Basin in Kisarazu City, Chiba Prefecture. Snails collected in Nirasaki and Kisarazu were exposed to 3 geographical strains of S. japonicum originating from Japan, China, and the Philippines. Both isolates of O. nosophora showed high susceptibility to the Japanese strain of S. japonicum (74.0% - 82.2%, for the Nirasaki isolate and 58.0% - 56.0% for the Kisarazu isolate) and low susceptibility to the Chinese strain (0.0% - 1.3% and 1.4% - 7.9% respectively). In contrast, the susceptibility of the snails to the Philippine strain was significantly different (P < 0.01) between the isolates (3.3% - 6.6% for the Nirasaki isolate and 31.9% - 75.9% for the Kisarazu isolate). To examine the differences in infectivity in detail, we conducted histological observations of snails exposed to the Philippine strain at 3 h, 1, 3, and 15 days after miracidial exposure. We found differences in the development of the parasite between the isolates of snails from early after exposure.

Suspected intestinal myiasis due to Dryomyza formosa in a Japanese schizophrenic patient with symptoms of delusional parasitosis.

A third-stage larva of Dryomyza formosa (Wiedemann) (Diptera: Dryomyzidae) was found in the fresh stool of a 27-year-old Japanese woman resident of Shiobara, 150 km north of Tokyo, on 16 November 1998. This is the first record of myiasis due to Dryomyza. Detection of this maggot (2cm long) by the patient herself was associated with her longstanding delusion of abdominal parasitosis as a symptom of chronic schizophrenia. Circumstantial evidence agreed with this being a genuine case of intestinal myiasis, apparently due to accidental ingestion of the insect, with no signs that the patient had contrived the report, nor that the maggot had invaded the stool post-defaecation. This case draws attention to the likelihood that some personality states are predisposed to noticing and reporting myiasis, when it occurs. We review other conditions (mental and physical) that are more prone to myiasis.

Malaria infection induces rapid elevation of the soluble Fas ligand level in serum and subsequent T lymphocytopenia: possible factors responsible for the differences in susceptibility of two species of Macaca monkeys to Plasmodium coatneyi infection.

The intraerythrocytic stage of the simian malaria parasite Plasmodium coatneyi (CDC strain) was intravenously inoculated into two species of macaques with different susceptibilities to infection with this parasite, including four Japanese macaques (Macaca fuscata) and three cynomolgus macaques (M. fascicularis). The Japanese macaques infected with P. coatneyi developed severe clinical manifestations similar to those of severe human malaria and eventually became moribund, while the infected cynomolgus macaques, natural hosts of the parasite, exhibited no severe manifestation of disease except anemia and finally recovered from the infection. In the infected Japanese macaques, peripheral CD4(+) and CD8(+) T-cell populations were markedly decreased and fragmentation of chromosomal DNA in peripheral blood mononuclear cells was detected during the terminal period of infection, suggesting that apoptotic cell death was responsible at least in part for the T lymphocytopenia. Furthermore, soluble Fas ligand levels in sera of the infected Japanese macaques increased gradually to a markedly high level of 28. 83 +/- 10.56 pg/ml (n = 4) when the animals became moribund. On the other hand, none of the infected cynomolgus monkeys exhibited either T lymphocytopenia or elevated soluble Fas ligand level. These findings suggest that differences in immune response between the two species of macaque tested accounted for the contrasting outcomes after infection with the same isolate of malarial parasite, and in particular that a profound T lymphocytopenia due to Fas-derived apoptosis played a role in the fatal course of malaria in the infected Japanese macaques.

Human antibody isotype responses to Schistosoma japonicum egg antigen.

Schistosoma japonicum-infected subjects from Hubei province of China were investigated to determine the class and subclass of the antibody response to soluble egg antigen (SEA), using an enzyme-linked immunosorbent assay. The subjects were 50 acute and 55 chronic cases. In acute cases, the mean OD values for IgA, IgE and IgG3 were very high, while the positive ratios of IgA and IgE were only 78% and 74%, respectively. The positive ratios of IgG, IgM, IgG1, IgG3 and IgG4 were all above 90%. In chronic cases, the mean OD values for IgG, IgG3 and IgG4 were very high, and the positivity rates of IgG, IgG1, IgG3 and IgG4 were all above 90%. Comparing the two study groups, the mean OD values of IgM, IgA, IgE were higher in acute cases than those of chronic cases (p < 0.0001), while the mean OD values of IgG, IgG4 were higher in chronic cases than in acute cases (p < 0.05). The mean OD values of IgG3 in both groups were high and those of IgG2 in both groups were low.

Cloning of chicken slow muscle troponin T and its sequence comparison with that of human.

