• Title/Summary/Keyword: Local isolate

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Pathogenicity of a Local Isolate of Chicken Anemia Agent for Chickens and Prevalence of Antibody in Chicken Flocks (국내분리 chicken anemia agent의 닭에 대한 병원성과 야외계군의 항체 보유상황)

  • 김선중
    • Korean Journal of Poultry Science
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    • v.18 no.3
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    • pp.141-150
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    • 1991
  • A local isolate of chicken anemia agent (CAA), isolate 89-69. was tested for pathogenicity for chickens. When chickens from a specific pathogen free (SPF) flock were inoculated intramuscularly with the isolate at one day old, all the chickens showed severe anemia at 14 to 18 days post inoculation(DPI) and returned to normal at 25DPI, Some of the inoculated chickens (27∼33%) died between 13 to 17 DPI's with lesions of severe aplasia of bone marrow and thymic atrophy. In chickens kept in contact with inoculated chickens, some of the chickens had anemia at 25 and 28 DPI's. Virus could be reisolated from inoculated as well as in contact chickens till 21 DPI. Antibodies to CAA could be detected in all inoculated and in contact chickens when tested at 42 DPI by the indirect fluorescent antibody method. When chickens from a different SPF flock were inoculated at one day old, degrees of anemia, both in frequency of incidence and severity, were low These chickens were proved partly to have antibodies to CAA when tested for hatchmates. In a survey for antibodies to CAA in field chicken flocks, one out of 7 flocks(14%) aged 3 to 10weeks was antibody positive whereas 19 out of 20 flocks(95%) over 20 weeks of age were positive. Altogether 29 out of 39 flocks (74%) were antibody positive.

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Solid Substrate and Submerged Culture Fermentation of Sugar Cane Bagasse for the Production of cellulase and Reducing Sugars by a Local Isolate, Aspergillus terreus SUK-1

  • Wan Mohtar, Yusoff;Massadeh, Muhannad Illayan;Kader, Jalil
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.770-775
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    • 2000
  • Several process parameters were studied to ascertain the effect on degradation of sugar cane bagasse in relation to the production of cellulase enzyme and reducing sugars by Solid Substrate Fermentation (SSF) and Submerged Culture Fermentation (SCF) of Aspergillus terreus SUK-1. The effect of air-flow rate (0-1.3 v/v/m), of different ratios of substrate weight to liquid volume (1:6, 1:10, 1:20, and 1:30 w/v, g/ml), scale-up effect (10, 20, and 100 times of 1:10 ration, w/v) and the effect of temperature (30, 40, 50, and $60^{\circ}C$) in SSF were studied. Air-flow rate of 1.0 v/v/m gave the highest enzyme activity (FPase 0.25 IU/ml, CMCase 1.24 IU/ml) and reducing sugars concentration (0.72 mg/ml). Experiment using 1:10 ratio (w/v) was found to support maximum cellulase activity (FPase 0.58 IU/ml, CMCase 1.97 IU/ml) and reducing sugar concentration (1.23 mg/ml). Scaling-up the ratio of 1:10(w/v) by a factor of 20 gave the highest cellulase activity (FPase 0.71 IU/ml, CMCase 2.25 IU/ml) and reducing sugar concentration (3.67 mg/ml). The optimum temperature for cellulase activity and reducing sugar production was $50^{\circ}C$(FPase 0.792 IU/ml, CMCase 2.25 IU/ml and 3.85 mg/ml for reducing sugar concentration). For SCF, the activity of cellulase enzyme and reducing sugar concentration was found to be lower than that obtained for SSF. The highest cellulase activity obtained in SCF was 50% lower than the highest cellulase activity in SSF, while for reducing sugar concentration, the highest concentration obtained in SCF was 90% lower than that obtained in SSF.

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Optimization of Extraction of Fpase from the Fermented Bran of Aspergillus niger in Solid State Fermentation

  • Chandra, M. Subhosh;Reddy, B. Rajasekhar;Choi, Yong-Lark
    • Journal of Applied Biological Chemistry
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    • v.51 no.4
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    • pp.155-159
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    • 2008
  • A local isolate of Aspergillus niger was cultivated under optimal growth conditions on wheat bran in solid state fermentation. Filter paperase from fermented bran was separately extracted with different solvents to test the recovery of the enzyme. Among solvents tested, distilled water served as the best leachate, thus the conditions were further optimized with distilled water. After two washes of fermented bran with distilled water for 1.5 h each under stationary conditions at 1 g wheat bran: 5 mL distilled water, the maximum recovery of 13.5 $Ug^{-1}$ of wheat bran was obtained.

