Echinococcosis/hydatidosis in western Iran

In the present study, 115 stray dogs (56 males and 59 females, mixed breed), 86 golden jackal (Canis aureus, 42 males and 44 females), 60 red foxes (Vulpes vulpes, 33 males and 27 females), and three female wolves (Canis lupus) were examined for Echinococcus granulosus infection, as well as, 32,898 sheep, 10,691 goats, 15,779 cattle and 659 buffaloes for hydatid infection from five provinces in western Iran during 3 years (1997-2000). Meanwhile fertility rates of different types and forms of cysts isolated from infected animals and the viability of protoscolices were also determined. Results indicated that 19.1% of the dogs, 2.3% of the golden jackals and 5% of the red foxes were infected with Echinococcus granulosus. 11.1% of the sheep, 6.3% of the goats, 16.4% of the cattle and 12.4% of the buffaloes were also found to be infected with hydatid cyst. The cysts isolated from liver and lungs of the sheep show higher fertility rate than the cysts of liver and lungs of goats, cattle and buffaloes.     

A study on intestinal helminthes of dogs, foxes and jackals in the western part of Iran

Human infection especially with helminth parasites is an emerging health issue, as the human environment is increasingly shared with infected animals, either pets or wild life. In this survey, the intestinal content of 83 stray dogs, 22 red foxes and 10 golden Jackals collected from the West Azarbaijan, Kordestan and Kermanshah provinces in the west of Iran, were studied for the presence of helminth parasites. The percentage of different species recovered from these animals is listed as follows: From stray dogs: Toxocara canis (6.02%), Toxascaris leonina (32.53%), Ancylostoma caninum (3.61%), Oxynema sp. (1.35%), Rictularia affinis (12.05%), Taenia hydatigena (53.01%), Taenia ovis (7.23%), Taenia multiceps (4.82%), Echinococcus granulosus (13.25%), Dipylidium caninum (38.55%), Mesocestoides lineatus (26.50%) and Macracanthorhynchus hirudinaceus (4.82%). From red foxes: T. canis (4.54%), T. leonina (31.82%), A. caninum (4.54%), Uncinaria stenocephala (13.64%), Oxynema sp. (9.09%), R. affinis (54.54%), Strongyloides sp. (4.54%), Physaloptera sp. (4.54%), T. hydatigena (9.09%), E. granulosus (4.54%), D. caninum (9.09%), M. lineatus (81.82%), Joyeuxiella pasqalei (27.27%), Diplopylidium nolleri (4.54%), M. hirudinaceus (22.72%) and Macracanthorhynchus sp. (9.09%). From golden jackals: T. canis (10%), T. leonina (30%), R. affinis (50%), T. hydatigena (10%), D. caninum (20%), M. lineatus (70%), J. pasqalei (30%.), Alaria canis (10%), M. hirudinaceus (30%) and Macracanthomynchus sp. (10%).     

Evaluation of the Efficacy of Atovaquone Alone or in Combination with Azithromycin against Acute Murine Toxoplasmosis

Mice were infected intraperitoneally with 10 000 tachyzoites of Toxoplasma gondii (RH) strain and, 24 h later, were treated orally for 10 days with atovaquone and azithromycin, either alone or in combination. Evaluation of the efficacy of the drugs was performed by microscopic examination of smears prepared from the organs of the mice, and by subinoculation of visceral and brain suspensions from surviving mice into healthy mice at the end of the experiments. It was found that 58%, 83% and 100% of the mice survived after administration of 75, 150 or 200 mg/kg per day of azithromycin, respectively. Moreover, 8%, 17% and 25% of the mice survived after treatment with atovaquone at 20, 50 or 100 mg/kg per day, respectively. No synergistic or additive effects of combinations of atovaquone and azithromycin were observed. However, azithromycin did not eradicate the parasite from the brain and viscera of the infected mice, whereas atovaquone at 20, 50 and 100 mg/kg per day removed the parasite from viscera and at 100 mg/kg per day eradicated the parasite from the brain of infected mice. The combinations of atovaquone and azithromycin failed to completely eradicate the parasite from the brain and viscera of infected mice.     

