Secretion pattern of canine amniotic stem cells derived extracellular vesicles

Extracellular vesicles (EVs) derived from stem cells (SCs) have regenerative potential and the possibility of being used in treating chronic diseases. EVs present lower risk of tumorigenicity and easily to isolation and storage. Therefore, this research aims to compare the morphological characteristics of the EVs (up to 150nm) derived from stem cells obtained from canine amniotic membranes in different passages during the in vitro culture. For this, cells from the amniotic membranes were isolated, cultured, and characterized. In order to answer our aim, the number of cells was normalized at each passage to generate conditioned media for EVs separation. The cells were differentiated into adipogenic, chondrogenic, and osteogenic tissue, to characterize these cells as mesenchymal stem cells (MSC). Moreover, flow cytometry analysis was performed and showed that the MSC were positive for CD90, CD105 and negative for CD34, CD45, mesenchymal and hematopoietic markers, respectively. For EVs analysis, MSC in different passages (P0-P2) were culture until 80% of confluence, then the medium was replaced by EVs depleted medium. After 48h, culture medium was collected and centrifuged to separate EVs, followed by nanoparticle tracking analysis. The EVs were also characterized by western blot and transmission electron microscopy (TEM). EVs were positive for Alix and negative for Cytochrome C as well as presented the traditional cup-shape by transmission electronic microscopy. Our results demonstrated that the concentration in the different passages was increased in P0 compared to P1 and P2 (p<0.05). No differences were found in EVs size (P0=132nm, P1=130nm and P2=120nm). Together, these results demonstrate that P0 of MSC is enriched of EVs when compared to later passages, suggesting that this passage would be the best to be applied in pre-clinical tests. Despite that, more studies are necessary to identify the EVs content and how the cells will respond to treatment with them.(AU)

Myelodysplastic syndrome progression to acute myeloid leukemia in a cat FeLV seroreactive

Background: Acute myeloid leukemia (AML) is a group of non-lymphoid hematological tumors characterized by aberrant proliferation and/or decreased apoptosis of a clone of non-mature cells, resulting in the accumulation of immature blast cells in the bone marrow and peripheral blood. It is considered rare, as it represents 10% of neoplasms of hematopoietic origin. However, it is known that felines seroreactive for FIV and FeLV are more predisposed and reports of this type of leukemia in cats in the literature are scarce. Thus, the objective of this study was to evaluate the blood and bone marrow of a cat seroreactive for FeLV that presented with myelodysplastic syndrome that progressed to acute myeloid leukemia. Case: A 6-year-old male mixed-breed cat, neutered, seroreactive for FeLV, showed apathy, weight loss, and pale mucous membranes. Initial peripheral blood smear evaluation revealed hypochromic normocytic anemia, leukopenia, neutropenia, lymphopenia, and thrombocytosis with many macropackets and giant platelets. Based on this blood picture, a long-spectrum antimicrobial therapy with amoxicillin and clavulanate [Clavulin® BD - 25 mg/kg, every 12 h] was started. Granulocyte colony stimulating factor used filgrastim (rHu G-CSF) [Fiprina® - 5 µg/kg, SC, every 48 h] and appetite stimulant mirtazapine [Mirtz® - 2 mg/cat, orally, every 48 h] were used to correct leukopenia and nutritional status, respectively. Follow-up blood smear evaluation on the 30th day showed persistence of the hematological changes noticed earlier. A bone marrow puncture was performed, and immunosuppressive therapy with prednisolone [Predsim® - 4 mg/kg, orally, every 24 h] was initiated. The aspirated material showed increased cellularity for age, decreased myeloid:erythroid ratio, and 39.8% of blasts of myeloid origin. An average of 17.7 megakaryocytes were observed per field (10x magnification). Bone marrow cytological evaluation suggested acute myeloid leukemia with dysmegakaryocytopoiesis. After the diagnosis, the examinations were repeated monthly, and there was still intense leukopenia. However, in view of the stable clinical status and leukopenia with neutropenia, treatment for leukemia was not instituted and only supportive treatment was administered when necessary. Eight months after the diagnosis, clinical status had worsened, and unlike the earlier hemograms, global leukocyte count had increased with predominant lymphocytosis (95% of the total leukocytes) with atypical lymphocytes. The cat died a few days later. Discussion: Bone marrow evaluation is indicated when peripheral blood cell abnormalities are present and cannot be explained in the context of the clinical history. In the present report, the bone marrow aspirate was hypercellular (cellularity above 75%); however, intense leukopenia was observed in the peripheral blood. In myelodysplastic syndromes, it is common for the bone marrow to be normal to hypercellular, which occurs when there is a greater production of myeloid or erythroid cell lines in response to the loss, destruction, or consumption of cells. Despite this, cytopenias may be present in the peripheral blood, since the defective cells undergo apoptosis and die before being released into the circulation, characterizing inefficient hematopoiesis. The diagnosis of acute leukemia comprises a variety of hematopoietic neoplasms that are complex and unique. Each acute leukemia subtype has defining characteristics that affect the prognosis and treatment of each animal.