Experimental Pathogenesis of Pullorum Disease with the Local Isolate of Salmonella enterica serovar. enterica subspecies Pullorum in Pullets in Bangladesh

  • Haider, M.G.;Chowdhury, E.H.;Khan, M.A.H.N.A.;Hossain, M.T.;Rahman, M.S.;Song, H.J.;Hossain, M.M.
    • Korean Journal of Poultry Science
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    • v.35 no.4
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    • pp.341-350
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    • 2009
  • The research work was carried out to study the pathogenesis covering the clinical signs, gross and histopathological lesions in different organs, and reisolation and identification of the organisms after experimental infection with the local isolate of Salmonella enterica serovar. enterica subspecies (S.) Pullorum at different time interval of the experiment during the period February 2006 to December 2006. One hundred pullets (seronegative to S. Pullorum of 12 weeks age were purchased and divided into 5 (A, B, C, D and E) groups and each group consisted of 20 birds. Four groups (A, B, C and D) were infected orally with a dose of $10^6\;CFU$, $10^7\;CFU$, $2{\times}10^7\;CFU$, $10^8\;CFU$ of S. Pullorum, respectively, and one group (E) was treated as uninfected control. The used methods were necropsy and histopathology, culture of bacteria, staining and biochemical test of Salmonella. Five birds from each group were randomly selected and sacrificed $1^{st}$ week, $2^{nd}$, $3^{rd}$ and $4^{th}$ weeks of post infection (PI). From all the groups, the bacteriological samples (crop, liver, lung, heart, spleen, bile duodenum, ceca and blood) were collected with pre enriched in buffered peptone water in sterile poly bags. Liver, lungs, heart, spleen, intestine, etc. were collected in 10% buffered-formalin for histopathological examination. No clinical signs, gross and histopathological lesions were found in control group and no S. Pullorum was reisolated. Clinical sign of experimentally infected with S. Pullorum in pullets were loss of appetite (100%), slight depression (75%), ruffled feathers (85%), diarrhea (60%) and loss of weight (100%) in chickens. The feed intake and body weight at different weeks after PI differed significantly (p<0.01) among the groups. Grossly, the highest recorded lesion was button-like ulcer in the ceca (80%) and the lowest was white nodules in lungs (1.25%). S. Pullorum were reisolated from crop (91.25%), liver (91.25%), lung (83.75%), heart (71.25%), spleen (87.75%), bile (33.25%), duodenum (92.50%), ceca (97.50%) and from different group of infection (61.25%). The highest microscopic findings were intestinal and cecal mucosa and submucosa exhibited infiltration of mononuclear cells and congestion (96.25%), and the lowest finding was nodule formation in the lungs (3.75%). The pattern of the disease production by local isolate of S. Pullorum in Bangladesh is almost similar with other isolates in different countries.

Bioprocess Development for Production of Alkaline Protease by Bacillus pseudofirmus Mn6 Through Statistical Experimental Designs

  • Abdel-Fattah, Y.R.;El-Enshasy, H.A.;Soliman, N.A.;El-Gendi, H.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.4
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    • pp.378-386
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    • 2009
  • A sequential optimization strategy, based on statistical experimental designs, is employed to enhance the production of alkaline protease by a Bacillus pseudofirmus local isolate. To screen the bioprocess parameters significantly influencing the alkaline protease activity, a 2-level Plackett-Burman design was applied. Among 15 variables tested, the pH, peptone, and incubation time were selected based on their high positive significant effect on the protease activity. A near-optimum medium formulation was then obtained that increased the protease yield by more than 5-fold. Thereafter, the response surface methodology(RSM) was adopted to acquire the best process conditions among the selected variables, where a 3-level Box-Behnken design was utilized to create a polynomial quadratic model correlating the relationship between the three variables and the protease activity. The optimal combination of the major medium constituents for alkaline protease production, evaluated using the nonlinear optimization algorithm of EXCEL-Solver, was as follows: pH of 9.5, 2% peptone, and incubation time of 60 h. The predicted optimum alkaline protease activity was 3,213 U/ml/min, which was 6.4 times the activity with the basal medium.

