Effect of Physiologically Relevant Heat Shock on Development,Apoptosis and Expression of Some Genes in Buffalo (Bubalus bubalis) Embryos Produced In Vitro

For investigating the effects of physiologically relevant heat shock, buffalo oocytes/embryos were cultured at 38.5°C (control) or were exposed to 39.5°C (Group II) or 40.5°C (Group III) for 2 h once every day throughout in vitro maturation (IVM), fertilization (IVF) and culture (IVC). Percentage of oocytes that developed to 8‐cell, 16‐cell or blastocyst stage was lower (p < 0.05) and the number of apoptotic nuclei was higher (p < 0.05) for Group III > Group II > controls. At both 8–16‐cell and blastocyst stages, relative mRNA abundance of stress‐related genes HSP 70.1 and HSP 70.2 and pro‐apoptotic genes CASPASE‐3, BID and BAX was higher (p < 0.05) in Groups III and II than that in controls with the exception of stress‐related gene HSF1. Expression level of anti‐apoptotic genes BCL‐XL and MCL‐1 was also higher (p < 0.05) in Groups III and II than that in controls at both 8–16‐cell and blastocyst stages. Among the genes related to embryonic development, at 8–16‐cell stage, the expression level of GDF9 was higher (p < 0.05) in Group III than that in controls, whereas that of GLUT1, ZAR1 and BMP15 was not significantly different among the three groups. At the blastocyst stage, relative mRNA abundance of GLUT1 and GDF9 was higher (p < 0.05) in Group II than that in controls, whereas that of ZAR‐1 and BMP15 was not affected. The results of this study demonstrate that exposure of buffalo oocytes and embryos to elevated temperatures for duration of time that is physiologically relevant severely compromises their developmental competence, increases apoptosis and affects stress‐, apoptosis‐ and development‐related genes.     

Buffalo (Bubalus bubalis) ES Cell–Like Cells are Capable of In Vitro Skeletal Myogenic Differentiation

When buffalo embryonic stem (ES) cell–like cells that expressed surface markers SSEA‐4, TRA‐1‐60, TRA‐1‐81, CD9 and CD90 and intracellular markers OCT4, SOX2 and FOXD3, as shown by immunofluorescence, and that expressed REX‐1 and NUCLEOSTEMIN as confirmed by RT‐PCR, were subjected to suspension culture in hanging drops in absence of LIF and buffalo foetal fibroblast feeder layer support, they differentiated to form three‐dimensional embryoid bodies (EBs). Of 231 EBs examined on Day 3 of suspension culture, 141 (61.3 ± 3.09%) were of compact type, whereas 90 (38.4 ± 3.12%) were of cystic type. The cells obtained from EBs were found to express NF‐68 and NESTIN (ectodermal lineage), BMP‐4 and α‐skeletal actin (mesodermal lineage), and α‐fetoprotein, GATA‐4 and HNF‐4 (endodermal lineage). When these EBs were cultured on gelatin‐coated dishes, they spontaneously differentiated to several cell types such as epithelial‐ and neuron‐like cells. When EBs were cultured in the presence of 1 or 2% DMSO or 10^?8 m or 10^?7 m retinoic acid for 25 days, ES cells could be directed to form muscle cell–like cells, the identity of which was confirmed by expression of α‐actinin by immunofluorescence and of MYF‐5, MYOD and MYOGENIN genes by RT‐PCR. MYOD was first detected on Day 10 in both treatment groups and on Day 15 in controls, whereas MYOGENIN was first detected on Day 10, Day 15 and Day 25 in the presence of retinoic acid, in the presence of DMSO and in controls, respectively. The present study demonstrates the ability of buffalo ES cell–like cells to undergo directed differentiation to cells of skeletal myogenic lineage.     

