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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.
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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.
Assuntos
Criopreservação , Técnicas de Cultura Embrionária , Bovinos , Animais , Mercaptoetanol/farmacologia , Criopreservação/veterinária , Fertilização in vitro , Vitrificação , BlastocistoRESUMO
Background: The use of conventional artificial insemination (AI) in sheep production is usually associated with lower fertility rates when frozen semen is used. Cooled ram semen has been an alternative over frozen semen due to the higher viability, seminal quality and fertility rates following AI. The semen preservation process promotes sperm cell modifications similar to capacitation (capacitation-like) that causes cell damage affecting viability and seminal quality, but such effects are unclear for cooled semen. The aim of this study was to determine the status of sperm cell capacitation (CA) and acrosome reaction (AR) during ram semen processing and cooling under different extenders, dilution factors, and aerobiosis conditions as a function of storage time at 5o C. Materials, Methods & Results: Two consecutive ejaculates per day per male were collected from 2 adult rams by artificial vagina at 48-72 h intervals, in three replications. After macro- and microscopic evaluations, semen was segregated into groups under 3 extenders (Tris-egg yolk or TY, citrate-egg yolk or CY, skimmed milk or SM), 2 dilution factors (1 x 109 or Bi, 100 x 106 or Mi cells/mL), and 2 aerobiosis conditions (aerobic or A, semi-anaerobic or SA). Diluted semen was cooled to 5ºC and stored for up to 72 h, with evaluations every 24 h. Aliquots of fresh ejaculates and of each cooled diluted subgroup, according to extender, dilution, and aerobiosis, were collected at times T0 and T72 for determination of acrosome status and membrane integrity by the chlortetracycline (CTC) and trypan blue-Giemsa stainings, respectively. No differences were detected in sperm cell motility (M) and motility vigor (V) between fresh and diluted semen. After cooling, a significant decrease in M was observed after 48 h in CY and SM compared with fresh semen and 0 h of cooling, while V started to decrease after 24 h in CY compared with TY. Likewise, M/V from different dilutions and aerobic conditions decreased more significantly after 48 and 24 h of cooling, respectively. The sperm capacitation status did not show differences in the proportion of non-capacitated (NCA), CA and AR sperm cells between TY, CY, and SM extenders (NCA: 75.0%, 71.3%, 74.0%; CA: 15.7%, 17.2%, 15.9%; AR: 9.3%, 11.5%, 10.2%) or between Bi and Mi dilutions (NCA: 74.0%, 72.9%; CA: 15.9%, 16.6%; AR: 10.1%, 10.5%), respectively. However, differences (P < 0.05) were observed between A and SA aerobic conditions, with CA (17.0% vs. 15.5%) and AR (11.9% vs. 8.7%) rates being higher in A than SA, respectively, with no differences in NCA (71.1% vs. 75.8%), irrespective of the storage time. Sperm cell viability decreased after 48 h, especially in CY (P < 0.05). Discussion: Ram sperm cells can suffer irreversible damage due to thermal shock during cooling. Egg yolk-based extenders provide phospholipids and cholesterol to protect the sperm cell membrane during the thermal shock caused by the change in temperature. In this study, sperm cells had irreversible decreases in M/V, with increase in acrosome and plasma membrane damage after cooling to 5ºC. The largest and smallest decreases in M and V over time were observed in the CY and TY extenders, respectively. In addition to the extender type, the semen preservation method and storage time promoted changes in the capacitation status, AR and in sperm cell viability, which per se were associated with a decrease in semen fertility. In fact, the proportions of CA and/or AR sperm cells gradually increased over time after dilution and storage at 5ºC, with a negative correlation between sperm cell viability and M/V over time. In summary, extender and cooling time affected mostly M/V, while aerobiosis condition and dilution factor were more associated with acrosome status and sperm survival, with the extender having less impact on the acrosome status as a function of time.
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Animais , Masculino , Preservação do Sêmen/métodos , Preservação do Sêmen/veterinária , Preservação de Tecido/métodos , Ovinos , Análise do Sêmen/veterinária , Sobrevivência Celular , Técnicas de Diluição do Indicador , AerobioseRESUMO
A 20-month-old miniature mixed-breed heifer weighing 200 kg was presented with a history of dysphagia, sialorrhea and dyspnea after being fed with food residues. At home it was treated with intravenous hydration, antibiotic combination, diuretics and AINEs. Additionally, one failed passage of oro-ruminal tube and several ruminal trocarizations were performed. At clinical examination the animal showed signs compatible with digestive obstruction, severe dyspnea and shock. Complete blood count showed leukocytosis with neutrophilia, left shift and lymphocytosis. Serum chemistry tests showed hyperlactatemia, hypokalemia, hypocalcemia, hyperglycemia and azotemia. Acid-base analysis indicated severe metabolic acidosis and respiratory alkalosis. Esophagoscopy was performed, during which the patient went into shock and died. In necropsy a complete middle third esophageal obstruction with rupture by a carrot was identified.
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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.
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Doenças dos Cavalos , Útero , Aneuploidia , Animais , Colômbia , Feminino , Cavalos , Mosaicismo , Cromossomo XRESUMO
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.
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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.
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Agapornis , Doenças das Aves/diagnóstico , Doenças Ósseas Metabólicas/veterinária , Ossificação Heterotópica/veterinária , Dermatopatias Genéticas/veterinária , Animais , Doenças das Aves/patologia , Doenças Ósseas Metabólicas/diagnóstico , Doenças Ósseas Metabólicas/patologia , Feminino , Ossificação Heterotópica/diagnóstico , Ossificação Heterotópica/patologia , Dermatopatias Genéticas/diagnóstico , Dermatopatias Genéticas/patologiaRESUMO
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
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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.