Cloning and Expression of D-Threonine Aldolase from Ensifer arboris NBRC100383

D-erythro-cyclohexylserine (D chiral unnatural β-hydroxy amino acid expected for the synthesis of drug for AIDS treatment. To develop a continuous bioconversion system with whole cell biocatalyst of D-threonine aldolase (D genes for the D-erythro-CHS production, D-threonine aldolase gene was amplified from Ensifer arboris 100383 by direct PCR amplication using two degenerated oligonucleotide primers designed based on genomic sequence of Shinorhizobium meliloti Sequence analysis of the cloned DNA fragment revealed one open-reading frame of 1059 bp and 386 amino acids. This putative D-TA gene was cloned into NdeI and EcoRI (pEnsi His-tag sequence or BamHI (pEnsi-DTA[2]) sequence of the pET21(a) vector. The expression level of the cloned gene was extremely overexpressed by E. coli BL21(DE3) transformed with pEnsi-DTA[1] compared to E. coli BL21(DE3) transformed with pEnsi-DTA[2]. When the cells expressing the wild used for D-TA enzyme activity, 12 mM glycine was successfully detected in HPLC analysis. Moreover, the whole cells harbouring the recombinant D-TA was able to synthesize D-erythro of 0.6 mg/ml in a batch reaction.

Effect of Pectinase on the Physico-Chemical Properties of Juice from Pawpaw (Carica papaya) Fruits

A procedure for the preparation of clarified Pawpaw Juice was developed. About 750ml Pawpaw pulp was measured into 2 measuring cylinders A & B of capacity 1 litre heated to 400C, cooled to 200C. 30mls pectinase was added into cylinder A, while 30mls distilled water was added into cylinder B. Enzyme treated sample (A) was allowed to digest for 5hours after which it was heated to 900C for 15 minutes to inactivate the enzyme. The heated sample was cooled and with the aid of a mucillin cloth the pulp was filtered to obtain the clarified pawpaw juice. The juice was filled into 100ml plastic bottles, pasteurized at 950C for 45 minutes, cooled and stored at room temperature. The sample treated with 30mls distilled water also underwent the same process. Freshly pasteurized sample was analyzed for specific gravity, titratable acidity, pH, sugars and ascorbic acid. The remaining sample was then stored for 2 weeks and the above analyses repeated. There were differences in the results of the freshly pasteurized samples and stored sample in pH and ascorbic acid levels, also sample treated with pectinase yielded higher volumes of juice than that treated with distilled water.

Virulent-GO: Prediction of Virulent Proteins in Bacterial Pathogens Utilizing Gene Ontology Terms

Prediction of bacterial virulent protein sequences can give assistance to identification and characterization of novel virulence-associated factors and discover drug/vaccine targets against proteins indispensable to pathogenicity. Gene Ontology (GO) annotation which describes functions of genes and gene products as a controlled vocabulary of terms has been shown effectively for a variety of tasks such as gene expression study, GO annotation prediction, protein subcellular localization, etc. In this study, we propose a sequence-based method Virulent-GO by mining informative GO terms as features for predicting bacterial virulent proteins. Each protein in the datasets used by the existing method VirulentPred is annotated by using BLAST to obtain its homologies with known accession numbers for retrieving GO terms. After investigating various popular classifiers using the same five-fold cross-validation scheme, Virulent-GO using the single kind of GO term features with an accuracy of 82.5% is slightly better than VirulentPred with 81.8% using five kinds of sequence-based features. For the evaluation of independent test, Virulent-GO also yields better results (82.0%) than VirulentPred (80.7%). When evaluating single kind of feature with SVM, the GO term feature performs much well, compared with each of the five kinds of features.

