ß-Mercaptoethanol in culture medium improves cryotolerance of in vitro-produced bovine embryos.

The objective of this study was to investigate the effects of adding ß-mercaptoethanol (ßME) to culture medium of bovine in vitro-produced (IVP) embryos prior to or after vitrification on embryo development and cryotolerance. In Experiment I, Day-7 IVP blastocysts were vitrified and, after warming, cultured in medium containing 0, 50 or 100 µM ßME for 72 h. Embryos cultured in 100 µM ßME attained higher hatching rates (66.7%) than those culture in 0 (47.7%) and 50 (52.4%) µM ßME. In Experiment II, IVP embryos were in vitro-cultured (IVC) to the blastocyst stage in 0 (control) or 100 µM ßME, followed by vitrification. After warming, embryos were cultured for 72 h (post-warming culture, PWC) in 0 (control) or 100 µM ßME, in a 2 × 2 factorial design: (i) CTRL-CTRL, control IVC and control PWC; (ii) CTRL-ßME, control IVC and ßME-supplemented PWC; (iii) ßME-CTRL, ßME-supplemented IVC and control PWC; or (iv) ßME-ßME, ßME-supplemented IVC and ßME-supplemented PWC. ßME during IVC reduced embryo development (28.0% vs. 43.8%) but, following vitrification, higher re-expansion rates were seen in ßME-CTRL (84.0%) and ßME-ßME (87.5%) than in CTRL-CTRL (71.0%) and CTRL-ßME (73.1%). Hatching rates were higher in CTRL-ßME (58.1%) and ßME-ßME (63.8%) than in CTRL-CTRL (36.6%) and ßME-CTRL (42.0%). Total cell number in hatched blastocysts was higher in ßME-ßME (181.2 ± 7.4 cells) than CTRL-CTRL (139.0 ± 9.9 cells). Adding ßME to the IVC medium reduced development but increased cryotolerance, whereas adding ßME to the PWC medium improved embryo survival, hatching rates, and total cell numbers.

Chemical activation of mammalian oocytes and its application in camelid reproductive biotechnologies: A review.

Mammalian oocyte activation is a critical process occurring post-gamete fusion, marked by a sequence of cellular events initiated by an upsurge in intracellular Ca2+. This surge in calcium orchestrates the activation/deactivation of specific kinases, leading to the subsequent inactivation of MPF and MAPK activities, alongside PKC activation. Despite various attempts to induce artificial activation using distinct chemical compounds as Ca2+ inducers and/or Ca2+-independent agents, the outcomes have proven suboptimal. Notably, incomplete suppression of MPF and MAPK activities persists, necessitating a combination of different agents for enhanced efficiency. Moreover, the inherent specificity of activation methods for each species precludes straightforward extrapolation between them. Consequently, optimization of protocols for each species and for each technique, such as PA, ICSI, and SCNT, is required. Despite recent strides in camelid biotechnologies, the field has seen little advancement in chemical activation methods. Only a limited number of chemical agents have been explored, and the effects of many remain unknown. In ICSI, despite obtaining blastocysts with different chemical compounds that induce Ca2+ and calcium-independent increases, viable offspring have not been obtained. However, SCNT has exhibited varying outcomes, successfully yielding viable offspring with a reduced number of chemical activators. This article comprehensively reviews the current understanding of the physiological activation of oocytes and the molecular mechanisms underlying chemical activation in mammals. The aim is to transfer and apply this knowledge to camelid reproductive biotechnologies, with emphasis on chemical activation in PA, ICSI, and SCNT.

Inter-software reliability and agreement for follicular and luteal morphometric and echotextural ultrasonographic parameters in beef cattle.