Achado acidental de mumificação fetal em cadela cardiopata / Accidental finding of mummified fetus in bitch with cardiac disease

A mumificação fetal é um processo que ocorre após a morte do feto, que já tem mineralização da matriz óssea do esqueleto. O mecanismo exato deste fenômeno não é totalmente conhecido, entretanto, sabe-se que após a morte fetal ocorre rápida absorção dos fluidos uterinos e fetais juntamente com a involução uterina. As membranas fetais e placentárias desidratadas aderem ao feto formando uma massa seca e dura. Apesar de ser considerada incomum, já foi descrita em várias espécies de animais. É de rara ocorrência em cadelas e gatas. O objetivo deste artigo foi relatar o diagnóstico acidental de mumificação fetal em uma cadela com sinais de alteração cardíaca. Uma cadela de 11 anos apresentando sinais de insuficiência cardíaca congestiva foi submetida a radiografia torácica para avaliação da cardiopatia. No exame radiográfico foi observado acidentalmente a presença de um feto mumificado na cavidade abdominal cranial direita. A paciente foi submetida ao tratamento clínico para estabilização e controle dos sinais e apresentou, posteriormente, uma rápida recuperação clínica. O animal não apresentava sinais ou complicações decorrentes da mumificação fetal. Visto isso e para não a submeter aos riscos cirúrgicos, a tutora optou por não realizar o procedimento cirúrgico para retirada do feto. O presente relato descreve um caso clínico atípico de mumificação em cadela, pois não houve procedimento terapêutico específico e isso não interferiu com a boa qualidade de vida da paciente.

Fetal mummification occurs after the death of the fetus, which already has mineralization the skeleton bone matrix. The exact mechanism of this phenomenon is not fully understood; however, it is known that after fetal death a fast absorption of uterine and fetal fluids take place along with uterine involution. Dehydrated fetal and placental membranes adhere to the fetus forming a dry, hard mass. Although considered unusual, it has been described in various species of animals. It is rare in dogs and cats. The purpose of this paper was to report the accidental diagnosis of fetal mummification in a bitch with signs of cardiac alteration. An 11-year-old bitch showing signs of congestive heart failure was admitted to a veterinary clinic and it was requested to perform chest radiographs for heart evaluation. Radiography accidentally revealed the presence of mummified fetus in the right cranial abdominal cavity. The patient was submitted to clinical treatment for stabilization and control of the disease signs and clinical recovery rapidly occurred. The animal showed no signs or complications due to fetal mummification and had a good quality of life. Once the dog was clinically normal, and not to submit it to surgical risks, the owner chose not to perform the surgical procedure to remove the mummified fetus.

Eversão da cartilagem da terceira pálpebra em um gato / Eversion of the third eyelid cartilage in a cat

Background: Eversion of the cartilage of the third eyelid is a rare congenital disease in cats. It is caused by the anterioreversion of the cartilage edge of the third eyelid. Clinical signs may be associated with secondary keratoconjunctivitis,third eyelid gland protrusion, and ocular surface irritation. The diagnosis is made by ophthalmic examination, and treatment consists of surgical resection of the everted cartilage portion. The goal of the present study was to report a case ofeversion of third eyelid cartilage in a cat, given that it is an unusual abnormality in this animal species, and an importantdifferential diagnosis to be considered in the disorders of the third eyelid.Case: A 6-year-old neutered female Persian cat was presented with a presumptive diagnosis of protrusion of the thirdeyelid gland, history of ocular irritation, and epiphora in the left eye. The disorder had been intermittently present sincethe animal was 1-year-old, with spontaneous disappearance after approximately 15 days. The owner related the reappearance of the disorder to stressful situations, with no previous history of trauma or other ocular alteration. During theophthalmic examination, suspended solute was observed through biomiscroscopic examination in both eyes, as well asan increase in volume of the third eyelid in the left eye, without other changes. A thorough examination, under generalanesthesia, indicated the protruding volume of the cartilage of the everted third eyelid. The third eyelid was pleated inits upper portion, demonstrating that the cartilage of the third eyelid was folded instead of following the curvature of theocular surface. Under general anesthesia, the cartilage was partially removed through two parallel incisions on the bulbarconjunctival surface, divulsioning 5 mm in length in the vertical portion of the cartilage in a ‘T’ shape, and separating the...

Eversão da cartilagem da terceira pálpebra em um gato / Eversion of the third eyelid cartilage in a cat

