Cloning of a novel phytase from an anaerobic rumen bacterium,Mitsuokella jalaludinii,and its expression in Escherichia coli

The full length phytase gene of Mitsuokella jalaludinii was successfully cloned and was found to be 1 047 bp in length, with 348 amino acids, and was designated as PHY7 phytase gene. A comparison of the sequence of PHY7 phytase gene of M. jalaludinii with various microbial phytase gene sequences showed that it was not similar to those from other bacteria except Selenomonas ruminatium, thus suggesting that they may both express a new class of phytase. The PHY7 phytase gene was subsequently subcloned into bacterial expression vector, p ET32 a, for expression in Escherichia coli strain Rosetta-gami. Expression of the recombinant phytase gene was optimised and characterised. The recombinant phytase was estimated to be approximately 55 k Da by SDS-PAGE analysis. The recombinant phytase exhibited optimum activity at 55°C, p H 4.5 and showed good p H stability from p H 3.5 to 5.5(78% relative activity). Metal ions such as Ca2+, Mg2+, and K+ were found to exert significant stimulatory effect on the recombinant phytase activity while Cu2+, Fe3+, and Zn2+ greatly inhibited the enzyme activity. The recombinant phytase showed moderate resistance to trypsin proteolysis, but susceptible to pepsin proteolysis. The results of the study showed that several characteristics of recombinant phytase were slightly different from the native enzyme. Unfavourable characteristics such as reduced p H stability and metal ion effects should be taken into consideration during feed enzyme formulation.     

Brightness of venous blood in South American camelids: implications for jugular catheterization

Objective  To compare the brightness of South American camelid venous blood to that of Equidae.
Study design  Prospective clinical evaluation.
Animals  Twelve South American camelids (eight llamas, four alpacas), eight horses and ponies (control group).
Methods  Appropriately sized catheters were placed in the jugular vein of each animal under local anaesthesia. The blood spilt before the catheter was capped was caught on a white tile. A sample of blood was drawn for blood-gas analysis. The brightness of the blood (both on the tile and in the syringe) was matched to a colour chart (1 = darkest red, 8 = brightest red) by a single observer under bright light conditions. Packed cell volume (PCV) and partial pressure of oxygen (PvO₂) in the blood were also measured on the syringe blood. Normally distributed data were compared using a two tailed t -test, and non-normally distributed data were compared using a Mann–Whitney U -test. Significance was set at p  < 0.05.
Results  Camelid venous blood was significantly brighter red than that of horses and ponies both on the white tile ( p  = 0.0003) and in the syringe ( p  = 0.0001). PCV was significantly lower in camelids (32 ± 4%) compared with horses (37 ± 5%). Partial pressure of oxygen values were similar between groups.
Conclusions and clinical relevance  Jugular venous blood in alpacas and llamas is significantly brighter red than that of horses. Colour should not be used as a sole determinant of venous or arterial catheterization in this species.     

Lactose Synthase Components in Milk: Concentrations of α-Lactalbumin and β1,4-Galactosyltransferase in Milk of Cows from Several Breeds at Various Stages of Lactation

It is believed that milk production is determined by the number and activity of mammary secretory cells. Secretory activity, as assessed by milk volume, depends on secretion of the major osmole in milk, lactose, which is produced by lactose synthase. The amount of either of the two proteins in lactose synthase may regulate milk production. The objective of this study was to determine whether the concentrations in milk of the two components of lactose synthase, α‐lactalbumin (α‐LA) and β1,4‐galactosyltransferase (B4GALT), were related to genetic background, stage of lactation, breed or parity of dairy cows. α‐Lactalbumin and B4GALT concentrations were measured by ELISA and by enzyme assays, respectively, from single milk samples. Two herds with a total of 279 cows were used in the analysis. One herd contained Ayrshire, Brown Swiss, Holstein and Jersey cows; the second herd contained two groups of cows; Holsteins selected for high milk production and Holsteins with 1960s genetics. The α‐LA concentration in milk was greater in Jerseys and Ayrshires than in Holsteins and Brown Swiss. However, no difference in α‐LA concentration was observed in milk from high and low genetic merit cows in the Minnesota herd or among different genetic backgrounds in the Illinois herd. β1,4‐Galactosyltransferase concentrations were similar for all groups that were analyzed. α‐Lactalbumin concentrations were positively correlated with milk protein concentration, milk fat concentration and lactose concentration. β1,4‐Galactosyltransferase concentration in milk exhibited a strong positive correlation with number of days in milk. Although the concentration of B4GALT increased as lactation progressed, the values did not show any correlation with persistency of lactation or late lactation milk production. In conclusion, this survey shows that the two components of lactose synthase are each correlated to protein concentration and individually correlated to the concentration of other milk components and stage of lactation.     

