BALB/c mice infected with a low virulent strain of Trypanosoma brucei gambiense IL3253 were treated intraperitoneally（ip）with either Melarsoprol （Mel-B, obtained from WHO，Geneva）or phosphate-buffered saline（PH 7.8） containing 1％ glucose （PSG） as controls. The mice were subsequently monitored regularly for parasites by direct microscopic examination of their tail blood or buffy coat and by polymerase chain reaction（PCR）．Suratex®（AccuPharma，NY）， an　assay　that　detects　circulating trypanosome parasite antigen，was also used．Mel-B was found to be an effective drug for treatment against T.b. gambiense and at the end of the first treatment schedule of 3 series of 3 injections at 7 days post- infection（DPI），all treated mice were negative for parasites even by PCR, while all the control animals were positive. Suratex®（AccuPharma, NY）was found to be inappropriate in the serodiagnosis of trypanosomosis due to T.b. gambiense in mice as it gave high levels of false positives. At 34 DPI, all the BALB/c mice were sacrificed and 0.5ml of their blood injected ip into individual SCID mice. The SCID mice were likewise monitored for parasites or for evidence of infection by microscopic examination and PCR for up to 115 DPI. None of the SCID mice injected with blood from BALB/c previously treated with Mel-B showed any parasite either in their tail blood or buffy coat over the entire sampling period, though some of these mice were shown to harbor an infection by PCR at some time over the sampling interval. This experiment affirms the importance of the various techniques used in the diagnosis of trypanosomosis. A repeated　negative　 PCR　test　in　combination　with　clinical　and conventional microscopical examination may be a useful diagnostic regimen in the diagnosis of trypanosomosis.

抄録全体を表示

We have simplified a non-surgical procedure as an alternative to surgically obtained samples to identify transgenic mice harboring SAG-1 transgene. The protocol involves the use of a small amount of oral wash containing enough oral different cells including epithelial cells serving as a sufficient source of DNA for two-step polymerase chain reaction（PCR）analysis．Oral wash is boiled and aqueous phase is applied directly to PCR cocktail containing an outer pair of primers for the first round amplification. Afterward, a small amount of the first round product is added to PCR cocktail containing an inner pair of primers for the second round reaction. The procedure provides outcomes consistantly matching those of tissue biopsy one. This protocol can be repeated many times with minimal stress to mice. This technique is reliable, does not require protease digestion and phenol-chloroform extraction，is more humane，and can be completed within 4 hr. This technique is an alternative to the tissue biopsy procedure to determine the SAG-1 transgenic status in mice.

抄録全体を表示

In a survey for sarcocysts in muscle tissues obtained from 142 water buffaloes（Bubalus bubalis）, 92(64.8%) carcasses had sarcocysts. Macroscopic and two forms of microscopic cysts, the spindle-shaped or fusiform cysts commonly occurring in the muscles of the esophagus, throat and limbs, and the globular to oval cysts which were the dominant form in the diaphragm and cervical muscle tissue were noted. Ultrastructural analysis of macroscopic and microscopic cysts and their cyst wall revealed two distinct species of Sarcocystis infecting Philippine water buffaloes. These are the macroscopic species, Sarcocystis fusiformis which has been previously reported in the country possessing highly-dendritic cauliflower-like projections emanating from the primary cyst wall, with annulated microfibrils and numerous electron dense granules; and the newly redescribed Sarcocystis levinei (Dissanaike and Kan 1978; Huong, Dubey and Uggla 1997b) exhibiting a cyst wall with numerous, minute hair-like villar protrusions with expanded or dome-shaped base, an intermediate finger-like, and distal tapering segments which at some points join to form conical tufts． Our findings represent the first report of S. levinei in Philippine water buffaloes supported with ultrastructural analysis of the sarcocyst and its cyst wall, and likewise refute earlier published reports that all microscopic cysts in Philippine water buffaloes are developing forms of S. fusiformis．

抄録全体を表示

Examination of muscle tissues of carcasses of 370 imported Brahman breed Cattle slaughtered in three different abattoirs in Metro Manila revealed an infection rate of 10.8%. Three morphologically different microscopic sarcocysts were detected; the spherical and radially striated or hirsute cysts with thick cyst wall (Type1); the spherical to oval cysts exhibiting thinner cyst wall（Type2） compared to Type1; and the spindle-shaped to elongate cysts with prominent compartmentalized arrangement of zoites separated by septae. Sarcocysts morphology and their host location suggest sarcocystis hominis and Sarcocystis cruzi as the most likely etiologic species. The infections noted may either be local or imported in origin. In the absence of any documented studies on local or imported bovine sarcocystosis in the country to date，these initial findings are valuable. However, for future studies on ultrastructural analysis of the sarcocysts and the cyst wall to confirm species identification, and experimental exposure studies to determine the probable definitive host(s) of Sarcocystis species are necessary.

抄録全体を表示

The gene encoding P15 of Cryptosporidium parvum (C. parvum) sporozoites was amplified from Mito strain isolated from calf in Japan, and inserted into the thymidine kinase gene of vaccinia virus LC16mO strain under the control of the early-late promoter for the vaccinia virus 7.5-kilodalton (kDa) polypeptide. The P15 expressed by recombinant vaccinia virus in mammalian cells (RK13) had apparent molecular masses of 18-22 kDa. The vaccinia virus expressed P15 was glycosylated and transported to the surface of infected cells. Mice inoculated with the recombinant vaccinia virus produced high titers of antibodies against sporozoites or merozoites of C. parvum. Our next step will be to examine the ability of the recombinant P15 as a potential subunit vaccine to control cryptosporidiosis in animals.

抄録全体を表示