Background: Eversion of the cartilage of the third eyelid is a rare congenital disease in cats. It is caused by the anterioreversion of the cartilage edge of the third eyelid. Clinical signs may be associated with secondary keratoconjunctivitis,third eyelid gland protrusion, and ocular surface irritation. The diagnosis is made by ophthalmic examination, and treatment consists of surgical resection of the everted cartilage portion. The goal of the present study was to report a case ofeversion of third eyelid cartilage in a cat, given that it is an unusual abnormality in this animal species, and an importantdifferential diagnosis to be considered in the disorders of the third eyelid.Case: A 6-year-old neutered female Persian cat was presented with a presumptive diagnosis of protrusion of the thirdeyelid gland, history of ocular irritation, and epiphora in the left eye. The disorder had been intermittently present sincethe animal was 1-year-old, with spontaneous disappearance after approximately 15 days. The owner related the reappearance of the disorder to stressful situations, with no previous history of trauma or other ocular alteration. During theophthalmic examination, suspended solute was observed through biomiscroscopic examination in both eyes, as well asan increase in volume of the third eyelid in the left eye, without other changes. A thorough examination, under generalanesthesia, indicated the protruding volume of the cartilage of the everted third eyelid. The third eyelid was pleated inits upper portion, demonstrating that the cartilage of the third eyelid was folded instead of following the curvature of theocular surface. Under general anesthesia, the cartilage was partially removed through two parallel incisions on the bulbarconjunctival surface, divulsioning 5 mm in length in the vertical portion of the cartilage in a ‘T shape, and separating the...(AU)

Pancitopenia induzida em um cão pelo uso concomitante de fármacos mielotóxicos / Pancytopenia in a dog induced by concomitant use of myelotoxic drugs

Background: The indiscriminate use of drugs is an issue in Veterinary Medicine, as it has serious consequences for theanimals. Many drugs are myelotoxic and cause a decrease in the production of blood cells, which may be irreversible insome cases. The present work reports a case of pancytopenia induced by the concomitant use of myelotoxic drugs (estrogen, metamizole and phenobarbital) in a dog and describes findings on myelotoxicity, hematological alterations andtreatment success.Case: A 7-year-old Lhasa Apso bitch was referred to the Veterinary Hospital of Federal University of Paraná, Curitibacampus, with hematuria and a history of treatment with phenobarbital [2 mg/kg twice a day (bis in die, BID)], metamizole[25 mg/kg 3 times a day (ter in die, TID)], and use of estrogen hormone (estradiol cypionate). At physical examination, theanimal was normohydrated and exhibited normal palpable lymph nodes, pale mucous membranes, galactorrhea, and a bodytemperature of 36°C. A complete blood count including reticulocyte count and a total plasma protein (TPP) exam wererequested. The results revealed pancytopenia (18% hematocrit, 1,400 total leucocytes/µL, and 22,000 reticulocytes/µL).An abdominal ultrasound exam did not detect any relevant alterations. In view of the results obtained, medullary aplasiawas suspected. A bone marrow aspiration was performed. A myelogram revealed a decrease in cellularity (erythrocyticand granulocytic hypoplasia), with presence of rare erythroid and granulocytic precursors. The diagnosis was medullaryaplasia. The animal was treated, and the evolution of the hematological alterations was monitored. The treatment consistedof administration of erythropoietin (100UI/kg subcutaneously every 48 h), prednisone (2 mg/kg BID), Leucogen (3 mg/kg BID), interferon (0.2 IU/kg BID) and Eritrós Dog Tabs [1 tablet once a day (semel in die, SID)]. After 5 days of treatment, the...

Pancitopenia induzida em um cão pelo uso concomitante de fármacos mielotóxicos / Pancytopenia in a dog induced by concomitant use of myelotoxic drugs

Background: The indiscriminate use of drugs is an issue in Veterinary Medicine, as it has serious consequences for theanimals. Many drugs are myelotoxic and cause a decrease in the production of blood cells, which may be irreversible insome cases. The present work reports a case of pancytopenia induced by the concomitant use of myelotoxic drugs (estrogen, metamizole and phenobarbital) in a dog and describes findings on myelotoxicity, hematological alterations andtreatment success.Case: A 7-year-old Lhasa Apso bitch was referred to the Veterinary Hospital of Federal University of Paraná, Curitibacampus, with hematuria and a history of treatment with phenobarbital [2 mg/kg twice a day (bis in die, BID)], metamizole[25 mg/kg 3 times a day (ter in die, TID)], and use of estrogen hormone (estradiol cypionate). At physical examination, theanimal was normohydrated and exhibited normal palpable lymph nodes, pale mucous membranes, galactorrhea, and a bodytemperature of 36°C. A complete blood count including reticulocyte count and a total plasma protein (TPP) exam wererequested. The results revealed pancytopenia (18% hematocrit, 1,400 total leucocytes/µL, and 22,000 reticulocytes/µL).An abdominal ultrasound exam did not detect any relevant alterations. In view of the results obtained, medullary aplasiawas suspected. A bone marrow aspiration was performed. A myelogram revealed a decrease in cellularity (erythrocyticand granulocytic hypoplasia), with presence of rare erythroid and granulocytic precursors. The diagnosis was medullaryaplasia. The animal was treated, and the evolution of the hematological alterations was monitored. The treatment consistedof administration of erythropoietin (100UI/kg subcutaneously every 48 h), prednisone (2 mg/kg BID), Leucogen (3 mg/kg BID), interferon (0.2 IU/kg BID) and Eritrós Dog Tabs [1 tablet once a day (semel in die, SID)]. After 5 days of treatment, the...(AU)