Microbial and chemical sources of phosphorus supply modulate the yield and chemical composition of essential oil of rose-scented geranium (Pelargonium species) in sodic soils

Rose-scented geranium (Pelargonium sp.) is a highly valued aromatic crop. Its growth is limited by soil salinity and sodicity stress. Arbuscular mycorrhizal (AM) fungus, phosphate-solubilizing bacteria (PSB), and P fertilizers may enhance the growth and secondary metabolism in geranium plants. In this context, a pot experiment was conducted to study the effects of PSB, AM fungi (Glomus intraradices), and P fertilizer on the yield, chemical composition of essential oil, and mineral element acquisition of geranium. The dry matter yield of shoot and essential oil yield, and mineral element (P, K, Ca, Mg, Na, Fe, Cu, and Zn) uptake in shoot tissues of geranium were significantly increased by the inoculation with AM fungi, co-inoculation with AM fungi and PSB, and P fertilization as compared to control. While the co-inoculation of geranium with AM fungi and PSB significantly enhanced the content of the monoterpenes such as citronellol, geraniol, geranial, and a sesquiterpene (10-epi-γ eudesmol), the P fertilization only enhanced the content of a sesquiterpene, 10-epi-γ eudesmol in the volatile oil. We conclude that the co-inoculation of PSB and AM fungi could be the best natural alternative to phosphate fertilizers to enhance the yield and quality of essential oil from geranium plants grown in sodic soils.     

Effect of Biogas Digested Liquid on CH4 and N2O Flux in Paddy Ecosystem

Biogas production generates digested slurry as a byproduct. It can be used as a fertilizer especially after its conversion into digested liquid. A pot based study was conducted in order to evaluate the effect of the application of digested liquid on CH₄ and N₂O flux, and plant biomass in paddy. Analysis revealed that digested liquid treated soils released more CH₄ compared to ammonium sulphate and the control. Ammonium sulphate treated soil emitted the highest N₂O whereas digested liquid application decreased its emission significantly. Further, the cumulative emission over 101 d of the experiment was found to be higher for CH₄ (16.9 to 29.9 g m⁻²) compared to N₂O (−49.3 to 18.9 mg m⁻²) for all treatments. Digested liquid application had positive impact on plant variables such as panicle number and weight of panicles. This study suggests that digested liquid application significantly decrease N₂O emission and increase CH₄ emission possibly due to affecting the availability of organic C in the soil to microbial activity for methanogenesis. Another possibility for enhancing CH₄ emission by following biogas digested liquid could be attributed to the increase in plant biomass.     

Immune responses to haemorrhagic septicaemia (HS) vaccination in <Emphasis Type="Italic">Trypanosoma evansi</Emphasis> infected buffalo-calves

To assess the immunosuppressive effect of Trypanosoma evansi infection in buffalo-calves on immune responses to heterologous antigen, the study was planned to examine the responses of haemorrhagic septicaemia vaccination in simultaneously and previously (80 days before vaccination) T. evansi-infected buffalo-calves. Eight buffalo-calves were divided into three groups. Buffalo-calves of group A (n = 3) were previously (80 days before primary vaccination with haemorrhagic septicaemia [HS] vaccine) infected with T. evansi (1 × 10⁷ tryps.calf⁻¹; sc) and that of group B (n = 3) were infected with T. evansi (1 × 10⁷ tryps.calf⁻¹; sc) on the day of primary vaccination with HS vaccine. Two healthy uninfected control calves given only HS vaccine were kept in group C. All the buffalo-calves were given a booster dose of vaccine 21 days post-primary vaccination (PPV). Twenty eight days PPV, animals of group A were given trypanocidal quinapyramine prosalt at 6.66 mg kg⁻¹. Immunosuppressive effect of T. evansi infection was evident from day 7 PPV with HS vaccine. The effect was more pronounced in previously T. evansi-infected buffalo-calves as compared with simultaneously infected buffalo-calves. Group A buffalo-calves appeared to have recovered from the immunosuppressive effect after 28 days post-trypanocidal treatment as observed by humoral and cell-mediated immune responses. Immunosuppressive effect to HS vaccination was observed in T. evansi-infected buffalo-calves, and trypanocidal therapy enabled the calves to mount the responses similar to uninfected controls.     

Assessment of DNA Damage during In Vitro Development of Buffalo (Bubalus bubalis) Embryos: Effect of Cysteamine

