Effect of Supplementation of Taurine or Trehalose in Extender on Immunolocalization of Tyrosine Phosphoproteins in Buffalo and Cattle (Karan Fries) Cryopreserved Spermatozoa

The present study assessed the effects of incorporation of Taurine or Trehalose in extender on immunolocalization of tyrosine phosphoproteins, Cryocapacitation and other sperm quality parameters (motility, viability and membrane integrity) in post‐thawed sperm from Buffalo (Murrah) and Cattle (Karan Fries). Six ejaculates from six individual bulls from both species were chosen at random and split into four aliquots: one aliquot without dilution (fresh sample), another diluted in egg yolk tris‐citrate (EYTC) extender and the rest of aliquots with EYTC dilution supplemented with taurine (50 mm ) or trehalose (100 mm ), respectively, and cryopreserved. Following cryopreservation, semen were thawed and assessed for standard semen quality parameters. Extent of capacitation in cryopreserved spermatozoa was measured by inducing in vitro acrosome reaction followed by dual staining. Immunolocalization of tyrosine phosphoproteins was carried out by immunocytochemistry using primary antibody clone pT‐154 (anti‐phosphotyrosine antibody) and FITC‐conjugated secondary antibody. Immunofluorescent signals were analysed for level of protein tyrosine phosphorylation in spermatozoa. Post‐thaw semen evaluation showed supplementation of taurine or trehalose to EYTC extender significantly (p < 0.05) increased motility, viability and membrane integrity of spermatozoa in both species. Percentage of cryocapacitated spermatozoa was significantly (p < 0.05) higher in cattle as compared to buffalo and degree of cryocapacitaion of spermatozoa decreased significantly (p < 0.05) upon supplementation of additives in both the species. It was also found that tyrosine phosphoproteins were localized differentially in fresh and cryopreserved spermatozoa. Supplementation of taurine or trehalose to freezing extender changed the localization of tyrosine phosphoproteins in cryopreserved spermatozoa similar to fresh in both the species. The results obtained clearly indicated that supplementation of taurine or trehalose to EYTC prior to cryopreservation improves Buffalo and Cattle sperm quality in terms of cryocapacitation and immunolocalization of tyrosine phosphoproteins during freezing–thawing process.     

An efficient method for the purification and characterization of nematicidal azadirachtins A,B, and H,using MPLC and ESIMS

Azadirachtin A enriched concentrate containing 60% active ingredient (a.i.) was prepared from the methanolic extract of the de-fatted neem (Azadirachta indica A. Juss) seed kernels. Azadirachtins A, B, and H, the three major bioactive constituents of neem seed kernel, were purified from this methanolic concentrate by employing reverse phase medium-pressure liquid chromatography (MPLC), using methanol-water solvent system as an eluant. The three pure azadirachtin congeners thus obtained were characterized by their unique mass spectral fragmentation, using electrospray probe in positive ion mode (ESI). All three azadirachtins exhibited nematicidal and antifungal activities. Azadirachtin B was the most effective against the reniform nematode Rotylenchulus reniformis (EC(50) 96.6 ppm), followed by Azadirachtin A (119.1 ppm) and H (141.2 ppm). At 200-ppm concentration, the test compounds caused 50-65% mortality of Caenorhabditis elegans nematode. Azadirachtin H showed the highest activity against the phytophagous fungi Rhizoctonia solani (EC(50) 63.7 ppm) and Sclerotium rolfsii (EC(50) 43.9 ppm), followed by B and A. The isolation of pure azadirachtins A, B, and H directly by MPLC purification from its concentrate and their characterization by ESIMS are unique and less time-consuming.     

Active root distribution pattern of <Emphasis Type="Italic">Hevea brasiliensis</Emphasis> determined by radioassay of latex serum

The active root distribution pattern of mature rubber (Hevea brasiliensis Muell. Arg.) up to a lateral distance of 250 cm from the tree and to a soil depth of 90 cm was studied in an oxisol by employing ³²P soil injection technique in Kerala, the state which accounts for 83% of rubber cultivation in India. The trees were aged 18 years and grown at a spacing of 4.9 × 4.9 m. The extent of absorption of applied ³²P by the tree from various placements was assessed by radio assay of leaf and latex serum. Latex serum registered higher counts and variability was less compared to leaf indicating the suitability of latex serum as a potential source for radio assay for ³²P studies in rubber. The results revealed that rubber is a surface feeder with 55% of the root activity confining to the top 10 cm of soil layer. Root activity declined with increasing depths and the concentration of physiologically active roots at 90 cm depth was only 6%. A more or less uniform distribution of root activity was noticed with respect to lateral distance indicating more extensive spread of lateral roots. Concentration of physiologically active roots in the surface layer suggests the possibility for competition under intercropped situation in mature plantations.     