Pupillary membrane persistence in a feline

Background: Pupillary membrane persistence (PMP) is a congenital abnormality, which is not usually reported in felines.It is characterized by remnants of the fetal membrane that persist as filamentous tissue across the pupil. In general, thischange does not cause any clinical signs. However, the filaments may either attach to the cornea and cause small opacitiesin it or attach to the lens and cause cataracts. In most cases, there is no visual impairment, so treatment is not required.This report aims to describe a case of PMP in a domestic cat diagnosed at the Veterinary Hospital of the State Universityof Santa Cruz (HV-UESC).Case: A two-and-a-half-year-old mixed-breed castrated male cat was brought to the HV-UESC with dermatological complaints. Upon physical examination, the animal was alert with a body temperature, heart, and respiratory rate within thenormal parameters for the feline species. The lymph nodes were non-reactive, and the coloration of the oral mucosa wasnormal. There was no ophthalmic complaint from the owner, nor any loss of visual acuity. In addition, the animal hadmoderate pruritus, redness, and alopecia in the region of the ears, head, neck, chest, and back. Bristle samples were collected for an optical microscope analysis and an infestation with lice (Felicola subrostratus) was confirmed. An endectocidecontaining selamectin (15 mg; single application every 30 days) was prescribed. During physical examination, filamentoustissue crossing from iris to iris through pupil was observed in both eyes. The eyelid, corneal, and pupillary reflexes werewithin normal ranges. An ophthalmic evaluation did not identify conjunctival hyperemia or episcleral vessel congestion,and the eyelid, corneal, and pupillary reflexes were determined to be within the normal range. A slit-lamp biomicroscopydid not detect any anterior chamber alteration besides the filamentous tissue previously mentioned...

Pupillary membrane persistence in a feline

Background: Pupillary membrane persistence (PMP) is a congenital abnormality, which is not usually reported in felines.It is characterized by remnants of the fetal membrane that persist as filamentous tissue across the pupil. In general, thischange does not cause any clinical signs. However, the filaments may either attach to the cornea and cause small opacitiesin it or attach to the lens and cause cataracts. In most cases, there is no visual impairment, so treatment is not required.This report aims to describe a case of PMP in a domestic cat diagnosed at the Veterinary Hospital of the State Universityof Santa Cruz (HV-UESC).Case: A two-and-a-half-year-old mixed-breed castrated male cat was brought to the HV-UESC with dermatological complaints. Upon physical examination, the animal was alert with a body temperature, heart, and respiratory rate within thenormal parameters for the feline species. The lymph nodes were non-reactive, and the coloration of the oral mucosa wasnormal. There was no ophthalmic complaint from the owner, nor any loss of visual acuity. In addition, the animal hadmoderate pruritus, redness, and alopecia in the region of the ears, head, neck, chest, and back. Bristle samples were collected for an optical microscope analysis and an infestation with lice (Felicola subrostratus) was confirmed. An endectocidecontaining selamectin (15 mg; single application every 30 days) was prescribed. During physical examination, filamentoustissue crossing from iris to iris through pupil was observed in both eyes. The eyelid, corneal, and pupillary reflexes werewithin normal ranges. An ophthalmic evaluation did not identify conjunctival hyperemia or episcleral vessel congestion,and the eyelid, corneal, and pupillary reflexes were determined to be within the normal range. A slit-lamp biomicroscopydid not detect any anterior chamber alteration besides the filamentous tissue previously mentioned...(AU)

Avaliação inicial da gestação equina por ultrassonografia 3D Relato de caso / Initial evaluation of equine pregnancy by 3D ultrasound Case report

O presente trabalho objetivou o acompanhamento da gestação inicial na égua entre os dias 12 a35 pós-ovulação por meio da ultrassonografia tridimensional (3D). Após o diagnóstico de gestação no dia12 pós-ovulação, a reprodutora foi examinada diariamente por palpação retal e ultrassonografia paraadquirir imagens no modo B e em 3D. Nas avaliações diárias e com as imagens construídas pela técnicade ultrassonografia tridimensional foi possível acompanhar o crescimento do concepto, a identificação doembrião propriamente dito e a diferenciação de estruturas e membranas (saco vitelino e saco alantoide)permitindo visualizações em 360 graus. Pode-se concluir que na técnica de processamento da imagemtridimensional as imagens geradas fornecem excelente material educacional e acadêmico, precisando demais estudos para permitir uma efetiva aplicação clínica na perda embrionária.(AU)

The present study aimed at monitoring the initial pregnancy in the mare between days 12 to 35after ovulation using three-dimensional (3D) ultrasonography. After the diagnosis of gestation on day 12after ovulation, the reproductive tract was examined daily by rectal palpation and ultrasonography toacquire images in B mode and in 3D. In the daily evaluations and with the images constructed by thetechnique of three-dimensional ultrasonography, it was possible to follow the growth of the concept, theidentification of the embryo proper and the differentiation of structures and membranes (yolk andallantoic sac) allowing visualizations in 360 degrees. It can be concluded that the 3D generated imagesprovide excellent educational and academic material however, more studies are needed to allow aneffective application in embryonic loss.(AU)

