Comparative Expression Analysis of Gametogenesis‐Associated Genes in Foetal and Adult Bubaline (Bubalus bubalis) Ovaries and Testes

This study was conducted to identify and analyse the expression of gametogenesis‐associated genes and proteins in foetal and adult buffalo gonads of both the sexes. Relative quantification of the genes was determined by qPCR and Western blotting. Immunohistochemistry was also performed for various gametogenesis‐associated proteins in foetal and adult gonads of both the sexes. We observed significantly (p < 0.05) increased expression of primordial germ cell‐specific, meiotic as well as genes associated with oocyte maturation and development in foetal ovaries as compared to the adult ones. However, significantly (p < 0.05) increased expression of proteins associated with oocyte maturation like GDF9 and ZP4 was found in adult ovaries, indicating temporal regulation of mRNA translation during oogenesis. Meiotic genes showed significantly (p < 0.05) increased expression in adult testes as compared to foetal testes and ovaries, indicating onset of meiosis at a later stage in spermatogenesis. In general, the expression of primordial germ cell‐associated as well as meiotic genes was higher in adult testes, indicating the increased biological activity in the organ. Immunohistochemistry revealed localized expression of gametogenesis‐associated proteins in ovarian follicles and seminiferous tubules of testes, while the surrounding somatic tissues were devoid of these proteins. The study gives an understanding of the sequential and temporal events of gene expression as well as mRNA translation during male and female gametogenesis. It could also be concluded that follicles and seminiferous tubules are the functional units of the female and male gonads, respectively, and their function could be enhanced by appropriate chemical and genetic intervention of the somatic tissue immediately surrounding them. This assumes importance in the context that buffalo attains sexual maturity at an older age of 2–3 years and have smaller ovaries with lesser number of primordial follicles in comparison with cattle, which is suggested to be the main reason of their poor breeding performance.     

Expression of inhibins, activins, insulin-like growth factor-I and steroidogenic enzymes in the equine placenta

In this study, the expression patterns of inhibins, activins, insulin-like growth factor-I (IGF-I) and steroidogenic enzymes in equine placentae recovered during the latter two-thirds of gestation were examined. Concentrations of inhibin A and inhibin pro-alphaC in endometrial and fetal placental tissue homogenates were very low during the period examined, whereas these tissues contained high concentrations of activin A. In both maternal endometrial and fetal placental tissues, activin A levels decreased as pregnancy progressed. Expression of inhibin alpha-subunit was not observed in the placenta using either immunohistochemistry or in situ hybridization. Inhibin/activin betaA-subunit and its mRNA were confined to maternal endometrial glands, whereas immunopositive betaB-subunit was not detected in either endometrial glands or microcotyledons. Cytochrome P450 side chain cleavage enzyme was detected by immunohistochemistry in both endometrial glands and microcotyledons, whereas cytochrome P450 17alpha-hydroxylase/lyase was absent in these tissues. Immunopositive signals for 3beta-hydroxysteroid dehydrogenase and cytochrome P450 aromatase were localized in microcotyledons but not in endometrial glands. Immunohistochemistry revealed that IGF-I was highly expressed in microcotyledons around Day 130, and decreased as pregnancy progressed. Changes in the expression of IGF-I were correlated with the number of PCNA positive cells in the placenta. The present study demonstrated the presence and localized the site of expression of activin, IGF-I and steroidogenic enzymes in equine placental tissues during the latter two-thirds of gestation; the results suggest that activin and IGF-I may be involved in the regulation of placental development.     

In vitro inhibitory effect of trichostatin A on canine grade 3 mast cell tumor

Mast cell tumor (MCT) is one of the most prevalent neoplasms that affect skin and soft tissue in dogs. Because mast cell tumors present a great variety of clinical appearance and behavior, their treatment becomes a challenge. Trichostatin A (TSA), an antifungal antibiotic, has shown inhibitory effects on the proliferation and induction of apoptosis in various types of cancer cells. In order to evaluate the potential of trichostatin A as a therapeutic drug, cells of grade 3 MCT were cultured and treated with concentrations of 1 nM to 400 nM of TSA. MTT assay and trypan blue exclusion assays were performed to estimate cell growth and cell viability, and cell cycle analysis was evaluated. TSA treatment showed a reduction in numbers of viable cells and an increase of cell death by apoptosis. The cell cycle analysis showed an increase of hypodiploid cells and a reduction of G0/G1 and G2/M –phases. According to these results, trichostatin A may be an interesting potential chemotherapeutic agent for the treatment of canine MCT.     

