Characterization of recombinant phytase of Klebsiella sp. and the influence of novel 3-phytase on mineral solubility in broiler diets under an in vitro digestion assay.

Phytate (inositol hexaphosphate) is the major storage form of phosphorus (P) in nature, and phytases catalyze the hydrolysis of P from phytate and the formation of inositol phosphate isomers. In this study, a bacterium that produces phytase was isolated in a phytase screening medium. The bacterium was identified as Klebsiella sp. using phenotypic and molecular techniques. The PhyK phytase gene was successfully amplified from the genome, inserted into the pET-21a (+) vector, and expressed as a recombinant protein in E. Coli BL21. The efficiency of a laboratory phytase (Lab-Ph, PhyK phytase) was determined and compared with a commercial phytase (Com-Ph, Quantum Blue 40P phytase, AB Vista) under an in vitro digestion assay. The native signal peptide effectively facilitated the translocation of the protein to the periplasmic space of E. Coli BL21, resulting in the proper folding of the protein and the manifestation of desirable enzyme activity. The Lab-Ph displayed the temperature and pH optima at 50 °C and 5 respectively. In addition, the Lab-Ph was inactivated at 80 °C. Under an in vitro digestion assay condition, Lab-Ph improved the P solubility coefficient in broiler diets. In comparison, the Com-Ph significantly increased the P solubility coefficient even when compared with the Lab-Ph. In summary, this study has shown that Lab-Ph possesses the necessary biochemical properties to be used in various industrial applications. However, Lab-Ph is extremely sensitive to heat treatment. The Lab-Ph and Com-Ph under an in vitro digestion assay improved the solubility coefficient of P in the broiler diet.

Molting method alternative and detection of estrogen receptors by immunohistochemical methods on molted layers.

Three experiments were designed to evaluate the effects of different molting methods on the growth performance, detection of estrogen receptors (ERs), and immunohistochemical properties of some tissues in pre-, during, and post-molting of layers. In experiment 1, 302 Hy-line W-36 were reared from 75 to 76 weeks. In experiment 2, a total of 252 Hy-line W-36 were randomly allocated to 7 groups, with 6 replications and 6 birds in each. Hens fed in the E and D groups reached 30% of loss weight sooner (P < 0.05). Egg production was also sooner stopped in the FW group than in other ones (P < 0.05). In experiment 3, after inducing molting, each group in experiment 2 was divided into 2 groups with and without Humulus lupulus (Hop). The E group feed intake was increased, as compared to groups F and C (P < 0.05). A significant increase in egg weight was found by applying Hop and molting methods (P < 0.05). Days for return to the initial egg production and 10% egg laying were significantly decreased in the birds fed by Hop in the E and D groups (P < 0.05). The W-D, N-D, W-E, and N-E groups sooner returned to 50% egg laying, in comparison to other treatments (P < 0.05). Plasma estrogen and ERs were decreased by the molting programs, as compared with pre-molting; however, Hop increased their post-molting. After the molting period, egg production and ERs were increased significantly, as compared with the pre-molting period. To conclude, the white button mushroom residual, through decreasing ERs, could be used successfully for forced molting, and Hop could lead to a good performance by increasing ERs in the second laying cycle.

Approaches to determine the efficiency of novel 3-phytase from Klebsiella pneumoniae and commercial phytase in broilers from 1 to 14 d of age.

This study aimed to evaluate the effects of a laboratory 3-phytase (the expression of the phyK gene, Lab-Phy) and a commercial 6-phytase (Quantum Blue 40 P, Com-Phy) alone and in combination (corn-soy-based diets) in broilers. A total of 400, day-old Ross 308 male broilers were randomly assigned to 5 treatments with 10 replicate cages (8 chicks/cage) for a 14-day trial. Experimental treatments included the positive control (0.95% Ca and 0.48% nonphytate phosphorus (nPP), PC), negative control (0.90% Ca and 0.22% nPP, NC), and NC which was supplemented with Lab-Phy 250 FTU/kg and Com-Phy 250 FTU/kg alone or in combination of Lab-Phy 125 FTU/kg and Com-Phy 125 FTU/kg. The inclusion of Lab-Phy in the NC diet significantly improved the P and Ca content in the tibia compared to the NC group. Moreover, the inclusion of Com-Phy alone and in combination with Lab-Phy in the NC diet significantly increased the P and Ca content in the tibia compared to the Lab-Phy. The mRNA expression of NaPi-IIb was upregulated in the duodenum by the reduction of nPP and downregulated by the inclusion of any phytase, whereas other nutrient transporters were not influenced by the reduction of nPP or the addition of phytase in the small intestine mucosa. Broilers receiving the NC diet obtained the lowest body weight (BW) and body weight gain (BWG) at 8 to 14 and 1 to 14 d of age. The NC group showed the lowest villi height and surface area, Newcastle disease (ND) antibody titer, and digestibility of nutrients compared to the PC group at 14 d of age. Supplementing the NC diet with the Lab-Phy and Com-Phy individually, or in combination tended to improve BW, BWG, tibia characteristics, villi characteristics, ND, and retained CP and P, and apparent ileal digestibility of CP, P, methionine, and threonine. The present research indicated that the studied traits by the combination of phytases were slightly better than the average of the 2 individually, suggesting there might be some value in combining the laboratory and commercial phytases.

Effect of a phytogenic feed additive on performance, ovarian morphology, serum lipid parameters and egg sensory quality in laying hen.

This present study was conducted to evaluate the effects of dietary inclusion of 4, 8 and 12 g kg(-1) phytogenic feed additives mixture on performance, egg quality, ovary parameters, serum biochemical parameters and yolk trimethylamine level in laying hens. The results of experiment have shown that egg weight was increased by supplementation of 12 g kg(-1) feed additive whereas egg production, feed intake and feed conversion ratio (FCR) were not significantly affected. There were no significant differences in egg quality parameters by supplementation of phytogenic feed additive, whereas yolk trimethylamine level was decreased as the feed additive level increased. The sensory evaluation parameters did not differ significantly. No significant differences were found in serum cholesterol and triglyceride levels between the treatments but low- and high-density lipoprotein were significantly increased. Number of small follicles and ovary weight were significantly increased by supplementation of 12 g kg(-1) feed additive. Overall, dietary supplementation of polyherbal additive increased egg weigh, improved ovary characteristics and declined yolk trimethylamine level.