Polymerase chain reaction amplification of 16S-23S spacer region for rapid identification of Salmonella serovars

Polymerase chain reaction (PCR) was used to amplify the spacer regions between the 16S and 23S genes of rRNA genetic loci of Salmonella serovars for their rapid identification. These genetic loci revealed a significant level of polymorphism in length across the species/serovar lines. When the 16S-23S spacer region amplification products were subjected to agarose electrophoresis, the patterns observed could be used to distinguish all the serovars of Salmonella tested. Unique elements obtained in amplification products were mostly clustered at serovar level, although certain genus-specific patterns were also observed. On the basis of the results obtained, the amplification of 16S-23S ribosomal spacer region could suitably be used in a PCR-based identification method for Salmonella serovars.     

Concentrations of substance P and prostaglandin E2 in synovial fluid of normal and abnormal joints of horses

OBJECTIVE: To correlate substance P content of synovial fluid with prostaglandin E2 content, radiographic evidence of joint abnormality, and anatomic location of the joint for normal and osteoarthritic joints of horses. SAMPLE POPULATION: Synovial fluid from 46 normal joints in 21 horses and 16 osteoarthritic joints in 10 horses. PROCEDURE: Normal and osteoarthritic joints were identified by clinical and radiographic examination, by response to nerve blocks, during scintigraphy or surgery, or by clinicopathologic evaluation. Substance P and prostaglandin E2 contents of synovial fluid were determined by radioimmunoassay. Radio-graphs of joints were assigned a numeric score reflecting severity of lesions. Joints were assigned a numeric score reflecting anatomic location. RESULTS: Median concentrations of substance P and prostaglandin E2 were significantly increased in osteoarthritic joints, compared with normal joints. A significant correlation was found between concentrations of substance P and prostaglandin E2 in synovial fluid, but a correlation was not detected between substance P concentration in synovial fluid and anatomic location of the joint or between radiographic scores of osteoarthritic joints and concentrations of substance P or prostaglandin E2. CONCLUSIONS AND CLINICAL RELEVANCE: A correlation existed between concentrations of substance P and prostaglandin E2 in synovial fluid obtained from normal and osteoarthritic joints. However, content of substance P in synovial fluid cannot be predicted by the radiographic appearance of the joint or its anatomic location. Substance P and prostaglandin E2 may share an important and related role in the etiopathogenesis of osteoarthritis, lending credence to the importance of neurogenic inflammation in horses.     

Immune Responses to Intermediate Strain IBD Vaccine at Different Levels of Maternal Antibody in Broiler Chickens

Tropical Animal Health and Production -     

Biological activity and characterization of nucleopolyhedrovirus isolates ofSpodoptera litura

Geographical isolates ofSpodoptera litura (Fabricius) (Noctuidae: Lepidoptera) nucleopolyhedrovirus (SpltNPV), collected from different parts of India and maintained at Tamil Nadu Agricultural University, were compared for their biological activity and subjected to Restriction Endonuclease (REN) analysis. Neonate and second instar bioassay studies revealed similarity in biological activity as shown by the overlapping fiducial limits of LC₅₀ values. However, there were differences in yield among isolates: significantly higher yields were obtained from isolates UAS and CBE than from the BARC isolate. REN analysis of the four isolates withPst I,Hind III,Bam HI andEco RI enzymes indicated genotypic variation among the isolates. Based on the commonality of the bands, the isolates could be broadly divided into two groups: isolates AU and CBE formed one group, and the other group comprised UAS and BARC based on genetic relatedness. http://www.phytoparasitica.org posting Nov. 21, 2005.     

Behavioral hormoligosis in oviposition preference of Bemisia tabaci on cotton

The study on behavioral hormoligosis in oviposition preference in Bemisia tabaci (Genn.) on cotton was conducted, at Entomological Research Farm, Punjab Agricultural University (PAU), Punjab, India, during 2001 crop season and repeated in the same season. Multiple-choice test was followed for conducting the experiment. Quinalphos (250, 375, and 500), carbaryl (625, 938, and 1250), acephate (750, 1125, and 1500), endosulfan (438, 656, and 875), and fenvalerate (25, 38, and 50 g ai/ha) were repeatedly sprayed on potted plants of American cotton (var. LH-1556). The impact of these insecticides was evaluated in term of oviposition preference by B. tabaci to treated plants. Also, it investigated changes in biochemical components of treated cotton leaves and the correlation with oviposition preference. The results revealed that, fenvalerate treated plants were more preferred by whitefly for oviposition. Maximum number of eggs was observed on fenvalerate treated plants, 38, 50, and 25 g/ha (39.3, 37.3, and 36.1 eggs/leaf, respectively) followed by acephate 1500 g/ha (26.9 eggs/ leaf) compared with untreated control (14.1 eggs/leaf). Almost similar trend of results was observed in the repeated experiment. The results obtained from biochemical studies revealed that all the insecticidal treatments caused reduction in total sugars compared with untreated control except fenvalerate and low dose of quinalphos. All insecticides caused increase in total free amino acids and brought significant changes in total phenols and pH value of treated plants. These results have confirmed the behavioral hormoligosis in oviposition preference that induced by fenvalerate and acephate in B. tabaci, which may be one of the causes behind its resurgence on plants repeatedly treated with these insecticides.     