Expression and localization of angiopoietin family in corpus luteum during different stages of oestrous cycle and modulatory role of angiopoietins on steroidogenesis,angiogenesis and survivability of cultured buffalo luteal cells

The objective of this study was to document the expression and localization of angiopoietin (ANGPT) family members comprising of angiopoietin (ANGPT1 and ANGPT2), and their receptors (Tie1 and Tie2) in buffalo corpus luteum (CL) obtained from different stages of the oestrous cycle, and the modulatory role of ANGPT1 and ANGPT2 alone or in combinations on progesterone (P₄) secretion and mRNA expression of phosphotidylinositide‐3kinase‐protein kinase B (PI3K‐AKT), phosphoinositide‐dependent kinase (PDK), protein kinase B (AKT), Bcl2 associated death promoter (BAD), caspase 3 and von willebrand factor (vWF) in luteal cells obtained from midluteal phase (MLP) of oestrous cycle in buffalo. Real‐time RT‐PCR (qPCR), Western blot and immunohistochemistry were applied to investigate mRNA expression, protein expression and localization of examined factors whereas, the P₄ secretion was assessed by RIA. The mRNA and protein expression of ANGPT1 and Tie2 was maximum (p < .05) in mid luteal phase (MLP) of oestrous cycle. The ANGPT2 mRNA and protein expression was maximum (p < .05) in early luteal phase, decreased in MLP and again increased in late luteal phase of oestrous cycle. ANGPT family members were localized in luteal cells and endothelial cells with a stage specific immunoreactivity. P₄ secretion was highest (p < .05) with 100 ng/ml at 72 hr when luteal cells were treated with either protein alone. The mRNA expression of PDK, AKT and vWF was highest (p < .05) and BAD along with caspase 3 were lowest (p < .05) at 100 ng/ml at 72 hr of incubation period, when cultured luteal cells were treated with either protein alone or in combination. To conclude, our study explores the steroidogenic potential of angiopoietins to promote P₄ secretion, luteal cell survival and angiogenesis through an autocrine and paracrine actions in buffalo CL.     

Relationship between rice grain amylose and pasting properties for breeding better quality rice varieties

One hundred and sixty‐seven rice varieties/laudraces from 23 different rice‐growing countries were used for studying the relationship between the amylose content (AC %) and paste viscosity parameters (RVU) through Pearson correlation. AC and paste viscosity parameters were measured using Auto Analyser and Rapid Visco Analysers respectively. Based on all five RVA pasting properties, i.e. peak viscosity (PV), minimum viscosity (MV), breakdown (BD), final viscosity (FV) and setback (SB), the simple paired t‐test (at P = 0.05) was carried out to identify varieties that showed non‐significant differences (similar) from the check variety, separately for each of the four AC groups, e.g. Waxy rice AC (0–8%, n = 21) –‘IR29’ (Philippines); Low AC (8.1–16%, n = 58) –‘Koshihikari’ (Japan); Intermediate AC (16.1–24%, n = 64) –‘Local Sadri’ (Iran); high AC (>24.1%, n = 24) –‘Kasalath’ (India). Significant variation among the rice varieties for AC and pasting properties were observed. Means of the AC and the viscosity parameters including PV, MV, BD, FV and SB were 15.8, 416.1, 180.3, 235.8, 380.9 and 200.6 RVU respectively. Significant correlations existed between AC and paste viscosity parameters at AC group level. The paste viscosity parameters, i.e. PV, BD and SB are important within an AC group as far as cooking quality is concerned. In the low AC group, ‘Koshihikari’ (check) was most preferred by Japanese consumers, which was used for identifying 16 varieties that showed non‐significant differences for pasting properties with it. Some of the important Japanese varieties were ‘Todorokiwase’, ‘Owarihatamochi’, ‘Sasanishiki’, ‘Reimei’ and ‘Kinuhikari’. These varieties had good cooking quality with low AC, high PV and BD with low SB. In contrast, the Iranian cultivar ‘Local Sadri’ with good cooking quality had intermediate levels of AC and SB while recording high PV and BD. The varietal diversity for pasting properties available within each AC group may allow the breeder to make effective selections for improving the cooking quality according to consumer choice.     

