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Proteomic Analysis of Global Changes in Protein Expression During Exposure of Gamma Radiation in Bacillus sp. HKG 112 Isolated from Saline Soil

  • Gupta, Anil Kumar;Pathak, Rajiv;Singh, Bharat;Gautam, Hemlata;Kumar, Ram;Kumar, Raj;Arora, Rajesh;Gautam, Hemant K.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.6
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    • pp.574-581
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    • 2011
  • A Gram-positive bacterium was isolated from the saline soils of Jangpura (U.P.), India, and showed high-level of radiation-resistant property and survived upto 12.5 kGy dose of gamma radiation. The 16S rDNA sequence of this strain was examined, identified as Bacillus sp. strain HKG 112, and was submitted to the NCBI GenBank (Accession No. GQ925432). The mechanism of radiation resistance and gene level expression were examined by proteomic analysis of whole-cell extract. Two proteins, 38 kDa and 86.5 kDa excised from SDS-PAGE, which showed more significant changes after radiation exposure, were identified by MALDI-TOF as being flagellin and S-layer protein, respectively. Twenty selected 2-DE protein spots from the crude extracts of Bacillus sp. HKG 112, excised from 2- DE, were identified by liquid chromatography mass spectrometry (LC-MS) out of which 16 spots showed significant changes after radiation exposure and might be responsible for the radiation resistance property. Our results suggest that the different responses of some genes under radiation for the expression of radiation-dependent proteins could contribute to a physiological advantage and would be a significant initial step towards a fullsystem understanding of the radiation stress protection mechanisms of bacteria in different environments.

Effect of feeding of blend of essential oils on methane production, growth, and nutrient utilization in growing buffaloes

  • Yatoo, M.A.;Chaudhary, L.C.;Agarwal, N.;Chaturvedi, V.B.;Kamra, D.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.5
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    • pp.672-676
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    • 2018
  • Objective: An experiment was conducted to study the effect of a blend of essential oils (BEO) on enteric methane emission and growth performance of buffaloes (Bubalus bubalis). Methods: Twenty one growing male buffaloes (average body weight of $279{\pm}9.3kg$) were divided in to three groups. The animals of all the three groups were fed on a ration consisting of wheat straw and concentrate mixture targeting 500 g daily live weight gain. The three dietary groups were; Group 1, control without additive; Group 2 and 3, supplemented with BEO at 0.15 and 0.30 mL/kg of dry matter intake (DMI), respectively. Results: During six months feeding trial, the intake and digestibility of dry matter and nutrients (organic matter, crude protein, ether extract, neutral detergent fibre, and acid detergent fibre) were similar in all the groups. The average body weight gain was tended to improve (p = 0.084) in Group 2 and Group 3 as compared to control animals. Feeding of BEO did not affect feed conversion efficiency of the animals. The calves of all the three groups were in positive nitrogen balance with no difference in nitrogen metabolism. During respiration chamber studies the methane production (L/kg DMI and L/kg digestible dry matter intake was significantly (p<0.001) lower in Group 2 and Group 3 as compared to control animals. Conclusion: The results indicated that the BEO tested in the present study have shown potential to reduce enteric methane production without compromising the nutrient utilization and animal performance and could be further explored for its use as feed additive to mitigate enteric methane production in livestock.

The Effects of Dietary Turkish Propolis and Vitamin C on Performance, Digestibility, Egg Production and Egg Quality in Laying Hens under Different Environmental Temperatures

