Erythrocytic antioxidant defense, lipid peroxides level and blood iron, zinc and copper concentrations in dogs naturally infected with Babesia gibsoni

Babesiosis is a common tick borne disease of dogs in tropical and subtropical regions of the world caused by different species of Babesia. The present study aimed to examine erythrocyte lipid peroxide and erythrocytic antioxidant levels in dogs with clinical babesiosis, caused by Babesia gibsoni, and impact of the disease on blood iron, zinc and copper levels. The study was conducted on 10 naturally occurring cases of canine babesiosis with the history of tick infestation, erratic pyrexia, and prolonged illness. Microscopic examination of Giemsa stained peripheral blood smears confirmed B. gibsoni infection in the erythrocytes. Six apparently healthy dogs of different age, sex and breeds, brought for either health checkup or vaccination were used for comparison. Levels of erythrocytic antioxidant enzymes were significantly (P<0.01) higher in sick dogs than those of cytologically negative dogs (catalase: 0.192+/-0.024 units/mg Hb vs 0.074+/-0.004 units/mg Hb; superoxide dismutase: 0.014+/-0.0009 units/mg Hb vs 0.006+/-0.0008 units/mg Hb and lipid peroxide: 6.01+/-0.30 nmol MDA/mg Hb vs 1.89+/-0.10 nmol MDA/mg Hb). The levels of blood micronutrients were significantly low in these dogs (iron: 89.87+/-8.12 microg/g vs 126.44+/-14.65 microg/g; zinc: 3.67+/-1.85 microg/g vs 5.62+/-1.83 microg/g and copper: 0.55+/-0.63 microg/g vs 0.65+/-0.04 microg/g). The study demonstrated oxidative damage in dogs naturally infected with B. gibsoni. Low level of blood iron, zinc and copper seems to have an additional role in the genesis of anaemia and oxidative stress.     

Changes in plasma hormones profile and liver function in cows naturally exposed to lead and cadmium around different industrial areas

The present study was carried out to assess the endocrine status and liver function in adult cows reared in polluted environment around different industrial units in India.The effect on endocrine system was examined by determination of plasma level of thyroid hormones, thyroxin (T4) (n=269) and triidothyronin (T3) (n=269), stress hormone cortisol (n=266), and reproductive hormones such as estradiol (n=84) and progesterone (n=84) in cows (>3 years) reared around different polluted industrial and non-polluted areas.The respective blood lead and cadmium concentration was also determined in all the cows.The mean plasma levels of both T3 and T4 were significantly (P<0.05) higher around lead zinc smelter (2.43+/-0.26 and 41.1+/-2.9nmol/L) and closed lead cum operational zinc smelter (1.81+/-0.16 and 42.4+/-6.2nmol/L), where the mean blood lead level (0.86+/-0.06 and 0.51+/-0.09mug/ml) was also significantly higher than that of cows (0.07+/-0.01mug/ml) from unpolluted areas.Regression analysis of data from 269 cows revealed a significant (P<0.01) positive correlation between the blood lead and plasma T3 (r=0.287) and T4 (r=0.173).The correlation between thyroidal hormones and the blood cadmium concentration (r=-0.079 and -0.48; P>0.05) was not significant.Plasma cortisol level had also a non-significant (P>0.05) correlation (r=-0.092) with blood lead level.However, the mean cortisol level (4.02+/-1.96nmol/L) of cows in phosphate rock mining areas was significantly (P<0.05) higher than that of controls (1.98+/-0.70nmol/L). The mean plasma estradiol level was significantly (P<0.05) higher in cows around closed lead cum operational zinc smelter (47.1+/-19.5pg/ml) than that of the control animals (21.8+/-3.9pg/ml) and in rest of the areas, the difference did not reach the statistical significance (P>0.05).The serum biochemical analysis in 36 cows around lead-zinc smelter with the highest mean blood lead level (0.86+/-0.06mug/ml) amongst all the industrial/urban areas surveyed, and in 15 animals from non-polluted areas revealed a significant positive correlation between blood lead and serum ALT (alanine transaminase) (r=0.688, P<0.01) and AST (aspartate transaminase) (r=0.390, P<0.01) and a negative correlation with serum total lipids (r=-0.337, P<0.05), total protein (r=-0.449, P<0.01) and albumin(r=-0.662, P<0.01). It is concluded from the study that the natural exposure to lead in polluted environments disturbs the endocrine profile and the higher blood lead level alters serum biochemical parameters indicative of liver functions.     

