Chronic pancreatits in goats: first report in South America

This study reports the clinical and laboratory profile of an adult Saanen goat with chronic pancreatitis admitted in the Cattle and Small Ruminant Practice. The main complaint was loss of weight in the absence of dysphagia and anorexia. Physical examination showed normal vital function, cachexia, body condition score equal (ECC) to 2 (1 to 5), and polyphagia. Parasitological examination of feces and radial immunodiffusion for caprine arthritis encephalitis presented negative results. Laboratory exams showed fasting hyperglycemia, glucosuria, ketonuria and aciduria. Serum amylase activity was 10.5U/L, lower than the values obtained from two healthy animals kept in the CBPR (21.2U/L and 52.2U/L), once reference values for amylase in goats are not available. Insulin assessment, however, was not carried out because there are no laboratories in Brazil that work with goat insulin. After two episodes of bronchopneumonia, the animal was euthanized and necropsied. Hystopathological examination of the pancreas showed serious chronic-active pancreatitis, with marked acinar fibrosis and atrophy associated to rarefaction of islets of Langerhans. Besides, there were ductal hyperplasia with irregularities, and mucoid metaplasia. Thus, clinical, laboratorial and histopathological findings indicate that the animal presented primary chronic pancreatitis, compromising the endocrine and exocrine pancreas.Trabalho apresentado ao 9º Congresso Brasileiro de Buiatria, Goiânia, 2011

Profile of microRNAs present in semen and embryos and their relationship to fertility

Dentre outros fatores, a fertilidade depende da qualidade espermática (QE; características morfofuncionais) e dos componentes espermáticos moleculares. Devido à QE nem sempre estar associada à fertilidade, o estudo dos componentes moleculares ganha cada vez mais destaque. Os microRNAs, reguladores pós-transcricionais, estão presentes nos espermatozoides e desempenham funções importantes na espermatogênese, na maturação espermática e no desenvolvimento embrionário. Entretanto, os mecanismos pelos quais estes regulam a fertilidade masculina são pouco conhecidos. Portanto, o objetivo dos estudos aqui presentes foi investigar o papel dos miRNAs na determinação e regulação da fertilidade. No primeiro estudo, partidas de sêmen criopreservado de touros Aberdeen Angus (Bos taurus) com alta (54,3±1,0%; AF; n=3) e baixa fertilidade (41,5±2,3%; BF; n=3) foram submetidas às avaliações da QE; à produção in vitro e coleta de embriões com uma célula, duas células e blastocistos; e à avaliação do perfil de 380 miRNAs. Os genes-alvo dos miRNAs diferentemente abundantes nos espermatozoides e nos embriões dos touros AF e BF foram investigados. O diâmetro, número de células e taxa de proliferação dos blastocistos também foram analisados. Ainda, neste estudo, foi validado um novo modelo para provar que o espermatozoide é capaz de entregar miRNAs ao embrião. Para a análise estatística, foram empregadas as análises de variância (ANOVA) e de Qui-Quadrado. Quando não mencionado, o nível de significância foi de 5%. Dentre os miRNAs avaliados, o miR-216b foi menos abundante nos espermatozoides (P=0,08) e zigotos (P<0,05) dos touros AF. O gene-alvo deste miRNA, KRAS, associado à proliferação celular, foi mais abundante (P<0,05) nos embriões de duas células dos touros AF assim como a taxa de primeira clivagem e o número de células dos blastocistos que foram maiores neste grupo. Ainda, provou-se que o miR-216b é entregue pelo espermatozoide ao embrião. No segundo estudo, seis touros da raça Nelore (Bos indicus) foram utilizados, sendo três submetidos ao estresse térmico testicular (heat stress; HS). O sêmen foi coletado sete dias antes e 21 dias após o estresse. A QE foi avaliada seguida pela análise do perfil de 380 miRNAs espermáticos e exossomais. Os dados foram analisados por ANOVA. O nível de significância foi de 5%. HS apresentou 21 miRNAs espermáticos diferentemente abundantes, mas não apresentou diferença no conteúdo de miRNAs exossomais. Os miR-126-5p e -146a apresentaram menor abundância nos espermatozoides HS assim como apresentaram menor abundância nos espermatozoides dos touros BF do primeiro estudo. Já o miR-216b não apresentou diferença neste segundo estudo. Assim, com base nos dois estudos, podemos concluir que 1) os miRNAs são importantes para a fertilidade; 2) existem miRNAs importantes para a fertilidade que são alterados por injúrias à espermatogênese; e 3) o miR-216b, entregue pelo espermatozoide ao embrião, é importante para o desenvolvimento embrionário inicial regulando de maneira diferente embriões de touros de AF e BF; provando que miRNAs espermáticos possuem papel na regulação e determinação da fertilidade. Estes estudos possuem grande importância na geração de conhecimentos sobre os miRNAs espermáticos e apresentam resultados pioneiros que confirmam que o espermatozoide bovino contribui com mais que o DNA ao desenvolvimento embrionário.Bulls fertility relies of many factors. In concern to sperm cells, it depends of sperm quality (SQ; morphofunctional features) and sperm molecular components. Due to the fact thatsperm quality is not always related with fertility, the effect of many molecular factors on fertility has been intensively studied. miRNAs, which have post transcriptional action, are present on sperm cells and are important to spermatogenesis, sperm maturation and embryo development. However, the mechanisms by which they regulate male fertility are not completely understood. Thus, the objective of the present studies was to investigate the role of miRNAs on fertility regulation. On the first study, frozen-thawed semen batches of high (54.3±1.0%; HF; n=3) and low fertility (41.5±2.3%; LF; n=3) Aberdeen Angus (Bos taurus) bulls were analyzed in concern to SQ; were used in embryo in vitro production to produce and collect embryos of one cell, two cells and blastocysts; and were analyzed to the profile of 380 sperm miRNAs. The target genes of the miRNAs that presented different abundance levels on sperm cells and embryos of HF and LF bulls were investigated. Blastocysts diameter, cell number and proliferation index were also evaluated. Besides, in this study, we validated a new model to prove that some miRNAs are delivered by sperm cell to embryo. Data were analyzed by analyzes of variance (ANOVA) and Chi-Square. Signicante level was 5% unless otherwise stated. Among miRNAs evaluated, miR- 216b presented less abundance on HF sperm cells (P=0.08) and HF zygotes (P<0.05). miR- 216b target gene, KRAS, that is related with cell proliferation, presented high abundance (P<0.05) on HF two cells embryos such as the first clivage rate and the number of cells on blastocysts that were greater on this group. We also showed that miR-216b is delivered by sperm cells to embryos. On the second study, we used six Nelore (Bos indicus) bulls. Three bulls were submitted to testicular heat stress (HS group). Semen was collected three days before and 21 days after heat stress. Samples were analyzed in concern to SQ and to the profile of 380 miRNAs on sperm cells and on exossomes. Data were analyzed by ANOVA. Signicante level was 5%. HS presented 21 sperm miRNAs diferently abundant, but miRNAs from exossomes were not different between the HS and control groups. miR-126-5p and -146a presented less abundance on HS sperm cells and on BF bulls sperm cells of the first study. miR-216b was not different in the second study. Thus, we can conclude that 1) miRNAs are important to fertility; 2) there are miRNAs that are important to fertility and can be altered by spermatogenesis injuries; and 3) miR-216b is delivered by sperm cells to embryos and is important to initial embryo development regulating HF and LF embryos differently; showing that sperm miRNAs are important molecules for regulation and establishment of fertility. The studies here presented generate knowledge upon sperm miRNAs importance and present novel results that confirm that bull sperm cells contribute with more than DNA to embryo development

