Purification of an alpha amylase from Aspergillus flavus NSH9 and molecular characterization of its nucleotide gene sequence

In this study, an alpha-amylase enzyme from a locally isolated Aspergillus flavus NSH9 was purified and characterized. The extracellular α-amylase was purified by ammonium sulfate precipitation and anion-exchange chromatography at a final yield of 2.55-fold and recovery of 11.73%. The molecular mass of the purified α-amylase was estimated to be 54 kDa using SDS-PAGE and the enzyme exhibited optimal catalytic activity at pH 5.0 and temperature of 50 °C. The enzyme was also thermally stable at 50 °C, with 87% residual activity after 60 min. As a metalloenzymes containing calcium, the purified α-amylase showed significantly increased enzyme activity in the presence of Ca2+ ions. Further gene isolation and characterization shows that the α-amylase gene of A. flavus NSH9 contained eight introns and an open reading frame that encodes for 499 amino acids with the first 21 amino acids presumed to be a signal peptide. Analysis of the deduced peptide sequence showed the presence of three conserved catalytic residues of α-amylase, two Ca2+-binding sites, seven conserved peptide sequences, and several other properties that indicates the protein belongs to glycosyl hydrolase family 13 capable of acting on α-1,4-bonds only. Based on sequence similarity, the deduced peptide sequence of A. flavus NSH9 α-amylase was also found to carry two potential surface/secondary-binding site (SBS) residues (Trp 237 and Tyr 409) that might be playing crucial roles in both the enzyme activity and also the binding of starch granules. © 2018, Springer-Verlag GmbH Germany, part of Springer Nature

Trace Mineral Sources and Rosemary Oil in the Diet of Brown Laying Hens: Egg Quality and Lipid Stability

ABSTRACT Two experiments were carried out to evaluate the effects of rosemary oil (RO) and trace mineral sources (MS) on the internal quality and lipid stability of brown layer eggs. The treatments consisted of diets supplemented with two trace mineral sources (inorganic or organic) and three levels (0, 100, or 200 mg kg-1) of rosemary oil (RO), and three egg storage times. Eggs were stored at a controlled temperature (CT; 25.0ºC) in Experiment I and under refrigeration (RT; 5.0ºC) in Experiment II. The following parameters were analyzed on days 0 (fresh), 15 and 30 of storage: malonaldehyde level (MDA), egg weight (EW), Haugh unit (HU), yolk index (YI), albumen and yolk pH, raw yolk color (RYC), and egg weight loss. Data were analyzed according to completely randomized design in a 2x3x3 factorial arrangement (MS x RO x storage time). In Experiment I, there was an interaction between treatments for EW, HU and ALBp. Dietary OTM inclusion improved the results for all analyzed variables. The addition of 200 mg kg-1 RO reduced MDA and increased HU, YI and RYC. In experiment II, 200 mg kg-1 of RO in the diet improved HU. The internal quality of eggs stored both at CT and under RT is adversely affected by increasing storage periods, but this effect can be minimized by the dietary supplementation of OTM and 200 mg kg-1 rosemary oil. The lipid stability of eggs stored at CT improves with the supplementation of OTM and 200 mg kg-1 rosemary oil, but not of eggs stored under refrigeration

Variation in the Branching Pattern of the Superficial Palmar Arch

In this article we describe a rare variation in the superficial palmar arch (SPA) encountered during routine cadaveric dissection. SPA was formed by the superficial branches of ulnar and radial arteries which provided a proper digital branch to the ulnar side of the little finger and three common palmar digital branches to the medial four digits. Additionally a first common digital artery was seen to emerge from the radial side of SPA which further divided into the arteria princeps pollicis and arteria radialis indicis. We also found a small communicating branch between the arteria princeps pollicis and the radial artery in the anatomical snuff box. Such arterial variations in the palm due to alteration in the developmental sequence remains a crucial issue in the reconstructive hand surgeries, especially while dealing with the innovative microsurgical procedures, where these varied patterns act as pivotal points around which successful results of various advanced surgical procedures revolve

