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.     

Spontaneous regression of intracutaneous cornifying epitheliomata in a dog

The clinical and histopathological features are described of a case of multicentric intracutaneous cornifying epitheliomata in a four-year-old German shepherd dog. Multiple lesions developed over a one-and-a-half year observation period. Eighteen were surgically removed, many of the others spontaneously regressed. Novel histopathological features included prominent epithelial invasion of surrounding dermis and a high mitotic rate suggestive of squamous cell carcinoma, and the apparent origin of several tumours in hair shaft epithelium.     

Oxygen Concentration and Cysteamine Supplementation During In vitro Production of Buffalo (Bubalus bubalis) Embryos Affect mRNA Expression of BCL‐2, BCL‐XL,MCL‐1, BAX and BID

This study examined the effects of O₂ concentration (5% vs 20%) during in vitro maturation (IVM), fertilization (IVF) and culture (IVC) or supplementation of IVM and IVC media with cysteamine (50 and 100 μm , respectively; IVM, IVF and IVC carried out in 20% O₂), on blastocyst rate and relative mRNA abundance of some apoptosis‐related genes measured by real‐time qPCR in immature and in vitro‐matured buffalo oocytes and in embryos at 2‐, 4‐, 8‐ to 16‐cell, morula and blastocyst stages. The blastocyst rate was significantly higher (p < 0.05) while the percentage of TUNEL‐positive cells was significantly lower (p < 0.05) under 5% O₂ than that under 20% O₂. The mRNA expression of anti‐apoptotic genes BCL‐2 and MCL‐1 was significantly higher (p < 0.05) and that of pro‐apoptotic genes BAX and BID was lower (p < 0.05) under 5% O₂ than that under 20% O₂ concentration at many embryonic stages. Following cysteamine supplementation, the blastocyst rate and the relative mRNA abundance of BCL‐XL and MCL‐1 was significantly higher (p < 0.05) and that of BAX but not BID was lower (p < 0.05) at many stages of embryonic development, although it did not affect the percentage of TUNEL positive cells in the blastocysts significantly. The mRNA expression pattern of these genes during embryonic development was different in 5% vs 20% O₂ groups and in cysteamine supplemented vs controls. At the 8‐ to 16‐cell stage, where developmental block occurs in buffalo, the relative mRNA abundance of BCL‐2 and MCL‐1 was highest under 5% O₂ concentration and that of BAX and BID was highest (p < 0.05) under 20% O₂ concentration. These results suggest that one of the mechanisms through which beneficial effects of low O₂ concentration and cysteamine supplementation are mediated during in vitro embryo production is through an increase in the expression of anti‐apoptotic and a decrease in the expression of pro‐apoptotic genes.     

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.     

Accuracy of selected neurological clinical tests in diagnosing MRI-detectable forebrain lesion in dogs

This retrospective case study aims to evaluate the accuracy of menace response, response to nasal stimulation and proprioceptive placing in diagnosing forebrain lesion in dogs. A total of 145 client-owned dogs investigated by magnetic resonance imaging study of the brain between December 2017 and June 2019 were evaluated. Seventy-one dogs with no magnetic resonance imaging-detectable intracranial and significant cerebrospinal fluid abnormality or recent history of seizure (<48 h) served as controls. Binary regression analysis was performed to determine the sensitivity, specificity and likelihood ratios of each selected test. Older age at presentation was a significant risk factor for the presence of a forebrain lesion. Menace (62.5%) and proprioceptive deficits (40.5%) were common findings in all dogs. They were also significantly associated with the presence of forebrain abnormality. Moreover, they were more sensitive (77.3% and 82.2%, respectively) and specific (50.0% and 62.5%, respectively) when applied to dogs aged 6 years or older. Nonetheless, all of these tests' likelihood ratios, and thus reliability are poor. These neurological tests are commonly employed for diagnosing forebrain disease in dogs, yet are not highly accurate in diagnosing forebrain abnormality. Clinicians should interpret these clinical test results along with the patient history when designing a diagnostic plan.     

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.     

Equine wound management: are there significant differences in healing at different sites on the body?