Medium optimization for keratinase production by a local Streptomyces sp. NRC 13S under solid state fermentation

  • Shata, Hoda Mohamed Abdel Halim;Farid, Mohamed Abdel Fattah
    • Journal of Applied Biological Chemistry
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    • v.56 no.2
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    • pp.119-129
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    • 2013
  • Thirteen different Streptomyces isolates were evaluated for their ability to produce keratinase using chicken feather as a sole carbon and nitrogen sources under solid state fermentation (SSF). Streptomyces sp. NRC 13S produced the highest keratinase activity [1,792 U/g fermented substrate (fs)]. The phenotypic characterization and analysis of 16S rDNA sequencing of the isolate were studied. Optimization of SSF medium for keratinase production by the local isolate, Streptomyces sp. NRC13S, was carried out using the one-variable-at-a-time and the statistical approaches. In the first optimization step, the effect of incubation period, initial moisture content, initial pH value of the fermentation medium, and supplementation of some agro-industrial by-products on keratinase production were evaluated. The strain produced about 2,310 U/gfs when it grew on chicken feather with moisture content of 75% (w/w), feather: fodder yeast ratio of 70:30 (w/w), and initial pH 7 using phosphate buffer after 8 days. Based on these results, the Box-Behnken design and response surface methodology were applied to find out the optimal conditions for the enzyme production. The corresponding maximal production of keratinase was about 2,569.38 U/gfs.

Immunosuppressive effects and pathogenicity of a Korean isolate of reticuloendotheliosis virus in chickens (Reticuloendotheliosis virus의 닭에 대한 면역억제효과와 병원성)

  • Han, Myung-guk;Kim, Sun-joong
    • Korean Journal of Veterinary Research
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    • v.40 no.2
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    • pp.311-323
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    • 2000
  • Immunosuppressive effects of reticuloendotheliosis virus (REV) infection in chickens were investigated. Primary antibody responses to Newcastle disease virus (strain B1) and sheep red blood cells were significantly low in chickens inoculated with the local isolate 89-74 of REV compared to those of uninfected chickens. In chickens infected with REV strain T or 89-74, blastogenesis of spleen cells and peripheral blood lymphocytes (PBL) to concanavalin A (Con A) was severely suppressed. When specific pathogen free (SPF) chickens were inoculated with the isolate, the suppressive effect was observed up to 7 weeks of age while, in the contact infected chickens, the suppression was absent. Similar suppressive effects were observed in chickens inoculated with REV strain T at 2, 3 and 4 weeks of age. When spleen cells or PBL from uninfected chickens were co-cultured with spleen cells or PBL from chickens infected with REV at 1 day-old or 2 week-old, the blastogenesis of the normal cells was suppressed. The suppressive effect of PBL from REV-infected chickens on normal lymphocytes was abrogated by the treatment with trypsin. However the suppressive activity of the REV-infected PBL was not influenced at removing machrophage from the cell suspension by incubation in plastic petri dishes. In addition to the immunosuppression, chickens infected with the REV isolate showed abnormal feather development (nakanuke), anemia, paralysis and retarded growth. Three out of 11 chickens inoculated with the isolate at day-old died between 6 and 9 weeks of age by bacterial infections.

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Fabrication of Biogenic Antimicrobial Silver Nanoparticles by Streptomyces aegyptia NEAE 102 as Eco-Friendly Nanofactory