Long‐term selection using a single trait criterion,non‐destructive deformation,in White Leghorns: Effect over time on genetic parameters for traits related to egg production

Although non‐destructive deformation is relevant for assessing eggshell strength, few long‐term selection experiments are documented which use non‐destructive deformation as a selection criterion. This study used restricted maximum likelihood‐based methods with a four‐trait animal model to analyze the effect of non‐destructive deformation on egg production, egg weight and sexual maturity in a two‐way selection experiment involving 17 generations of White Leghorns. In the strong shell line, corresponding to the line selected for low non‐destructive deformation values, the heritability estimates were 0.496 for non‐destructive deformation, 0.253 for egg production, 0.660 for egg weight and 0.446 for sexual maturity. In the weak shell line, corresponding to the line selected for high non‐destructive deformation values, the heritabilities were 0.372, 0.162, 0.703 and 0.404, respectively. An asymmetric response to selection was observed for non‐destructive deformation, egg production and sexual maturity, whereas egg weight decreased for both lines. Using non‐destructive deformation to select for stronger eggshell had a small negative effect on egg production and sexual maturity, suggesting the need for breeding programs to balance selection between eggshell traits and egg production traits. However, the analysis of the genetic correlation between non‐destructive deformation and egg weight revealed that large eggs are not associated with poor eggshell quality.     

Comparison of behavioral responses to a novel environment between three teleosts,bluegill <Emphasis Type="Italic">Lepomis macrochirus</Emphasis>, crucian carp <Emphasis Type="Italic">Carassius langsdorfii</Emphasis>, and goldfish <Emphasis Type="Italic">Carassius auratus</Emphasis>

To investigate the emotional reactivity of fish in a novel environment, the swimway test was developed. The swimway apparatus consists of a shaded start chamber and an open, illuminated swimway. Fish were first introduced and habituated to the start chamber. A door partitioning the start chamber from the swimway was then opened, and behavioral responses of the fish in the apparatus were measured. By using the swimway test, behavioral responses to a novel environment of bluegill Lepomis macrochirus, crucian carp Carassius langsdorfii, and goldfish Carassius auratus were quantified and compared. The emotional reactivities in blue gill were found to be the lowest and crucian carp the highest, indicating bluegill are relatively active or ‘bold’, and that the crucian carp are relatively passive or ‘shy’, in a novel environment. It is suggested that the swimway test is applicable to examning inter-species differences in relative emotional reactivity or boldness in a simplified novel situation.     

Prothrombin time and activated partial thromboplastin time using a point‐of‐care analyser (Abaxis VSpro®) in Bennett's wallabies (Macropus rufogriseus)

There are few reports of coagulation times in marsupial species. Blood samples collected from 14 Bennett's wallabies (Macropus rufogriseus) under anaesthesia during routine health assessments were analysed for prothrombin time (PT) and activated partial thromboplastin time (aPTT) using a point‐of‐care analyser (POC) (Abaxis VSPro®). The wallabies had an aPTT mean of 78.09 s and median of 78.1 s. The PT for all wallabies was greater than 35 s, exceeding the longest time measured on the POC. Although PT was significantly longer, aPTT was similar to the manufacturer's domestic canine reference range.     

Diversification of Multipurpose Plant, <Emphasis Type="Italic">Perilla Frutescens</Emphasis>

The traditional Asian crop, Perilla frutescens has multiple uses. There are specialized cultivars for seed oil and for medicinal use, as well as wild/weedy forms growing in various habitats. Based on selective characteristics of leaf odor, anthocyanin pigmentation, seed hardness and seed diameter, the diversity of this species was investigated to clarify the intraspecific differentiation. P. frutescens was divided into five groups by the combination of three qualitative characteristics: leaf odor, anthocyanin pigmentation and seed hardness. Most of the plants cultivated for oil belonged to one group, while medicinal plants belonged to three other groups. Wild/weedy forms were in the last group. The five groups could not be distinguished by seed diameter. Though the plants cultivated for oil tended to have larger seeds than the medicinal and wild/weedy plants, there was no boundary either between the two crops, or among various phenotypes of P. frutescens.