  • Seven, Pinar Tatli
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.8
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    • pp.1164-1170
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    • 2008
  • In this study, the effects of propolis and vitamin C (L-ascorbic acid) supplementation in diets were investigated on feed intake (FI), body weight (BW), body weight gain (BWG), feed conversion rate (FCR) and digestibility and on egg production and qualities (weight, mortality, shell thickness) in laying hens exposed to heat stress. A total of 150 Hyline White Leghorn, aged 42 weeks, hens was divided into five groups of 30 hens. Chicks were randomly divided into 1 positive control, 1 control and 3 treatment groups. The chicks were kept in cages in temperature-controlled rooms at $22^{\circ}C$ for 24 h/d (positive control, Thermoneutral, TN group) or $34^{\circ}C$ for 9 h/d from 08.00-17.00 h followed by $22^{\circ}C$ for 15 h (control, heat stress, HS group) and fed a basal diet or basal diet supplemented with vitamin C (250 mg/kg of L- ascorbic acid/kg of diet) or two levels of propolis (2 and 5 g of ethanol extracted propolis/kg of diet). Increased FI (p<0.05) and improvement in FCR (p<0.05), hen day egg (p<0.05) and egg weight (p<0.05) were found in Vitamin C and propolis-supplemented laying hens reared under heat stress conditions. Mortality rate was higher in the control group than TN, vitamin C and propolis groups (p<0.05). Digestibility of dry matter, organic matter, crude protein and ether extract improved with increasing of both dietary vitamin C and propolis (p<0.05). Vitamin C or propolis supplementation did not affect either the percentage shape index, yolk index or haugh unit and albumen index (p>0.05). However, the egg shell thickness and egg shell weight appeared to be increased in Vitamin C and propolis groups in comparison to HS group birds (p<0.05). In conclusion, dietary supplementation of laying hens with anti-oxidants (vitamin C and propolis) can attenuate heat stress-induced oxidative damage. These positive effects were evidenced by increased growth performance and digestibility, improvement of egg shell thickness and egg weight in comparison to non-supplemented birds. Moreover, supplementation with propolis (5 g/kg diet) was the most efficient treatment.

Effects of first feed intake time on growth performance, nutrient apparent metabolic rate and intestinal digestive enzyme activities in broilers

  • Wang, J.S.;Guo, T.Y.;Wang, Y.X.;Li, K.X.;Wang, Q.;Zhan, X.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.6
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    • pp.899-904
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    • 2018
  • Objective: This experiment studied the effects of first feed intake time post-hatch on growth performance, nutrient apparent metabolic rate and intestinal digestive enzyme activities in broilers. Methods: Two thousand five hundred and twenty LingNan Yellow broilers were randomly allotted to seven treatments with six replicates of 60 each. The only experimental factor was the first feed intake time which was 18, 24, 30, 36, 42, 48, and 54 hours after hatching. The whole experiment lasted for 21 days. Results: During the whole period, the 30 h treatment had the best body weight and average daily gain (p<0.05), followed by the 24 h group performance optimization. Also, the 30 h group was observed to have the best apparent metabolic rate for ether extract (p<0.05) and crude protein (p<0.05) and the highest activities of amylase, lipase and trypsin in small intestine. And the 24 h group was second only to the 30 h group in terms of the above two measures. Conclusion: These results indicated that the appropriate first feeding time of LingNan Yellow broilers was 24 to 30 hours after hatching.

Inhibitory Effect of Hizikia fusiformis Solvent-Partitioned Fractions on Invasion and MMP Activity of HT1080 Human Fibrosarcoma Cells

  • Lee, Seul-Gi;Karadeniz, Fatih;Oh, Jung Hwan;Yu, Ga Hyun;Kong, Chang-Suk
    • Preventive Nutrition and Food Science
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    • v.22 no.3
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    • pp.184-190
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    • 2017
  • Matrix metalloproteinases (MMPs) are endopeptidases that take significant roles in extracellular matrix degradation and therefore linked to several complications such as metastasis of cancer progression, oxidative stress, and hepatic fibrosis. Hizikia fusiformis, a brown algae, was reported to possess bioactivities, including but not limited to, antiviral, antimicrobial, and anti-inflammatory partly due to bioactive polysaccharide contents. In this study, the potential of H. fusiformis against cancer cell invasion was evaluated through the MMP inhibitory effect in HT1080 fibrosarcoma cells in vitro. H. fusiformis crude extract was fractionated with organic solvents, $H_2O$, n-BuOH, 85% aqueous MeOH, and n-hexane (n-Hex). The non-toxicity of the fractions was confirmed by MTT assay. All fractions inhibited the enzymatic activities of MMP-2 and MMP-9 according to the gelatin zymography assay. Cell migration was also significantly inhibited by the n-Hex fraction. In addition, both gene and protein expressions of MMP-2 and -9, and tissue inhibitor of MMPs (TIMPs) were evaluated by reverse transcription-polymerase chain reaction and Western blotting, respectively. The fractions suppressed the mRNA and protein levels of MMP-2, MMP-9 while elevating the TIMP-1 and TIMP-2, with the $H_2O$ fraction being the least effective while n-Hex fraction the most. Collectively, the n-Hex fraction from brown algae H. fusiformis could be a potential inhibitor of MMPs, suggesting the presence of various derivatives of polysaccharides in high amounts.

