An improved method for a rapid determination of phytase activity in animal feed.
ABSTRACT The current direct colorimetric assay for phytase activity in feeds has interference from high P background and other factors. Our objective was to develop a rapid and reliable spin column method to accurately determine phytase activity in feed ingredients or complete diets. After the feed sample was extracted by stirring in 0.2 M citrate buffer, pH 5.5, for 30 min at room temperature, the oily layer of the supernatant fraction was removed by passing through an acrodisc syringe filter (0.45-microm HT Tuffryn membrane, Gelman Laboratory, Ann Arbor, MI). The filtrate was then loaded onto a spin column (MW cutoff 30,000, Millipore, Bedford, MA) to remove free phosphate before the phytase activity assay. Compared with the direct assay, this new procedure improved both accuracy and reproducibility. When diets contained phytase at 0 to 1,500 U/kg (as fed), the CV for multiple assays of the same samples (n = 6) by the new method ranged from 1 to 6% compared with 28 to 39% by the direct method. A linear relationship was found between the added phytase activity in practical diets and the analyzed activity by the new method (r2 = 0.99; P < 0.01). In conclusion, the spin column method is an improved assay for phytase activity in animal feed, and may be used for quality control of phytase supplementation.
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ABSTRACT: Inter-simple sequence repeats (ISSR) were used to study the genetic diversity of Haplocladium microphyllum in China. Of the 100 primers, 11 produced clear and reproducible ISSR bands, and the di-nucleotide accounted for 72.7% of those primers. The primer UBC840 generated the largest number of polymorphic bands (12), followed by UBC841, UBC881, which amplified 11 and 10 polymorphic bands, respectively. Using the 11 primers, 85 polymorphic bands out of a total of 110 were generated from 28 individuals of 4 populations sampled from Zhejiang and Jiangsu provinces of China. Nei's gene diversity (He) and Shannon index (I) revealed a high level of genetic diversity with He = 0.2331 and I = 0.3581. The coefficient of gene differentiation (Gst) and gene flow (Nm) were 0.38 and 0.8157, respectively. The UPGMA tree illustrated that populations from QZ and JH were closely related, while the population from NJ was found to be the most diverse. Our results also provided an optimized method for evaluating genetic diversity of H. microphyllum using ISSR markers which was useful for further investigation.Biochemical Systematics and Ecology 08/2014; 55:107–111. · 1.17 Impact Factor
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ABSTRACT: Phytase producing thirteen microbial strains were identified in poultry soil sample collected at Sathyamangalam, Erode district, Tamilnadu. Among thirteen isolates, two of them were the best of phytases enzyme producer and identified as Aspergillus fumigatus and Aspergillus terreus. Under solid substrate culture condition Aspergillus fumigatus was screened for phytase production on agricultural by-products. The results showed that wheat bran with sugarcane molasses was the best substrate for production of phytase. Phytase purified by non chromatography method, the purified enzyme has an apparent molecular weight 64kDa. Keywords: Phytase, Aspergillus fumigatus and Aspergillus terreus. INTRODUCTION Phytase is widespread in nature, occurring in plants, microorganisms, as well as in some animals. Microbial Phytase activity was most frequently detected in fungi. Phytic acid was discovered as early as in 1872 by Pfeffer and a first note on Phytase can be found in the literature in 1907. Phytase belongs to the group of phosphoric monoester hydrolases; it catalyzes the hydrolysis of myo-inositol hexakisphosphate to inorganic monophosphate and lower phosphoric esters of myo-inositol, or in some cases to free myo-inositol (Vats and Banerjee, 2004 and Howson and Davies, 2005). Shieh and Ware (1968) isolated 30 microorganisms from soil for extracellular Phytase activity. All Phytase producers were filamentous fungi, 28 of them belonged to the genus Aspergillus, one species belonging to Penicillium and on to Mucor. Most of the isolates produced only intracellular Phytase (Ghada et al., 2011). Similarly A. niger strains was best producers of extracellular Phytase while bacterial cultures produced only intracellular enzyme confirmed by (Volfov et al., 1994 and Sabu et al., 2005). Phytase production by yeasts was also studied. Saccharomyces cerevisiaewas found to produce Phytase (Nayini and Markakis, 2008) and further Candida tropicalis, Torulopsis candida, Debaryomycescastelii, Kluyveromycesfragilis and Schwanniomycescastellii were able to hydrolyze Phytate (Lambrechts et al., 1993). S. castellii, which showed the highest extracellular Phytase activity, was chosen for the further study (Lambrechts et al., 1993). There are two main areas where Phytase can be used. The first one is Phytate elimination in feed and food industries, and the second one a preparation of special myo-inositol phosphates as tools for biochemical investigation. The main reason for removal of Phytate from food and feed is its indigestibility for monogastric animals including man (Simell et al., 1989 and Traylor et al., 2001). Undigested Phytate together with part of additionalGlobal J. of Mod. Biol. & Tech. 01/2012;
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ABSTRACT: This study was conducted to investigate the effects of phytase in diets containing cottonseed and soybean meal (CS) on growth performance, feed utilization and digestibility of protein and phosphorus in juvenile olive flounder (initial body weight 2.5 g), Paralichthys olivaceus. Four experimental diets replacing 0%, 30%, 30% and 40% fish meal protein with CS in equal proportion were formulated to be isonitrogenous and isocaloric (designated as CS0, CS30, CS30+P, CS40+P, respectively). Phytase of 1,000 FTU/kg was supplemented in diets CS30+P and CS40+P. Three groups of fish (25 fish per group) were fed one of the experimental diets for 10 weeks. No significant differences were observed in growth performance of fish groups except for the CS40+P diet. Apparent digestibility coefficients of protein and phosphorus in fish fed phytase-containing diets were significantly higher than those of fish fed the CS0 diet. Serum cholesterol was significantly reduced in fish fed the CS-containing diets. Antioxidant activities in the diets and liver of fish were significantly increased with the increment of dietary CS. Gossypol was only detected and found in liver of the fish fed the CS-containing diets. The findings suggest that supplementation of microbial phytase could improve the apparent digestibility of protein and phosphorus in juvenile olive flounder fed the CS-containing diets.Asian Australasian Journal of Animal Sciences 09/2008; 21(9). · 0.56 Impact Factor