Avaliação inicial da gestação equina por ultrassonografia 3D – Relato de caso / Initial evaluation of equine pregnancy by 3D ultrasound – Case report

O presente trabalho objetivou o acompanhamento da gestação inicial na égua entre os dias 12 a35 pós-ovulação por meio da ultrassonografia tridimensional (3D). Após o diagnóstico de gestação no dia12 pós-ovulação, a reprodutora foi examinada diariamente por palpação retal e ultrassonografia paraadquirir imagens no modo B e em 3D. Nas avaliações diárias e com as imagens construídas pela técnicade ultrassonografia tridimensional foi possível acompanhar o crescimento do concepto, a identificação doembrião propriamente dito e a diferenciação de estruturas e membranas (saco vitelino e saco alantoide)permitindo visualizações em 360 graus. Pode-se concluir que na técnica de processamento da imagemtridimensional as imagens geradas fornecem excelente material educacional e acadêmico, precisando demais estudos para permitir uma efetiva aplicação clínica na perda embrionária.

The present study aimed at monitoring the initial pregnancy in the mare between days 12 to 35after ovulation using three-dimensional (3D) ultrasonography. After the diagnosis of gestation on day 12after ovulation, the reproductive tract was examined daily by rectal palpation and ultrasonography toacquire images in B mode and in 3D. In the daily evaluations and with the images constructed by thetechnique of three-dimensional ultrasonography, it was possible to follow the growth of the concept, theidentification of the embryo proper and the differentiation of structures and membranes (yolk andallantoic sac) allowing visualizations in 360 degrees. It can be concluded that the 3D generated imagesprovide excellent educational and academic material however, more studies are needed to allow aneffective application in embryonic loss.

Increased quality of in natura and cryopreserved semen of water buffaloes supplemented with saturated and unsaturated fatty acids from the palm oil industry

Ruminant energy supplementation with vegetable oils or fats has been standing out worldwide and oil palm processing has been receiving growing interest. This study assessed the effect of supplementation with saturated and unsaturated fatty acids from the palm oil industry on the lipid profile of seminal plasma and of the sperm membrane, as well as on the morphological and functional characteristics of raw and cryopreserved buffalo semen. Twelve purebred Murrah bulls (Bubalus bubalis) were assigned to the experimental groups and fed diets for 120 days with no added lipids (CONT, four bulls), or with an extra amount of 3% lipids from crude palm oil (PALM, four bulls), or from palm oil deodorizer distillate (PODD, four bulls). Semen was collected and cryopreserved every 15 days. The lipid composition of membranes and semen quality were determined after collections. Lipid supplementation did not impact feed intake (P>0.05). Diet enrichment with PALM increased the linoleic acid (C18:2,6) in seminal plasma. Lipid supplementation did not increase the polyunsaturated fatty acids in the sperm membrane composition, but significantly increased the lignoceric acid (C24:0). Cryopreserved semen of the supplemented bulls presented higher progressive motility (60.2 vs. 67.9 vs. 65.2%; P 0.05) and sperm viability detected by eosin-nigrosin staining (61.1 vs. 69.4 vs. 67.8%; P 0.05). Palm oil reduced major sperm defects in both raw (12.2 vs. 9.3 vs. 13.2%; P 0.0001) and cryopreserved semen (12.4 vs. 9.4 vs. 11.2%; P 0.0001). The lipids added to the diet did not impact the population of spermatozoa with intact plasma and acrosomal membranes (PI-/PSA-), but significantly increased the percentage of spermatozoa with high mitochondrial potential (25.6 vs. 31.5 vs. 32.0%; P=0.008). The results suggest that lipid supplementation based on crude palm oil or palm oil deodorizer distillate can be safely used to feed buffalo bulls and may increase sperm attributes related to male fertility.

Increased quality of in natura and cryopreserved semen of water buffaloes supplemented with saturated and unsaturated fatty acids from the palm oil industry

Ruminant energy supplementation with vegetable oils or fats has been standing out worldwide and oil palm processing has been receiving growing interest. This study assessed the effect of supplementation with saturated and unsaturated fatty acids from the palm oil industry on the lipid profile of seminal plasma and of the sperm membrane, as well as on the morphological and functional characteristics of raw and cryopreserved buffalo semen. Twelve purebred Murrah bulls (Bubalus bubalis) were assigned to the experimental groups and fed diets for 120 days with no added lipids (CONT, four bulls), or with an extra amount of 3% lipids from crude palm oil (PALM, four bulls), or from palm oil deodorizer distillate (PODD, four bulls). Semen was collected and cryopreserved every 15 days. The lipid composition of membranes and semen quality were determined after collections. Lipid supplementation did not impact feed intake (P>0.05). Diet enrichment with PALM increased the linoleic acid (C18:2,6) in seminal plasma. Lipid supplementation did not increase the polyunsaturated fatty acids in the sperm membrane composition, but significantly increased the lignoceric acid (C24:0). Cryopreserved semen of the supplemented bulls presented higher progressive motility (60.2 vs. 67.9 vs. 65.2%; P 0.05) and sperm viability detected by eosin-nigrosin staining (61.1 vs. 69.4 vs. 67.8%; P 0.05). Palm oil reduced major sperm defects in both raw (12.2 vs. 9.3 vs. 13.2%; P 0.0001) and cryopreserved semen (12.4 vs. 9.4 vs. 11.2%; P 0.0001). The lipids added to the diet did not impact the population of spermatozoa with intact plasma and acrosomal membranes (PI-/PSA-), but significantly increased the percentage of spermatozoa with high mitochondrial potential (25.6 vs. 31.5 vs. 32.0%; P=0.008). The results suggest that lipid supplementation based on crude palm oil or palm oil deodorizer distillate can be safely used to feed buffalo bulls and may increase sperm attributes related to male fertility.(AU)

