Signalling Events and Associated Pathways Related to the Mammalian Sperm Capacitation

Capacitation is a biological phenomenon occurring prior to fertilization and is a multiple event process. Many physiological and biochemical changes takes place during the process; these changes are related to lipid composition of membrane, intracellular modulation of ion concentration, protein phosphorylation, sperm movement and membrane permeability. These events occur when the sperm is exposed to the new environment of ion concentration in the female reproductive tract. Ions such as bicarbonate and calcium facilitate capacitation by activating adenylyl cyclase, thus initiating protein kinase A (PKA) signalling cascade. Extracellular‐regulated kinase pathway is activated by ligand binding to the membrane receptors and intracellular activation by reactive oxygen species (ROS). Activation of these pathways leads to the phosphorylation of different proteins, which is associated with events such as capacitation, hyperactivation and acrosome reaction that are essential for successful fertilization. Extensive studies were carried out on protein phosphorylation in relation to capacitation, but its role still remains ambiguous.     

Infiltrating Foxp3+ Regulatory T cells and Histopathological Features in Canine Classical and Spermatocytic Seminomas

In humans, regulatory T (T reg) cells are known to play a critical role in both the regulation of immune homoeostasis and the progression of cancer. However, there is little information about the identification, characterization and the function of T reg cells in canine tumours. We identified T reg cells in 28 canine seminoma samples using a Forkhead box P3 (Foxp3) antibody and investigated the relationship between T reg cell infiltration and histopathological features of classical and spermatocytic seminomas (SE and SS, respectively). The Foxp3 protein showed nuclear immunostaining in infiltrating lymphocytes, and Foxp3+ cells were diffused or focally distributed in seminoma tissues. Foxp3+ cells were frequently present in the SS histotype, in seminomas that showed no evidence of tumour cell invasion into the vessels and in seminomas showing a diffuse growth pattern with three cell types. Neither the SE/SS histotype nor the histopathological features of the tumour correlated with Foxp3+ cell counts. These results indicate that Foxp3+ T reg cells may be associated with a less malignant histological phenotype or may not play a critical role in the immune response of canine seminomas. Moreover, Foxp3+ T reg cells may be associated with SS seminoma, but further studies, involving a larger number of samples, are required to better understand whether these cells play a critical role in the immune response in canine seminomas. This is the first report to demonstrate the characteristics of T reg cell infiltration in canine seminoma.     

Activation of Zona‐Free Buffalo (Bubalus bubalis) Oocytes by Chemical or Electrical stimulation,and Subsequent Parthenogenetic Embryo Development

Parthenogenetic activation using zona‐free oocytes offers an alternative model that could be applied to develop protocols for the activation of reconstructed embryos for cloning. The aim of this study was to compare the efficacy of different methods for the activation of zona‐free buffalo oocytes in terms of their effects on the developmental competence of parthenogenetic embryos. The effects of zona removal on parthenogenetic activation and in vitro developmental competence of metaphase II oocytes were also examined. All activation methods were followed by incubation of 2 mm 6‐dimethylaminopurine (6‐DMAP) for 4 h. Out of three different pulse strengths (1.2, 2.1 or 3.3 kV/cm) used, 2.1 kV/cm resulted in the highest blastocyst rate (25.3%). On comparing different chemical agents and electric pulse, highest blastocyst rate was observed for calcium ionophore (CaI) (28.6%) followed by ethanol (25.0%), electric pulse (22.5%) and combined CaI and ethanol treatment (16.7%) although differences among them were not significant. Furthermore, a significantly reduced developmental potential was observed in zona‐free oocytes when compared to zona‐intact ones up to the blastocyst stage (44.3% vs 27.1%). In conclusion, zona‐free buffalo oocytes can be successfully activated for parthenogenetic development using chemical or electrical stimulation. Out of different agents examined, CaI followed by 6‐DMAP resulted in the highest blastocyst rate.     

