Amelioration of bleomycin induced pulmonary fibrosis by administration of Salvianolic acid B in mice.

Pulmonary fibrosis is the end­stage manifestation of wide range of respiratory diseases and during pulmonary fibrosis, pulmonary inflammation and epithelial­mesenchymal transition (EMT) play important roles. Salvianolic acid B (SAB) from the herb Salviae miltiorrhiza has been reported to possess an excellent anti­inflammatory, antifibrotic and antioxidant activity. The present study aimed to investigate the ameliorative effect of SAB on bleomycin induced pulmonary fibrosis in mice. Adult albino mice were divided as SHAM/control group (saline alone), BLM group (bleomycin @ 1mg/kg intratracheally once) and SAB groups (BLM challenged once and SAB administration in three dosages @ 5, 10 and 15 mg/kg intraperitoneally daily for 30 days). Lungs wet/dry ratio and protein concentration in bronchoalveolar lavage fluid, MPO activity, oxidative stress markers, hydroxyproline assay, levels of inflammatory cytokines (TNF­α, IL­6 and TGF­ß1), NF­κB activity, histopathology, immunostaining (E­cadherin, vimentin and alpha ­smooth muscle actin) and ultrastructural changes were studied. SAB showed anti­inflammatory and anti­fibrotic effects through inhibition of inflammatory cell infiltration, alveolar structure disruption, and collagen deposition and the expression of several fibrogenic cytokines. SAB also up­regulate E­cadherin and down­regulated vimentin and alpha­smooth muscle actin expression. In conclusion, Salvianolic acid B is effective in alleviating the BLM induced lung fibrosis through suppression of oxidative stress, inflammation, histological, ultrastructural changes and EMT.

Bacillus circulans MTCC 7906 aided facile development of bioconjugate nano-silica alkaline protease formulation with superlative dehairing potential.

The tannery industries utilize environmentally hazardous chemicals to achieve dehairing of animal hides, which causes enormous waterbed pollution & high TDS load. Alkaline protease enzyme for dehairing can be an effective solution to resolve the environmental problems of the tannery industry waste. However, stable, cost-efficient and eco-benign formulations of alkaline protease need to be developed for commercial applications in the tannery industry. This works aimed at development of a nano-formulation of the enzyme alkaline protease (AKP) as a bioconjugate nano silica-alkaline protease enzyme (BC-SiNP-AKP). This work reports one pot green synthesis of the BC-SiNP-AKP bionanoconjugate complex which included both biotemplating and immobilization of the AKP on to the synthesized silica nanoparticles from cell-free extracts of Bacillus circulans grown in potato peel based medium. Among the cell free crude, acetone concentrated and purified sols of the enzyme AKP, acetone precipitated enzyme sol was found to be best for the biological SiNP synthesis and formation of BC-SiNP-AKP conjugate. The BC-SiNP-AKP had size ranging from 100 to 200 nm with crystalline morphologies varying from spherical, tubular to laminated crystallites. The developed bioconjugate formulation displayed 1.7-fold increase in the enzyme activity post nano-conjugation with superlative dehairing potential on goat skin. The optimized parameters for dehairing were found to be as temperature 37 °C for 24 h of incubation and with enzyme to buffer ratio (2: 50 mL). Thereafter, the dehaired skin was assessed for its histopathological effects, which were found to be safe without any deteriorative changes. The developed formulation is environmentally congenial for its use as depilating agent for animal hides in terms of being green, single pot and cost effective synthesis.

Assessment of antigenic specificity of polyclonal antisera raised against Avibacterium paragallinarum by ELISA.

Lack of availability of commercial antibodies against whole-cell antigen or an antigenic epitope of Avibacterium paragallinarum (Av. paragallinarum) has hindered the development of novel immunoassays for the diagnose infectious coryza (IC). In this study, we raised polyclonal antisera against Av. paragallinarum and evaluated its antigenic-specificity using enzyme linked immunosorbent assay (ELISA). We standardized antigen coating concentration(s), antibody detection limit, and optimal range of dilutions of primary antisera and secondary conjugated antibody. Our results show the development of antigen-specific antibody response in rabbits following repeated antigenic exposure with 0.5% formalinized antigen over a period of four weeks. Further, we showed its possible applicability in detection of pathogens in tissues by immunohistochemistry for confirmatory disease diagnosis and disease pathogenetic study.