A full-length cDNA coding for chicken slow muscle troponin T (TnT) was for the first time isolated from a cDNA library of 10-day-old embryos, using an RT-PCR product of chicken slow muscle TnT. It showed about 60% homology for chicken fast and slow muscle TnTs and 75.2% for human slow muscle TnT. The 16 amino acid sequence found in the carboxyl terminus of human slow muscle TnT was absent in the chicken slow muscle TnT. The 5E-5A-7E sequence found in the amino terminal region of chicken slow muscle TnT was partly similar to the counterpart of human slow muscle TnT, but not to those of chicken fast and cardiac muscle TnTs. With this report of chicken slow muscle TnT, cDNA information on chicken TnTs of all three striated muscles was completed following those of human TnTs.

Analysis of dystrophin in muscular diseases by two-dimensional gel electrophoresis using agarose gels in the first dimension.

We analyzed dystrophin in case of normal control, Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD) and infectious muscular disease using two-dimensional gel electrophoresis and immunoblotting with 3 monoclonal dystrophin antibodies: Dys 1, a mid-rod-domain antibody; Dys 2, a C-terminal-domain antibody; and Dys 3, an N-terminal-domain antibody. In cases of normal control, a clearly separated doublet of bands was observed for Dys 1 and 3 at molecular weights 400 and 420 kDa. The isoelectric point was between pH approximately 5.7-approximately 5.9, similar to that for the myosin heavy chain. In one DMD case, a single faint band was observed for Dys 2. BMD presented a single-band pattern for each antibody. Infectious diseases cases showed 3- to 5-band patterns for Dys 1 and single or no bands for Dys 2 and 3. The pI of the Dys 1 band was almost identical. These results suggest coexistence of normal dystrophin and its proteolytic products, both containing triple helical segment, and show that two-dimensional gel electrophoresis may be applicable in the analysis of dystrophin in muscular disease.

Persistent expression of tissue-specific troponin T isoforms in transplanted chicken skeletal muscle.

This study attempted to investigate the expression of skeletal muscle troponin T isoforms in chicken reared for six months after muscle transplantations of breast muscle into leg muscle, leg muscle into breast muscle, and slow muscle into breast and leg muscles of the same animal. The regenerated muscle after transplantation was studied by histological observation, two-dimensional SDS-polyacrylamide gel electrophoresis, and immunoblotting with anti-troponin T antibodies. Persistent expression of troponin T isoforms specific to donor tissue was observed in the regenerated muscle, and compared with their expression in the normal developing muscles. During the regeneration, the cells grew up and expressed troponin T isoforms in a manner similar to that in normal developing muscles, and on around the 178th day after the transplantation, the regenerated muscle expressed the adult type troponin T isoforms. Based on the troponin T isoforms expressed in the transplants, we consider that one type of skeletal muscle has some inherent potential to grow in and coexist with other types for a long term.

Detection and characterization of muscle-specific nuclear proteins.

To identify nuclear proteins related to muscle tissue specificity, we tried to prepare antibodies recognizing muscle-specific nuclear proteins. Taking advantage of the autoimmunity of some nuclear proteins, we prepared an antiserum against chick muscle nuclear proteins by injecting protein components of the nuclei isolated from chick breast muscles into breast muscle. Three proteins, named p30, p32, and p37, were detected with the antiserum in a two-dimensional SDS-PAGE pattern of the isolated nuclei. P30 and p32 were not detected in the nuclei of liver, brain, cardiac muscle, or slow type skeletal muscle (anterior latissimus dorsi). They were detected in those of fast type skeletal muscle (pectoralis major and semitendinosus) and smooth muscle (gizzard) at all developmental stages examined. On serial fractionation of muscle cell nuclei, they were detected in a fraction obtained after DNase I treatment of the sample, suggesting that the proteins weakly bind to chromatin. A homology search of amino acid sequences showed that there is no known protein similar to p32.

Expression pattern of skeletal muscle troponin T isoforms is fixed in cell lineage.

The expression of fast-muscle-type troponin T isoforms in chicken skeletal muscles was studied by two-dimensional SDS-polyacrylamide gel electrophoresis and immunoblotting. According to the pattern of troponin T isoform expression, chicken fast muscle was classified into two groups: One group expressed breast-fast-muscle-type troponin T in addition to leg-fast-muscle-type troponin T, the other expressed only leg-fast-muscle-type troponin T. To the former group belong breast and wing fast muscles and some of the back fast muscles, and to the latter group belong the fast muscles in leg, abdomen, and neck. Transplantation of breast muscle into leg was performed in order to change the physical environment and to investigate the mechanism of isoform expression. Histological observation of the transplant revealed severe degeneration of muscle cells, followed by differentiation of myoblasts in which breast-muscle-type troponin T was eventually expressed. The results showed that the pattern of troponin T isoform expression is primarily fixed in the cell lineage, although nerves modulate it.