Antipathogenic Activity of Lactobacillus plantarum Isolated from Pickled Mulberry Leaf (뽕잎 장아찌로부터 분리된 Lactobacillus plantarum 균주의 유해균 증식 억제 활성)

  • Park, Eun-Hee;Kim, Myoung-Dong
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.163-170
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    • 2016
  • Strains of lactic acid bacteria were isolated from a variety of fermented foods collected in Korea. The strain L2167 showed a strong antipathogenic activity against Bacillus cereus, Listeria monocytogenes, Salmonella Typhimurium, Staphylococcus aureus, and Staphylococcus epidermidis. L2167 was identified as Lactobacillus plantarum by sequence analysis of its 16S rRNA gene. Scanning electron microscopy revealed rough and wrinkled morphology of B. cereus, L. monocytogenes, S. Typhimurium, S. aureus, and S. epidermidis cell membranes after treatment with a crude cell extract of L. plantarum L2167, indicating that Lactobacillus plantarum L2167 might destroy the cell membrane of pathogenic bacteria. The optimal temperature and initial medium pH for Lactobacillus plantarum L2167 growth were 35℃ and 5.5, respectively. Lactobacillus plantarum L2167 was more sensitive to NaCl than Lactobacillus plantarum KCTC21004, used as a control strain. Lactobacillus plantarum L2167 is expected to be developed as a prominent starter strain for efficient inhibition of growth of pathogens.

Effect of temperature on fractional precipitation for paclitaxel purification

  • Jeon, Soon-Yim;Kim, Jin-Hyun
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.355-359
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    • 2005
  • Fractional precipitation is a simple, efficient method for pre-purifying paclitaxel from plant cell cultures. The storage temperature of factional precipitation was optimized in terms of the yield and purity of paclitaxel with a fixed methanol concentration in water (61.5%, v/v), paclitaxel content in the crude extract (0.5%, w/v), and storage time (3 days). The greatest yield $({\sim}84%)$ was obtained with storage at a constant temperature $(0^{\circ}C)$ for 3 days. Conversely, the highest purity $({\sim}79%)$ was obtained with stepwise reduction in temperature over 3 days. For a constant storage temperature, the maximum purity $({\sim}67%)$ was obtained at $0^{\circ}C$. There was little difference in the yield of paclitaxel between -20 and $12^{\circ}C$. This pre-purification process serves to minimize solvent usage, and the size and complexity of the high-performance liquid chromatography (HPLC) operation required for paclitaxel purification.

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Purification of Bacillus sp. β-Mannanase and Separation of Xanthan Gum Hydrolysate by Chromatography Methods (Bacillus sp. 유래 β-Mannanase의 정제 및 Chromatography에 의한 Xanthan Gum 가수분해물의 분리)

  • 박귀근
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.4
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    • pp.562-566
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    • 2003
  • A $\beta$-mannanase of Bacillus sp. was purified by DEAE Sephacel ion exchange column chromatography. The specific activity of the purified enzyme was 17.41 units/mg protein, representing an 84.74-folds purification of the original crude extract. For the separation of two types of hydrolysates by the action of purified $\beta$-mannanase, carbon column chromatography, sephadex G-25 column chromatography and thin layer chromatography were accomplished. Main hydrolysates were D.P value 5 and 7 containing of low D.P values. By the method of FACE (Fluorophore Assisted Carbohydrate Electrophoresis), two types of hydrolysates were identified to homo type.

