An indirect enzyme linked immunosorbent assay for the detection of bovine antibodies to multiple Leptospira serovars

An indirect enzyme linked immunosorbent assay was developed for the detection of bovine antibodies to multiple pathogenic Leptospira serovars, including canicola, copenhageni (represents icterohaemorrhagiae), grippotyphosa, hardjobovis, pomona, and sejroe. The antigen utilized in this assay was a sonicated mixture of equal parts of killed whole cells of each of the 6 serovars named above. A mouse monoclonal antibody against bovine immunoglobulin (Ig)G₁ that was conjugated with horseradish peroxidase was used for detection of bound antibodies. This assay was evaluated with sera (n = 3107) that were microscopic agglutination test (MAT)-negative (at a 1:100 dilution) for each of the 6 serovars listed above and sera (n = 601) that were MAT-positive (at a 1:100 dilution) for 1, or any combination of the 6 listed serovars. In addition, sera from serial weekly bleedings of cows, which were individually experimentally infected with serovars hardjobovis, copenhageni, grippotyphosa, or canicola, were also tested in this assay.

At an optimal cut-off point determined by receiver operating characteristic (ROC) curve analysis, the relative sensitivity and specificity of the assay were 93.5% (95% confidence interval = 91.2% to 95.3%) and 94.7% (95% confidence interval = 93.9% to 95.5%), respectively. This assay was able to detect antibody in the sera of animals experimentally infected with serovar hardjobovis as early as 1 week postinoculation

     

Comparative morphometrical studies on the skull bones of barking deer (Muntiacus muntjak) and sambar deer (Rusa unicolor)

The present study reports data on the skull bone morphometry of barking and sambar deer. The skulls of adult barking deer (n = 6) and sambar deer (n = 6) of either sex (n = 3 males and n = 3 females) were collected from the Aizawl Zoological Park, Aizawl, Mizoram, India, with official permission from the Government of Mizoram. Anatomically, barking and sambar deer's skulls were elongated, pyramid-like, dolichocephalic and consisted of thirty-two cranial and facial bones. The cranial bones were eleven (three single and four paired), comprising of occipital, sphenoid, ethmoid, frontal, interparietal, parietal and temporal. The facial bones were twenty-one (one single and ten were paired), consisting of the maxilla, premaxilla (incisive), palatine, pterygoid, nasal, lacrimal, zygomatic (malar), vomer, turbinates, mandible and hyoid. In the present study, altogether 41 different measurements were taken morphologically and 6 different indices were applied. The obtained morphometrical parameters were significantly (p < .01, p < .05) higher in males than females of both species. Species wise, all obtained parameters were higher in sambar deer than barking deer. The obtained 41 different skull parameters and 6 indices showed statistically significant differences (p < .01 and p < .05) between both sexes of barking and sambar deer; however, practically these differences were meagre. The present morphometrical study on the skull of both species can help the wildlife professionals and zoo veterinarians determine the sex of these animals and differentiate it from other domestic and wild small ruminants for solving veterolegal cases. This study's findings will also motivate and assist other comparative studies with various domestic and wild small ruminants.     

Agronomic fortification of rice grains with secondary and micronutrients under differing crop management and soil moisture regimes in the north Indian Plains

Although the System of Rice Intensification (SRI) has been reported to produce higher paddy (Oryza sativa L.) yields with better-quality grains, little research has addressed the latter claim. This study investigated the interactive effects of rice cultivation methods with different irrigation schedules and plant density on the uptake and concentration of sulfur (S), zinc (Zn), iron (Fe), manganese (Mn) and copper (Cu) in the grain and straw of two rice cultivars during two rainy seasons in the northern plains of India. As the two seasons differed in amounts of rainfall, there were impacts of soil moisture differences on nutrient uptake. Plots with SRI cultivation methods enhanced the grain uptake and concentrations of S, Zn, Fe, Mn and Cu by 36, 32, 28, 32 and 63%, respectively, compared to conventional transplanting (CT). Under SRI management, the highest concentrations of S, Zn and Cu in the grain and straw occurred with irrigation intervals scheduled at 3 days after disappearance of ponded water (DADPW; 3_D), whereas Fe and Mn concentrations in the grain and straw were higher with irrigation at 1 DADPW (1 _D ) compared with plots under 3 _D or 5 DADPW (5 _D ). The higher nutrient uptakes were also manifested in higher grain yield in 1 _D and 3 _D plots (by 9 and 6%, respectively) compared with 5 _D . Wider spacing (25 × 25 cm) compared with closer spacing (20 × 20 cm) significantly increased yield and the uptake and concentrations of all the said nutrients in the grains. When comparing the performance of two cultivars, the total uptakes of Zn, Fe, Mn and Cu in both grain and straw were significantly more in Hybrid 6444 than the improved variety Pant Dhan 4. Overall, SRI crop management compared to CT practices led to more biological fortification of rice grains with respect to S and the four micronutrients studied, giving a concomitant yield advantage of about 17% on average in this region.     

