Production of Lambs After the Transfer of Fresh and Cryopreserved in vitro Produced Embryos from Prepubertal Lamb Oocytes and Unsorted and Sex-sorted Frozen-thawed Spermatozoa

Cumulus-oocyte complexes from hormone-stimulated 3-4-week-old (n=43) and 6-7-week-old (n=12) prepubertal lambs were matured in vitro and incubated with unsorted, or X- or Y-spermatozoa separated with a high-speed cell sorter (SX MoFlo)frozen-thawed. Presumptive zygotes were then cultured to the blastocyst stage, and transferred to recipients fresh or after cryopreservation (frozen). Oocyte cleavage was higher (p <0.05) with unsorted (515/926, 55.6%) than X- or Y-spermatozoa (261/672, 38.8% and 229/651, 35.2%, respectively) and blastocyst formation (% zygotes) by Day 9 of in vitro culture was lower (p <0.05) for X- (102/261, 39.1%) than unsorted spermatozoa (249/515, 48.3%), but did not differ between Y-spermatozoa (103/229, 45.0%) and unsorted spermatozoa, or between X- and Y-spermatozoa (p >0.05). For fresh embryos, survival to term was 50.0% (3/6) for unsorted, 0.0% (0/6) for X- and 16.7% (1/6) for Y-spermatozoa-derived embryos (p >0.05), and for frozen embryos was 4.0% (2/50) for unsorted, 9.1% (2/22) for X- and 2.9% (1/34) Y-spermatozoa-derived embryos (p >0.05). Of the two lambs born from X-spermatozoa-derived embryos, one was female (50%), and from the two Y-spermatozoa-derived lambs, both were male (100%), demonstrating that lambs can be produced after the transfer of fresh and cryopreserved IVP embryos derived from prepubertal lamb oocytes and frozen-thawed sex-sorted sperm.     

Device Applications of Side-Chain Ferroelectric Liquid Crystalline Polymer Films

Side-chain ferroelectric liquid crystalline polymers are currently used in a number of applications, including displays and electrical sensors. Comparisons with existing technologies and materials indicate that relative to ceramics, such polymers have lower figures of merit but offer greater durability in sensor applications.     

Impact of Distillery Effluent on Soil Carbon and Nitrogen Dynamics in a Vertisol of Central India

Distilleries produce a huge quantity of effluents, popularly known as spent wash (SW), which when bio-methanated produce post-methanation effluents (PME). A field experiment on soybean–wheat system was conducted for five consecutive years in a Vertisol of central India to evaluate the effect of distillery effluent (DE) on soil carbon and nitrogen dynamics. Ten treatment combinations consisting of control, 100% NPK + Farmyard Manure (FYM), and graded level of SW and PME were applied. Total carbon content of soil increased significantly with applications of FYM and DE. SW was found superior in enhancing carbon content of soil in comparison to PME. Farmyard Manure contributed more carbon toward the recalcitrant pool, whereas DE contributed more carbon toward the active and slow pool. Nitrogen (N) availability was significantly improved with the application of DE. Balanced application of DE may act as amendment for increasing C and N stocks in Vertisol.     

Thymoquinone enhances the activities of enzymes related to energy metabolism in peripheral leukocytes of diabetic rats

The aim of this study was to examine the effect of glycemic control using thymoquinone (TQ) on energy metabolism related enzymes in leukocytes of streptozotocin (STZ)-induced diabetic rats. The treatment of both TQ and insulin commenced 4 weeks after induction of diabetes. Plasma glucose, cholesterol and triglycerides levels were significantly reduced after TQ treatment, whereas immunoreactive insulin (IRI) showed significant increase. The activities of malate dehydrogenase (MDH) in cytosolic and mitochondrial fractions of peripheral blood leukocytes were significantly higher in rats treated with TQ and insulin as compared to that in diabetic controls. On the other hand the activities of lactic dehydrogenase (LDH) showed no significant changes between groups. ML ratio (cytosolic MDH/LDH specific activity ratio) was restored to those in the control rats. The results of this study demonstrate that TQ significantly increased insulin level and the activities of cytosolic and mitochondrial MDH in leukocytes of STZ-diabetic rats.     

Isolation and molecular characterisation of Neospora caninum in cattle in New Zealand

AIM: To isolate Neospora caninum from the brains of naturally infected cattle and use molecular techniques to characterise the isolates.

METHODS: Neospora caninum tachyzoites were isolated in Vero cell culture from the brains of a cow and two calves. The isolates were characterised using polymerase chain reaction (PCR) methods, DNA sequencing, an immunofluorescent anti-body test (IFAT), transmission electron microscopy (TEM), and immunohistochemistry (IHC). The brains of the three cattle were subjected to histopathological examination. A pathogenicity study was conducted in 120 BALB/c mice.

RESULTS: Neospora caninum tachyzoites were isolated from all three cases and first observed in vitro between 14 and 17 days post-inoculation. Parasites were sub-cultured and maintained in Vero cell culture for more than 6 months. PCR products were generated for all three isolates, using two different primers. Sequencing of the PCR products and a subsequent BLAST search identified the isolates as N. caninum. In addition, the isolates tested positive using IFAT and IHC, and ultrastructure revealed by TEM was characteristic of N. caninum. Histopathological examination revealed lesions characteristic of N. caninum in 1/3 brains. In the pathogenicity study using BALB/c mice, the mortality rate was 3–7%.

CONCLUSION: This was the first successful isolation of N. caninum in New Zealand confirmed using molecular characterisation tests.     

Evaluation of different diagnostic methods for diagnosis of Lumpy skin disease in cows

Viral isolation, polymerase chain reaction (PCR), dot blot hybridization (DBH), and indirect enzyme-linked immunosorbent assay (iELISA) were used for the diagnosis of lumpy skin disease in clinically infected, fevered, and apparently normal dairy cows. Lumpy skin disease virus (LSDV) was isolated from skin biopsies and blood samples collected from clinically infected cows in percentages of 72% and 20%, respectively. The virus recovered from blood samples collected from fevered cows in percentage of 33.3%. Both PCR and DBH detected viral DNA in 100% of skin biopsies collected from clinically infected cows whereas the detection rates in blood samples collected from clinically infected animals were 100% and 84% using PCR and DBH, respectively. Viral DNA was detected in blood samples collected from fevered cows using PCR and DBH in percentages of 77.8% and 66.6%, respectively. Only 19.1% of blood samples collected from in-contact cows was positive for both of PCR and DBH. Detection rates of antibodies against LSDV using iELISA in serum samples collected from clinically infected and fevered cows were 56% and 11.1%, respectively, whereas all in-contact cows had no antibodies against the virus.