Effects of the cryopreservation process on dog sperm integrity

Sperm cryopreservation has become an indispensable tool in reproductive biology. However, frozen/thawed semen has a short lifespan due to loss of sperm cell integrity. To better understand which sperm cell structures are compromised by the cryopreservation process and apoptosis markers, the sperm of five healthy mature dogs was analyzed in this study. Analysis was performed after collection, cooling, and thawing via computer assisted sperm analyzer (CASA) and evaluation of membrane fluidity and permeability, phosphatidylserine translocation (Annexin V), membrane integrity, mitochondrial membrane potential, membrane lipid peroxidation (LPO) and activity of the apoptotic markers caspases 3 and 7 by flow cytometry. Cryopreservation decreased total and progressive motility and the percentage of rapid sperm (P < 0.01). Damage to sperm cells was confirmed by Annexin V (P <0.01), indicating that capacitation-like changes were induced by the cryopreservation procedures. An increase in sperm membrane fluidity was also noted in frozen/thawed samples (P < 0.01). Plasma and acrosomal cell membranes were affected (P < 0.01), with decreases in the subpopulation displaying high membrane potential (P < 0.01). Membrane LPO was increased in thawed sperm compared to cooled sperm (P < 0.05) but was not different from that in fresh sperm. No differences were observed in caspase 3 and 7 activity after cooling, freezing, or thawing. In conclusion, total and progressive motility, plasma membrane integrity and mitochondrial membrane potential suffered from the deleterious effects caused by cryopreservation, unlike the activity of caspases that remained stable during the freezing process.

Effects of the cryopreservation process on dog sperm integrity

Sperm cryopreservation has become an indispensable tool in reproductive biology. However, frozen/thawed semen has a short lifespan due to loss of sperm cell integrity. To better understand which sperm cell structures are compromised by the cryopreservation process and apoptosis markers, the sperm of five healthy mature dogs was analyzed in this study. Analysis was performed after collection, cooling, and thawing via computer assisted sperm analyzer (CASA) and evaluation of membrane fluidity and permeability, phosphatidylserine translocation (Annexin V), membrane integrity, mitochondrial membrane potential, membrane lipid peroxidation (LPO) and activity of the apoptotic markers caspases 3 and 7 by flow cytometry. Cryopreservation decreased total and progressive motility and the percentage of rapid sperm (P < 0.01). Damage to sperm cells was confirmed by Annexin V (P <0.01), indicating that capacitation-like changes were induced by the cryopreservation procedures. An increase in sperm membrane fluidity was also noted in frozen/thawed samples (P < 0.01). Plasma and acrosomal cell membranes were affected (P < 0.01), with decreases in the subpopulation displaying high membrane potential (P < 0.01). Membrane LPO was increased in thawed sperm compared to cooled sperm (P < 0.05) but was not different from that in fresh sperm. No differences were observed in caspase 3 and 7 activity after cooling, freezing, or thawing. In conclusion, total and progressive motility, plasma membrane integrity and mitochondrial membrane potential suffered from the deleterious effects caused by cryopreservation, unlike the activity of caspases that remained stable during the freezing process.(AU)

Bovine abortion associated with Staphylococcus aureus infection: characterization of S. aureus strain isolated from fetal tissues / Aborto bovino associado à infecção por Staphylococcus aureus: caracterização da cepa de S. aureus isolada a partir de tecidos fetais

Staphylococcus aureus is a gram-positive bacterium, commonly found colonizing the skin and mucous membranes of humans and animals. This report describes a case of fetal loss associated with S. aureus infection in a cow. A six-month old, crossbred male bovine fetus from a beef farm was submitted for necropsy. At gross examination fibrinous pleuropneumonia was observed. Histologically, lesions were restricted to the lungs and consisted of marked multifocal to coalescing areas of inflammatory infiltrate of neutrophils, abundant fibrin exudation, necrosis of bronchiolar epithelium and numerous aggregates of coccoid bacteria. Lung and abomasal fluid bacterial culture yielded pure culture of S. aureus, which was characterized as a multidrug resistant strain. Molecular analysis indicated that the studied strain presented several genes of virulence factors including toxic shock syndrome toxin-1 (tst), staphylococcal enterotoxin type A (sea), Panton-Valentine leukocidin (pvl), alpha-hemolysin (hla) and delta-hemolysin (hld). This report documents an infrequent case of fetal loss in cattle due to infection with a highly virulent S. aureus strain.(AU)

