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Isolation, purification and anti-bacterial property of calprotectin from bovine neutrophil
2016
Imani, Mahdi | Tukmechi, Amir
BACKGROUND: It is believed that bovine neutrophils contain several peptides and protein which exhibit antimicrobial activity against microorganisms such as fungi and bacteria. Objectives: The purpose of this study was to isolate and purify a potential antibacterial protein from bovine neutrophil and test its anti bacterial activity. Methods: Neutrophils were isolated from bovine blood using dextran sedimentation and centrifugation on Ficoll-Hypaqe. Cell viability is examined by trypan blue dye exclusion method. Protein extract was then dialyzed and applied onto ion exchange chromatography for further purification, and its potential cytostatic activity was examined against Staphylococcus aureus, Aeromonas hydrophila and Yersinia ruckeri. Results: Viability of isolated neutrophil was over 95%, chromatographic results and SDS-PAGE analysis exhibited that neutrophil cytosolic proteins were fractionated so that the purification of P7/P23 was almost 60% in the first step and purification was completed in the second phase. Using calibration curve according to molecular mass markers, the relative molecular mass of P7 and P23 determined as 7 kDa and 23 kDa, respectively. Also, results showed that this protein has antibacterial activity and has higher bactericidal activity against Y. ruckeri. Conclusions: It could be concluded that purification of P7/P23 sustains its biological activity and has wide range antibacterial activity. Moreover, taking all data into account may suggest that the cytostatic activity of neutrophil, to some extent, results from P7/P23 protein which is abundant in the cytosole.
Afficher plus [+] Moins [-]Isolation and antibiogram of Salmonella spp. from duck and pigeon in Dinajpur, Bangladesh
2016
Md. Moshiur Rahman | Md. Mostafizer Rahman | Mirza Mienur Meher | Md. Safikul Islam Khan | AKM Mostafa Anower
Objective: This research work was conducted for isolation of Salmonella spp. from duck and pigeon and assessment of antibiotic sensitivity patterns of the isolated Salmonella spp.Materials and methods: A total of 48 duck samples comprising of liver (n=16) spleen (n=16) and intestinal content (n=16) were collected from 16 ducks (8 sick and 8 dead). Similarly, 42 pigeon samples comprising of liver (n=14), spleen (n=14) and intestinal contents (n=14) were collected from 14 pigeons (7 sick and 7 dead). The samples were collected from the selected Duck and Pigeon farms at three Upazillas (sub-districts) in Dinajpur district, Bangladesh. The samples were subjected for isolation and identification of Salmonella spp. following standard bacteriological examinations such as cultural and morphological characteristics, biochemical properties and motility test. Commercially available antibiotic discs were used for the assesment of antibiotic resistant patterns by disc diffusion method.Results: Salmonella spp. could be isolated from 39.58% (n=19/48) duck and 28.57% (n=12/42) pigeon samples. The growth of colony with different characteristics in Brilliant Green agar, Nutrient agar, Salmonella-Shigella agar and Mac Conkey agar indicated positive results. Dextrose and mannitol were fermented and in Triple Sugar Iron (TSI) slant media, the isolates produced red color, yellow in butt with slightly black color. Positive result was found in Indole and Methyl Red media. The isolated Salmonella organisms were found to be highly sensitive to Azithromycin, Ciprofloxacin and Levofloxacin. Conclusion: Pigeons and ducks found in the study area may be the carrier of Salmonella spp. The farm should be checked periodically to know the status of Salmonella infection. Further study might be isolation and identification of Salmonella organism with detail genomic analysis and drug resistance. [J Adv Vet Anim Res 2016; 3(4.000): 386-391]
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