Disposition kinetics and dosage regimen of levofloxacin on concomitant administration with paracetamol in crossbred calves

The disposition kinetics of levofloxacin was investigated in six male crossbred calves following single intravenous administration, at a dose of 4 mg/kg body weight, into the jugular vein subsequent to a single intramuscular injection of paracetamol (50 mg/kg). At 1 min after the injection of levofloxacin, the concentration of levofloxacin in plasma was 17.2 ± 0.36 µg/ml, which rapidly declined to 6.39 ± 0.16 µg/ml at 10 min. The drug level above the MIC₉₀ in plasma, was detected for up to 10 h. Levofloxacin was rapidly distributed from blood to the tissue compartment as evidenced by the high values of the distribution coefficient, α (17.3 ± 1.65 /h) and the ratio of K₁₂/K₂₁ (1.83 ± 0.12). The values of AUC and Vd_area were 12.7 ± 0.12 µg.h/ml and 0.63 ± 0.01 l/kg. The high ratio of the AUC/MIC (126.9 ± 1.18) obtained in this study indicated the excellent antibacterial activity of levofloxacin in calves. The elimination half-life, MRT and total body clearance were 1.38 ± 0.01 h, 1.88 ± 0.01 h and 0.32 ± 0.003 l/kg/h, respectively. Based on the pharmacokinetic parameters, an appropriate intravenous dosage regimen for levofloxacin would be 5 mg/kg repeated at 24 h intervals when prescribed with paracetamol in calves.     

Subcutaneous pharmacokinetics and dosage regimen of cefotaxime in buffalo calves (Bubalus bubalis)

The pharmacokinetics and dosage regimen of cefotaxime following its single subcutaneous administration (10 mg/kg) were investigated in buffalo calves. Plasma and urine samples were collected over 10 and 24 h post administration, respectively. Cefotaxime in plasma and urine was estimated by microbiological assay technique using E. coli as test organism. The pharmacokinetic profiles fitted one-compartment open model. The peak plasma levels of cefotaxime were 6.48 ± 0.52 µg/ml at 30 min and the drug was detected upto 10 h. The absorption half-life and elimination half-life were 0.173 ± 0.033 h and 1.77 ± 0.02 h, respectively. The apparent volume of distribution and total body clearance were 1.17 ± 0.10 l/kg and 0.45 ± 0.03 l/kg/h, respectively. The urinary excretion of cefotaxime in 24 h, was 5.36 ± 1.19 percent of total administrated dose. A satisfactory subcutaneous dosage regimen for cefotaxime in buffalo calves would be 13 mg/kg repeated at 12 h intervals.     

Genetic diversity of bitter gourd (Momordica charantia L.) genotypes revealed by RAPD markers and agronomic traits

Bitter gourd or bitter melon (Momordica charantia L.) is considered as minor cucurbitaceous vegetable in spite of having considerable nutritional and medicinal properties. Although some reports on genetic diversity based on morphological characterization are available, no work has been conducted to estimate genetic diversity using molecular markers in this crop. In the present study, 38 genotypes of M. charantia including few commercially cultivars collected from different parts of India based on agro-ecological zones were analysed for diversity study both at morphological and molecular levels. Genomic DNA was extracted from young healthy leaves following the procedure of Doyle and Doyle [Doyle, J.J., Doyle, J.L., 1990. A rapid DNA isolation procedure from small quantity of fresh leaf material. Phytochem. Bull. 119, 11–15]. Pair-wise comparison of genotypes was calculated as per the procedure of Jaccard [Jaccard, P., 1908. Nouvelles recherches sur la distribution florale. Bull. Soc. Vaud. Sci. Nat. 44, 223–270]. Dendrogram was constructed using the unweighted pair group method with arithmetic averages (UPGMA) and the computation for multivariate analysis was done using the computer programme NTSYS-pc Version 2.0 [Rohlf, F.J., 1998. NTSYS-pc Numerical Taxonomy and Multivariate Analysis System, Version 2.01. Exeter Software, Setauket, NY, USA]. Diversity based on yield related traits and molecular analysis was not in consonance with ecological distribution. Among 116 random decamer primers screened 29 were polymorphic and informative enough to analyse these genotypes. A total of 208 markers generated of which 76 (36.50%) were polymorphic and the number of bands per primer was 7.17 out of them 2.62 were polymorphic. Pair-wise genetic distance (GD) based on molecular analysis ranged from 0.07 to 0.50 suggesting a wide genetic base for the genotypes. The clustering pattern based on yield related traits and molecular variation was different.