Schisandrin B co-treatment ameliorates the impairment on mitochondrial antioxidant status in various tissues of long-term ethanol treated rats

The effect of schisandrin B (Sch B) on long-term ethanol-induced oxidative stress in various rat tissues was investigated. Long-term ethanol treatment increased reactive oxygen metabolites (ROM) level in plasma. The ethanol-induced oxidative stress was assessed by mitochondrial glutathione and α-tocopherol levels, antioxidant enzyme activities, malondialdehyde (mtMDA) production and heat shock protein (Hsp) 25/70 levels. Liver was most susceptible to oxidative stress with a significant increase in mtMDA production. Long-term Sch B treatment enhanced mitochondrial antioxidant status in a tissue non-specific manner. Sch B co-treatment ameliorated the alterations in plasma ROM levels, mtMDA production and Hsp 25/70 expression in rat tissues.     

Mating and dispersal behaviour of Scirpophaga incertulas and Chilo suppressalis (Lepidoptera; Pyralidae) in relation to resistance management for rice transformed with Bacillus thuringiensis toxin genes

Information on the mating and dispersal behaviour of the rice stem borers Scirpophaga incertulas and Chilo suppressalis is necessary for the design of resistance management strategies for rice varieties genetically engineered with toxin genes from Bacillus thuringiensis (Bt). We investigated whether S. incertulas and C. suppressalis move from the site of adult eclosion before mating to determine the likelihood that random mating will occur between adults eclosing in fields of Bt rice and refuge fields of non-Bt rice. Observations were made under laboratory, screenhouse and field conditions. Almost all S. incertulas males and females flew out of sight after cuticle expansion and hardening, as did most C. suppressalis. Approximately 15% of C. suppressalis females called and mated within 3?m of the site of eclosion, and approximately 5% of C. suppressalis males mated within 5?m of the site of eclosion. These results suggest that random mating is likely to occur between S. incertulas eclosing in Bt rice fields and nearby refuge fields. Although a small proportion of C. suppressalis did not move far before mating, random mating of moths from Bt and nearby refuge fields is also likely to occur in this species if Bt-resistant moths are rare.     

A Novel Pretreatment Method of Lignocellulosic Material as Adsorbent and Kinetic Study of Dye Waste Adsorption

Sulphuric acid-modified bagasse has been used as low-cost adsorbent for the removal of methylene blue (MB) dye from aqueous solution. In order to remove organic compounds that contribute to chemical oxygen demand (COD), pretreatment with thorough washing of adsorbent using boiling distilled water was performed instead of conventional washing using distilled water at room temperature only. This has resulted in the highest efficiency of color removal of 99.45% and COD reduction of 99.36% for MB dye solution at pH 9. Effects of initial pH, dye concentration, adsorbent dosage, temperature, and contact time have been studied. The adsorption of MB dye was pH dependent. Langmuir and Freundlich isotherm models were tested on the adsorption data. The kinetic experimental data were analyzed using pseudo-first order, pseudo-second order, and the intraparticle diffusion model in order to examine the adsorption mechanisms. The adsorption process followed the Langmuir isotherm as well as the Freundlich isotherm and pseudo-second-order kinetic model. The process was found to be endothermic in nature.     

Introduction and elimination of Bovine Viral Diarrhoea Virus in a commercial beef herd: a case study

Routine Bovine Viral Diarrhoea Virus (BVDV) monitoring of a commercial beef herd in southern New South Wales over a 10-year period provided an opportunity to assess the impact of the introduction of BVDV on that herd. BVDV antibody testing provided strong evidence that the herd was initially free of BVDV (2009–2011). Testing from 2012 suggested BVDV had been introduced into the herd and this was confirmed in 2015 with the identification of persistently infected (PI) animals. Having become established in the herd, the owners then set out to eliminate BVDV from the herd. Antigen testing aimed at identifying PI animals revealed BVDV was already absent from the herd. Subsequent antibody testing confirmed that the herd was now free from BVDV. Despite the incursion of BVDV in this herd, there was little measurable impact on reproductive performance (pregnancy rates), although suspected increased calf losses from birth to calf marking were reported. This is the first time such self-clearance has been documented as part of a longitudinal study under Australian conditions.     

Long‐Term Outcome after Arthroscopic Debridement of Distal Phalanx Extensor Process Fragmentation in 13 Horses

Objective— To report long-term outcome after arthroscopic removal of fragmentation of the extensor process of the distal phalanx in horses.
Study Design— Case series.
Animals— Adult horses (n=13).
Methods— Medical records (2003–2004) of horses that had arthroscopic debridement of fragmentation of the extensor process of the distal phalanx were reviewed. Inclusion criteria included: lameness localized to the foot, fragmentation of the extensor process of the distal phalanx debrided arthroscopically, and a follow-up period of ≥4 years.
Results— Of the 13 horses, lameness was resolved in 11 (85%) initially but distal interphalangeal joint pain recurred in 2 (15%) within 1 year of surgery. Three (23%) other horses were retired because of lameness at other sites during the follow-up period resulting in 46% (6/13) being not lame and in full work after 4 years. Substantial changes were identified at surgery in the contralateral joint of 7 horses, even when fragments were only present unilaterally.
Conclusions— Arthroscopic debridement of fragmentation of the extensor process of the forelimb distal phalanx has a good short-term prognosis for resolution of lameness and return to work but a more guarded prognosis for long-term soundness.
Clinical Relevance— These results allow for more accurate prognostication preoperatively and may support early debridement of fragmentation of the extensor process of the distal phalanx.