This study aimed to compare the inter-software and inter-observer reliability and agreement for the assessment of follicular and luteal morphometry and echotexture parameters in beef crossbreed females (3/8 Bos taurus indicus and 5/8 Bos taurus taurus). B-mode and color Doppler ultrasonographic ovarian images were obtained at specific time points of estradiol-progesterone-based protocols for timed artificial insemination (TAI). Sonograms were analyzed by two observers using a licensed (IASP1) and an open access (IASP2) software package. A total of 292 snap-shot sonograms were analyzed for morphometric parameters and 504 for echotexture parameters. inter-software reliability was judged moderate to excellent (ICC or CCC=0.73-0.98), whereas inter-observer reliability for morphometric parameters was deemed good to excellent (ICC or CCC=0.82-0.98). A small percentage (up to 10.95 %) of measured parameters fell outside the limits of inter-software and inter-observer agreement. For echotexture parameters, inter-software reliability varied widely (ICC or CCC=0.16-0.95) based on the size of regions of interest (ROI), while inter-observer reliability ranged from moderate to excellent (ICC or CCC= 0.71-0.97). The highest inter-software reliability for pixel value and heterogeneity value was observed for the corpus luteum (ICCs=0.81-0.95; P>0.05), followed by the peripheral follicular antrum (ICCs=0.75-0.78; P<0.05). However, lower reliability was determined for the follicular wall (ICCs=0.08-0.33; P<0.0001) and perifollicular stroma (ICCs=0.16-0.46; P<0.05). In conclusion, both software packages showed high reproducibility for morphometric measurements, while echotexture measurements were more challenging to replicate based on ROI sizes. Caution is advised when selecting ROI sizes for echotexture measurements in bovine ovaries.

In vitro and in vivo embryo production efficiency in Flemish and Holstein donor females.

The aim of this study was to compare embryo production efficiency in Flemish and Holstein donor females using ovum pick-up and in vitro fertilization (OPU-IVF) or in vivo production (superovulation; SOV) procedures. The study was conducted using a split-plot design, with eight Flemish and eight Holstein non-lactating cycling females. Females were subjected to ten weekly OPU/IVF sessions and/or two SOV/embryo collections sessions at a 63-day interval, for a total of 160 OPU-IVF and 32 SOV sessions. Mean numbers of follicles and corpora lutea, and cumulus-oocyte complex (COC) recovery rates were similar between breeds after the OPU and SOV sessions. However, Flemish donors yielded better quality grade II COCs (301, 41.9%) than Holstein females (609, and 202, 33.1%). Also, cleavage and blastocyst rates, and the total number and the mean number of viable embryos obtained after OPU-IVF were higher in Flemish (49.6% and 11.8%, and 63 and 11.8 per donor, respectively) than in Holstein (32.8% and 7.2%, and 34 and 7.2 per donor, respectively) females. Flemish females were also more efficient in yielding viable embryos after SOV (111, 7.3 per donor) than Holstein (48, 3.3 per donor) females. Overall, Flemish donor females had better responses to OPU-IVF or SOV procedures than Holstein counterparts. Irrespective of the breeds, SOV procedures were more efficient than OPU-IVF in yielding more viable embryos, under the conditions of this study. Both reproductive procedures were useful tools for the genetic conservation of the Flemish cattle breed in Southern Brazil.

Rhabdoid melanoma in a harpy eagle (Harpia harpyja).

A 28-year-old male harpy eagle (Harpia harpyja) with a history of anorexia, hyporexia, lethargy, and progressive weight loss was found dead and submitted for post-mortem examination. Gross findings include dark brown discolouration of testes and lungs; the testes were bilaterally enlarged, glistening brown-grey to blackish in appearance, firm, smooth, and multilobulated. The lungs contained a mass with similar features to the testicles, irregularly shaped with multiple nodules. Histology of testis showed round, polygonal and pleomorphic cells, containing melanin pigments and a typical eosinophilic vacuole in their cytoplasm and with severe pleomorphism. An immunohistochemistry panel with Melan-A, vimentin, CK AE1/AE/3, MUM-1 and CD-68 were performed, yielding a positive reaction for Melan-A and vimentin. The morphology of the tumour cells, the presence of melanin pigment and the immunoreactivity for Melan-A and vimentin by the cells led to a diagnosis of rhabdoid melanoma. This is the first case of this pathology in the testis with lung metastasis in a harpy eagle.

Invasive leiomyosarcoma involving the tongue of an Arabian mare.

A 10 year-old endurance Arabian mare was presented with an invasive, firm, multilobulated mass at the left ventral face of the tongue. The mare had a history of four months of ptyalism, dysphagia, severe halitosis, and dysmastication. The Evaluation of the horse included physical and oral examination, complete blood count and serum biochemistry profile, computed tomography evaluation of the head, and histopathology of the mass. The computerized tomography (CT) head scan showed the homogeneous mass in the oral cavity confirming the degree of invasion on the tongue, as well as a small mineral focus at the left dorsal mandibular border and increased density of the submandibular lymph nodes. At necropsy, gross findings in the oral cavity revealed an irregular, nodular and partially ulcerated mass at the level of the body of the tongue in the ventral aspect, with a size of approximately 10 × 15 × 8 cm. Histologically, the tongue revealed a neoplastic process of mesenchymal muscle origin. The tumor cells showed distinctive morphological and architectural patterns in some areas. An immunohistochemistry panel was done yielding a positive reaction for Calponin and Desmin, confirming diagnostics of as a soft tissue leiomyosarcoma in the tongue. Immunohistochemistry results, in combination with histopathologic morphology, were suggestive of a leiomyosarcoma originating in the perivascular wall tissue. Oral leiomyosarcoma are very rare in veterinary medicine, and to the authors' knowledge, this is the first case report to utilize histopathology, immunohistochemistry and imaginology to describe oral leiomyosarcoma in a horse.

