Present study aimed to elucidate the immunosuppressive effect of prednisolone on Naegleria fowleri infection in mice. N. fowleri was cultured in CGVS medium (Willert and Le Ray, 1973). White female mice, weighing about 18 g, used for experiments were divided into five groups; untreated control group, prednisolone treated groups (before, during and after infection), and only prednisolone treated group. In the prednisolone treated group, the hormone was injected intramuscularly 5 doses of 10 mg/kg every other day. According to designated time of treatment, each mouse was challenged with 1 x 10(5) N. fowleri intranasally. Changes of body weights, clinical manifestations and number of dead mouse were observed. Brain and lung tissues of dead mice were cultured in the non-nutrient agar (Kasprzak and Mazur, 1972), or stained with hematoxylin-eosin for the examination of histopathological changes. Results of the experiment are summarized as follows: Mortality among the prednisolone treated groups was higher than that in untreated control group, and among the treated groups, the pretreated group showed shorter survival time. Body weights among untreated control mice showed no significant increase, however, treated groups of mice showed the decrease during the administration and recovery of the weights were observed at 2 to 3 days after the completion of treatment. In the treated control groups, the infected mice began to show the pathologic findings 5 days after infection while the untreated mice began to show the findings 8 days after infection. Tissue damages in brain and lung occurred due to virulence of amoeba were more severe among treated mice than that in untreated control group. The above mentioned results suggest that the treatment with prednisolone weaken the resistance of mice against N. fowleri infection, and probably induce more severe primary amoebic meningoencephalitis. Especially severe pathological findings were shown in pre-treated group, compared with untreated group.
parasitology-protozoa ; Naegleria fowleri ; meningoencephalitis ; brain ; prednisolone

The role of passive cell-mediated transfer of immunity against primary amoebic meningoencephalitis(PAME) in mice was studied. Naegleria fowleri, ITMAP 359, were cultured in CGVS medium. The ICR mice used were six week-old males of average weight of 15 g. Immunization was done by three intraperitoneal injections of l x 10(6) N. fowleri trophozoites at the interval of one week. Splenocytes were obtained from normal and immune mice spleens, and 1 x 10(7) cells were administered intraperitoneally into mice 3 days before challenge infection. Mice were infected intranasally with 7 x 10(4) N. fowleri trophozoites in a 3 microliter suspension under secobarbiturate anesthesia. Transplants of normal or immune splenocytes seem to alter the pattern of the PAME development. The splenocytes transferred from immune mice reduced the mortality rate in the N. fowleri infected mice, as compared with the mice transferred with the same number of normal splenocytes or without splenocyte. The blastogenic response of the splenocytes to both lipopolysaccharide and concanavalin A was elevated on day 7 after infection the mice transinoculated with immune splenocytes. The serum antibody titers in the mice transferred with immune splenocytes were increased gradually from day 7 up to day 20 after infections by mean of ELISA. It is suggested that the transfer of splenocytes from immunized mice conferred immunity against N. fowleri infection.
parasitology-protozoa ; Naegleria fowleri ; meningoencephalitis ; brain ; immunology

To elucidate the effect of splenectomy on the development of experimental primary amoebic meningoencephalitis in mice, the death rate and survival time of mice infected intranasally with Naegleria fowleri trophozoites 5 x 10(4) cultivated in CGVS medium were compared according to the age when splenectomy was done, and post-operation until experimental infection. Immunodiffusion was undergone to detect the presence of serum antibody due to N. fowleri infection in mice. Polyacrylamide gel electrophoresis was done to compare the protein fractions of mouse serum in each experimental groups. In experiment I, splenectomy was done 3 weeks and infection 4 weeks after birth, the death rate of control, sham operated and splenectomized group were 100 percent, 85 percent and 95 percent, and the mean survival time after infection 7.3 days, 7.5 days and 7.8 days, respectively. In experiment II, splenectomy was undergone 3 weeks and infection 6 weeks after birth, the death rate of control, sham operated and splenectomized group were 95 percent, 95 percent and 95 percent , and the mean survival time after infection 12.1 days, 11.5 days and 11.5 days, respectively. In experiment III, splenectomy was done 5 weeks and infection 6 weeks after birth, the death rate of control, sham operated and splenectomized group were 95 percent, 90 percent and 95 percent, and the mean survival time after infection 8.1 days, 8.3 days and 8.5 days, respectively. By Ouchterlony immunodiffusion, anti-N. fowleri antibody in the serum of mouse with primary amoebic meningoencephalitis was detected against a N. fowleri antigen, which was prepared by ultrasonication of N. fowleri trophozoites, each reacting two lines of precipitation. The patterns of serum fractions by polyacrylamide gel electrophoresis were different between control and sham operated groups from splenectomized group in fraction II, III and V, the sera of which were collected after N. fowleri infection. This results may be summarized as that splenectomy has no effect on the development of primary amoebic meningoencephalitis in mice.
parasitology-protozoa ; Naegleria fowleri ; meningoencephalitis ; brain ; immunology ; spleen ; brain