Alteration in important quality traits and antioxidant activities in Brassica oleracea with Ogura cybrid cytoplasm

In vegetable brassicas, cytoplasmic male sterility (CMS) system is the most preferred mechanism for hybrid seed production and Ogura cybrid cytoplasm is the only source used widely. Effects of the alien cybrid cytoplasm on important quality traits need to be understood for their effective use in breeding programme. In analysing 38 Ogura cybrid cytoplasm‐based cauliflower CMS lines, it was revealed that introgression of Ogura cytoplasm resulted in significant alteration in important quality traits. Cupric reducing antioxidant activity and ferric reducing ability of plasma values were increased up to 5–10 times in different genotypes. Among 38 analysed CMS lines, concentration of plant pigments such as anthocyanin, total chlorophylls and ascorbic acid was reduced in 21, 17 and 32 genotypes, respectively. However, the concentration of total carotenoids and β‐carotene was elevated in most of the CMS lines and total carotenoid was increased up to 20 times in the CMS line, Ogu13‐85‐2A. The results indicate the role of nuclear–cytoplasmic interaction and mitochondrial genome in determining concentration of different quality traits.     

Relative potencies of natural estrogens on vitellogenin and choriogenin levels in the Indian freshwater spotted snakehead, <Emphasis Type="Italic">Channa punctata</Emphasis>: in vivo and in vitro studies

The relative efficacies of three natural estrogens viz., estrone (E₁), estradiol-17β (E₂) and estriol (E₃) to induce synthesis of vitellogenin (Vg) and choriogenin (Chg) were assessed in primary hepatocyte cultures of the Indian freshwater spotted snakehead, Channa punctata. Hepatocytes were isolated from the spotted snakehead liver by a non-enzymatic protocol. Optimum culture conditions were standardized for ensuring their viability and functioning. Isolated hepatocytes were cultured for 48 h for monolayer formation and then exposed to various concentrations (0.001–10 μM) of the three estrogens. Competitive homologous ELISAs, developed and validated for spotted snakehead Vg and Chg were employed to determine the amounts of these two proteins secreted into the culture medium after 48 h of incubation. The results reveal that although all the three estrogens were effective in inducing the production of Vg and Chg in a dose-dependent manner, there were differences in their relative potencies. Of three estrogens, E₁ was the least potent and could induce synthesis of Vg and Chg only at a minimum concentration of 0.5 μM; whereas significant levels of both the proteins were quantified in culture medium by exposing the hepatocytes to E₂ or E₃ even at a concentration of 0.001 μM. All three estrogens were effective in inducing synthesis of Vg and Chg in vivo also. These results suggest the possibility of employing the above in vitro experimental design to monitor the presence of estrogens/estrogen-like chemicals in natural waters, which could interfere with the estrogen receptor system of fish. This study further points to the possibility of using Chg, in addition to Vg, as a parameter for screening various chemicals for their estrogenic activity.     

Studies on pesticides based on coumarin. III. Synthesis and antifungal activity of substituted 4-methyl-coumarins and related compounds

A series of substituted 4-methylcoumarins was synthesised and the members tested for their toxicity towards mycelial growth of seven phytopathogenic fungi in culture. Rhizoctonia solani, Alternaria alternata and Fusarium solani exhibited maximum sensitivity to these compounds whereas Pythium aphanidermatum, Colletotrichum falcatum, Drechslera oryzae and Macrophomina phaseolina were relatively less sensitive. 6-Ethyl-3-n- propyl-7-hydroxy4-methylcoumarin ( I ) was relatively toxic towards all fungi except C. falcatum, P. aphanidermatum and M. phaseolina. The 6-n-butyl ( III ) and 6-(1, 1, 3, 3-tetramethylbutyl) ( VI ) derivatives were highly toxic to R. solani with EC₅₀, values of lμg ml^?1.     

