Immobilization of Urease onto Modified Egg Shell Membrane through Cross Linking

Background:Immobilization is an approach in industry to improve stability and reusability of urease. The efficiency of this technique depends on the type of membrane and the method of stabilization. Methods:The PEI-modified egg shell membrane was used to immobilize urease by absorption and glutaraldehyde cross-linking methods. The membranes were characterized by FTIR and AFM, and Nessler method was applied to measure the kinetic of the immobilized enzymes. Finally, the storage stability (6 °C for 21 days) and reusability (until enzyme activity reached to zero) of the immobilized enzymes were investigated. Results:Based on FTIR, three new peaks were observed in both the absorption- (at 1389.7, 1230.8, and 1074.2 cm^-1) and the cross-linking (at 1615-1690, 1392.7, 1450 cm^-1) immobilized enzymes. The surface roughness of the native membrane was altered after PEI treatment and enzyme immobilization. The optimal pH of cross-linking immobilized enzymes was shifted to a more neutral pH, while it was alkaline in adsorption-immobilized and free enzymes. The reaction time decreased in all immobilized enzymes (100 min for free enzyme vs. 60 and 30 min after immobilizing by adsorption and cross-linking methods, respectively). The optimal temperature for all enzymes was 70 °C and they had a higher K_m and a lower V_max than free enzyme. The stability and reusability of urease were improved by both methods. Conclusion:Our findings propose these approaches as promising ways to enhance the urease efficiency for its applications in industries and medicines. Key Words: Egg shell, Immobilization, Polyethylenimine, Urease     

Rumen development and growth of Balouchi lambs offered alfalfa hay pre- and post-weaning

The consumption of solid feed is essential for successful transition from a pre-ruminant to a functional digestive tract. Lambs fed starter rations containing highly fermentable carbohydrates often experience dramatic changes in concentrations of rumen and blood metabolites. The optimal amount of roughage required in the diet of pre-ruminant animals is still unclear. The objective of this study was to determine the effect of feeding alfalfa hay on performance and rumen development in young Balouchi lambs. In a completely randomized design, 30 lambs were fed one of three experimental diets consisting of a control, without alfalfa hay (C), a diet containing 7.5% alfalfa hay (A1), and a diet containing 15% alfalfa hay (A2). Lambs fed A1 and A2 diets had lower dry matter intake during the pre-weaning period (P?P?=?0.02), but feed conversion ratio and average daily gain were not affected by feeding alfalfa hay. Concentration of beta-hydroxybutyric acid was higher in C compared with the A1 and A2 groups (P?P?=?0.04) and increased thickness of muscular layer (P?=?0.05). We concluded that including 15% alfalfa hay in the starter diet could reduce thickness of the keratinization layer and increase muscularity of rumen wall without adverse effects on growth and performance of newborn lambs.     

Study of nano-fiber cellulose production by Glucanacetobacter xylinum ATCC 10245

Bacterial Celluloses (BC) are gaining importance in research and commerce due to numerous factors affecting the bacterial cellulose characteristics and application in different industries. The aim of the present study was to produce bacterial cellulose in different media using different cultivation vessels. Bacterial cellulose was produced by static cultivation of Glucanacetobacter xylinum ATCC 10245 in different culture media such as Brain Heart Agar, Luria Bertani Agar /Broth, Brain Heart Infusion, Hestrin-Schramm and medium no. 125. Cultivation of bacterium was conducted in various culture vessels with different surface area. The cellulose membrane was treated and purified with a 0.1 M NaOH solution at 90 degreesC for 30 min and dried by a freeze- drier at -40 degreesC to obtain BC. The prepared bacterial cellulose was characterized by scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy and X-ray diffraction (XRD). The amount of produced BC was related directly to the surface area of culture vessels.     

Protozoa of a rotating biological contactor treatment plant in Mexico

A protozoological analysis of the biofilm developed on the discs of an RBC unit was performed using light microscopy; the species found belonged to 2 phyla: Sarcomastigophora with 23, and Ciliophora with 30 species. All isolates were free-living with the exception of Tritrichomonas fecalis. Most abundant species were: Euglena gracilis, Paramecium multimicronucleatum, Tetrahymena pyriformis, Polytoma uvella, Didinium nasutum, Bodo caudatus, Vorticella microstoma, Cyclidium glaucoma, Sathrophilus agitatus, Antophysa vegetans, and Urothrica farcta. Ciliates predominated over the flagellates in number and diversity. According to their locomotion most abundant species were free-swimming, followed by the attached, and crawling forms. In relation to their nutrition most frequent species were bacterivorous, saprozoic, and carnivorous. Values of the saprobic index calculated for each sampling station indicated that α-mesosaprobic conditions prevailed along the contactor with a zone of overlap with polysaprobic conditions at station IV. The RBC received an influent flow that varied from 1.5 to 13.5 L s^?1 and a soluble BOD₅ which range from 46 to 170 mg L^?1. The contactor studied showed an overall BOD₅ removal of 86% and a COD removal of 74%. DO levels influenced the values obtained for the saprobic index and the kind of protozoan populations and communities present. The biological findings (indicator species of the saprobic system), clearly reflected and agreed with the physicochemical results obtained simultaneously; the results showed that the system performed efficiently.     

Effect of forage inclusion and particle size in diets of neonatal lambs on performance and rumen development

A slaughter experiment was conducted to determine the effects of alfalfa particle size on rumen morphology and performance of lambs. Twenty‐four Balouchi lambs aged 21 days (9.1 ± 1.1 kg) were randomly fed control (diet without alfalfa hay; CON) and mixed rations containing 15% finely ground (FINE; 2 mm) and 15% coarsely chopped alfalfa hay (LONG; 3 to 4 cm). After a 63 days feeding period, nine animals (three per treatment) were slaughtered to obtain ruminal tissue samples for morphological analyses. Alfalfa particle size did not affect (p > 0.05) papillae density, height, width, epithelium depth and surface area. Coarse alfalfa decreased the stratum corneum and increased (p < 0.05) muscle depth compared with fine and control diets. Neither DNA content and nor RNA concentration of rumen tissue was affected by feeding different diets. Forage particle size did not affect the blood concentration of glucose, urea nitrogen (BUN), beta‐hydroxybutyric acid (BHBA) and non‐esterified fatty acids (NEFA). Dry matter intake and feed conversion ratio were higher for control diet; however, there were no significant differences between treatments for average daily gain. These data suggest that coarse alfalfa significantly reduces the stratum corneum and increases muscularity of rumen wall and tended to better feed conversion ratio.