Staphylococcus aureus é uma bactéria gram-positiva, comumente encontrada colonizando a pele e as membranas mucosas de humanos e animais. O presente relato descreve um caso de aborto bovino associado à infecção por S. aureus. Um feto bovino, macho, cruzado, com seis meses de idade gestacional proveniente de uma fazenda de gado de corte foi submetido para a necropsia. Pleuropneumonia supurativa foi observada na avaliação macroscópica. Histologicamente as lesões encontravam-se restritas aos pulmões e eram representadas por infiltrado inflamatório acentuado, multifocal a coalescente de neutrófilos, acentuada exsudação de fibrina, necrose do epitélio bronquiolar e numerosos agregados bacterianos cocoides. A cultura bacteriana de fragmento de pulmão e líquido do abomaso revelou o crescimento puro de S. aureus, que foi caracterizado como uma cepa multirresistente a drogas. Análises moleculares indicaram que a cepa estudada apresentava vários fatores de virulência, incluindo toxina 1 da síndrome do choque tóxico (TSST-1), enterotoxina estafilocócica tipo A (sea), leucocidina Panton-Valentine (pvl), hemolisina alfa (hla) e hemolisina delta (hld). O presente relato documenta um caso infrequente de aborto bovino devido à infecção por uma cepa altamente virulenta de S. aureus.(AU)

Effect of different doses of dexamethasone on labor in Santa Inês ewes and its influence on the onset of labor and the expulsion of fetal membranes / Efeito de diferentes doses de dexametasona para a indução do parto em ovelhas Santa Inês e sua influência sobre as características do parto

The induction of labor aims to concentrate births to follow up better the parturient and the first care to the neonates. However, even if the labor induction technique with dexamethasone administration has been successfully described since the late 1970s, few studies report the technique of birth development and neonatal vitality in Santa Inês sheep. This study aimed to evaluate the efficiency of dexamethasone use in two doses (8 and 16mg) in labor induction of Santa Inês ewes at 145 days of gestation and to evaluate its effects on the birth characteristics. In this study, 58 ewes were used, raised in an extensive system in the experimental farms of UFBA, with confirmation pregnancy after fixed-time artificial insemination or controlled breeding. These female ewes were separated into three groups according to the dose of dexamethasone administered (G1 = 0mg, G2 = 8mg, and G3 = 16mg). From these births, 79 lambs were born. This study analyzed the period from induction of labor to birth, fetal presentation at birth, the weight of the placenta, and the period for placenta expulsion. The data were analyzed by the Statistical Analysis System (SAS v.9.1.3®, 2002), and the significance level considered for all analyzes was 5%. Births of induced groups occurred on average at 48.4±22.17 hours after induction, while the females with non-induced labor gave birth 131.96±41.9 hours on average after the placebo application (P<0.05), confirming the efficiency of both doses for induction of labor. The period from induction to birth did not differ (P>0.05) between the doses used. There were no differences in delivery about the fetal static relation, time to placental attachment, and weight. With this study, it can be concluded that the induction at 145 days of gestation with eight or 16mg of dexamethasone is a useful technique and does not alter the labor in Santa Inês sheep.(AU)

A indução do parto visa concentrar os nascimentos para melhor acompanhamento das parturientes e primeiros cuidados aos neonatos. Contudo, mesmo que a técnica de indução de parto, com administração de dexametasona, tenha sido descrita com sucesso desde o final da década de 70, existem estudos escassos que relatam a influência desta técnica sobre o parto em ovinos da raça Santa Inês. Dessa forma, o objetivo do estudo foi avaliar a eficácia da dexametasona em duas doses (8 e 16mg), para a indução do parto de ovelhas Santa Inês com 145 dias de gestação e avaliar os seus efeitos nas características de desencadeamento e finalização do parto. Para este estudo foram utilizadas 58 ovelhas, criadas em sistema extensivo nas fazendas experimentais da UFBA, com prenhez confirmada após inseminação artificial em tempo fixo ou monta controlada. Essas fêmeas foram separadas em três grupos, de acordo com a dose de dexametasona administrada (G1 = 0mg, G2 = 8mg e G3 = 16mg). Destes partos nasceram 79 cordeiros. Foram avaliados o período em horas da indução do parto aos nascimentos, a apresentação fetal ao nascimento, assim como o peso da placenta e o período para o delivramento. Os dados foram analisados pelo pacote estatístico Statistical Analysis System (SAS v.9.0®, 2002) sendo considerado para todas as análises o nível de significância de 5%. Os nascimentos dos grupos induzidos ocorreram em média com 48,4±22,1 horas após a indução, enquanto que as fêmeas com parto não induzido pariram em média 131,96±41,9 horas após aplicação do placebo (P<0,05), confirmando a eficácia de ambas as doses para indução do parto. O período da indução até o parto não diferiu (P>0,05) entre as doses utilizadas. Não ocorreram diferenças no parto em relação à estática fetal, tempo para o delivramento e peso da placenta nos diferentes grupos. Com este estudo, conclui-se que a indução de parto em ovelhas aos 145 dias de gestação com oito e 16 mg de dexametasona é uma técnica eficaz e que não altera o trabalho de parto nas ovelhas da raça Santa Inês.(AU)

