N-Dimethylamino Succinamic Acid (B-Nine) as a Potential Aid in Improvement of Medicago sativa L.

The influence of N-dimethylamino succinamic acid (B-nine) on lucerne plants cv. IGFRI-244 was investigated. B-nine (a growth regulant) was sprayed as a foliar sprays, employing 10, 100, 250, 500, 1000 and 5000 ppm alongwith water spray as control, and the effects were observed on plant growth, flowering, nutritive constituents and seed yield of the crop. Plant height was reduced from 6.19 to 26.85 % in various concentrations of B-nine being highest retardation in 5000 ppm, whereas 250 ppm produced more branches and leaves with dense foliage in comparison to control and other treatments. Delayed flowering by 2–15 days with significant increase in seed yield and 1000 seed weight B-nine bein maximum seed yield in 250 ppm. However, total seed yield was increased by 4.16 to 24.76 per cent over unsprayed control plants. Among the treated levels of B-nine, 250 ppm also gave higher values for Carbohydrate, IVTDMD in shoot of lucerne plants with considerable decreased in 5000 ppm. NDF condent decreased progressively with increased concentration of B-nine but always lesser than the value of IVTDMD.     

Choosing optimal design depth for surface irrigation systems

The surface irrigation system design was formulated as a mathematical programming problem. The minimum cost of a furrow irrigation system for a hypothetical case was calculated for different design depths (25, 51, 76, 102 and 127 mm). The crop yields and net returns were simulated for the given design depths. A design (depletion) depth of 51 mm was found optimal under the given conditions.     

Differentiation of Pasteurella multocida isolates from cases of atrophic rhinitis in pigs from Zimbabwe by RAPD and ribotyping

Atrophic rhinitis in pigs is rarely reported in Southern Africa. To determine the relationship between Pasteurella multocida clones from clinical cases of atrophic rhinitis, twenty-one strains were characterised by selected phenotypic and genotypic methods. Biochemical analysis classified 18 strains as P. multocida subspecies multocida, whilst the remainder were grouped into separate unassigned biotypes. Capsular groups A (16/21) and D (l/21) were found among the isolates by PCR. Four ribotype patterns were obtained following HpaII ribotyping, whilst random amplification of polymorphic DNA (RAPD) revealed three main clusters. However, subclusters were also noted for each RAPD cluster. Our results indicate that RAPD offers a better discrimination of strains than ribotyping and that none of the phenotypic characters were directly related to the genotypic clusters.     

Recombinant LipL32 antigen-based single serum dilution ELISA for detection of canine leptospirosis

A recombinant antigen-based single serum dilution enzyme-linked immunosorbent assay (ELISA) was developed to measure the specific antibody activity in sera of dogs with leptospirosis. The recombinant antigen developed and used in the assay was specific for the pathogenic serovars of Leptospira. A linear relationship was found to exist between the predicted antibody titres at a single working dilution of 1:1000 and the corresponding observed serum titres as determined by the standard serial-dilution method. Regression analysis was used to determine a standard curve from which an equation can be derived that allows demonstration of the mentioned correlation. The equation was then used to convert the corrected absorbance readings of the single working dilution directly into the predicted ELISA antibody titres. The assay was proved to be sensitive, specific and accurate as compared to the standard microscopic agglutination test (MAT).     

Residues of fluoroquinolone drugs in the cloacal gland and other tissues of Japanese quail

1. In this study we investigated the residues of fluoroquinolone drugs (ciprofloxacin and pefloxacin) in the cloacal gland (a site of foam synthesis) and other tissues such as breast muscle, testes, brain, kidney and plasma. 2. Fifty-four healthy male Japanese quail were selected at random from a flock, maintained under uniform husbandry conditions and divided into three groups, each of 18 birds. Group I (control) received 1 ml vehicle (normal saline 0.9% (w/v) NaCl) daily for 12 d through the intraperitoneal route. Birds of groups II and III received ciprofloxacin and pefloxacin by the same route at the rate of 10 and 12 mg/kg body weight, respectively, every day for a similar period. 3. Birds from each group were killed, at 1, 5 and 10 d after the cessation of treatment, to collect the cloacal gland together with other tissues that were analysed for residual drugs. 4. Cloacal gland retained the maximum drug residues of ciprofloxacin (60%) and pefloxacin (80%) on d 10 compared with that on d 1 after drug withdrawal. The drug residues were found 60 and 80% in ciprofloxacin and pefloxacin groups, respectively, in the cloacal gland tissue even on d 10 after withdrawal of the treatment. 5. In the ciprofloxacin-treated group, all tissues except cloacal gland contained very small amounts of the drug residues on d 10 after treatment ended. In the pefloxacin group the cloacal gland, breast muscle and kidney retained a fairly high amount of drug even on d 10 after treatment ceased. No residues of pefloxacin were detectable in testes and brain throughout. 6. In conclusion, the cloacal gland in Japanese quail acted as the largest sink for the fluoroquinolone drugs. Ciprofloxacin was more widely distributed in different tissues and persisted for a shorter period than pefloxacin.     