Abstract Trunk/body wounds heal rapidly with prominent Contraction while wounds on the limb commonly fail to heal or heal slowly by centripetal epithelialization. often with insignificant contraction. Chronic exuberant granulating wounds on the limbs heal well after grafting from donor sites on the trunk. Indolent wounds are less common but may granulate significantly following moist wound-healing management. Sarcoid transformation is an increasingly important cause of healing failure. Sarcoid transformations on the trunk are commonly verrucose while those on the limb are usually aggressive and fibroblastic. The primary objective of wound management should be to encourage rapid progression from acute inflammation to repair without intervention of chronic inflammation which is a significant factor in the pathophysiology of wound healing failure. Wounds fail to heal because there is disruption of the normal delicate balance of growth factors and inflammatory mediators. Wounds should be managed in such a way as to restore the balance of healing processes without damaging any of the cells involved in healing. Resumen Las heridas localizadas en el cuerpo o tronco curan rapidamente con contracción elevada, mientras que las de las extremidades no curan o curan lentamente por epitelización centrípeta, a menudo con contracción insignificante. Las heridas crónicas con tejido de granulación exhuberantes en las extremidades curan bien después de injertos a partir de áreas del tronco. Las heridas indolentes son menos frecuentes pero pueden granular significativamente después de un manejo húmedo de la curacion de la herida. La transformación sarcoidal es una causa cada vez más frecuente de fracaso de la curación. Las transformaciones sarcoidales en el tronco son generalmente de tipo verrucoso mientras que las de las extremidades son normalmente agresivas y fibroblásticas. El objetivo principal del manejo de heridas tendría que ser la estimulación de una progresión rápida de una inflamación aguda a la resolución sin intervención de inflamación crónica, que es un factor significativo en la patofisiología del fracaso en la curación de heridas. Las heridas no curan porque se produce una alteración en el frægil equilibrio normal de factores de crecimiento y mediadores inflamatorios. Las heridas tendrian que ser manejadas de forma que se restaure el equilibrio del proceso curativo sin dañar ninguna de las células implicadas en la curacion. [Knottenbelt, D.C. Equine wound management: are there significant differences in healing at different sites on the body? (Manejo de heridas equinas: existen diferencias significativas en la curación en diferentes areas cutaneas?). Veterinar.): Dermatology 1997; 8 : 273–290] Zusammenfassung Wunden am Rumpf heilen schnell und mit deutlicher Kontraktion, während sich Wunden an den Gliedmassen oft nur geringfügig zusammenziehen und entweder gar nicht oder nur langsam vom Rande her epithelisieren. Chronische, stark wuchernde Wunden an den Gliedmassen heilen nach Gewebetransplantation vom Rumpf zufriedenstellend. Indolente Wunden sind weniger häfig, können aber nach benetzender Wundversorgung deutlich granulieren. Sarkoide Transformation ist ein wichtiger Grund für Wundheilungstörungen. Sarkoide Transformationen am Rumpf sind gewöhnlich warzig, jene an den Gliedmassen üblicherweise aggressiv und fibroblastisch. Das wichtigste Ziel der Wundversorgung sollte es sein, eine schnelle Entwicklung von akuter Entzündung zur Heilung zu fördern, ohne in die chronische Entzündung einzugreifen, die ein massgeblicher Faktor in der Störung der Windheilung darstellt. Der Grund für eine nichtheilende Wunde ist eine Störung des enipfindlichen Gleichgewichts zwischen Wachstumsfaktoren und Entzündungsmediatoren. Wundversorgung soll das Gleichgewicht des Heilungsprozesses wiederherstellen, ohne Zellen zu beschädigen, die zur Wundheilung beitragen. [Knottenbelt, D.C. Equine wound management: are there significant differences in healing at different sites on the body? (Wundversorgung beim Pferd: Sind signifikante Heilungsunterschiede an verschiedenen Körperstellen vorhanden?). Veterinary Dermatology 1997; 8 : 273–290] Résumé Les plaies affectant le tronc cicatrisent rapidement avec une contraction proéminente tandis que les plaies des membres cicatrisent ma1 ou lentement avec une épithélialisation centripète, et souvent avec une contraction insignifiante. Les plaies présentant une granulation chronique exhubérante sur les membres cicatrisent bien après greffe à partir d'un fragment prélevé sur le tronc. Les plaies atones sonl moins fréquentes, mais peuvent présenter un tissu de granulation significatif après application de pansements humides. La transformation en sarcoïde est une cause de plus en plus importante d'échec de cicatrisation. Les transformations en sarcoïdes sur le tronc sont fréquemment verruqueuses, alors que celles localisées sur les membres sont généralement agressives et fibroblastiques. Le but premier du traitement d'une plaie est de promouvoir la progression rapide d'une inflammation aigüe en cicatrisation sans passage par l'inflammation chronique qui est un facteur significatif dans la pathophysiologie de l'échec de cicatrisation des plaies. Les plaies ne cicatrisent pas parce qu'il y a un déséquilibre de la balance délicate entre facteurs de croissance et médiateurs de l'inflammation. Les plaies doivent être traitées de manière à restaurer l'équilibre des processus de cicatrisation sans altération des cellules impliquées dans l'inflammation. [Knottenbelt, D.C. Equine wound management: are there significant differences in healing at different sites on the body? (Traitement des plaies équines: y-a-t-il des différences significatives de cicatrisation en fonction de la topographie corporelle?). Veterinary Dermatology 1997; 8 : 273–290]