Trace Mineral Sources and Rosemary Oil in the Diet of Brown Laying Hens: Egg Quality and Lipid Stability

<div><p>ABSTRACT Two experiments were carried out to evaluate the effects of rosemary oil (RO) and trace mineral sources (MS) on the internal quality and lipid stability of brown layer eggs. The treatments consisted of diets supplemented with two trace mineral sources (inorganic or organic) and three levels (0, 100, or 200 mg kg-1) of rosemary oil (RO), and three egg storage times. Eggs were stored at a controlled temperature (CT; 25.0ºC) in Experiment I and under refrigeration (RT; 5.0ºC) in Experiment II. The following parameters were analyzed on days 0 (fresh), 15 and 30 of storage: malonaldehyde level (MDA), egg weight (EW), Haugh unit (HU), yolk index (YI), albumen and yolk pH, raw yolk color (RYC), and egg weight loss. Data were analyzed according to completely randomized design in a 2x3x3 factorial arrangement (MS x RO x storage time). In Experiment I, there was an interaction between treatments for EW, HU and ALBp. Dietary OTM inclusion improved the results for all analyzed variables. The addition of 200 mg kg-1 RO reduced MDA and increased HU, YI and RYC. In experiment II, 200 mg kg-1 of RO in the diet improved HU. The internal quality of eggs stored both at CT and under RT is adversely affected by increasing storage periods, but this effect can be minimized by the dietary supplementation of OTM and 200 mg kg-1 rosemary oil. The lipid stability of eggs stored at CT improves with the supplementation of OTM and 200 mg kg-1 rosemary oil, but not of eggs stored under refrigeration.</p></div

Performance and morphometry of the intestinal mucosa of laying hens fed diets containing xylanase

The objective of this study was to evaluate the effect of dietary energy level reduction and xylanase inclusion on the performance and on intestinal mucosa morphometry of two- to six-week-old laying hens. In total, 400 Hy-line W36 laying hens were distributed according to a completely randomized design in 2 x 2 factorial arrangement (energy level x inclusion of xylanase), totaling four treatments with 10 replicates of 10 birds per experimental unit. The following treatments were evaluated: positive control (balanced diet); positive control + xylanase; negative control (diet with of 100 kcal ME reduction /kg); negative control + xylanase. Body weight, weight gain, feed conversion ratio, uniformity and livability were not influenced by diets with metabolizable energy reduction and xylanase inclusion; however, the addition of xylanase to the diets resulted in shallower crypts depth and greater villus:crypt ratio in the ileum. The energy reduction of the diet associated with the supplementation of xylanase did not influence performance, but increased the feed intake of 2- to 6-week-old laying hens and increased villus height in the ileum of 6-wk-old hens. Xylanase reduces crypt depth in the ileum of 6-week-old hens

Adding value to the meat of spent laying hens manufacturing sausages with a healthy appeal

The aim of this study was to evaluate the viability of the use of spent laying hens' meat in the manufacturing of mortadella-type sausages with healthy appeal by using vegetable oil instead of animal fat. 120 Hy-line® layer hens were distributed in a completely randomized design into two treatments of six replicates with ten birds each. The treatments were birds from light Hy-line® W36 and semi-heavy Hy-line® Brown lines. Cold carcass, wing, breast and leg fillets yields were determined. Dry matter, protein, and lipid contents were determined in breast and leg fillets. The breast and legg fillets of three replicates per treatment were used to manufacture mortadella. After processing, sausages were evaluated for proximal composition, objective color, microbiological parameters, fatty acid profile and sensory acceptance. The meat of light and semi-heavy spent hens presented good yield and composition, allowing it to be used as raw material for the manufacture of processed products. Mortadellas were safe from microbiological point of view, and those made with semi-heavy hens fillets were redder and better accepted by consumers. Values for all sensory attributes were evaluated over score 5 (neither liked nor disliked). Both products presented high polyunsaturated fatty acid contents and good polyunsaturated to saturated fatty acid ratio. The excellent potential for the use of meat from spent layer hens of both varieties in the manufacturing of healthier mortadella-type sausage was demonstrated