Regulation of B cell tolerance by the lupus susceptibility gene Ly108

The susceptibility locus for the autoimmune disease lupus on murine chromosome 1, Sle1z/Sle1bz, and the orthologous human locus are associated with production of autoantibody to chromatin. We report that the presence of Sle1z/Sle1bz impairs B cell anergy, receptor revision, and deletion. Members of the SLAM costimulatory molecule family constitute prime candidates for Sle1bz, among which the Ly108.1 isoform of the Ly108 gene was most highly expressed in immature B cells from lupus-prone B6.Sle1z mice. The normal Ly108.2 allele, but not the lupus-associated Ly108.1 allele, was found to sensitize immature B cells to deletion and RAG reexpression. As a potential regulator of tolerance checkpoints, Ly108 may censor self-reactive B cells, hence safeguarding against autoimmunity.     

Association mapping for pre-harvest sprouting tolerance in common wheat (Triticum aestivum L.)

Association mapping was undertaken in common wheat to identify markers associated with pre-harvest sprouting tolerance (PHST). For this purpose, a population of 242 wheat genotypes and 250 SSR markers were used. The population used consisted of diverse germplasm, which carried sufficient phenotypic variation for PHS for conducting association mapping. The population was found to be structured and stratified into 15 sub-populations; the tolerant and moderately tolerant wheat genotypes were distributed in all the sub-populations. This feature of the population along with other information on population structure was used in association mapping using both the available models, the general linear model (GLM) and the mixed linear model (MLM); hopefully, this minimized the rate of false positives. As many as 30 markers were found to be associated with PHST, 26 markers with GLM and 17 markers with MLM; 13 markers were detected using both the approaches. Only eight SSR markers associated with QTL for PHST were such, which were located within the marker intervals that were earlier reported to carry QTLs for PHST. The remaining 22 markers that were found to be associated with PHST could not be associated with any of the genomic regions known to carry QTLs for PHST, which are known to occur on all the 42 chromosome arms of wheat genome.     

Canine corneal fibroblast and myofibroblast transduction with AAV5

Objective The aims of this study were (1) to determine the efficacy of adeno‐associated vector serotype 5 (AAV5) for delivering gene therapy to canine corneal fibroblasts (CCFs) and myofibroblasts (CCMs) using enhanced green fluorescent protein (GFP) marker gene and (2) to evaluate the cytotoxicity of AAV5 to CCFs and CCMs using an in vitro model. Methods Healthy donor canine corneas were used to generate primary CCFs by growing cultures in minimal essential medium supplemented with 10% fetal bovine serum. Canine corneal myofibroblasts were produced by growing cultures in serum‐free medium containing transforming growth factor β1 (1 ng/mL). An AAV5 titer (6.5 × 10¹² μg/mL) expressing GFP under control of hybrid cytomegalovirus + chicken β‐actin promoters (AAV5‐gfp) was used to transduce CCF and CCM cultures. Delivered gene expression in CCFs and CCMs was quantified using immunocytochemistry, fluorescent microscopy, and real‐time PCR. Transduction efficacy of the AAV5 vector was determined by counting DAPI‐stained nuclei and EGFP‐positive cells in culture. Phase‐contrast microscopy, trypan blue, and dUTP nick end labeling (TUNEL) assays were used to determine the toxicity and safety of AAV5 in this canine corneal model. Results Topical AAV5 application successfully transduced a significant population of CCFs (42.8%; P < 0.01) and CCMs (28%; P < 0.01). Tested AAV5 did not affect CCF or CCM phenotype or cellular viability and did not cause significant cell death. Conclusions The tested AAV5 is an effective and safe vector for canine corneal gene therapy in this in vitro model. In vivo studies are warranted.