Purification and development of ELISAs for two forms of vitellogenin in Indian walking catfish, <Emphasis Type="Italic">Clarias batrachus</Emphasis> (L.)

Two forms of vitellogenin (Vg: Vg1 and Vg2) were purified from the plasma of estradiol-17β (E₂)-treated Indian walking catfish, Clarias batrachus, by gel filtration and adsorption chromatography. Native Vg1 and Vg2 had apparent molecular masses of 375 and 450 kDa, respectively, and both Vgs resolved into two similar major bands (95 and 67 kDa) in SDS-PAGE under reducing condition. Polyclonal antisera raised against each form of Vg were absorbed with a combination of hypophysectomized male catfish serum proteins and alternate Vg to ensure specificity. Immunological analyses verified the presence of Vg1 and Vg2 in the plasma of female catfish. Homologous ELISAs were developed for Vg1 and Vg2 using their respective harvested antisera, which exhibited the detection limit of 100 ng ml^?1 for Vg1 and 40 ng ml^?1 for Vg2, and low level of cross-reactivity (not parallel to the standard) was found with alternate Vg in each assay. Treatment of male catfish with E₂ induced both Vgs showing a proportionate ratio of Vg1 to Vg2 at 5.6:1. Plasma concentrations of both Vgs measured by ELISAs at different reproductive phases of field collected female catfish increased in accordance with the ovarian development, keeping the proportionate ratio of Vg1 to Vg2 at about 2:1 in fish undergoing vitellogenesis during prespawning period and 1:20 during spawning period, suggesting that Vg1 may be the major Vg to contribute in yolk formation, whereas Vg2, besides its role in yolk formation, may facilitate other physiological functions. The present study, thus, demonstrates the occurrence of two unequally synthesized Vgs in the catfish.     

Soil and geologic formations as antidotes for CO2 sequestration?

Rapid and far-reaching transitions are required to combat climate change and its impacts. Carbon capture and storage within mineral deposits is a promising solution to remove CO2 from the atmosphere. In-situ geological storage and ex-situ mineral sequestration are practically sufficient for sequestering all the anthropogenic CO2. Recent research reports that more than 95% of injected CO2 was mineralized into carbonates in two years by using in-situ geological approach, and mining wastes and secondary minerals were recycled as resources for ex-situ CO2 sequestration. However, geological activity is the major risk of in-situ storage, while high energy consumption and associated cost may limit the application of ex-situ carbonation. Significant technical breakthroughs of mineral and geological CO2 sequestration are therefore of vital importance to realize a “net-zero CO2 emissions” and even “carbon-negative” society.     

Observations on the epidemiology of bovine cryptosporidiosis in India

The significance of Cryptosporidium as a causative agent of diarrhea has been assessed in bovine for a period of 2 years. A total of 940 faecal samples (470 samples in each year) both from diarrhoeic and non-diarrhoeic bovine (0-12 months age) were examined during three different seasons (rainy season, summer and winter). Overall Cryptosporidium was detected in 17.46% and 18.04% cases in first and second year, respectively. Out of 50.21% diarrhoeic and 49.79% non-diarrhoeic cases Cryptosporidium was detected in 26.79% and 8.13% in first year and 27.49% and 8.59% in second year. Year did not have any significant effect on the occurrence of cryptosporidiosis in bovine during this study period. The prevalence of cryptosporidiosis, both in diarrhoeic (61.64%) and non-diarrhoeic (47.22%) cases was highest in 0-1-month age group (P<0.01). Such a high percentage of cryptosporidiosis in clinically asymptomatic animals indicated that the particular age group of animals might be reservoir for the parasite. During this study period highest prevalence was recorded in rainy season (27.55%) followed by summer (16.99%) and winter (8.71%) (P<0.01). A total of 166 positive cases were genotyped. Molecular characterization of bovine cryptosporidiosis has been carried out by PCR-RFLP analysis of SSU rRNA gene and results indicated that Cryptosporidium parvum mainly responsible for diarrhea in bovine in India.     