Comet assay was used in the present study to examine DNA damage to buffalo oocytes and embryos during in vitro culture. Embryos were produced in vitro from oocytes obtained from slaughterhouse ovaries in presence of cysteamine (IVM and IVC media supplemented with 50 and 100 μm , respectively) or in its absence (controls). Compared to controls, cysteamine supplementation increased (p < 0.01) cleavage rate and proportion of oocytes that developed to 8‐ to 16‐cell stage. The incidence of DNA damage was lower (p < 0.01) in cysteamine group than that in controls at 8‐ to 16‐ (19.3 ± 4.24 vs 72.0 ± 5.22%) but not in 2‐cell stage embryos (11.7 ± 5.63 vs 20.8 ± 5.49%) or in mature oocytes (5.3 ± 3.43 vs 10.3 ± 4.73%). The tail length, which indicates magnitude of DNA damage, was shorter (p < 0.01) in cysteamine group than in controls in mature oocytes (25.5 ± 0.5 vs 36.0 ± 0.71 pixels) and 8‐ to 16‐cell stage (49.2 ± 1.64 vs 152.7 ± 1.28 pixels) but not in 2‐cell stage embryos (36.3 ± 1.54 vs 36.4 ± 0.75 pixels). Also, exposure of oocytes/embryos to UV radiation or H₂O₂ caused extensive DNA damage. In conclusion, these results suggest that oocytes/embryos suffer from DNA damage during progress of in vitro culture, which can be partly ameliorated by cysteamine supplementation of culture media.     

Influence of soil variability on single and competitive interaction of ammonium and potassium: experimental study on seven different soils

Journal of Soils and Sediments - Ammonium and potassium are nutrients that generally co-exist together in soils in municipal wasteland. The main objective of this study is to investigate the...     

Novel quantitative trait loci for yield and yield related traits identified in Basmati rice (Oryza sativa)

Increasing crop productivity is one of the prime goals of crop breeding research. Rice grain yield is a complex quantitative trait governed by polygenes. Although several QTLs governing grain yield traits have been reported and limited attempts have been made to map QTLs for grain yield parameters in Basmati rice. A population from the cross Sonasal and Pusa Basmati 1121 comprising 352 RILs was generated through the single seed descent method. A total of 12 QTLs governing yield and yield-related traits were mapped on six chromosomes, namely, 1, 2, 3, 7, 8 and 9, of which five QTLs were novel. We identified a novel and robust epistatic QTL (qPH1.1 and qPL1.1) governing plant height and panicle length, flanked by the markers RM5336-RM1 on chromosome 1. The gene encoding brassinosteroid insensitive 1-associated receptor kinase 1 precursor is the putative candidate gene underlying this epistatic QTL. Another novel QTL, qNT3.1, governing tiller number was bracketed to a region of .77 Mb between the markers RM15247 and RM15281 on chromosome 3. Of the 57 annotated gene models, Os03g0437600 encoding alpha/beta-fold hydrolase, a homologous to AtKai2 is a putative candidate gene underlying the novel QTL qNT3.1. The other QTLs such as qDFF1.1 governing days to 50% flowering co-localizes with the gene Ghd7, QTL for plant height qPH1.2 co-localizes with the gene sd1, the QTLs for panicle length co-localizes with FUWA and DEP2, the QTL for tiller number co-localizes with OsRLCK57 and QTLs for thousand-grain weight co-localize with the major gene GS3. The QTLs identified in the current study can be effectively used in marker-assisted selection for developing Basmati rice varieties with a higher yield.     

Effect of biochar on CH4 and N2O emission from soils vegetated with paddy

Biochar is believed to have positive impact on soil properties and plant yield. Due to the presence of C, it can also enhance CH₄ emission in paddy soils. On the other hand, ammonium sulphate can decrease CH₄ emission due to negative impacts on methanogenesis. Keeping these points in view, a pot experiment was conducted to determine the effect of biochar along with ammonium sulphate on CH₄ and N₂O emission from paddy soil. Analysis revealed that biochar treated soils released more CH₄ compared to untreated. Ammonium sulphate treated soil emitted the highest N₂O whereas biochar addition decreased its emission significantly. Further, total emission was found to be higher for CH₄ (16.9–34.7 g/m²) in comparison to N₂O (?0.05 to 0.02 g/m²) for all treatments. Biochar application has positive impact on plant variables such as panicle number and weight of panicles. This study suggests that biochar application significantly decrease N₂O emission and increase CH₄ emission possibly due to affecting the availability of organic C in the soil to microbial activity for methanogenesis. Another possibility for enhancing CH₄ emission by following biochar could be attributed to the increase in plant biomass.     

Estimates of covariance components and genetic parameters for growth,average daily gain and Kleiber ratio in Harnali sheep

Tropical Animal Health and Production - The aim of the present study was to estimate the (co)variance components and genetic parameters for various growth traits (weight at birth (BWT) and 3 (WT3),...