Cyanide and Heavy Metal Concentration of Fermented Cassava Flour (Lafun) Available in the Markets of Ogun and Oyo States of Nigeria

Fermented cassava flours (lafun) sold in Ogun and Oyo States of Nigeria were collected from 10 markets for a period of two months and analysed to determine their safety status. The presence of trace metals was due to high vehicular movement around the drying sites and markets. Cyanide and moisture contents of samples were also determined to assess the adequacy of fermentation and drying. The result showed that sample OWO was found to have the highest amount of 16.02±0.12mg/kg cyanide while the lowest was found in sample OJO with 10.51±0.10mg/kg. The results also indicated that sample TVE had the highest moisture content of 18.50±0.20% while sample OWO had the lowest amount of 12.46±0.47%. Copper and lead levels were found to be highest in TVE with values 28.10mg/kg and 1.1mg/kg respectively, while sample BTS had the lowest values of 20.6mg/kg and 0.05mg/kg respectively. High value of cyanide indicated inadequate fermentation.

Evolutionary Distance in the Yeast Genome

Whole genome duplication (WGD) increased the number of yeast Saccharomyces cerevisiae chromosomes from 8 to 16. In spite of retention the number of chromosomes in the genome of this organism after WGD to date, chromosomal rearrangement events have caused an evolutionary distance between current genome and its ancestor. Studies under evolutionary-based approaches on eukaryotic genomes have shown that the rearrangement distance is an approximable problem. In the case of S. cerevisiae, we describe that rearrangement distance is accessible by using dedoubled adjacency graph drawn for 55 large paired chromosomal regions originated from WGD. Then, we provide a program extracted from a C program database to draw a dedoubled genome adjacency graph for S. cerevisiae. From a bioinformatical perspective, using the duplicated blocks of current genome in S. cerevisiae, we infer that genomic organization of eukaryotes has the potential to provide valuable detailed information about their ancestrygenome.

Improvement of Milk Production with Half Day Milking; a Case Study of Communal Goat Housing in Sukorejo, Yogyakarta, Indonesia

The case study was conducted to show the effect of  milking method in goat called half day milking on the milk production and the growth of kids. Data were collected by  interviewing farmers and investigating goat production in the  communal goat housing from June 2008 to May 2009. The interview  was conducted to collect data about goat management. The  observations were conducted on 10 goats, which were selected based  on the uniformity of age, number of kid born/goat and the milking method in practice. The samples were divided into two groups; those  were full 3 months nursing and half day milked goats (in this group the kids were separated from goat during the previous night milking  and then the kids were allowed to suck the goat during the day). The result showed that the communal goat housing had 138 goats and 25% of the farmers milked the goat. The implementation of half day milking increased the milk production significantly (P

Coastal Resource Management: Fishermen-s Perceptions of Seaweed Farming in Indonesia

Seaweed farming is emerging as a viable alternative activity in the Indonesian fisheries sector. This paper aims to investigate people-s perceptions of seaweed farming, to analyze its social and economic impacts and to identify the problems and obstacles hindering its continued development. Structured and semi-structured questionnaires were prepared to obtain qualitative data, and interviews were conducted with fishermen who also plant seaweed. The findings showed that fishermen in the Laikang Bay were enthusiastic about cultivating seaweeds and that seaweed plays a major role in supporting the household economy of fishermen. However, current seaweed drying technologies cannot support increased seaweed production on a farm or plot, especially in the rainy season. Additionally, variable monsoon seasons and long marketing channels are still major constraints on the development of the industry. Finally, capture fisheries, the primary economic livelihood of fishermen of older generations, is being slowly replaced by seaweed farming.

The Appropriate Time Required for Newborn Calf Camel to Get Optimal Amount of Colostrums Immunoglobulin (IgG) with Relation to Levels of Cortisol and Thyroxin

A major challenge in camel productivity is the high mortality rate of camel calves in the early stage due to the lack of colostrums. This study investigates the time required for the calves to obtain the optimum amount of the immunoglobulin (IgG). Eleven pregnant female camels (Camelus Dromedarus) were selected randomly and variant in age and gestation. After delivery, 7 calves were obtained and used for this investigation. Colostrum samples were collected from mothers immediately after parturition. Blood samples were obtained from the calves as follow: 0 day (before suckling), 24, 48, 72, 96, 120 and 144 hours, 2nd, 3rd, and 4th weeks post suckling. Blood serum and colostrums whey were separated and used to determine IgG concentration, total protein and concentration of Cortisol and Thyroxin. The results showed high levels of IgG in camel colostrums (328.8 ± 4.5 mg / ml). The IgG concentration in serum of calves was the highest within 1st 24 h after suckling (140.75 mg /ml), and then declined gradually reached lower level at 144 h (41.97 mg / ml). The average turnover rate (t 1/2) of serum IgG in the all cases was 3.22 days. The turnover of ranged from 2.56 days for calves have values of IgG more than average and 7.7 days for those with values below average. In spite of very high levels of thyroxin in sera of new born the results showed no correlation between cortisol and thyroxin with IgG levels.