Segmental Cervical Aplasia in a Colombian Creole Mare with Mosaic X-Chromosome Aneuploidy.

A 4-year-old Colombian Creole mare was presented for diagnosis because the external orifice of her cervix was not detectable when a uterine lavage as therapy for uterine fluid accumulation was attempted. Clinical and ultrasonographic evaluation of the genital tract revealed that ovaries were of normal size and showed structures suggestive of regular ovarian activity. However, granular free-floating fluid material distending the uterus was detected by ultrasound. Upon vaginal examination, the normal external cervical morphology was not evident. The vagina ended in a blind bag with a small papilla with no evident external cervical os. Cytology of uterine fluid obtained by transvaginal aspiration showed findings compatible with mucometra. Cytogenetic analysis revealed an abnormal karyotype (63,X and 64,XX both 45% and 65,XXX 10%). A diagnosis of congenital segmental cervical aplasia was proposed possibly related to the mosaicism detected. To our knowledge, this is the first case of this reproductive pathology in a mare with regular ovarian activity and confirmed aneuploidy in mosaic form of the X sex chromosome.

Bilateral osteoma cutis in a Peach-Faced Lovebird (Agapornis roseicollis).

An osteoma is an infrequent tumor documented in avian species. An adult female Peach-Faced Lovebird (Agapornis roseicollis) with a history of previous trauma was examined due to the presence of bilateral hard and yellowish-white masses in the radio-cubital humerus junction. Histopathological dermal examination revealed a non-neoplastic process of mesenchymal origin, characterized by the formation of well-differentiated trabecular bone, multiple areas of medullary bone and loose connective tissue and coagulation of the necrosis foci. Based on the histological findings and the medical history, the masses were diagnosed as bilateral secondary osteoma cutis. To our knowledge, this is the first report of this pathology with an acute course in this exotic pet bird. The previous trauma could be the initiating cause.

Primer registro de Scleroderma bovista (Boletales, Sclerodermataceae) para Colombia / First record of Scleroderma bovista (Boletales, Sclerodermataceae) for Colombia

Se describe e ilustra por primera vez a Scleroderma bovista para Colombia, un hongo gasteroide hallado en el campus de la Universidad Pedagógica Nacional (Bogotá), departamento de Cundinamarca. Se aporta información sobre su distribución, ecología y sustrato de crecimiento. Así, el género Scleroderma queda representado en el país por cuatro especies: S. albidum, S. areolatum, S. citrinum y S. bovista.

Scleroderma bovista is described and illustrated for the first time in Colombia. Scleroderma bovista is a gasteroid fungi found on the campus of the Universidad Pedagógica Nacional (Bogotá), department of Cundinamarca. Information on distribution, ecology and growth substrate is provided. Thus, the genus Scleroderma is represented in the country by four species: S. albidum, S. areolatum, S. citrinum y S. bovista

Laternea pusilla (PHALLALES, PHALLACEAE) UNA NUEVA ESPECIE PARA COLOMBIA / Laternea pusilla (PHALLALES, PHALLACEAE) a New Species for Colombia

RESUMEN Se registra por primera vez a Laternea pusilla para Colombia, un macrohongo que prolifera en un bosque húmedo montano bajo (bh-MB) de los Cerros Orientales de Bogotá, departamento de Cundinamarca. La especie es descrita e ilustrada con fotografías y se aporta información sobre su distribución, ecología y sustrato de crecimiento. Así, el género Laternea queda representado en el país por dos especies, L. triscapa y L. pusilla.

ABSTRACT Laternea pusilla is recorded for the first time in Colombia. Laternea pusilla is a macrofungi that occurs on a lower mountain humid rainforest (bh-MB) of the eastern hills of Bogota city, department of Cundinamarca. The species is described and illustrated with photographs, and information on distribution, ecology and growth substrate is provided. Thus, the genus Laternea is represented in the country by two species, L. triscapa and L. pusilla.