QTL identification for molecular breeding of fibre yield and fibre quality traits in jute

In jute (Corchorus olitorius), quantitative trait loci (QTL) analysis was conducted to study the genetics of eight fibre yield traits and two fibre quality traits. For this purpose, we used a mapping population consisting of 120 recombinant inbred lines (RILs) and also used a linkage map consisting of 36 SSR markers that was developed by us earlier (Das et al. 2011). The RIL population was derived from the cross JRO 524 (coarse fibre) × PPO4 (fine fibre) following single seed descent. Using single-locus analysis involving composite interval mapping, a total of 21 QTLs were identified for eight fibre yield traits whereas for fibre quality (fibre fineness), only one QTL was detected. The QTL for fibre fineness explained 8.31–10.56% of the phenotypic variation and was detected in two out of three environments. Using two-locus analysis involving QTLNetwork, as many as 11 M-QTLs were identified for seven fibre yield traits (excluding top diameter) and one M-QTL was identified for fibre fineness which accounted for 4.57% of the phenotypic variation. For six fibre yield traits, we detected 16 E-QTLs involved in nine QQ epistatic interactions. For fibre fineness, four E-QTLs involved in two QQ epistatic interactions and for fibre strength, six E-QTLs involved in three QQ epistatic interactions were identified. Eight out of the 11 M-QTLs observed for the fibre yield traits were also involved in QE interactions; for fibre fineness and fibre strength, no QE interactions were observed.     

Characterization of shiga toxin producing (STEC) and enteropathogenic Escherichia coli (EPEC) in raw yak (Poephagus grunniens) milk and milk products

Thirty-one shiga toxin-producing (STEC) and 6 enteropathogenic Escherichia coli (EPEC) were isolated from 87 raw yak milk and 63 'churpi' samples. Of 18 stx(1) positive isolates (48.6%), 14 carried stx(1c) (77.7%). Subtyping of 28 stx(2) positive isolates (75.7%) revealed the presence of stx(2c) (9, 32.1%), stx(2d) (3, 10.7%), stx(2e) (1, 3.57%) and stx(2f) (3, 10.7%) variants. Furthermore, intimin (eaeA), enterohaemolysin (ehxA), autoagglutinating adhesin (saa), iha (adherence conferring protein), efa1 (EHEC factor for adherence), bundle forming pilli (bfpA) and toxB (type III secreted protein encoded on LEE Island, similar to toxin B of Clostridium difficile) genes were detected in 14, 16, 12, 4, 3, 2 and 2 isolates, respectively. Univariate and multivariate analysis depicted that both stx(1) and stx(2) or their variants were more likely to occur in isolates from Arunachal Pradesh (p<0.04) rather than Sikkim. Dendogram constructed on the basis of RAPD and ERIC PCR profile distributed the STEC and EPEC isolates in separate clusters irrespective of their sources and serotypes. The STEC and EPEC isolates exhibited resistance against erythromycin, amikacin, azithromycin, amoxicillin, ampicillin+cloxacillin, cephalothin, furazolidone, gentamicin, kanamycin, streptomycin and tetracycline. This is the first ever report on occurrence and characterization of STEC and EPEC isolated from yak milk and milk products.     

Synthesis and properties of β-d-glucosidasephenolic copolymers as analogues of soil humic-enzyme complexes

Several β-d-glucosidase-phenolic copolymers were synthesized and three examined in detail: those containing l-tyrosine, pyrogallol and resorcinol. These copolymers were similar to naturallyoccurring soil humic-enzyme complexes in many ways: E₄/E₆ ratios, C, H, N and S content and IR spectra. The enzyme activity of the copolymers showed varying degrees of resistance to proteolysis, organic solvents, and storage at high temperatures. All immobilized enzymes had increased K_m values and decreased V_max values in comparison with soluble β-d-glucosidase (9.3 mM, 190μmol p-nitrophenol mg^?1h^?1); the β-d-glucosidase-resorcinol copolymer was the most active (10.5 mM, 104μmol p-nitrophenol mg^?1h^?1). β-d-Glucosidase activity was completely resistant to protease when the copolymer was fixed to bentonite clay but V_max values were reduced further (e.g. β-d-glucosidase-resorcinol-bentonite complex, 58.5μmol p-nitrophenol mg^?1 h^?1). On addition to soil, soluble β-d-glucosidase was rapidly inactivated (38% loss in 3 days, 80% loss in 21 days) whereas β-d-glucosidase-resorcinol/pyrogallol and β-d-glucosidase-L-tyrosine copolymers were comparatively stable (no loss in 9 days, 25% and 44% loss in 21 days). It is suggested that the copolymerization of enzyme during humic matter formation is a major factor leading to the stabilisation of soil enzymes and that adsorption and entrapment are comparatively insignificant.     