Biological Characterization of the New Invasive Brine Shrimp Artemia franciscana in Tunisia: Sabkhet Halk El-Menzel

Endemic Artemia franciscana populations can be found throughout the American continent and also as an introduced specie in several country all over the world, such as in the Mediterranean region where Artemia franciscana was identified as an invasive specie replacing native Artemia parthenogenetica and Artemia salina. In the present study, the characterization of the new invasive Artemia franciscana reported from Sabkhet Halk El-Menzel (Tunisia) was done based on the cysts biometry, nauplii instar-I length, Adult sexual dimorphism and fatty acid profile. The mean value of the diameter of non-decapsulated and decapsulated cysts, chorion thickness and naupliar length is 235.8, 226.3, 4.75 and 426.8 μm, respectively. Sexual dimorphism for adults specimen showed that maximal distance between compound eyes, diameter for compound eyes, length of first antenna and the abdomen length compared to the total body length ratio, are the most important variables for males and females discrimination with a total contribution of 62.39 %. The analysis of fatty acid methyl esters profile of decapsulated cysts resulted in low levels of linolenic acid (LLA, C18:3n-3) and high levels of eicosapentaenoic acid (EPA, C20:5n-3) with 3.11 and 11.10 %, respectively. Low quantity of docosahexaenoic acid (DHA, 22:6n-3) was also observed with 0.17 mg.g-1 dry weight.

Experimental and Theoretical Investigation of Rough Rice Drying in Infrared-assisted Hot Air Dryer Using Artificial Neural Network

Drying characteristics of rough rice (variety of lenjan) with an initial moisture content of 25% dry basis (db) was studied in a hot air dryer assisted by infrared heating. Three arrival air temperatures (30, 40 and 500C) and four infrared radiation intensities (0, 0.2 , 0.4 and 0.6 W/cm2) and three arrival air speeds (0.1, 0.15 and 0.2 m.s-1) were studied. Bending strength of brown rice kernel, percentage of cracked kernels and time of drying were measured and evaluated. The results showed that increasing the drying arrival air temperature and radiation intensity of infrared resulted decrease in drying time. High bending strength and low percentage of cracked kernel was obtained when paddy was dried by hot air assisted infrared dryer. Between this factors and their interactive effect were a significant difference (p

Spatial Variability of Some Soil Properties in Mountain Rangelands of Northern Iran

In this paper spatial variability of some chemical and physical soil properties were investigated in mountain rangelands of Nesho, Mazandaran province, Iran. 110 soil samples from 0-30 cm depth were taken with systematic method on grid 30×30 m2 in regions with different vegetation cover and transported to laboratory. Then soil chemical and physical parameters including Acidity (pH), Electrical conductivity, Caco3, Bulk density, Particle density, total phosphorus, total Nitrogen, available potassium, Organic matter, Saturation moisture, Soil texture (percentage of sand, silt and clay), Sodium, Calcium, magnesium were measured in laboratory. Data normalization was performed then was done statistical analysis for description of soil properties and geostatistical analysis for indication spatial correlation between these properties and were perpetrated maps of spatial distribution of soil properties using Kriging method. Results indicated that in the study area Saturation moisture and percentage of Sand had highest and lowest spatial correlation respectively.