Effect of Tannin and Species Variation on In vitro Digestibility, Gas, and Methane Production of Tropical Browse Plants

  • Gemeda, Belete Shenkute;Hassen, A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.2
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    • pp.188-199
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    • 2015
  • Nineteen tanniferous browse plants were collected from South Africa to investigate their digestibility, gas production (GP) characteristics and methane production. Fresh samples were collected, dried in forced oven, and ground and analyzed for nutrient composition. In vitro GP and in vitro organic matter digestibility (IVOMD) were determined using rumen fluid collected, strained and anaerobically prepared. A semi-automated system was used to measure GP by incubating the sample in a shaking incubator at $39^{\circ}C$. There was significant (p<0.05) variation in chemical composition of studied browses. Crude protein (CP) content of the species ranged from 86.9 to 305.0 g/kg dry matter (DM). The neutral detergent fiber (NDF) ranged from 292.8 to 517.5 g/kg DM while acid detergent fiber (ADF) ranged from 273.3 to 495.1 g/kg DM. The ash, ether extract, non-fibrous carbohydrate, neutral detergent insoluble nitrogen, and acid detergent insoluble nitrogen and CP were negatively correlated with methane production. Methane production was positively correlated with NDF, ADF, cellulose and hemi-cellulose. Tannin decreased GP, IVOMD, total volatile fatty acid and methane production. The observed low methanogenic potential and substantial ammonia generation of some of the browses might be potentially useful as rumen manipulating agents. However, a systematic evaluation is needed to determine optimum levels of supplementation in a mixed diet in order to attain a maximal depressing effect on enteric $CH_4$ production with a minimal detrimental effect on rumen fermentation of poor quality roughage based diet.

Black gram (Vigna Mungo L.) foliage supplementation to crossbred cows: effects on feed intake, nutrient digestibility and milk production

  • Dey, Avijit;De, Partha Sarathi;Gangopadhyay, Prabir Kumar
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.2
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    • pp.187-191
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    • 2017
  • Objective: An experiment was conducted to examine the effect of dietary supplementation of dried and ground foliage of black gram (Vigna mungo L.) on feed intake and utilization, and production performance of crossbred lactating cows. Methods: Eighteen lactating crossbred (Bos taurus${\times}$Bos indicus) cows (body weight $330.93{\pm}10.82kg$) at their second and mid lactation (milk yield $6.77{\pm}0.54kg/d$) were randomly divided into three groups of six each in a completely randomized block design. Three supplements were formulated by quantitatively replacing 0, 50, and 100 per cent of dietary wheat bran of concentrate mixture with dried and ground foliage of black gram. The designated supplement was fed to each group with basal diet of rice straw (ad libitum) to meet the requirements for maintenance and milk production. Daily feed intake and milk yield was recorded. A digestion trial was conducted to determine the total tract digestibility of various nutrients. Results: The daily feed intake was increased (p<0.05) with the supplementation of black gram foliage. Although the digestibility of dry matter, organic matter, crude protein, and ether extract did not vary (p>0.05), the fibre digestibility was increased (p<0.05), which ultimately improved (p<0.05) the total digestible nutrients content of composite diet. Although, the average milk yield (kg/animal/d) and composition did not differ (p>0.05) among the groups, milk yield was increased by 10 per cent with total replacement of wheat bran in concentrate mixture with of black gram foliage. The economics of milk production calculated as feed cost per kg milk yield (INR 10.61 vs 7.98) was reduced by complete replacement of wheat bran with black gram foliage. Conclusion: Black gram foliage could be used as complete replacement for wheat bran in concentrate mixture of dairy cows in formulating least cost ration for economic milk production in small holders' animal production.