Development of NILs from heterogeneous inbred families for validating the rust resistance QTL in peanut (Arachis hypogaea L.)

Heterogeneous inbred families segregating for rust resistance were identified from the two crosses involving susceptible (TAG 24 and TG 26) and resistant (GPBD 4) varieties of peanut. Rust‐resistant (less than score 5) and rust‐susceptible (more than score 5) plants were identified in each HIF and evaluated under rust epiphytotic conditions. The set of plants belonging to the same HIF, but differing significantly in rust resistance, not in other morphological and productivity traits, was regarded as near‐isogenic lines (NILs). Largely, rust‐resistant NILs had GPBD 4‐type allele, and susceptible NILs carried either TAG 24 or TG 26‐type allele at the three SSR loci (IPAHM103, GM1536 and GM2301) linked to a major genomic region governing rust resistance. Comparison of the remaining genomic regions between the NILs originating from each of the HIFs using transposon markers indicated a considerably high similarity of 86.4% and 83.1% in TAG 24 × GPPBD 4 and TG 26 × GPBD 4, respectively. These NILs are useful for fine mapping and expression analysis of rust resistance.     

Cross-amplification of EST-derived markers among 16 grass species

The availability of a large number of expressed sequence tags (ESTs) has facilitated the development of molecular markers in members of the grass family. As these markers are derived from coding sequences, cross-species amplification and transferability is higher than for markers designed from genomic DNA sequences. In this study, 919 EST-based primers developed from seven grass species were assessed for their amplification across a diverse panel of 16 grass species including cereal, turf and forage crops. Out of the 919 primers tested, 89 successfully amplified DNA from one or more species and 340 primers generated PCR amplicons from at least half of the species in the panel. Only 5.2% of the primers tested produced clear amplicons in all 16 species. The majority of the primers (66.9%) were developed from tall fescue and rice and these two species showed amplification rate of 41.6% and 19.0% across the panel, respectively. The highest amplification rate was found for conserved-intron scanning primers (CISP) developed from pearl millet (91%) and sorghum (75%) EST sequences that aligned to rice sequences. The primers with successful amplification identified in this study showed promise in other grass species as demonstrated in differentiating a set of 13 clones of reed canary grass, a species for which very little genomic research has been done. Sequences from the amplified PCR fragments indicated the potential for the transferable CISP markers for comparative mapping purposes. These primer sets can be immediately used for within and across species mapping and will be especially useful for minor grass species with few or no available molecular markers.     

Influence of Ovulation Status,Seasonality and Embryo Transfer Method on Development of Cloned Porcine Embryos

To improve pig cloning efficiency, the present study evaluated the effect of ovulation status, seasonality and embryo transfer (ET) method on in vivo development of cloned porcine embryos. Cloned embryos were transferred to surrogate mothers on the same day of somatic cell nuclear transfer. In pre‐ovulation stage (PO), pregnancy rate (PR) and delivery rate (DR) were 36.3% and 9.4%, respectively. In post‐ovulation stage, 22.7% PR and 2.1% DR were recorded (both PR and DR are significantly higher in PO). When ET was performed during winter (December–February), spring (March–May), summer (June–August) and autumn (September–November), the PRs were 13.4%, 37.3%, 24.6% and 51.0%, while DRs were 0%, 12.7%, 4.3% and 7.8%, respectively. The highest PRs were recorded in autumn groups. However, DRs were significantly lower in autumn (7.8%) group compared with spring (12.7%) group. The PR was the lowest and no piglets were born in winter group, which might be because of the effect of low temperature during ET. To overcome the low PR in winter group, 0.25 ml straws were used for ET to minimize exposure time of embryos to ambient temperature. The straw ET group showed significantly higher PR in the winter group (23. 9%) compared with the conventional catheter‐loading group (7.7%). We suggest that using PO recipient and ET in spring is the best condition for pig cloning. In addition, alternative method to reduce cold shock during ET in winter is necessary.     

Identification of quantitative trait loci for protein content, oil content and oil quality for groundnut (Arachis hypogaea L.)