Effect of testicular tissue lysate on developmental competence of porcine oocytes matured and fertilized in vitro

The objective of the present study was to investigate the effect of testicular tissue lysate (TTL) on developmental competence of germinal vesicle (GV) stage porcine oocytes. Two types of TTL were prepared through repeated freeze–thaw in liquid nitrogen, one from whole testicular tissue (wTTL) and other from either of four different sections of testes, namely just beneath the tunica albuginea (TA), from the transitional area between the seminiferous cord/tubules and the mediastinum testis (TR) and from the intermediate area (parenchymal tissue origin) and CE (cauda epididymis origin). The whole or section‐wise TTL treatments were given for 44 hr during in vitro maturation (IVM). Oocyte maturation was done in either of the two media, namely defined (high‐performance basic medium for porcine oocyte maturation, commercially available) and serum containing (TCM199). After maturation, oocytes were co‐incubated with fresh spermatozoa for 6 hr and then transferred to embryo culture media. Treatment of GV stage oocytes with wTTL (1 mg/ml) increased the cleavage and morula percentage rate (69.23 ± 6.23 and 48.15 ± 6.77, respectively) than that of their control (58.33 ± 8.08 and 32.54 ± 5.53, respectively) in defined media, and in serum‐containing media, cleavage and morula percentage rate were almost equal in both treatment (54.56 ± 7.79 and 34.70 ± 6.78, respectively) and control (59.52 ± 8.21 and 38.52 ± 6.54, respectively). However, effect of wTTL was not significant. In case of section‐wise TTL supplements, TR section significantly (p < .01) improved cleavage and morula rate (58.43 ± 7.98 and 36.14 ± 6.89, respectively) followed by TA. In conclusion, present study indicates that IVM, in vitro fertilization and in vitro culture of embryo are improved in the presence of TTL, particularly its TR section. Further study is expected to reveal the principal components of TTL which may prove useful for IVM.     

Phosphorus uptake and use efficiency in different varieties of bread wheat (Triticum Aestivum L)

Thirty spring wheat varieties were evaluated and classified into eight different groups on the basis of their grain yield performance and phosphorus (P) uptake using Metroglyph analysis. Significant variability was observed for grain and biomass yield, plant height, P content in grain and straw, total P uptake and phosphorus harvest index and P use efficiency traits. Varieties WH 711 and PBW 343 exhibited high grain yield as well as high P uptake (HGY-HP). WH 283 and UP 2425 with high index score of 19 and 16 respectively, constituted the high grain yield-medium P uptake (HGYMP) group. Both these varieties, though had similar grain yield of 5348 kg/ha, but WH 283 (12.64 kg/ha) utilized much lower P as compared to UP 2425 (16.94 kg/ha). Moreover, WH 283 (81.64) also showed higher values for phosphorus harvest index (PHI) than UP 2425 (67.88%). P uptake of WH 283 was comparable with that of Raj 3765 (10.78 kg/ha) and grouped into high grain yield and low P uptake (HGY-LP) group. The grain yield performance of these two varieties with a relatively low P uptake is reflected in their high index score for P use efficiency thus, earmarking them for low P regimes. Variety HW 2006, despite low grain yields of 4665 kg/ha had high index score of 16 due to its higher value for Phosphorus Biological (PBER) and Economic Yield (PEER) Efficiency Ratio as it has effected least (7.18 kg/ha) P mobilization. In addition high P translocation in the grain was also observed for this variety. Inter-mating of genotypes like HW 2006, UP 2338 and HW 2016 with those belonging to HGY-HP (PBW 343 and WH 711) and HGY-LP (Raj 3765 and WH 283) would be an ideal strategy to develop the cultivars for efficient phosphorus use.     

Conventional and contemporary approaches for drought tolerance rice breeding: Progress and prospects

Drought is becoming a major threat to rice farming across the globe owing to the depletion of water tables in rice-growing belts. Drought affects rice plants at multiple stages, causing damage at morphological and physio-biochemical levels, leading to severe losses that exceed losses from all other stresses. The amalgamation of conventional breeding methods with modern molecular biology tools and biometrical methods could help accelerate the genetic gain for drought tolerance in rice. Many drought-tolerance traits with genetic determinants have been identified and exploited for tolerance rice variety breeding. The integration of genome-wide association study and genomic selection tools with speed breeding shortened the breeding cycle and aided in rapid improvement of genetic gain. In this review, we emphasized the progress made through classical breeding as well as the limitations and usefulness of current genomic methods in improving drought tolerance. We briefly addressed methods for identifying genetic determinants for drought tolerance and deploying them through genomics-assisted breeding programmes to develop high-yielding drought-tolerant rice cultivars.     

