Crocin (bioactive compound of Crocus sativus L.) potently restores REM sleep deprivation-induced manic- and obsessive-compulsive-like behaviors in female rats.

Rapid-eye movement (REM) sleep deprivation (SD) can induce manic-like behaviors including hyperlocomotion. On the other hand, crocin (one of the main compounds of Crocus sativus L. or Saffron) may be beneficial in the improvement of mental and cognitive dysfunctions. Also, crocin can restore the deleterious effects of SD on mental and cognitive processes. In this study, we investigated the effect of REM SD on female rats' behaviors including depression- and anxiety-like behaviors, locomotion, pain perception, and obsessive-compulsive-like behavior, and also, the potential effect of crocin on REM SD effects. We used female rats because evidence on the role of REM SD in modulating psychological and behavioral functions of female (but not male) rats is limited. REM SD was induced for 14 days (6h/day), and crocin (25, 50, and 75 mg/kg) was injected intraperitoneally. Open field test, forced swim test, hot plate test, and marble burying test were used to assess rats' behaviors. The results showed REM SD-induced manic-like behavior (hyperlocomotion). Also, REM SD rats showed decreased anxiety- and depression-like behavior, pain subthreshold (the duration it takes for the rat to feel pain), and showed obsessive compulsive-like behavior. However, crocin at all doses partially or fully reversed REM SD-induced behavioral changes. In conclusion, our results suggested the possible comorbidity of OCD and REM SD-induced manic-like behavior in female rats or the potential role of REM SD in the etiology of OCD, although more studies are needed. In contrast, crocin can be a possible therapeutic choice for decreasing manic-like behaviors.

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.

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.