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Excretory/secretory proteins of Haemonchus contortus (HcESPs) intermingle comprehensively with host immune cells and modulate host immune responses. In this study, H contortus ES antigen named as elongation factor 1 alpha (HcEF‐1α) was cloned and expressed. The influences of recombinant HcEF‐1α on multiple functions of goat peripheral blood mononuclear cells (PBMCs) were observed in vitro. Immunoblot analysis revealed that rHcEF‐1α was recognized by the serum of goat infected with H contortus. Immunofluorescence analysis indicated that rHcEF‐1α was bound on surface of PBMCs. Moreover, the productions of IL‐4, TGF‐β1, IFN‐γ and IL‐17 of cells were significantly modulated by the incubation with rHcEF‐1α. The production of interleukin IL‐10 was decreased. Cell migration, cell proliferation and cell apoptosis were significantly increased; however, nitric oxide production (NO) was significantly decreased. The MHC II molecule expression of cells incubated with rHcEF‐1α was increased significantly, whereas MHC‐I was not changed as compared to the control groups (PBS control and pET32a). These findings indicated that rHcEF‐1α protein might play essential roles in functional regulations of HcESPs on goat PBMC and mediate the immune responses of the host during host‐parasite relationship. [ABSTRACT FROM AUTHOR]

Galectins are glycan-binding proteins that are widely expressed and distributed in mammalian tissues as well as cells of innate and adaptive immune responses. CD4+ T-helper cells differentiate into effector subsets in response to cytokines. T helper 9 cells are one of the recently described subsets of effector T cells that are relatively new and less studied. In this study, galectin domain containing protein from Haemonchus contortus (Hc-GDC) was cloned, expressed in pET32a, and immunoblotting was performed. Localization of recombinant (r)Hc-GDC on outer and inner surface of H. contortus worm and binding with goat Peripheral Blood Mononuclear cells (PBMCs) were performed using immunofluorescence assay. Moreover, effects of rHc-GDC on proliferation, apoptosis, cell migration, and the nitric oxide production in goat PBMCs were evaluated. Furthermore, modulatory effects of rHc-GDC on production of Th1, Th2, and Th9 cells were evaluated by flowcytometry and on interferon gamma, interleukin (IL)-4 and IL-9 were evaluated by quantitative real-time polymerase chain reaction. The results demonstrated that rHc-GDC was successfully cloned, expressed in expression vector as well as in the gut surface of adult H. contortus worm and successful binding with PBMCs surface were observed. Immunoblotting results revealed that rHc-GDC is an important active protein of H. contortus excretory and secretory products. Moreover, the interaction of rHc-GDC with host cells increased the production of Th2, Th9 cells, IL4, IL-9, PBMC proliferation, nitric oxide, and cell migration. No effects of rHc-GDC were observed on PMBC apoptosis, production of Th1 cells, and secretions of IFN-γ and IL-10 cytokines. These findings indicate that recombinant GDC protein from H. contortus modulates the immune functions of goat PBMCs and has the potential to enhance protective immunity by inducing T helper-9-derived IL-9 in vitro. [ABSTRACT FROM AUTHOR]

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Serine/threonine-protein phosphatases (STPs), as integral constituents of parasitic excretory/secretory proteins, are assumed to be released during the host–parasite interactions. However, knowledge about these phosphatases and their immunoregulatory and immune protective efficiencies with host peripheral blood mononuclear cells (PBMCs) is scant. In this study, an open reading frame of STP from Haemonchus contortus designated as HcSTP-1 was amplified and cloned using reverse-transcription-polymerase chain reaction (RT-PCR) method. The 951-bp nucleotides sequence was encoded to a protein of 316 amino acid residues, conserved in characteristics motifs GDXHG, GDYVDRG, GNHE, HGG, RG, and H. The HcSTP-1 protein was detected at approximately 35 kDa as recombinant protein fused in an expression vector system and resolved on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Immunohistochemically, HcSTP-1 was found to be localized in both male and female adult worm sections. Using immunofluorescence assay, the binding activity of rHcSTP-1 was confirmed on surface of goat PBMCs, which resulted in expression of multiple cytokines and various immunoregulatory activities in vitro. The RT-PCR results showed that mRNA level of interleukin-2, TGF-β1, IFN-γ, and IL-17 (with 10 µg/ml) was upregulated and IL-10 was decreased. However, IL-6 showed no change after PBMCs incubated with rHcSTP-1 protein. Further functional analysis showed that migratory activity of cells, intracellular nitrite production (NO), and apoptotic efficiency of PBMCs were elevated at significant level, whereas the proliferation of goat PBMCs and monocytes-associated major histocompatibility complex (MHC)-I and MHC-II expressions were decreased significantly at concentration-dependent fashion. Our results showed that the HcSTP-1 protein engaged in vital suppressive regulatory roles on host immune cells, which might represent a potential molecular target for controlling H. contortus infection in future. [ABSTRACT FROM AUTHOR]