Alley cropping of maize with calliandra and leucaena in the subhumid highlands of Kenya: Part 2. Biomass decomposition, N mineralization, and N uptake by maize

A major challenge in developing agroforestry approaches that utilize tree-leaf biomass for provision of N to crops is to ensure synchrony between the N released from decomposing prunings and N demand by crops. A study was conducted in the subhumid highlands of Kenya to assess the rate of decomposition and mineralization of soil-incorporated Calliandra calothyrsus Meissner (calliandra) and Leucaena leucocephala (Lam.) de Wit (leucaena) tree biomass and maize roots (Zea mays L.) both in an alley cropping and a sole cropping system. The amount of mineralized N peaked four weeks after planting (WAP) maize in all the treatments during both seasons of 1995. Cumulative mineralized N at week 20 ranged from 114 to 364 kg N ha⁻¹ season⁻¹, the absolute control treatment giving the lowest and the prunings-incorporated treatments giving the highest amounts in the two seasons. Total N uptake by maize, ranging from 42 to 157 kg ha⁻¹ season⁻¹, was lowest in the 'alley-cropped, prunings-removed' treatments, and highest in the 'non alley-cropped-prunings-incorporated' treatments. The apparent N recovery rate by maize was highest in the fertilizer applied treatments in the two seasons. Decomposition rate constants (kD) ranged from 0.07 to 0.21 week⁻¹, and the rates among the different plant residues were as follows: leucaena < calliandra < maize roots. Nitrogen release rate constants (kN), ranging from 0.04 to 0.25 week⁻¹, followed a similar pattern as the rate of decomposition with leucaena releasing the highest amount of N followed by calliandra and lastly by maize roots. This revised version was published online in June 2006 with corrections to the Cover Date.     

Multipurpose tree prunings as a source of nitrogen to maize under semiarid conditions in Zimbabwe

When multipurpose-tree (MPT) prunings are used as a source of nitrogen to annual corps in agroforestry systems, it is important that high levels of N recovery are attained. In order to test the effect of pruning quality and method of pruning application on N-recovery rates, a field experiment was conducted using prunings of five MPT species and two methods of application (surface versus incorporation). There was an interaction of pruning quality and method of pruning application on N recovery and maize grain yield. For most MPT species used, incorporated prunings gave high nitrogen recovery compared to surface applied prunings. In order to achieve synchrony between N supply from prunings and N demand by a maize crop, relatively large amounts of N should be released into the soil before peak N demand by the maize crop. This could be achieved by incorporating adequate quantities of high quality prunings such as those ofCajanus cajan andLeucaena leucocephala. Florida Agricultural Experiment Station Journal Series Number R-05489.     

Litter dynamics of six multipurpose trees in a homegarden in Southern Kerala, India

Multipurpose trees, the integral components of homegardens, contribute significantly to the closed nutrient cycling processes and sustainability of the ecosystem. Although, the litter production and probable nutrient returns via litter in homegardens have been documented, quantification and characterization of the decomposition and bioelement release from the litter have received relatively little scientific attention. The objective of the present study is to explore the litter dynamics of six locally important multipurpose trees (Mangifera indica L., Artocarpus heterophyllus Lamk., Anacardium occidentale L., Ailanthus triphysa Dennst., Artocarpus hirsutus L. and Swietenia macrophylla L.), in an agroforestry homegarden in Southern Kerala, India. Litterfall and nutrient additions in the six species ranged from 383 to 868 g m⁻² yr⁻¹, nitrogen, 6.4 to 8.8, phosphorus, 0.17 to 0.42 and potassium, 1.1 to 2.8 g m⁻² yr⁻¹. The annual litter output in the homegarden was 425 kg with A. hirsutus, M. indica, A. heterophyllus and A. occidentale recording significantly higher litter and nutrient additions. Leaf litter decay studies revealed A. heterophyllus and A. occidentale to be the most labile litter species and S. macrophylla the most recalcitrant. The decay rate coefficients varied significantly among the species. Foliage decomposition rates related to the initial chemical composition of the litter revealed best correlation with lignin. NPK release was almost complete by the end of decay in all species inspite of the initial phases of accumulation observed for nitrogen and phosphorus. Two-way analysis of variance test revealed significant differences in the contents of the three elements as a function of species and time elapsed. Macronutrients were released in the order K>N/P. The higher rates of decay and nutrient turnover in M. indica, A. heterophyllus and A. occidentale foliage indicated the potential of using these species’ litter as nutrient inputs in agriculture while A. triphysa, A. hirsutus and S. macrophylla perform better as organic mulches taking a longer time for decay and hence nutrient release.