Studies on pesticides based on coumarin. II. Synthesis and antifungal activity of ten 2-alkyl-7, 8-dihydro-3-hydroxynaphtho[1,2-c]chromen-6-ones

A series of 2-alkyl-7, 8-dihydro-3-hydroxynaphtho[1,2-c]chromen-6-ones was synthesised by the condensation of ethyl 3, 4-dihydro-1-oxonaphthalene-2-carboxylate with substituted phenols in the presence of POCl₃. The compounds were characterised and tested for their toxicity towards the mycelial growth of seven phytopathogenic fungi in culture. Drechslera oryzae, Rhizoctonia solani and Colletotrichum falcatum exhibited maximum sensitivity to these compounds whereas Macro-phomina phaseolina, Fusarium solani, Alternaria alternata and Pythium aphanidermatum were less sensitive. 2-Ethyl-7, 8-dihydro-3-hydroxy-naphtho[1,2-c]chromen-6-ones possessed greatest toxicity with EC₅₀ values ranging from 0.2 to 2.5 μg ml^?1 against all fungi except A. alternata and P. aphanidermatum.     

Higher than optimum temperature under CO2 enrichment influences stomata anatomical characters in rose (Rosa hybrida)

There is little available information on the effects of temperature and CO₂ enrichment on stomata anatomical characteristics of plants. Effect of these two microclimates was studied on five rose (Rosa spp.) cultivars, viz. ‘First Red’ (used as check), ‘Arjun’, ‘Raktima’, ‘Raktagandha’ and ‘Pusa Pitamber’. Budded, single-stemmed rose cultivars having five lateral buds were grown in controlled environment growth cabinets under enriched CO₂ (1000 μmol mol⁻¹) and optimum (28/18 °C, T₀) or high (35/25 °C, T₁) temperature for 50 days. All observations were made on the abaxial leaf surface. Significant increases in stomatal density (68.7%), index (29.6%) and epidermal cell density (37.3%) were recorded in plants grown at high temperature over control with CO₂ enrichment. The cultivars responded differently in terms of length and width of guard cell and stoma (pore) under high temperature, however, the values averaged over treatments showed a significant reduction in these parameters. Further, number of stomata per leaf was higher (28.3%) in plants grown at high temperature, except First Red. A reduction in mean leaf area (26.7%) and dry mass (32.0%) was recorded at high rather than optimum temperature. The specific leaf area was maximum in Arjun (87%) while in First Red, a 14% reduction was noted at high temperature.     

Kinetics of Interferon-Gamma Production and its Comparison with Anti-Listeriolysin O Detection in Experimental Bovine Listeriosis

The kinetics of the production of interferon gamma (IFN-) in whole blood culture and its comparison with anti-listeriolysin O (ALLO) detection by ELISA were studied during oral infection of calves with Listeria monocytogenes. Culture filtrate antigen (CFA), listeriolysin O (LLO), and sonicated antigen (SA) were used to prime the peripheral blood mononuclear cells (PBMCs) and the plasma from orally infected calves. IFN- and ALLO appeared as early as day 7 of an oral infection. IFN- was detected earlier with LLO than with SA. The Max50 interleukin (IL-2) activity and IFN- estimated in the culture supernatant from PBMCs primed in vitro with different antigens of L. monocytogenes revealed high induction of IL-2 and IFN- by CFA, LLO and live antigen. IFN- assay and ALLO detection were used for testing cases of repeat breeding in dairy cattle. It appeared that detection of IFN- employing LLO can be used to diagnose listerial infections.     

Light-energy processing and freezing-tolerance traits in red spruce and black spruce: species and seed-source variation

Red spruce (Picea rubens Sarg.) and black spruce (Picea mariana (Mill.) B.S.P.) are genetically and morphologically similar but ecologically distinct species. We determined intraspecific seed-source and interspecific variation of red spruce and black spruce, from across the near-northern margins of their ranges, for several light-energy processing and freezing-tolerance adaptive traits. Before exposure to low temperature, red spruce had variable fluorescence (Fv) similar to black spruce, but higher photochemical efficiency (Fv/Fm), lower quantum yield, lower chlorophyll fluorescence (%), and higher thermal dissipation efficiency (qN), although the seed-source effect and the seed-source x species interaction were significant only for Fv/Fm. After low-temperature exposure (-40 degrees C), red spruce had significantly lower Fv/Fm, quantum yield and qN than black spruce, but higher chlorophyll fluorescence and relative fluorescence. Species, seed-source effect, and seed-source x species interaction were consistent with predictions based on genetic (e.g., geographic) origins. Multi-temperature exposures (5, -20 and -40 degrees C) often produced significant species and temperature effects, and species x temperature interactions as a result of species-specific responses to temperature exposures. The inherent physiological species-specific adaptations of red spruce and black spruce were largely consistent with a shade-tolerant, late-successional species and an early successional species, respectively. Species differences in physiological adaptations conform to a biological trade-off, probably as a result of natural selection pressure in response to light availability and prevailing temperature gradients.