Authors:S. H. Basagoudanavar, J. R. Rao, Swati Omanwar, A. K. Tiwari, R. K. Singh, R. S. Kataria and G. Butchaiah
A highly reproducible, dominant, monomorphic fragment of 473 base pair (bp) amplified from the genome of Trypanosoma evansi by arbitrary primer — polymerase chain reaction (AP-PCR) was labelled with digoxigenin and investigated for its potential as DNA probe. Dot-blot hybridisation of total genomic DNA with the probe proved useful in detecting bubaline, cameline and equine strains of T. evansi down to 10 pg of parasite template DNA. No cross-hybridisation was seen with Babesia bigemina, Theileria annulata and the bubaline host DNA. This probe may facilitate laboratory identification of T. evansi in developing countries, without the inherent risk associated with radioisotopes.
Authors:R. Ponnuswamy, A. Rathore, A. Vemula, R.R. Das, A.K. Singh, D. Balakrishnan, H.S. Arremsetty, R.B. Vemuri and T. Ram
The All India Coordinated Rice Improvement Project of ICAR-Indian Institute of Rice Research, Hyderabad organizes multi-location testing of elite lines and hybrids to test and identify new rice cultivars for the release of commercial cultivation in India. Data obtained from Initial Hybrid Rice Trials of three years were utilized to understand the genotype × environment interaction (GEI) patterns among the test locations of five different agro-ecological regions of India using GGE and AMMI biplot analysis. The combined analysis of variance and AMMI ANOVA for a yield of rice hybrids were highly significant for GEI. The GGE biplots first two PC explained 54.71%, 51.54% and 59.95% of total G + GEI variation during 2010, 2011 and 2012, respectively, whereas AMMI biplot PC1 and PC2 explained 46.62% in 2010, 36.07% in 2011 and 38.33% in 2012 of the total GEI variation. Crossover interactions, i.e. genotype rank changes across locations were observed. GGE biplot identified hybrids, viz. PAN1919, TNRH193, DRH005, VRH639, 26P29, Signet5051, KPH385, VRH667, NIPH101, SPH497, RH664 Plus and TNRH222 as stable rice hybrids. The discriminative locations identified in different test years were Coimbatore, Maruteru, VNR, Jammu, Raipur, Ludhiana, Karjat and Dabhoi. The AMMI1 biplot identified the adaptable rice hybrids viz., CNRH102, DRH005, NK6303, NK6320, DRRH78, NIPH101, Signet5050, BPH115, Bio452, NPSH2003, and DRRH83. The present study demonstrated that AMMI and GGE biplots analyses were successful in assessing genotype by environment interaction in hybrid rice trials and aided in the identification of stable and adaptable rice hybrids with higher mean and stable yields.
Authors:A. Niranjan, J. Barthwal, A. Lehri, D. P. Singh, R. Govindrajan, A. K. S. Rawat and D. V. Amla
Artemisia pallens L. (Compositae) is used in Indian traditional medicine to treat diabetes mellitus, jaundice, hysteria, body pain, and bacterial and fungal infections. A major cause of a variety of diseases is oxidative stress which is reduced by antioxidants such as polyphenols. These secondary metabolites are generally ubiquitous in plants and extensively used in the pharmaceutical, cosmetic, and food industries. In this study a simple and sensitive HPLC-UV-MS-MS-based method was developed for separation, identification, and quantification of polyphenols, for example gallic, protocatechuic, chlorogenic, caffeic, and ferulic acids, rutin, quercetin, and kaempferol. Amounts of polyphenols detected in 50% methanol-water extracts of the plant varied from 0.005% (kaempferol) to 0.24% (protocatechuic acid). Separation of the polyphenols was achieved on a reversed-phase C18 with a mobile phase prepared from 1% aqueous with acetic acid and acetonitrile at a flow rate of 0.6 mL min−1. The phenolic compounds were detected by UV absorption at 254 nm. The method was validated for linearity, accuracy, precision, LOD, LOQ, specificity, selectivity, and compound stability. Results from intra and inter-day validation (n = 6) showed the method was efficient and rapid. The optimized method was applied to extracts of A. pallens for identification and quantification of the polyphenols. The reference standards and their presence in A. pallens were confirmed by mass spectrometry.