This study is to verify the protective ability against experimental Naegleria meningoencephalitis by immunization with Naegleria fowleri in mice. Naegleria fowleri, strain 0359, and Naegleria gruberi, strain EGB, were used in this study, and cultured in CGVS medium axenically. Inbred BALB/c mice, weighing about 20 g, were immunized by three intraperitoneal injection of 1 x 10(6) N. fowleri trophozoites at the interval of one week. This N. fowleri trophozoites antigen was fixed with 5 percent formaldehyde. N. fowleri trophozoites from culture were homogenized with sonicator at 4C as monitored by phase contrast microscopy, and their membrane and cell content preparations were made for the immunization of mice. Their inoculation dose in volume was equivalent to the 1 x 10(6) trophozoites in each injection for immunization. And N. gruberi trophozoites, which was fixed with 5 percent formaldehyde, were also used for immunization. Mice were inoculated intranasally with 5 x 10(4) N. fowleri trophozoites in a 5 microliter suspension under anesthesia by as intraperitoneal injection of about l mg secobarbiturate. Nervousness, rotation or sluggish behaviour were observed in the mice which were infected with N. fowleri. Necrotic lesion was demonstrated in the anterior portion of brain, especially in the olfactory lobe. The inflammatory cell infiltration with numerous N. fowleri trophozoites was noticed. This pathological changes were more extensive in the control than in the experimental groups. Mice were dead due to experimental primary amoebic meningoencephalitis that developed between 8 days and 23 days after inoculation. Mortality rate of the mice was low in the immunized experimental group. Mean survival time, which is the survival duration of mice from the infection to death, was prolonged significantly in the immunized mice except in the mice immunized with N. fowleri membrane. Even in the mice immunized with N. gruberi, survival time was delayed. In summary, the effectiveness of immunization is demonstrated in terms of protective immunity against Naegleria meningoencephalitis in mice.
parasitology-protozoa ; Naegleria fowleri ; meningoencephalitis ; immunology ; mouse ; brain

Female BALB/c mice weighing 18-20 g were immunized by three injections of 1 x 10(6) Naegleria fowleri trophozoites intraperitoneally at the interval of one week 6 times for the pregnant mice and 3 times for the offspring mice. One week after immunization the mice were challenged intranasally with N. fowleri trophozoites 5 x 10(4) under secobarbital anesthesia. Experimental primary amoebic meningoencephalitis developed between day 7 and 16 after infection. All mice were dead due to amoebic meningoencephalitis in all experimental groups except in the offspring born to non-immune mothers. Mean of survival time, which is the duration of survival of mice from infection to death, was delayed in the groups of mice born to immune mothers, immune mice born to immune mothers. Active or passive protective immunity against N. fowleri infection was demonstrated in the immunized mice and mice born to immune mothers. But the effectiveness of immunization was greatly impaired in terms of mortality in the immune mice born to immune mothers when N. fowleri was infected intranasally.
parasitology-protozoa ; Naegleria fowleri ; meningoencephalitis ; brain ; immunology ; pregnancy

Animals ; Brain Diseases ; parasitology ; Child ; Female ; Humans ; Sparganosis ; Sparganum

The purpose of this study was to investigate whether sporulated Neospora caninum oocysts, which had been stored for 46 mo in a 2% sulfuric acid solution at 4 degrees C, remain morphologically viable and infective to gerbils (Meriones unguiculatus). Six gerbils were orally inoculated with doses of 400 or 1,200 oocysts. Two mo after inoculation, the animals did not show any clinical signs, had no histological lesions, and were seronegative for N. caninum at 1: 50 in an immunofluorescent antibody test. PCR using the brain from each gerbil did not reveal N. caninum specific DNA. We conclude that oocysts preserved for 46 mo are not infective, despite being morphologically intact.
Acids ; Animals ; Brain/parasitology/pathology ; Cattle/parasitology ; Coccidiosis/parasitology/pathology/*veterinary ; Feces/parasitology ; Female ; Gerbillinae/*parasitology ; Neospora/genetics/growth & development/*pathogenicity ; Oocysts/*growth & development ; Polymerase Chain Reaction/methods ; Refrigeration ; Virulence

