We investigated the feasibility of cryopreservation of zebrafish (Danio rerio) blastomeres and primordial germ cells (PGCs) by rapid freezing of dechorionated whole embryos at the blastula, gastrula and segmentation stages. Initially we examined the glass-forming properties and embryo toxicities of 5 cryoprotectants: methanol (MeOH), ethylene glycol (EG), dimethyl sulfoxide (DMSO), propylene glycol (PG), and 1,3-butylene glycol (1,3-BG). Embryos at the blastula and gastrula stages had high sensitivities to cryoprotectant toxicities and were fragile against mechanical damage. Thus the segmentation stage embryos, the PGCs of which were visualized by injecting green fluorescence protein-nos1 3'UTR mRNA, were frozen using solutions containing each cryoprotectant at 6 M (first trial) and 2 types of cryoprotectants at 3 M each (second trial). In the first trial, live PGCs were recovered from most of the embryos frozen with EG (about 2 cells/embryo); however, a few embryos had live PGCs when embryos were frozen with other cryoprotectants. In the second trial; a mixture of EG + PG better preserved the viability of PGCs in frozen embryos. Live PGCs were recovered from all embryos frozen with EG + PG (about 3 cells/embryo), and the survival rate of PGCs was estimated to be about 25% based on the number of live PGCs in fresh embryos (about 12 cells/embryo). The present study indicates that we can utilize rapid freezing of dechorionated whole embryos at the segmentation stage for the cryopreservation of PGCs.

While the C-terminal cytoplasmic tail of anion exchanger 1 (AE1, band 3) has been reported to possess important physiological roles, including one for proper membrane trafficking, its precise characteristics remain unclear. To clarify the overall structural consequences of the conserved sequence EL(K/Q)(L/C)LD(A/G)DD, containing the core binding sequence LDADD for carbonic anhydrase II, in the C-terminal region, we analyzed the membrane expression and turnover of bovine AE1 with a series of truncation and substitution mutations in HEK293 cells. Immunofluorescence microscopy and cell-surface biotinylation demonstrated that truncation mutants missing 18 C-terminal residues targeted the plasma membrane, but the one lacking the conserved region, by truncation of 28 amino acid residues, was retained inside the cells. Substitutions of Ala for Glusup(901), Leusup(902), Leusup(905), and Aspsup(906) in the sequence E901L(K/Q)(L/C)LDADD909 of bovine AE1 or those in the corresponding murine sequence also caused intracellular retention, though these mutants had half-lives comparable to that for wild-type AE1. These data demonstrate that the conserved amino acid residues Glusup(1), Leusup(2), Leusup(5), and Aspsup(6) in the EL(K/Q)(L/C)LD(A/G)DD region have essential structural consequences in stable expression of AE1 at the plasma membrane regardless of the ability in binding to carbonic anhydrase II of this region.

The level of antibody by ELISA,immunsosuppressive effect baes on the response of birds to Newcastle disease vaccination and damage to bursa of Fabricius by IBD vaccination were studied. The efficacy of six different IBD vaccination schedules were studied using intermediate and intermediate plus strains of vaccines either alone or in combination. In vaccinated groups, the sero-conversion of the vaccine virus was noticed during fourth week, reaching to the peak between eight to twelve weeks of age in different groups. Afterwards, there was a gradual decrease in the titres, by the end of 20th week (maximum period tested). There was no significant difference in the titres of different treatment groups. However, all the groups showed titres above protective level during the entire period of study. There was significant difference in bursa body weight (B-BW) ratios of vaccinated groups in comparison with control group. Histopathological studies of bursal sections revealed depletion of lymphoid follicles, presence of cystic spaces, edema and hemorrhages. The birds vaccinated with hot strain of IBD vaccine showed metaplastic changes, presence of foam cells with pronounced interfollicular fibrosis. The bursal scores were maximum in the groups vaccinated with hot strains of IBD vaccines.

Hyalomma ticks of either sex were collected from the apparently healthy cross-bred cattle. The ticks were dissected and their salivary glands were stained by methyl green pyronin stain for the detection of Theileria annulata sporozoite in their salivary glands. The prevalence of Theileria annulata infection in Hyalomma ticks was found to be 21.5%. No correlation was found between parasitemia of cattle and tick infection rate.