Very few efforts have been made to improve the nutritional quality of groundnut, as biochemical estimation of quality traits is laborious and uneconomic; hence, it is difficult to improve them through traditional breeding alone. Identification of molecular markers for quality traits will have a great impact in molecular breeding. An attempt was made to identify microsatellite or simple sequence repeat (SSR) markers for important nutritional traits (protein content, oil content and oil quality in terms of oleic acid, linoleic acid and oleic/linoleic acid ratio) in a mapping population consisting of 146 recombinant inbreed lines (RILs) of a cross TG26 × GPBD4. Phenotyping data analysis for quality traits showed significant variation in the population and environment, genotype × environment interaction and high heritability was observed for all the traits. Negative correlation between protein content and oil content, oleic acid and linoleic acid indicated their antagonistic nature. After screening >1000 SSR markers, a partial genetic linkage map comprising of 45 SSR loci on 8 linkage groups with an average inter-marker distance of 14.62 cM was developed. QTL analysis based on single marker analysis (SMA) and composite interval mapping identified some candidate SSR markers associated with major QTLs as well as several minor QTLs for the nutritional traits. Validation of these major QTLs using a wider genetic background may provide the markers for molecular breeding for improving groundnut for nutritional traits.     

Genetic diversity for grain nutrients contents in a core collection of finger millet (Eleusine coracana (L.) Gaertn.) germplasm

Finger millet is a promising source of micronutrients and protein besides energy and can contribute to the alleviation of iron (Fe), zinc (Zn) and protein malnutrition affecting women and preschool children in African and south-east Asian countries. The most cost effective approach for mitigating micronutrient and protein malnutrition is to introduce staple crop cultivars selected and/or bred for Fe, Zn and protein dense grain. Breeding finger millet for enhanced grain nutrients is still in its infancy. Analysis, detection and exploitation of the existing variability among the germplasm accessions are the initial steps in breeding micronutrient and protein-dense finger millet cultivars. Evaluation of finger millet core collection for grain nutrients and agronomic traits revealed a substantial genetic variability for grain Fe, Zn, calcium (Ca) and protein contents. The accessions rich in nutrient contents were identified and their agronomic diversity assessed. The accessions rich in Zn content have significantly higher grain yield potential than those rich in Fe and protein content. Grain nutrient-specific accessions and those contrasting for nutrient contents were identified for use in the strategic research and cultivar development in finger millet.     

Genetic mapping and quantitative trait locus analysis of resistance to sterility mosaic disease in pigeonpea [Cajanus cajan (L.) Millsp.]

Sterility mosaic disease (SMD), considered as the “green plague of pigeonpea” and caused by pigeonpea sterility mosaic virus (PPSMV) is one of the major biotic factors, which leads to heavy yield losses and hence poses a big challenge for pigeonpea production in the Indian subcontinent. Variability in the sterility mosaic pathogen revealed the occurrence of five different isolates in India. Among them, three distinct SMD isolates have been characterized, viz., Patancheru, Bangalore and Coimbatore. Molecular tools offer a viable option to tackle these biotic stresses via identification of the genomic regions associated with the trait such as SMD resistance. With an aim of identifying the gene(s)/QTLs linked with SMD resistance, two F₂ populations, i.e. ICP 8863 × ICPL 20097 (segregating for Patancheru SMD isolate) and TTB 7 × ICP 7035 (segregating for both Patancheru and Bangalore SMD isolates) were developed and F_2:3 families were phenotyped for resistance to respective isolate(s) of SMD. After screening over 3000 SSR markers on parental genotypes of each mapping population, intra-specific genetic maps comprising of 11 linkage groups and 120 and 78 SSR loci were developed for ICP 8863 × ICPL 20097 and TTB 7 × ICP 7035 populations, respectively. Composite interval mapping (CIM) based QTL analysis by using genetic mapping and phenotyping data provided four QTLs for Patancheru SMD isolate and two QTLs for Bangalore SMD isolate. Identification of different QTLs for resistance to Patancheru and Bangalore SMD isolates is an indication of involvement of different genes conferring the resistance to these two SMD isolates. One QTL namely qSMD4 identified within an interval of 2.8 cM on LG 7 explaining 24.72% of phenotypic variance, once it is validated in other genetic background, seems to be a promising QTL for use in marker assisted selection. In summary, this is the first study on development of intra-specific genetic maps and identification of QTLs for SMD resistance in pigeonpea.