Development of nano-silver alkaline protease bio-conjugate depilating eco-benign formulation by utilizing potato peel based medium.

A new bio-conjugate nano-silver enzyme conjugate complex (BC-nAg-Akp) was formulated containing alkaline protease (Akp). The present research involved synthesis of nAg particles in acetone concentrated enzyme sol using 0.005 M AgNO3 solution formed within interaction time of 24 h through photo catalysis. The BC-nAG-Akp composite exhibited 1.9-fold increase in enzyme activity. The formulation was characterized using techniques viz., SEM, SEM-EDS, TEM, and DLS spectroscopy. The TEM analysis revealed synthesis of silver nano rods with size dimensions ranging from 40 to 80 nm. Likewise, the mean hydrodynamic diameter was 114 nm with polydispersity index of 0.260 and had the largest diffusion constant of 4.28 × 108 amongst the three forms of the formulation (crude, acetone concentrated and partially purified) on DLS characterization. The SEM-EDS analysis showed occurrence of 18.32 and 3.79%weight and %atom of Ag element respectively. The prepared formulation was investigated for its dehairing performance. The ideal dehairing was achieved at 37 °C after 12 h of treatment. The histopathological studies revealed that complete dehairing with minimal rarefication was achieved and was found perform better compared to the commercial Akp and control (crude enzyme) formulations.

Histopathological and immunohistochemical approaches for the diagnosis of Pasteurellosis in swine population of Punjab.

AIM: Infectious porcine bronchopneumonia, caused by Pasteurella multocida, is a widespread disease of major economic significance. Thus, the aim of the present study was to diagnose swine Pasteurellosis using gross, histopathological, and immunopathological approaches in the swine population of Punjab and to compare the efficacy of immunohistochemical (IHC) techniques with conventional diagnostic techniques. MATERIALS AND METHODS: A total of 71 adult swine lung samples showing gross pneumonic changes were collected along with the associated lymph nodes to carry out the study. The collected samples were then processed for histopathological and IHC studies. RESULTS: Out of the total 71 lung samples, 26 samples were found to be suspected for Pasteurellosis as per the microscopic changes observed, and out of these 26 samples, 16 cases were confirmed to be positive for Pasteurellosis by IHC. Varied macroscopic changes noted in lungs were pneumonic patches with consolidation of many lobes, congestion, and focal hemorrhages. Main lesions associated with lymph nodes were its enlargement and hemorrhages. Histologically, the lung showed fibrinous and suppurative bronchopneumonia, multifocal suppuration, thickening of septa with fibrin combined with cellular infiltration and edema. The higher IHC expression of P. multocida was seen in the bronchial epithelium besides in alveolar and bronchial exudate. Moreover, on comparing the histopathological and IHC scores which were calculated on the basis of characteristic microscopic lesions and number of antigen positive cells, respectively, a significant positive correlation (r=0.4234) was found. CONCLUSION: It was concluded that swine population of Punjab is having P. multocida infection. The gross and histopathological lesions can be helpful in the preliminary diagnosis of Pasteurellosis but needs to be supplemented by other immunodiagnostic tests. Moreover, IHC technique proved to be a specific, reliable, precise, and rapid technique to supplement these conventional methods of diagnosis for Pasteurellosis.

Ascaridia galli induced ulcerative proventriculitis in a poultry bird.

Various possible causes of proventriculitis include virus, bacteria, fungus, protozoans, nematodes, biogenic amines and excessive copper sulphate. In the present case, parasites were found in the lumen of the proventriculus, gizzard and duodenum of a poultry bird. Characteristic features of the parasite were studied and confirmed as Ascaridia galli. An ulcerative proventriculitis evident as denuded superficial epithelium, sub-epithelial hemorrhages, infiltration of the inflammatory cells and fibrosis were seen at histopathology. Proventriculitis caused by A. galli has not been reported till date. Here, we report a case of ulcerative proventriculitis in a poultry bird caused by nematode, A. galli.