  • El-Naggar, Noura El-Ahmady;Abdelwahed, Nayera A.M.;Darwesh, Osama M.M.
    • Journal of Microbiology and Biotechnology
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    • v.24 no.4
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    • pp.453-464
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    • 2014
  • The current research was focused on the extracellular biosynthesis of bactericidal silver nanoparticles (AgNPs) using cell-free supernatant of a local isolate previously identified as a novel Streptomyces aegyptia NEAE 102. The biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102 was quite fast and required far less time than previously published strains. The produced particles showed a single surface plasmon resonance peak at 400 nm by UV-Vis spectroscopy, which confirmed the presence of AgNPs. Response surface methodology was chosen to evaluate the effects of four process variables ($AgNO_3$ concentration, incubation period, pH levels, and inoculum size) on the biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102. Statistical analysis of the results showed that the linear and quadratic effects of incubation period, initial pH, and inoculum size had a significant effect (p < 0.05) on the biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102. The maximum silver nanoparticles biosynthesis (2.5 OD, at 400 nm ) was achieved in runs number 5 and 14 under the conditions of 1 mM $AgNO_3$ (1-1.5% (v/v)), incubation period (72-96 h), initial pH (9-10), and inoculum size (2-4% (v/v)). An overall 4-fold increase in AgNPs biosynthesis was obtained as compared with that of unoptimized conditions. The biosynthesized silver nanoparticles were characterized using UV-VIS spectrophotometer and Fourier transform infrared spectroscopy analysis, in addition to antimicrobial properties. The biosynthesized AgNPs significantly inhibited the growth of medically important pathogenic gram-positive (Staphylococcus aureus) and gram-negative bacteria (Pseudomonas aeruginosa) and yeast (Candida albicans).

Isolation, Identification and Production of Salmonella Pullorum Coloured Antigen in Bangladesh for the Rapid Whole Blood Test

  • Hoque, M.M.;Biswas, H.R.;Rahman, L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.1
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    • pp.141-146
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    • 1997
  • Postmortem examination was conducted on 350 (three hundred and fifty) chickens. Related samples (Liver, heart, ovary, spleen, bone-marrow, and caecal junction) were collected. The appropriate materials from the samples were cultured into different media. A total 40(forty) isolates of salmonella pullorum and S. gallinarum were identified and preserved. Characterization of the isolates were done by cultural, morphological, biochemical, and serological tests. Salmonella pullorum antigen was prepared from the local isolate, standardized and tested. This antigen was used in the field for the detection of pullorum or fowl typhoid infection or carrier birds. The antigen consisted of suspension of Salmonella pullorum in 0.50 percent sodium chloride plus 1.5 percent sodium sulfate and inactivated with 1% formalin U.S.P. and standardized with McFarland scale iv or by pour plate method containing 800 million organisms per milliliter and stained by the addition of alcoholic crystal violet. Sterility, safety and potency were tested and found as good as other international antigens. The antigen was found to retain its quality for six months when preserved at room temperatures. The test was made by mixing one drop of the antigen with a drop of blood or a drop of serum, on a glass plate or white tile. The locally produced antigen was as good as antigens from Japan, Hungary, Holland and India. A serological study was conducted with the locally prepared antigen in different farms, and the incidence was 0-4% in government farms, 5-10% in commercial imported breeds and 0-3% in cross breed local farms respectively.

Sequence Analysis of E2 Glycoprotein from Indian Isolate of Classical Swine Fever Virus (CSFV)

  • Bajwa, Mehak;Verma, Ramneek;Deka, Dipak;Dhol, Gagandeep Singh;Barman, Nagendra Nath
    • Microbiology and Biotechnology Letters
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    • v.43 no.1
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    • pp.22-30
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    • 2015
  • CSF is a major concern for the swine industry, representing currently the most epizootically dangerous disease to the species. Numerous CSFV isolates with various degrees of virulence have already been isolated worldwide, ranging from low virulent strains that do not result in any apparent clinical signs to highly virulent strains that cause a severe per acute hemorrhagic fever with very high mortality. The molecular epidemiology of CSFVs has proven to be an essential tool for effective disease control and the development of safe and effective vaccines. Therefore, this study cloned and sequenced local CSFV isolates, and conducted a phylogenetic analysis based on the E2 glycoprotein encoding sequences.The RNA was extracted from PK15 cell culture passaged CSFV isolates, the cDNA prepared, and the complete E2 gene amplified with a product size of 1186 bp. The gelpurified PCR product was cloned into a pGEMT easy vector and the positive clone commercially sequenced. Aligning the nucleotide (1119 bp) and amino acid (373) sequences with 29 reference strains revealed nucleotide and amino acid sequence identities of 82.60-97.80% and 88.70-98.70%, respectively, indicating a higher mutation rate of the field CSFV strains. The phylogenetic analysis based on the complete E2 amino acid sequences also revealed a reliable differentiation of all the analyzed strains into specific genetic groups and subgroups, plus the local isolate (CSFV-E2) was found to cluster with the CSFV subgroup 2.2. Thus, the full-length E2 cds proved to be most suitable for a reliable and statistically significant phylogenetic analysis of CSFV isolates.