Antimicrobial activity of partially characterized analytes from Bacillus pumilus(B2)

This study was carried out to characterize the antimicrobial substance produced by the strain of Bacillus pumilus (B2) obtained from Novozymes Biologicals Inc. to compare its antimicrobial activity by agar well diffusion assay and bacteriocin activity assay via critical dilution method against seven different strains of Vibrio spp., specifically V. alginolyticus (A01), V. cholerae (C01), V. fluvialis (F01, F02), V herveyii (H), V. mimicus (M01), V. parahaemolyticus (P01) and V. vulnificus (V01, V02). All Vibrio spp. were isolated from the hemolymph and intestine of the white faeces disease‐infected moribund pacific white‐leg shrimp Litopenaeus vannamei (Boone 1931) and one strain (V. harveyi) from its diseased postlarva. The cell‐free neutralized supernatant (CFNS) of B2 showed moderate thermo‐stability being stable up to 70°C for 60 min with, however, reducing activity above 80°C for 20 min. B2 antimicrobials showed a stable activity within the pH ranging from 6 to 10 at room temperature and at 4°C, while residual antimicrobial activity of crude CFNS showed tolerance to salinity up to 7% of sodium chloride below 4°C. No B2 activity was obtained while exposed to proteolytic enzyme, such as proteinase k and pepsin, while its activity kept stable exposed to lipase. Initial B2 characterization for antimicrobial substance in CFNS revealed proteinaceous in nature owing to activity loss against proteolytic enzymes and no lipid moiety activity against lipase, which could be categorized as bacteriocin‐like inhibitory substance having potential application against several strains of Vibrio spp. in aquaculture.     

Ultrastructural demonstration of cilia and ciliary rootlets in mammalian uterine tube epithelium in different functional states.

Tissue from the infundibulum region of the uterine tube (oviduct) of guinea pigs, cattle, sheep, and swine was examined by electron microscopy. In all specimens, cilia and ciliary rootlets were present in variable numbers in the ciliated cells during both the follicular and the luteal phases of the estrous cycle. True degeneration of cilia was not evident during luteal phase or pregnancy. The ciliary rootlets in ruminant and nonruminant species had structural similarities to those described in people and rhesus monkeys. These organelles measured approximately 1 mum long in most species studied, but in the cow, the rootlets, extending downward into the cytoplasm from the proximal end of the basal body, reached a length of 2 mum. The rootlets had a cross-striation of thick and thin bands, the period measuring 40 to 60 nm. Mitochondria were closely associated with the rootlets. The rootlets usually formed a small angle to the axis of the cilium. These organelles probably function as anchoring or stabilizing structures for the motile cilia. Results indicated that the rootlets are more widely distributed in mammalian uterine tube cilia than previously postulated. Polyribosomes, microfilaments, microtubules, and electron-opaque fibrous granules were frequently seen in the cytoplasm of the ciliated cells. The presence of fibrous granules in close association with the basal bodies indicate that these granules have a role in the development of cilia and rootlets. Cilia and precursor fibrous granules were also seen in porcine fetal uterine tube epithelial cells.     

Effect of Nutrient Application on Growth,Metabolic and Enzymatic Activities of Rice Seedlings During Flooding Stress and Subsequent Re‐Aeration

The impact of submergence on the allometry, changes in metabolic activities and antioxidant enzymes during oxidative stress in four Indica rice varieties namely IR‐20, IR‐64 Sub1, Swarna Sub1 and Savitri Sub1 was studied. The differential response of flooding under clear and turbid water with different nutrient application schedules was also examined during and after 12 days of complete submergence. Submergence substantially reduced allometric parameters and the activities of antioxidant enzymes but increased the % change in chlorophyll, soluble sugars and malondialdehyde (MDA) across cultivars with drastic effects on IR‐20. Turbid water resulted in higher leaf senescence, lodging, higher depletion of chlorophyll and soluble sugars because of poor light transmission. Pre‐submergence N application resulted in higher lodging, leaf senescence and higher MDA level but depletion of chlorophyll and NSC leading to higher % change over prior to submergence. Basal P application reduced the senescence and lodging, whereas increased the antioxidant enzyme activities. Foliar spray of post‐submergence N with basal P improved the retention and regain of chlorophyll, soluble sugar and increased the dry matter, leaf area and root shoot ratio. Crop establishment could therefore be enhanced in areas where untimely flooding is anticipated by applying basal P and foliar spray of urea after desubmergence.     

