This study was aimed to identify and characterize Leishmania amastigote, and axenic form antigens. Two in vitro techniques were used to change leishmania parasite isolates from promastigote form to amastigotes and amastigote like (axenic) forms. The main strategy relied upon in vitro infection of murine macrophages cell line J774 with leishmania promastigote, at 37°C with 5% CO2, while the second technique relied upon the culture of promastigote at 37°C with low pH (5.5), and 5-10% CO2. Proteins were extracted and fractionated utilizing 12% Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS PAGE). Antigens were recognized using both immune dot blot and western blot procedures. PCR was performed for recognition of leishmania parasites in infected J774 macrophages. L. major was quicker in infectivity of macrophages cell line than L. donovani. Shared proteins ranging from 26-116 kDa were identified by SDS PAGE in all stages. Immune Dot-blot method showed positive outcomes, while western blot identified an exceptional antigen band of 16 kDa in amastigote, this unique band could be of value in diagnosis and vaccination of leishmaniasis. PCR results confirmed presence of both isolates demonstrating that coinfection is conceivable, and no indications of hereditary recombination at kinetoplast DNA (kDNA) were identified in macrophages simultaneously infected by L. major and L. donovani.
Avian influenza or bird flu is a common problem of domestic and wild birds. Some of its strains are able to cross the species barrier and cause infection in various members of class Mammalia. In view of relatively lesser efficacy of vaccines, antiviral therapies remain the only choice for the sustenance of mammals acquiring this highly devastating infection. This study is based on the evaluation of antiviral potential of methanol extracts of eleven selected Cholistani plants. The methanol extracts were prepared by using dried plants material followed by concentrating in a rotary evaporator and finally air dried before dissolving in nanopure water. The suspension was filter sterilized and subjected to in ovo antiviral assays. The allantoic fluids were harvested and haemagglutinin (HA) titers were determined. Among the eleven plants evaluated all methanol extracts were found effective against AIV H9N2 except S. baryosma extract. The medicinal plants O. compressa, N. procumbens, and S. surattense were found to be more effective than others and they retained HA titers at 0 after challenge. The next in order were extracts of O. esculentum, H. salicornicum and S. fruticosa which kept HA titers at 4, 8 and 16 respectively. The extracts of H. recurvum, P. antidotale, S. icolados and A. aspera were found less effective than above mentioned plant extracts and they kept the HA titers at 32, 64, 128 and 256 respectively. These results led us to conclude that the medicinal plants of Cholistan region are a rich source of antiviral agent(s) against AIV H9N2 and could be a source of cost effective alternate therapeutics.
Sudanese mucosal leishmaniasis (ML) is a rare clinical form of leishmaniasis and characterized by persistent ulcer of the oral and/or the nasal mucous membranes caused by Leishmania donovani. No data is available about the systemic and local immune responses in mucosal leishmaniasis. This study aimed to measure the systemic and the local cytokines responses of Sudanese ML patients compared to cured cutaneous leishmaniasis patients (Leishmanin skin test positive, LST+ve) and unexposed healthy controls (Leishmanin skin test negative, LST-ve). Six parasitological confirmed ML patients, 7 LST+ve, and 6 LST-ve were enrolled. Systemic Th-1 (IFN-γ and TNF-α), Th-2 (IL-10 and IL-13), Treg (TGF-β1), and inflammatory cytokines IL-6 and IL-8 concentration were measured in the supernatant of whole blood samples following stimulation with live L. donovani promastigotes using ELISA. Local intralesion IL-10, IFN-γ, and IL-13 expression was measured using Real Time PCR. A significant high concentrations of IFN-γ, TNFα, IL-10, TGFβ, IL-6, and IL-8 were detected in the supernatant of stimulated whole blood samples of ML patients compared with the LST+ve and LST-ve controls. Using Real Time-PCR and primers for various cytokines, a significant high expression of TH2 cytokines IL-10 and IL-13 mRNA was detected in contrast to a low TH1 cytokine IFN-γ mRNA in the mucosal lesion. There is a clear dichotomy in the cytokine response during Mucosal leishmaniasis. A significantly high TH1, inflammatory and Treg cytokines response is produced systemically, in contrast to a significant high TH2 cytokines response in the mucosal lesion.
The study investigates the latent pollution sources and most significant parameters that cause spatial variation and develops the best input for water quality modelling using principal component analysis (PCA) and artificial neural network (ANN). The dataset, 22 water quality parameters were obtained from Department of Environment Malaysia (DOE). The PCA generated six significant principal component scores (PCs) which explained 65.40 % of the total variance. Parameters for water quality variation are mainlyrelated to mineral components, anthropogenic activities, and natural processes. However, in ANN three input combination models (ANN A, B, and C) were developed to identify the best model that can predict water quality index (WQI) with very high precision. ANN A model appears to have the best prediction capacity with a coefficient of determination (R2) = 0.9999 and root mean square error (RMSE) = 0.0537. These results proved that the PCA and ANN methods can be applied as tools for decision-making and problem-solving for better managing of river quality.
Organophosphate insecticide spray poses potential threat of contamination of environmental components their accumulation in aquatic organisms. Although various physiological deficits associated with their exposure in fishes are documented, yet their retention in their edible muscle tissues has been poorly studied. In this context, the study was undertaken to ascertain the bioaccumulation of two organophosphate insecticide compounds (dimethoate and chlorpyrifos) in the muscles of juvenile Cyprinus carpio. The study could provide insight into the risks to human health associated with consuming contaminated fish flesh. The fishes exposed to various concentrations of dimethoate and chlorpyrifos in-vivo for 96 to ascertain the uptake and retention of these insecticides in the muscle. Results indicated that fish muscles accumulated the residues at all the concentrations with the recovery of 2.99% (0.032 ppm) of dimethoate exposed to LC50 concentrations. In contrast, the chlorpyrifos residues were found Below the Detection Level (BDL) in the fishes exposed to LC50 concentrations. The percentage bioaccumulation of dimethoate in fish muscle was 88.10%, and that of chlorpyrifos was BDL. The bio-concentration factor was dose-dependent and increased with increasing doses of both insecticides. The study invites attention to human health risk assessment in the regions where contaminated fish are consumed without scientific supervision.