Effect of different doses of dexamethasone on labor in Santa Inês ewes and its influence on the onset of labor and the expulsion of fetal membranes / Efeito de diferentes doses de dexametasona para a indução do parto em ovelhas Santa Inês e sua influência sobre as características do parto

The induction of labor aims to concentrate births to follow up better the parturient and the first care to the neonates. However, even if the labor induction technique with dexamethasone administration has been successfully described since the late 1970s, few studies report the technique of birth development and neonatal vitality in Santa Inês sheep. This study aimed to evaluate the efficiency of dexamethasone use in two doses (8 and 16mg) in labor induction of Santa Inês ewes at 145 days of gestation and to evaluate its effects on the birth characteristics. In this study, 58 ewes were used, raised in an extensive system in the experimental farms of UFBA, with confirmation pregnancy after fixed-time artificial insemination or controlled breeding. These female ewes were separated into three groups according to the dose of dexamethasone administered (G1 = 0mg, G2 = 8mg, and G3 = 16mg). From these births, 79 lambs were born. This study analyzed the period from induction of labor to birth, fetal presentation at birth, the weight of the placenta, and the period for placenta expulsion. The data were analyzed by the Statistical Analysis System (SAS v.9.1.3®, 2002), and the significance level considered for all analyzes was 5%. Births of induced groups occurred on average at 48.4±22.17 hours after induction, while the females with non-induced labor gave birth 131.96±41.9 hours on average after the placebo application (P<0.05), confirming the efficiency of both doses for induction of labor. The period from induction to birth did not differ (P>0.05) between the doses used. There were no differences in delivery about the fetal static relation, time to placental attachment, and weight. With this study, it can be concluded that the induction at 145 days of gestation with eight or 16mg of dexamethasone is a useful technique and does not alter the labor in Santa Inês sheep.(AU)

A indução do parto visa concentrar os nascimentos para melhor acompanhamento das parturientes e primeiros cuidados aos neonatos. Contudo, mesmo que a técnica de indução de parto, com administração de dexametasona, tenha sido descrita com sucesso desde o final da década de 70, existem estudos escassos que relatam a influência desta técnica sobre o parto em ovinos da raça Santa Inês. Dessa forma, o objetivo do estudo foi avaliar a eficácia da dexametasona em duas doses (8 e 16mg), para a indução do parto de ovelhas Santa Inês com 145 dias de gestação e avaliar os seus efeitos nas características de desencadeamento e finalização do parto. Para este estudo foram utilizadas 58 ovelhas, criadas em sistema extensivo nas fazendas experimentais da UFBA, com prenhez confirmada após inseminação artificial em tempo fixo ou monta controlada. Essas fêmeas foram separadas em três grupos, de acordo com a dose de dexametasona administrada (G1 = 0mg, G2 = 8mg e G3 = 16mg). Destes partos nasceram 79 cordeiros. Foram avaliados o período em horas da indução do parto aos nascimentos, a apresentação fetal ao nascimento, assim como o peso da placenta e o período para o delivramento. Os dados foram analisados pelo pacote estatístico Statistical Analysis System (SAS v.9.0®, 2002) sendo considerado para todas as análises o nível de significância de 5%. Os nascimentos dos grupos induzidos ocorreram em média com 48,4±22,1 horas após a indução, enquanto que as fêmeas com parto não induzido pariram em média 131,96±41,9 horas após aplicação do placebo (P<0,05), confirmando a eficácia de ambas as doses para indução do parto. O período da indução até o parto não diferiu (P>0,05) entre as doses utilizadas. Não ocorreram diferenças no parto em relação à estática fetal, tempo para o delivramento e peso da placenta nos diferentes grupos. Com este estudo, conclui-se que a indução de parto em ovelhas aos 145 dias de gestação com oito e 16 mg de dexametasona é uma técnica eficaz e que não altera o trabalho de parto nas ovelhas da raça Santa Inês.(AU)

Potential application of aminiotic stem cells in veterinary medicine

In regenerative medicine stem cell biology has become one of the most interesting and more often studied subject. The amniotic membrane is the innermost layer of the fetal membranes and is considered a potential tool to treat many pathologies. It is used because it can be collected from discarded fetal material and is a rich source of stem cells with high proliferation and plasticity ratio capable of proliferating and differentiate in vitro. We propose to elucidate the characteristics and potencial clinical application of cells derived of amniotic membrane in veterinary medicine.

Potential application of aminiotic stem cells in veterinary medicine

In regenerative medicine stem cell biology has become one of the most interesting and more often studied subject. The amniotic membrane is the innermost layer of the fetal membranes and is considered a potential tool to treat many pathologies. It is used because it can be collected from discarded fetal material and is a rich source of stem cells with high proliferation and plasticity ratio capable of proliferating and differentiate in vitro. We propose to elucidate the characteristics and potencial clinical application of cells derived of amniotic membrane in veterinary medicine.(AU)