Authors:S. L. Krishnamurthy, S. K. Sharma, D. K. Sharma, P. C. Sharma, Y. P. Singh, V. K. Mishra, D. Burman, B. Maji, B. K. Bandyopadhyay, S. Mandal, S. K. Sarangi, R. K. Gautam, P. K. Singh, K. K. Manohara, B. C. Marandi, D. P. Singh, G. Padmavathi, P. B. Vanve, K. D. Patil, S. Thirumeni, O. P. Verma, A. H. Khan, S. Tiwari, M. Shakila, A. M. Ismail, G. B. Gregorio and R. K. Singh
Genotype × environment (G × E) interaction effects are of special interest for identifying the most suitable genotypes with respect to target environments, representative locations and other specific stresses. Twenty-two advanced breeding lines contributed by the national partners of the Salinity Tolerance Breeding Network (STBN) along with four checks were evaluated across 12 different salt affected sites comprising five coastal saline and seven alkaline environments in India. The study was conducted to assess the G × E interaction and stability of advanced breeding lines for yield and yield components using additive main effects and multiplicative interaction (AMMI) model. In the AMMI1 biplot, there were two mega-environments (ME) includes ME-A as CARI, KARAIKAL, TRICHY and NDUAT with winning genotype CSR 2K 262; and ME-B as KARSO, LUCKN, KARSA, GOA, CRRI, DRR, BIHAR and PANVE with winning genotypes CSR 36. Genotypes CSR 2K 262, CSR 27, NDRK 11-4, NDRK 11-3, NDRK 11-2, CSR 2K 255 and PNL 1-1-1-6-7-1 were identified as specifically adapted to favorable locations. The stability and adaptability of AMMI indicated that the best yielding genotypes were CSR 2K 262 for both coastal saline and alkaline environments and CSR 36 for alkaline environment. CARI and PANVEL were found as the most discernible environments for genotypic performance because of the greatest GE interaction. The genotype CSR 36 is specifically adapted to coastal saline environments GOA, KARSO, DRR, CRRI and BIHAR and while genotype CSR 2K 262 adapted to alkaline environments LUCKN, NDUAT, TRICH and KARAI. Use of most adapted lines could be used directly as varieties. Using them as donors for wide or specific adaptability with selection in the target environment offers the best opportunity for widening the genetic base of coastal salinity and alkalinity stress tolerance and development of adapted genotypes. Highly stable genotypes can improve the rice productivity in salt-affected areas and ensure livelihood of the resource poor farming communities.
Authors:K. R. W. Oliveira, R. A. Sversut, A. K. Singh, M. S. Amaral and N. M. Kassab
The present study aimed to develop and validate an analytical method for determination of marbofloxacin (MAR) in veterinary chewable tablets. The isocratic reversed-phase chromatographic method was developed and validated using a Vertisep®, RP C18 column (150 mm × 4.6 mm, 5.0 μm). The mobile phase was composed of water–acetonitrile (55:45, v/v) with pH adjusted to 3.0 with ortho-phosphoric acid and a flow rate set at 0.4 mL/min. The proposed method was validated for linearity in a concentration range of 2.5 to 17.5 μg/mL with a correlation coefficient of 0.99991. The mean content of MAR found in chewable tablets was 104.40% with RSD below 2%. The accuracy expressed as average recovery of the proposed method was 98.74%, and the precision expressed as relative standard deviation among repeated analysis was 0.55%. The method has adequate sensitivity with detection and quantitation limits of 0.25 and 0.81 μg/mL, respectively. Based on the presented results and according to the ICH and AOAC guidelines on validation of analytical methods, the proposed method was considered precise, accurate with adequate sensitivity, and robust in the MAR quantitative analysis. Therefore, the method can be used in the quality control of chewable veterinary tablets containing MAR.