Extraction and Characterisation of Protein Fraction from Date Palm Fruit Seeds

Date palm (Phoenix dactylifera L.) seeds are waste streams which are considered a major problem to the food industry. They contain potentially useful protein (10-15% of the whole date-s weight). Global production, industrialisation and utilisation of dates are increasing steadily. The worldwide production of date palm fruit has increased from 1.8 million tons in 1961 to 6.9 million tons in 2005, thus from the global production of dates are almost 800.000 tonnes of date palm seeds are not currently used [1]. The current study was carried out to convert the date palm seeds into useful protein powder. Compositional analysis showed that the seeds were rich in protein and fat 5.64 and 8.14% respectively. We used several laboratory scale methods to extract proteins from seed to produce a high protein powder. These methods included simple acid or alkali extraction, with or without ultrafiltration and phenol trichloroacetic acid with acetone precipitation (Ph/TCA method). The highest protein content powder (68%) was obtained by Ph/TCA method with yield of material (44%) whereas; the use of just alkali extraction gave the lowest protein content of 8%, and a yield of 32%.

Investigation of Anti-Inflammatory, Antipyretic and Analgesic Effect of Yemeni Sidr Honey

Traditionally, Yemini Sidr honey has been reported to cure liver problems, stomach ulcers, and respiratory disorders. In this experiment, we evaluated Yemeni Sidr honey for its ability to protect inflammations caused by acetic acid and formalin -induced writhing, carrageenan and histamine-induced paw oedema in experimental rat model. Hyperpyrexia, membrane stabilizing activity, and phytochemical screening of the honey was also examined. Yemini Sidr Honey at (100, 200 and 500 mg/kg) exhibited a concentration dependant inhibition of acetic acid induced and formalin induced writhing, paw oedema induced by carrageenan & histamine, and hyperpyrexia induced by brewer's yeast, it also inhibited membrane stabilizing activity. Phytochemical screenings of the honey reveal the presence of flavonoids, steroid, alkaloids, saponins and tannins. This study suggested that Yemeni Sidr honey possess very strong antiinflammatory, analgesic and antipyretic effects and these effects would be a result of the phytochemicals present.

Prevention of Biofilm Formation in Urinary Catheter by Coating Enzymes/ Gentamycin/ EDTA

Urinary Tract Infections (UTI) account for an estimated 25-40% nosocomial infection, out of which 90% are associated with urinary catheter, called Catheter associated urinary tract infection (CAUTI). The microbial populations within CAUTI frequently develop as biofilms. In the present study, microbial contamination of indwelling urinary catheters was investigated. Biofilm forming ability of the isolates was determined by tissue culture plate method. Prevention of biofilm formation in the urinary catheter by Pseudomonas aeruginosa was also determined by coating the catheter with some enzymes, gentamycin and EDTA. It was found that 64% of the urinary catheters get contaminated during the course of catheterization. Of the total 6 isolates, biofilm formation was seen in 100% Pseudomonas aeruginosa and E. coli, 90% in Enterococci, 80% in Klebsiella and 66% in S. aureus. It was noted that the biofilm production by Pseudomonas was prolonged by 7 days in amylase, 8 days in protease, 6 days in lysozyme, 7days in gentamycin and 5 days in EDTA treated catheter.

Proximate and Mineral Composition of Chicken Giblets from Vojvodina (Northern Serbia)

Proximate (moisture, protein, total fat, total ash) and mineral (K, P, Na, Mg, Ca, Zn, Fe, Cu and Mn) composition of chicken giblets (heart, liver and gizzard) were investigated. Phosphorous content, as well as proximate composition, were determined according to recommended ISO methods. The content of all elements, except phosphorus, of the giblets tissues were determined using inductively coupled plasma-optical emission spectrometry (ICP-OES), after dry ashing mineralization. Regarding proximate composition heart was the highest in total fat content, and the lowest in protein content. Liver was the highest in protein and total ash content, while gizzard was the highest in moisture and the lowest in total fat content. Regarding mineral composition liver was the highest for K, P, Ca, Mg, Fe, Zn, Cu, and Mn, while heart was the highest for Na content. The contents of almost all investigated minerals in analysed giblets tissues of chickens from Vojvodina were similar to values reported in the literature, i.e. in national food composition databases of other countries.