Background: Toxoplasma gondii can infect almost all warm-blood animals including human beings. The high incidence and severe damage that can be caused by T. gondii infection clearly indicates the need for the development of a vaccine. T. gondii elongation factor 1-alpha (TgEF-1α) plays an important role in pathogenesis and host cell invasion for this parasite. The aim of this study was to evaluate the immune protective efficacy of a DNA vaccine encoding TgEF-1α gene against acute T. gondii infection in mice.Methods: A DNA vaccine (pVAX-EF-1α) encoding T. gondii EF-1a (TgEF-1α) gene was constructed and its immune response and protective efficacy against lethal challenge in BALB/c mice were evaluated.Results: Mice inoculated with the pVAX-EF-1α vaccine had a high level of specific anti-T. gondii antibodies and produced high levels of IFN-gamma, interleukin (IL)-4, and IL-17. The expression levels of MHC-I and MHC-II molecules as well as the percentages of both CD4(+) and CD8(+) T cells in mice vaccinated with pVAX-EF-1α were significantly increased (p < 0.05), compared with those in all the mice from control groups (blank control, PBS, and pVAXI). Immunization with pVAX-EF-1α significantly (p < 0.05) prolonged mouse survival time to 14.1 ± 1.7 days after challenge infection with the virulent T. gondii RH strain, compared with mice in the control groups which died within 8 days.Conclusions: DNA vaccination with pVAX-EF-1α triggered strong humoral and cellular responses and induced effective protection in mice against acute T. gondii infection, indicating that TgEF-1α is a promising vaccine candidate against acute toxoplasmosis. [ABSTRACT FROM AUTHOR]

Background: Encephalitozoon cuniculi is a microsporidian parasite commonly found in rabbits that can infect humans, causing encephalitozoonosis. Our objective in this study was to evaluate the seroprevalence of this parasite in rabbits and humans in China Methods: Overall, 300 serum samples each from clinically healthy rabbit and human were collected from three regions of China (Sichuan Province, Chongqing Municipality and Jilin Province) from January to September 2013 and tested for anti- E. Cuniculi antibodies using an ELISA. Results: An overall seroprevalence of E. cuniculi was recorded as 56/300 (18.76%) and 29/300 (9.76%) in rabbit and human sera, respectively. The seropositivity of rabbit samples collected from Jilin province was 41%, which was significantly higher (P<0.01) than Sichuan Province (9%) and Chongqing Municipality (6%). Three breeds of rabbit were used in the present study and antibody detection in Rex Rabbit was significantly (P<0.01) higher than Japanese White and New Zealand Rabbit. In human, Jilin province was more prevalent (18%) followed by Sichuan Province (6%) and Chongqing Municipality (5%). Conclusions The E. cuniculi was present and widespread among healthy rabbits and humans in China [ABSTRACT FROM AUTHOR]

This study aimed to determine the changes of cytokines, specific serum IgG and several parameters in chickens vaccinated with DNA vaccine encoding Eimeria brunetti apical membrane antigen-1 (EbAMA1) antigen. Two-week-old chickens were divided into five groups (four groups for experiment) randomly. Experimental groups of chickens were immunized with DNA vaccine while control group of chickens were injected with pVAX1 plasmid alone or TE buffer solution. All immunizations were boosted 2 weeks later. The EbAMA1 specific IgG antibody responses were measured at weeks 1-6 post-second immunizations and several parameters were also identified. The result showed that the antibody titers in chickens vaccinated with DNA vaccines were significantly different from those of the control groups 1 week after the second immunization and reached the maximum values 3 weeks post-second immunization. IFN-γ concentration was increased the highest level against EbAMA1 of all chickens vaccinated with vaccines up to 56-fold, follow by the specific IgG antibody levels were increased 10-17-fold compared with those of TE solution and plasmid (pVAX1) control chickens 1-6 weeks post-second immunization. In case of the levels of IL-10 and IL-17 was increased in experimental chickens with 4-5-fold. Even though it was statistically significant, TGF-β and IL-4 levels were higher in vaccinated than unvaccinated chickens. The results suggested that DNA vaccines encoding E. brunetti apical membrane antigen-1 (EbAMA1) could increase serum specific IgG antibody and cytokines concentration and could give protection against E. brunetti infection. [ABSTRACT FROM AUTHOR]