Inland saline aquaculture increased carbon accumulation rate and stability in pond sediments under semi-arid climate

Purpose

Similar to fresh- and brackish water aquaculture ponds, commercial shrimp farming in degraded saline areas holds the potential to bury carbon (C) in the sediments. However, studies on the mechanisms of sediment C dynamics and C-flux in response to inland saline aquaculture management practices are still scarce. Therefore, the objectives of the present study are to quantify the C burial rate in inland saline aquaculture ponds and assess the impact of inland saline aquaculture on sensitive C fractions in the bottom sediment of the ponds.

Materials and methods

The sediment samples (n?=?12 from each pond) were collected from six shrimp farming ponds (1000 m² area of each pond) of different ages. The sediment depth, sediment accumulation rate and the levels of total carbon (TC), total organic carbon (TOC) and sediment oxidizable organic carbon (SOC) and its different fractions were determined using standard procedures. The data were analysed by one-way analysis of variance (ANOVA), followed by the Duncan's multiple range test for comparing the means, and the Pearson correlation test was used to assess the relationship between the different pond sediment parameters and SOC content.

Results and discussion

The results revealed that the annual C accumulation rates varied from 902 to 1346 kg C ha^?1 year^?1 in 7-year-old earthen ponds (EPs) and bottom cemented ponds (BCPs), respectively. The sediment C fractions, including TC, TOC, SOC and its fractions (very labile, VLc; labile, Lc; less labile, LLc), and non-labile carbon (NLc)) were progressively increased over the pond age. The inland saline aquaculture practices over the years increased both active (AC) and passive carbon (PC) pools in the pond sediments, helped in the restoration and improvement of sediment quality and enhanced C sequestration potential of the sediments. Furthermore, a significant increase in the level of particulate organic carbon (POC) in BCPs justified that the non-ploughing practices at BCPs facilitated the formation of macro- and micro-aggregates, thereby increasing the C retention and stability of the pond sediments.

Conclusion

This study suggested that the shrimp farming ponds in semi-arid saline soils represented considerable C burial hotspots, enhanced the stable passive C pools and improved the sediment quality.

     

Optimum plant stand and nutrient doses for summer groundnut under check basin irrigation and drip fertigation in light black soils of peninsular Western India

Abstract

A field experiment was conducted at Research Farm of ICAR-Directorate of Groundnut Research, Junagadh for consecutive three summer seasons of 2013, 2014, and 2015 with the objectives of identifying optimum plant density and nutrient doses under check basin irrigation and drip fertigation for higher productivity and net returns. The treatments were; three plant densities viz., 3,33,333 plants/ha (100% of recommended plant density; P1); 4,16,666 plants/ha (125% of recommended plant density; P2), and 4,99,999 plants/ha (150% of recommended plant density;P3) in main plots, and three nutrient doses viz., 18.75–37.5–22.5 NPK kg ha^?1 (75% of recommended nutrient dose; F1), 25-50-30 NPK kg ha^?1 (100% of recommended nutrient dose; F2), and 31.25–62.5–37.5 NPK kg ha^?1 (125% of recommended nutrient dose; F3) in sub-plots, and replicated thrice. The same sets of treatments were tested under both check basin irrigation and drip fertigation. The data were analyzed using split plot design. Pod yield, haulm yield, and net returns were significantly higher with P3 as compared to P1 under check basin irrigation but only haulm yield was found significantly higher with P3 under drip fertigation. Under check basin irrigation, NH₄–N, NO₃–N, and available P and K in soil were found in the order P1?>?P2?>?P3 (p?<?0.05) while in case of drip fertigation, differences were significant only for available K which was significantly higher in P1 over both P2 and P3. Under check basin irrigation, F2 i.e., application of 100 percent of recommended nutrient doses, being at par with F3, significantly improved pod yield, haulm yield and net returns over that with F1 however, differences were not significant under drip fertigation. NH₄–N, NO₃–N and available P and K in soil under both the irrigation systems were in the order F3?>?F2?>?F1 (p?<?0.05).