Utilization Juice Wastes as Corn Replacement in the Broiler Diet

An experiment was conducted with 80 unsexed broilers of the Arbor Acress strain to determine the capability of a carrot and fruit juice wastes mixture (carrot, apple, manggo, avocado, orange, melon and Dutch egg plant) in the same proportion for replacing corn in broiler diet. This study involved a completely randomized design (CRD) with 5 treatments (0, 5, 10, 15, and 20% of juice wastes mixture in diets) and 4 replicates per treatment. Diets were isonitrogenous (22% crude protein) and isocaloric (3000 kcal/kg diet). Measured variables were feed consumption, average daily gain, feed conversion, as well as percentages of abdominal fat pad, carcass, digestive organs (liver, pancreas and gizzard), and heart. Data were analyzed by analysis of variance for CRD. Increasing juice wastes mixture levels in diets increased feed consumption (P

Concentration of Micro Minerals in Fiber Fraction of Forages

This study was carried out to evaluate concentration of micro minerals (Zn, Fe, Mn, Cu and Se) of forages and their distribution in fiber fraction (neutral detergent fiber/NDF and acid detergent fiber/ADF) in South Sumatra during dry and rainy seasons. Seven species of commonly forages namely Axonopus compressus, Panicum maximum, Pennisetum purpuphoides, Leucaena leucocephala, Centrocema pubescens, Calopogonium mucunoides and Acacia mangium were collected at native pasture during rainy and dry seasons. The results showed that micro minerals concentration of forages and their distribution in fiber fraction varied among species and season. In general, concentration of micro minerals was slightly higher in rainy season compared to dry season either in grass or legumes forages. In grass, concentration of Fe and Mn were above the critical level, while 33.3 %, 100 % and 16.7 % of evaluated grass were deficient in Zn, Cu and Se. Data on legume forages show that 75 % of legumes were deficient in Zn and Mn, 62.5 % deficient in Cu and 50 % deficient in Se. There was no species of legume deficient in Fe. Distribution of micro minerals in NDF and ADF were also significantly affected by species and season and depends on the kinds of element measured. Generally, micro minerals were associated in fiber fractions much higher during dry season compared to rainy season. Iron (Fe) and selenium (Se) in forages were the highest elements associated in NDF and ADF, while the lowest was found in Copper (Cu).

Investigation of Phytoextraction Coefficient Different Combination of Heavy Metals in Barley and Alfalfa

Two seperate experiments by barley and alfalfa were conducted to a 2×8 factorial completely randomised design, with four replicates. Factors were inoculation (M) with Gomus mosseae or uninoculation (M0) and seven levels of contaminants (Co, Cd, Pb and combinations) plus an uncontaminated control treatment (C). Heavy metals in plant tissues and soil were quantified by Inductively Coupled Plasma Optical Emission Spectrometer (ICP-OES) (Variant- Liberty 150AX Turbo). Phytoextraction coefficient of contaminants calculated by concentration of heavy metals in the shoot (mgkg-1) / concentration of heavy metals in soil (mgkg-1). In the barley, the highest rate of phytoextraction coefficient of Pb, Cd and Co was in M0Pb, M0PbCoCd and MCo, respectively (P

Antioxidant Responses to Different Exposure Regimes of Kazakhstan Light Crude Oil in Livers of Male Albino Rats

Biochemical investigations were carried out to assess the effect of different exposure regimes of Kazakhstan crude oil (KCO) on hepatic antioxidant defense system in albino rats. Contaminants were delivered under two different dosing regimes, with all treatments receiving the same total contaminant load by the end of the exposure period. Rats in regime A injected with KCO once at a dose of 6 ml/kg bw while in regime B injected multiply at a dose of 1.5 ml/kg bw on day 1, 3, 5 and 8. Antioxidant biomarkers were measured in hepatic tissue after 1, 3, 5 and 8 days. Significant induction was observed in serum aminotransferases (ALT, AST) (p

A New Method for Rapid DNA Extraction from Artemia (Branchiopoda, Crustacea)

Artemia is one of the most conspicuous invertebrates associated with aquaculture. It can be considered as a model organism, offering numerous advantages for comprehensive and multidisciplinary studies using morphologic or molecular methods. Since DNA extraction is an important step of any molecular experiment, a new and a rapid method of DNA extraction from adult Artemia was described in this study. Besides, the efficiency of this technique was compared with two widely used alternative techniques, namely Chelex® 100 resin and SDS-chloroform methods. Data analysis revealed that the new method is the easiest and the most cost effective method among the other methods which allows a quick and efficient extraction of DNA from the adult animal.