Authors:J.F.F. Anderson, M.C.G. Gerlin, R.A. Sversut, L.C.S. Oliveira, A.K. Singh, M.S. Amaral and N.M. Kassab
The objective of this study was to develop and validate an assay method for simultaneous determination of atenolol, furosemide, losartan, and spironolactone in pharmaceutical formulations. A reverse-phase high-performance liquid chromatography procedure was developed, using a Kinetex® C-18 column (100 mm × 4.6 mm, 2.6 μm). The mobile phase was composed of methanol—water (75:25 v/v, pH 3.0, adjusted with phosphoric acid), with a flow rate of 0.4 mL min−1. All drugs were separated in less than 5 min. The method was validated according to International Conference on Harmonization (ICH) and Association of Official Analytical Chemists (AOAC) guidelines. The method showed linearity in a concentration range of 0.75–12.0 μg mL−1 for atenolol (r = 0.9995), 0.30–12.00 μg mL−1 for furosemide (r = 0.9997), 0.45–12.00 μg mL−1 for losartan (r = 0.9995), and 0.45–12.0 μg mL−1 for spironolactone (r = 0.9999). The method also showed repeatability and precision. The three-day average intra-day precisions were 101.35 ± 0.74% for atenolol, 95.84 ± 1.44% for furosemide, 98.90 ± 1.16% for losartan, and 97.19 ± 0.18% for spironolactone. Similarly, the inter-day precisions were 101.34 ± 0.72% for atenolol, 95.84 ± 0.1.50% for furosemide, 98.90 ± 1.17% for losartan, and 97.19 ± 0.83% for spironolactone. The method accuracy was also tested and validated — in this case, the average recovery values were 100.18 ± 1.20% for atenolol, 99.83 ± 1.54% for furosemide, 100.07 ± 0.95% for losartan, and 99.94 ± 0.93% for spironolactone. Finally, the method was successfully applied in the simultaneous determination of atenolol, furosemide, losartan, and spironolactone in magisterial formulas, as well as in commercial pharmaceutical formulations.
Authors:B. Kumar, K.S. Hooda, R. Gogoi, V. Kumar, S. Kumar, A. Abhishek, P. Bhati, J.C. Sekhar, K.R. Yathish, V. Singh, A. Das, G. Mukri, E. Varghese, H. Kaur, V. Malik and O.P. Yadav
Maydis leaf blight (MLB), a serious foliar fungal disease of maize, may cause up to 40% losses in yield. The present studies were undertaken to identify the stable sources of MLB resistance, its inheritance study, and testing of MLB resistance linked markers from diverse background in the Indian adapted tropical maize genotypes. A set of 112 inbred lines were screened under artificially created epiphytotics conditions at three hotspot locations. Analysis across multi-locations revealed significant effects of genotypes and environments, and non-significant effects due to genotypes × environment interaction on disease incidence. A total of 25 inbred lines with stable resistance were identified across multi-locations. Inheritance of resistance was studied in six F1s and two F2s of resistant and susceptible parents. The null hypothesis of segregation of resistance and susceptible for mono and digenic ratios in two F2 populations was rejected by Chi-square test. The non-significant differences among the reciprocal crosses depicted the complete control of nuclear genome for MLB resistance. Partial dominance in F1s and normal distribution pattern in F2s of resistant and susceptible parents suggested polygenic nature of MLB resistance. Correlation studies in F2 populations exhibited significant negative correlation between disease score and days to flowering. Five simple sequence repeats (SSRs) markers, found associated to MLB resistance in different studies were unable to differentiate amongst MLB resistance and susceptible parents in our study. This emphasizes the need of fine mapping for MLB resistance in Indian germplasm. The identified stable sources of resistance and information on inheritance study can be used further in strengthening of resistance breeding against MLB.