To determine alteration of immune responses during visceral larva migrans (VLM) caused by Toxascaris leonina at several time points, we experimentally infected mice with embryonated eggs of T. leonina and measured T-helper (Th) cell-related serial cytokine production after infection. At day 5 post infection (PI), most larvae were detected from the lungs, spleen, intestine, and muscle. Expression of thymic stromal lymphopoietin (TSLP) and CCL11 (eotaxin) showed a significant increase in most infected organs, except the intestine. However, expression of the CXCL1 (Gro-alpha) gene was most highly enhanced in the intestine at day 14 PI. Th1-related cytokine secretion of splenocytes showed increases at day 28 PI, and the level showed a decrease at day 42 PI. Th2-related cytokine secretion of splenocytes also showed an increase after infection; in particular, IL-5 level showed a significant increase at day 14 PI, and the level showed a decrease at day 28 PI. However, levels of Th17-related cytokines, IL-6 and IL-17A, showed gradual increases until day 42 PI. In conclusion, Th1, Th2, and Th17-related cytokine production might be important in immune responses against T. leonina VLM in experimental mice.
Animals ; Brain/parasitology ; Cytokines/*metabolism ; Female ; Gene Expression Regulation ; Heart/parasitology ; Interleukins/*metabolism ; Intestines/parasitology ; Larva Migrans, Visceral/*immunology/parasitology ; Liver/parasitology ; Lung/parasitology/pathology ; Mice ; Mice, Inbred C57BL ; Muscles/parasitology ; Spleen/parasitology ; Th1 Cells/immunology ; Th17 Cells/immunology ; Th2 Cells/immunology ; Toxascaris/*immunology

Toxoplasmosis is an important zoonotic disease that can cause abortion in humans and animals. The aim of this study was isolation and subsequent genotyping of Toxoplasma gondii isolates in ovine aborted fetuses. During 2012-2013, 39 ovine aborted fetuses were collected from sheep flocks in Khorasan Razavi Province, Iran. The brain samples were screened for detection of the parasite DNA by nested PCR. The positive brain samples were bioassayed in Webster Swiss mice. The serum samples of mice were examined for T. gondii antibodies by IFAT at 6 weeks post inoculation, and T. gondii cysts were searched in brain tissue samples of seropositive mice. The positive samples were genotyped by using a PCR-RLFP method. Subsequently, GRA6 sequences of isolates were analyzed using a phylogenetic method. The results revealed that T. gondii DNA was detected in 54% (20/37, 95% CI 38.4-69.0%) brain samples of ovine aborted fetuses. In bioassay of mice, only 2 samples were virulent and the mice were killed at 30 days post inoculation, while the others were non-virulent to mice. The size of cysts ranged 7-22 µm. Complete genotyping data for GRA6 locus were observed in 5 of the 20 samples. PCR-RLFP results and phylogenetic analysis revealed that all of the isolated samples were closely related to type I. For the first time, we could genotype and report T. gondii isolates from ovine aborted fetuses in Khorasan Razavi Province, Iran. The results indicate that the T. gondii isolates are genetically related to type I, although most of them were non-virulent for mice.
Aborted Fetus/*parasitology ; Animals ; Brain/parasitology ; Genotype ; Iran ; Mice ; Phylogeny ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Sheep ; Sheep Diseases/*parasitology ; Toxoplasma/classification/*genetics/*isolation & purification/pathogenicity ; Toxoplasmosis, Animal/*parasitology

/A total of 69 patients of confirmed neurocysticercosis was followed serologically by ELISA up to 22 months after praziquantel treatment. The intervals and numbers of follow-up were variable by patients. Serially collected samples of serum and CSF were examined simultaneously for their specific IgG antibody levels by ELISA, using cystic fluid, saline extracts of bladder wall and scolex as antigen. Within 4 months after praziquantel treatment, the antibody levels were elevated temporarily in both serum and CSF in most patients. In some cases antibody levels exhibited steady declining tendency after the treatment. Concomitant administration of dexamethasone appeared to suppress the elevation of antibody levels. The rate of mean absorbance of antibody changed more in serum than in CSF. The rate of elevation was greater in antibodies to parenchymal antigens than that to cystic fluid, but absolute difference of antibody levels was greater in anitbody to cystic fluid. Previously negative samples for IgG antibody may become positive after praziquantel treatment, which could be used as a complementary tool(provocation test) in serodiagnosis. One month was considered to be sufficient interval for the follow-up test for that purpose. In the follow-up of up to 22 months, only few cases of chronic neurocysticercosis showed declining tendency of IgG antibody levels below negative range. During acute encephalitic attacks in chronic patients, IgG antibody to parenchymal antigen were elevated in CSF temporarily. These results indicated that serologic follow-up of every year was recommendable to differentiate the cured patients from chronic patients with slowly calcifying lesions.
parasitology-helminth-cestoda ; Taenia solium ; cysticercus ; brain ; immunology ; praziquantel-chemotherapy ; praziquantel