Bioactivity of antioxidants in extruded products prepared from purple potato and dry pea flours

Measuring antioxidant activity using a biologically relevant assay adds important evidence to aid in understanding the role of phytochemicals based on data from in vivo and chemical assays of extrusion processed purple potato and pea flours. A cellular antioxidant activity assay could provide biologically relevant information on bioactive compounds in raw as well as processed food products. The objective of this study was to investigate the complete phytochemical profiles, antioxidant activity, cellular antioxidant activity, and their contribution to bioactivity in purple potato flour, dry pea flour, raw formulations, and extrusion cooked products prepared with the above ingredients. The free fraction of extracts contributed 68, 64, and 88% to total phenolics, total antioxidant activity (ORAC value), and total flavonoids, respectively, in purple potato flour (PPF). Similarly, extracts in the free fraction contributed 87, 86, and 64% to total phenolics, total antioxidant activity (ORAC value), and total flavonoids, respectively, in dry pea flour (DPF). The amount of total phenolics and total flavonoids in purple potato flour and the antioxidant activity of PPF and DPF were comparable to published data. However, a higher amount in the total flavonoids and lower in the total phenolics of DPF were observed. Caffeic, p-coumaric, and ferulic acids were mostly observed in the bound extracts of raw formulations as in the extrudates, whereas chlorogenic acid was predominant in the free extracts. The extruded products had significantly higher (p < 0.05) content of total phenolics, ORAC antioxidant activity, and flavonoids, compared to the raw formulations. Extrusion processing increased the cellular antioxidant activity of the extrudates prepared from 35:65 and 50:50 PPF/DPF (w/w) of ingredients compared with control raw formulations in a dose-dependent manner. Increase of PPF significantly increased (p < 0.05) the cellular antioxidant activity of 35-50% PPF formulations.     

Potassium application reduces methane emission from a flooded field planted to rice

In a field study, potassium (K) applied as muriate of potash (MOP) significantly reduced methane (CH₄) emission from a flooded alluvial soil planted to rice. Cumulative emission was highest in control plots (125.34 kg CH₄ ha⁻¹), while the lowest emission was recorded in field plots receiving 30 kg K ha⁻¹ (63.81 kg CH₄ ha⁻¹), with a 49% reduction in CH₄ emission. Potassium application prevented a drop in the redox potential and reduced the contents of active reducing substances and Fe²⁺ content in the rhizosphere soil. Potassium amendment also inhibited methanogenic bacteria and stimulated methanotrophic bacterial population. Results suggest that, apart form producing higher plant biomass (both above- and underground) and grain yield, K amendment can effectively reduce CH₄ emission from flooded soil and could be developed into an effective mitigation option, especially in K-deficient soils.     

Long‐term effects of fertilizer and manure applications on soil quality and yields in a sub‐humid tropical rice‐rice system