Immunization of sheep against Fasciola gigantica with glutathione S-transferase

Glutathione S-transferase in Fasciola gigantica (FgGST) was isolated by affinity chromatography, by which highly purified enzyme was obtained. FgGST on the SDS-PAGE showed three protein bands ranging 24.5-26.5kDa. Glutathione S-transferase (GST) activity was determined by HABIG method. FgGST was evaluated as vaccine alone or in combination with either aluminum hydroxide or saponin in sheep against F. gigantica infection. ELISA was used for detection of anti-FgGST IgG. After vaccination, all sheep were challenged with 120 metacercaria of F. gigantica. The results indicated that anti-GST IgG was not elevated after challenge. All sheep were slaughtered 24-26 weeks after challenge. The results indicated that, although after second vaccination, antibody titers rose markedly in GST-Al(OH)(3) and GST-saponin groups, but declined 4 weeks after challenge. No correlation between anti-GST IgG titers and protection was observed. The highest fluke burden reduction was observed in the group vaccinated with GST-saponin (32%), but this reduction was not statistically significant in comparison with the control group.     

Expression of the herpes simplex virus type 2 glycoprotein D in baculovirus expression system and evaluation of its immunogenicity in guinea pigs

Background: Herpes simplex virus type 2 (HSV-2) is highly prevalent and major cause of genital herpes in humans. The life-long nature of infection and the increasing prevalence of genital herpes imply that vaccination is the best strategy for controlling the spread of infection and limiting HSV disease. HSV glycoprotein D (gD) is one of the most important viral immunogen which has an essential role in virus infectivity and induction of immune responses. Methods: HSV-2 DNA was extracted and used as template in polymerase chain reactions to amplify gD2 gene. The PCR product was confirmed by restriction enzyme analysis, cloned into a cloning vector and then sequenced. The Bac-to-Bac expression system was used to express HSV-2 gD in insect cells. The expressed protein was used as subunit vaccine to immunize guinea pigs after confirmation. Results: The expressed protein was confirmed with SDS-PAGE and Western-blot analysis. In Western-blot analysis, two major protein bands, with approximate molecular weights of 52-55 and 41-43 kDa corresponding to the glycosylated and non-glycosylated forms of gD2 protein, were observed, respectively. Immunization with the recombinant gD2 could elicit humoral responses in guinea pigs as measured by neutralization test and ELISA, and offered high protection against induced HSV-2 genital disease. Conclusion: The baculovirus expression of heterologous genes permits proper folding, post-translational modification and oligomerization in manners that are often identical to those that occur in mammalian cells. Expression of proteins under the control of the strong polyhedrin promoter, allowing high level protein production, can be used as subunit vaccine.     

Immune responses against excreted/secreted antigens of Toxoplasma gondii tachyzoites in the murine model

In the present study, excretory secretory antigens (ESA) of Toxoplasma gondii were evaluated in immunization of 8-10 week inbred female Balb/c mice. Tachyzoites of the parasite were cultured in cell-free incubation medium (RPMI-1640), and then supernatant of the medium was loaded on an ion-exchange chromatography column. Two fractions (ESA-F(1) and ESA-F(2)) were collected from the column. For immunization of the mice, 50 were allocated into 5 groups of 10. The first, second, third, and fourth groups were immunized, twice with total-ESA, ESA-F(1), ESA-F(2) or toxoplasma lysate antigen (TLA), respectively. The fifth group was selected as a negative control group (non-immunized). The virulent RH strain of Toxoplasma gondii was used to challenge. Delayed-type hypersensitivity responses (DTHs) were measured by intra-footpad injection measuring induration at timed intervals. Lymphocyte transformation tests (LTTs) were done on lymph node cells using [3H] thymidine incorporation as an indication of reactivity. Peritoneal macrophages from sensitized mice were stimulated and nitric oxide was measured by Griess method. The ESA-F(1) and ESA-F(2) fractions were separated on poly acrylamide gel electrophoresis (PAGE) and SDS-PAGE. ESA-F(1) had 4 bands on PAGE and 14 bands on SDS-PAGE. ESA-F(2) had one band on PAGE and two bands on SDS-PGE. Sensitized mice showed DTH and lymphocyte transformation responses to total-ESA, ESA-F(1), and ESA-F(2) and peritoneal macrophages produce nitric oxide following stimulation. In challenge experiments, all non-immunized mice died within 10 days, whereas immunized mice survived for longer time periods (P<0.05). The highest survival rate was observed in mice that immunized with ESA-F(2). We suggest that these antigens especially ESA-F(2) should be of value for the development of new strategies for immunization against toxoplasmosis.