The present study was planned to carry out a seroprevalence study of PPR antibodies in North West Frontier Province (NWFP) including Federally Administered Tribal Areas (FATA), using competitive-ELISA (c-ELISA) kit. A total of 4548 serum samples collected from 26 locations of the NWFP including. Eight hundred twenty eight samples were found positive for PPR antibodies yielding an overall seroprevalence of 18.20 percent in the small ruminant population included in the study. 337 (24.90%) serum samples from sheep and 491 (15.36%) from goats were found positive in PPR c-ELISA yielding a marginal difference in seroprevalence of PPR. The current situation of PPR in Pakistan could be regarded as of low magnitude but constant threat. Incidence may increase to the alarming level due to uncontrolled/ free movement within and from across boarders to the stocks of non-immunized susceptible population of sheep and goats in the country but production systems and economic realities constraint the effective animal movement control within and across the boarders. Based on the limited reports of outbreaks and uneven distribution of occurrence over the years of past decade, the best available options are high level of prevention and objective surveillance. Keeping in view the high risk of PPR, control strategy should be adopted and further study needs to be undertaken to ascertain the extent of PPR virus circulating in the population of goats and sheep in the country. Under existing production and marketing system, sanitation and preventive vaccination are two practicable options for control of PPR in Pakistan. Homologous PPR vaccine is now available which is a freeze dried live vaccine derived from tissue culture from attenuated PPR virus. The vaccine is intended for sheep and goats and is required to be injected to all animals above the age of 3 month. The vaccine may be repeated after one year of the primary vaccination in high risk areas. [ABSTRACT FROM AUTHOR]

The present study was conducted to check out the efficacy of various antiprotozoal drugs on bovine babesiosis, anaplasmosis and theleriosis. 38 buffaloes naturaly infected with anaplasma, babesia and theileria were selected and divided into four groups. Group A (10), B (10), C (08) and D (10) infected with anaplasmosis, babesiosis, theileriosis, and Infected Control (anaplasmosis (4), anaplasmosis + babesiosis (1 each), babesiosis (2) and theileriosis (2) respectively. Group A was further subdivided into A-I (5 animals with pure Anaplasmosis) and A-II (5 animals with mixed infection of Anaplasmosis and Babesiosis). Group-A was treated with a single dose of imidocarb dipropionate (3 mg/kg body weight). In Group A-I one animal recovered completely, 02 moderately while remaining 02 animals did not respond. The later 02 animals recovered completely after additional 02 doses of 4 mg/kg body weight. In case of group A-II complete recovery was recorded in one animal, moderate recovery in three animals while one animal showed no responsel. Group B was treated with Diminazene acceturate (3. 5 mg/kg body weight) administered as a single dose. 6 animals recovered completely, 02 moderately while 02 animals showed no response. Group C was treated with a single dose of Buparvaqune (2.5 mg/kg body weight), complete recovery was recorded in 4 animals, 2 animals showed no response, while 2 animals with severe infection died after 24 hrs. [ABSTRACT FROM AUTHOR]

ADP-ribosylation factors (ARFs) are members of the Ras-related small GTPase family involved in the vesicular trafficking regulation. Immunomodulatory effects of these proteinson host cell arenot being addressed yet. H. contortus small GTPase ADP-ribosylation 1 gene (HcARF1) was cloned and recombinant protein of HcARF1 (rHcARF1) was successfully expressed in Escherichia coli . Binding activity of rHcARF1 to goat PBMCs was confirmed by immunofluorescence assay (IFA) and its immunomudulatory effects on cytokine secretion, cell proliferation, cell migration and nitric oxide production (NO) were observed by co-incubation of rHcARF1. IFA results revealed that rHcARF1 could bind to the PBMCs. The interaction of rHcARF1 modulated the cytokine production, the production of IL-4, IL-10 and IL-17 was increased in a dose dependent manner, however, the IFN-γ production was significantly decreased. Cell migration and NO production were significantly increased by rHcARF1, whereas, rHcARF1 treatment significantly suppressed the proliferation of the PBMC in a dose dependent manner. Our findings showed that the rHcARF1 play important roles on the goat PBMCs., Competing Interests: CONFLICTS OF INTEREST The authors declare that there is no conflict of interest.

Histological and hematological disturbance caused by Arsenic containing water were studied in mice model. Animal were divided into four groups. Control group exposed to arsenic free distilled water and 3 treatmental group exposed to the arsenic containing water with 30, 150 and 300 ppb. Blood samples and organs were collected after 40 days. Histopathological results revealed mild to severe type of necrosis and degenerative changes in kidney and liver of arsenic feed animals. Kidney of the 300 ppb group showed severe type of necrosis and degenerative changes in distal and proximal tubules. The renocytes of proximal and distal tubules were showing hydropic and fatty degeneration. Due to degenerative changes cells were showing cytoplasmic vacoulation and cytoplasmic and nuclear blebbing. Glomeruli cells were contracted and increased the bowman's spaces. Varied degrees of changes were also observed in 30 and 150 ppb exposed group. Necrosis of hepatocytes and cytoplasmic blebbing were also observed. The sinusoidal spaces were expanded due to shrinkage and necrosis of hepatocytes. Spleenocytosis occurred in spleen and the parenchymal and mesenchymal cells were replaced by connective tissue. The lymphocytes were severely damaged by arsenic toxicity. White Blood Cells (WBCs), Red Blood Cells (RBCs) and hemoglobin level in control groups were in normal range where as level were significantly decreased with the increase dose of arsenic in the respective treatmental groups. The data was analyzed statistically and was found that significant was found among the group (p<0.05).