Caudal anaesthesia of the infraorbital nerve for diagnosis of idiopathic headshaking and caudal compression of the infraorbital nerve for its treatment,in 58 horses

Reasons for performing study: Idiopathic headshaking is often a facial pain syndrome, but a diagnostic protocol has not been described. In a previous study, caudal compression of the infraorbital nerve for treatment offered a fair success rate, but low case numbers and short follow‐up time were limitations. Objectives: To describe a diagnostic protocol for headshaking, examining the role of bilateral local analgesia of the posterior ethmoidal nerve (PET block). To report longer‐term follow‐up after surgery of the original cases and further cases and to determine whether changes to the technique influence success rates and complications. Methods: Records of horses that had undergone PET block and caudal compression surgery at 3 hospitals were reviewed. Modifications to the surgical technique included placing additional coils into the infraorbital canal and/or performing concurrent laser cautery of the nerve. Follow‐up information was obtained by telephone contact with owners. Results: The PET block was performed in 27 horses, with a positive result in 23 of 27 (85%). Surgery was performed in 58 horses. A successful outcome was initially achieved in 35 of 57 (63%) horses, but recurrence occurred between 9 and 30 months later in 9 (26%). Surgery was repeated in 10 of 31 (32%) horses. Final success rate, considering only response to the last performed surgery, was 28 of 57 (49%) horses with median follow‐up time of 18 months (range 2–66 months). Nose‐rubbing was reported post operatively in 30 of 48 (63%) horses. This resolved in all but 4 horses, which were subjected to euthanasia. Response to PET block or change in surgical technique did not appear to influence outcome or complications. Conclusions and potential relevance: The diagnostic protocol described is recommended for the investigation of headshakers. Caudal compression offers the best prognosis for a successful outcome compared with other treatments, for horses in which the only alternative is euthanasia. Surgical treatment of the disorder requires refinement, and the pathogenesis of the disorder requires investigation.     

Draft Genome Sequence of Potato Dry Rot Pathogen <Emphasis Type="Italic">Fusarium sambucinum</Emphasis> Fckl. F-4

Fusarium sambucinum is one of the most important causal agents that not only cause the dry rot disease of potato tubers in fields and stores worldwide but also capable of producing secondary metabolites toxic for people and animals. Here we present the first draft genome sequence of the strain (F-4) estimated to be around appx. 42.0 Mb. The genome has 12,845 protein coding genes with more than 35,900 exons and gene density of 3.13 per 10Kb. F. sambucinum is evolutionary more close to the F. graminearum among the Fusarium species complex. The genome sequence represents a valuable resource for understanding the pathogenecity and virulence factors, and their evolution within the complex and highly plastic genus Fusarium.     

Positive Effects of Melatonin Treatment on the Reproductive Performance of Young Border Leicester Rams Mated to Merino Ewes in Spring: Preliminary Observations

Poor reproductive performance of Merino ewe flocks when mated to Border Leicester rams during spring may be due to seasonality of the Border Leicester breed. Two approaches were taken to test this assumption. Six young (12 months old) or six mixed‐age (12, 24 and ≥36 months old) Border Leicester rams were either treated or not treated with melatonin implants (2 × 2 design) 6 weeks before the four groups of rams were each put with approximately 300 Merino ewes for an 8‐week mating period. Implants were inserted in early September (experiment 1). The second approach was to yard or not yard ewes and mixed‐age rams on several occasions during the first 3 weeks of the mating period (experiment 2). Pregnancy rate and twinning percentage were assessed by ultrasonography. In experiment 1, melatonin treatment in young rams increased (p < 0.001) pregnancy rate from 5.0% to 92.6%, but mixed‐age rams did not respond (90.7% vs 89.5% for melatonin and non‐melatonin treatments, respectively). Twinning rate was similar (p > 0.05) for ewes mated to either melatonin or non‐melatonin‐treated young rams (36.8% vs 40.0%, respectively), whereas melatonin significantly improved (p < 0.05) twinning rate in those ewes mated to mixed‐age rams (49.1% vs 36.1%). After 6 weeks of melatonin treatment, scrotal circumference was greater (p < 0.05) in both young and mixed‐aged rams than in untreated counterparts. In experiment 2, yarding of ewes and rams overnight on several occasions early in the mating period reduced (p < 0.001) pregnancy rate compared with non‐yarded counterparts (89.5% vs 65.5%). Twinning rate was not affected (37.7% vs 36.1%, respectively). In summary, melatonin treatment of Border Leicester rams significantly improved flock reproductive performance in spring due to improved pregnancy rates with young rams and improved litter size with mixed‐age rams.