Filix-mas a homeopathic drug consisting etherieal extract of male fern is used to remove the tape worms. The drug was tested for Moniezia expensa infection in sheep. Ten sheep naturally affected Moniezia expansa were used. Filix-mas 200 (10 _ 15 drops) was given three times a day orally to animals of the group A, while the sheep group B were kept as untreated control. Efficacy of the drug was observed by calculating faecal egg count reduction. It was found that Filix-mas could cure 100% monieziosis in sheep.

An overall prevalence of 20.07% true anoestrus and delayed puberty in cattle was recorded out of 2202 gynaecological cases presented to the clinical complex and different clinical camps held at different locations of Kashmir valley over a period of three years (2005_2008). The highest prevalence of true anoestrus in cows was recorded in the 1st parity when they attained approximately 3 years of age and during autumn season.

The study was conducted in 18 clinically healthy male dogs of about 1 year of age, weighing up to 12 kg and divided into three groups of 6 animals each. Bupivacaine alone, bupivacaine + hyaluronidase and hyperbaric bupivacaine were given epidurally at lumbosacral space @2 mg/kg. bwt. with a maximum dose of 3 ml in the animals of groups I, II and III, respectively. A significant increase (P0.01) in rectal temperature, heart rate and respiration rate was recorded at initial intervals of observation in all the groups and thereafter these values follow a declining trend in all groups consistently and reached to almost base line value at 120 min. of observation The alterations of glucose, serum urea nitrogen, aspartate aminotransferase and creatinine estimated at 1hr, 2hrs, 4hrs and 24 hrs were remained within physiological limits in all the groups.Therefore induction of regional anaesthesia with bupivacaine and its combination revealed no permanent alteration in the biochemical and physiological parameters hence it is safe and satisfactory anaesthesia.

Clinico-physiological effects of Xylazine - Ketamine were evaluated in 12 clinical cases of uremic buffalo calves having urolithiasis. In group A, Xylazine -Ketamine were used at the dose rate of 0.05 mg/kg body weight and 2.5 mg/kg body weight respectively to create regional spinal anesthesia at the lumbosacral space in 6 buffalo calves. In group B, Xylazine and Ketamine at the same dose rates were used intramuscularly in 6 buffalo calves. Analgesia was then recorded at different regions by the pin prick method and scored on a scale and motor incoordination, sedation, complete duration of anesthesia, complete recovery and physiological parameters (heart rate, respiration rate and rectal temperature) were evaluated in both the groups at various intervals of time throughout the duration of surgery of Tube cystotomy. It was found that the animals of group B achieved a safer physiological peak values than animals of group A.

A total of 253 samples from cattle (52), buffalo (51), sheep (50), goat (50) and chicken (50) were screened for the presence of Pasteurella multocida. Out of these, 17 samples were found positive by species specific PCR (PM-PCR) with ~460 bp amplified product. Overall prevalence of P. multocida was found to be 6.7%. Out of 17 PM-PCR positive samples, 12 (70%) were found positive by capsular type-B specific (HSB-PCR) and multiplex PCR assays with amplification of ~620 bp product in both the cases. A total of 5 isolates of P. multocida were obtained in 17 PM-PCR positive samples of which 4 were of serotype B:2 and 1 was of serotype A:3. The PM-PCR and HSB-PCR assays can be performed directly on clinical samples from animals with reproducible results without any non-specific amplification. Multiplex PCR further reduces the time for the species and type specific detection of P. multocida.

Survivability of Listeria monocytogenes in clay loam soil was examined at 30° C for a period of 35 days. The microorganism inoculated in soil steadily declined from 3.5 × 10 6 g -1 to 3.9 × 10 4 g -1 (P = 0.05) on 35 th days. Destruction of normal soil microflora by heat treatment resulted in marginal rise in 3.3 × 10 6 g -1 of L. monocytogenes to 4.2 × 10 6 g -1 during the first 7 days, followed by steady decline to 5.6 × 10 4 g -1 on 35 th days. Heat treated soil inoculated with lower concentration of L. monocytogenes (3.6 × 10 4 g -1 ) supported sharp rise in its population to 4.8 × 10 6 g -1 (P=0.05) during the first weak which gradually declined, but remained higher (6.1 × 10 4 g -1 ) than the initial load even on 35 th days. In non-sterile soil, the rise was less appreciable but L. monocytogenes maintained its levels (10 4 g -1 ) during the experimental period. Survivability of the microorganisms was also observed in the soil with added chicken manure, cattle manure or sheep/goat manure.