Widespread yield stagnation and productivity declines in the rice–rice cropping system have been reported and many of the associated issues are related to soil quality. A long‐term experimental study was initiated in 1969 to assess the impact of continuous cultivation of rice as a single crop grown in wet as well as dry seasons using varying levels of chemical fertilizer and manure applications on soil quality indicators (physical, chemical and biological), a sustainable yield index (SYI) and a soil quality index (SQI). The treatments comprised chemical fertilizers and farmyard manure (FYM) either alone or in combination viz. control, N, NP, NK, NPK, FYM, N+FYM, NP+FYM, NK+FYM and NPK+FYM, laid out in a randomized complete block design with three replications. Soil samples were collected after the wet season rice harvest in 2010 and were analysed for physical, chemical and biological indicators of soil quality. A SYI based on long‐term yield data and SQI using principal component analysis (PCA) and nonlinear scoring functions were calculated. Application of NPK fertilizers in combination with FYM significantly increased the average grain yield of rice in both wet and dry seasons and enhanced the sustainability of the system compared to the control and plots in receipt of fertilizers. The SYI for the control was higher in the wet season than in the dry one, whereas the reverse was true for NPK+FYM treatment. The value of the dimensionless SQI varied from 1.46 in the control plot to 3.78 in the NPK+FYM one. A greater SYI and SQI in the NPK+FYM treatment demonstrated the importance of using a chemical fertilizer in combination with FYM. For the six soil quality indicators selected as a minimum data set (MDS), the contribution of DTPA‐Zn, available‐N and soil organic carbon to the SQI was substantial ranging from 59.4 to 85.7 per cent in NPK+FYM and control plots, respectively. Thus, these soil parameters could be used to monitor soil quality in a subhumid tropical rice–rice system.     

Development of an in vitro oviduct epithelial explants model for studying sperm–oviduct binding in the buffalo

In this study, we developed an in vitro model for studying sperm–oviduct binding in the buffalo. Oviduct explants were prepared by overnight culture of epithelial cells in TCM‐199 medium under 5% CO₂ at 38.5 °C. Cryopreserved spermatozoa from buffalo bulls (n = 4) were incubated with the oviduct explants, and the sperm–oviduct explants complex was stained with JC‐1. The effect of sperm concentration (2, 3 and 4 million), size of the oviduct explants (<0.2, 0.2–0.3, 0.3–0.4 and >0.4 mm²) and time of incubation (1 hr and 4 hr) on binding index (BI—number of sperm bound to unit area of explants) was studied. No significant difference was observed in the BI among <0.2, 0.2–0.3 and 0.3–0.4 mm² size of explants; however, the BI decreased significantly (p < .05) when the size of explants exceeded 0.4 mm². The BI decreased significantly (p < .05) when the sperm concentration was increased to 4 million, while the duration of incubation did not have any significant effect on the BI. The interaction of bulls with explants size, sperm concentration and incubation time was not significant. The developed assay has the potential to be used as an in vitro model for studying sperm–oviduct binding in the buffalo.     

Genetic Dissection of Grain Size Traits Through Genome-Wide Association Study Based on Genic Markers in Rice

Grain size plays a significant role in rice, starting from affecting yield to consumer preference, which is the driving force for deep investigation and improvement of grain size characters. Quantitative inheritance makes these traits complex to breed on account of several alleles contributing to the complete trait expression. We employed genome-wide association study in an association panel of 88 rice genotypes using 142 new candidate gene based SSR (cgSSR) markers, derived from yield-related candidate genes, with the efficient mixed-model association coupled mixed linear model for dissecting complete genetic control of grain size traits. A total of 10 significant associations were identified for four grain size-related characters (grain weight, grain length, grain width, and length-width ratio). Among the identified associations, seven marker trait associations explain more than 10% of the phenotypic variation, indicating major putative QTLs for respective traits. The allelic variations at genes OsBC1L4, SHO1 and OsD2 showed association between 1000-grain weight and grain width, 1000-grain weight and grain length, and grain width and length-width ratio, respectively. The cgSSR markers, associated with corresponding traits, can be utilized for direct allelic selection, while other significantly associated cgSSRs may be utilized for allelic accumulation in the breeding programs or grain size improvement. The new cgSSR markers associated with grain size related characters have a significant impact on practical plant breeding to increase the number of causative alleles for these traits through marker aided rice breeding programs.