Profile of microRNAs present in semen and embryos and their relationship to fertility

Dentre outros fatores, a fertilidade depende da qualidade espermática (QE; características morfofuncionais) e dos componentes espermáticos moleculares. Devido à QE nem sempre estar associada à fertilidade, o estudo dos componentes moleculares ganha cada vez mais destaque. Os microRNAs, reguladores pós-transcricionais, estão presentes nos espermatozoides e desempenham funções importantes na espermatogênese, na maturação espermática e no desenvolvimento embrionário. Entretanto, os mecanismos pelos quais estes regulam a fertilidade masculina são pouco conhecidos. Portanto, o objetivo dos estudos aqui presentes foi investigar o papel dos miRNAs na determinação e regulação da fertilidade. No primeiro estudo, partidas de sêmen criopreservado de touros Aberdeen Angus (Bos taurus) com alta (54,3±1,0%; AF; n=3) e baixa fertilidade (41,5±2,3%; BF; n=3) foram submetidas às avaliações da QE; à produção in vitro e coleta de embriões com uma célula, duas células e blastocistos; e à avaliação do perfil de 380 miRNAs. Os genes-alvo dos miRNAs diferentemente abundantes nos espermatozoides e nos embriões dos touros AF e BF foram investigados. O diâmetro, número de células e taxa de proliferação dos blastocistos também foram analisados. Ainda, neste estudo, foi validado um novo modelo para provar que o espermatozoide é capaz de entregar miRNAs ao embrião. Para a análise estatística, foram empregadas as análises de variância (ANOVA) e de Qui-Quadrado. Quando não mencionado, o nível de significância foi de 5%. Dentre os miRNAs avaliados, o miR-216b foi menos abundante nos espermatozoides (P=0,08) e zigotos (P<0,05) dos touros AF. O gene-alvo deste miRNA, KRAS, associado à proliferação celular, foi mais abundante (P<0,05) nos embriões de duas células dos touros AF assim como a taxa de primeira clivagem e o número de células dos blastocistos que foram maiores neste grupo. Ainda, provou-se que o miR-216b é entregue pelo espermatozoide ao embrião. No segundo estudo, seis touros da raça Nelore (Bos indicus) foram utilizados, sendo três submetidos ao estresse térmico testicular (heat stress; HS). O sêmen foi coletado sete dias antes e 21 dias após o estresse. A QE foi avaliada seguida pela análise do perfil de 380 miRNAs espermáticos e exossomais. Os dados foram analisados por ANOVA. O nível de significância foi de 5%. HS apresentou 21 miRNAs espermáticos diferentemente abundantes, mas não apresentou diferença no conteúdo de miRNAs exossomais. Os miR-126-5p e -146a apresentaram menor abundância nos espermatozoides HS assim como apresentaram menor abundância nos espermatozoides dos touros BF do primeiro estudo. Já o miR-216b não apresentou diferença neste segundo estudo. Assim, com base nos dois estudos, podemos concluir que 1) os miRNAs são importantes para a fertilidade; 2) existem miRNAs importantes para a fertilidade que são alterados por injúrias à espermatogênese; e 3) o miR-216b, entregue pelo espermatozoide ao embrião, é importante para o desenvolvimento embrionário inicial regulando de maneira diferente embriões de touros de AF e BF; provando que miRNAs espermáticos possuem papel na regulação e determinação da fertilidade. Estes estudos possuem grande importância na geração de conhecimentos sobre os miRNAs espermáticos e apresentam resultados pioneiros que confirmam que o espermatozoide bovino contribui com mais que o DNA ao desenvolvimento embrionário.Bulls fertility relies of many factors. In concern to sperm cells, it depends of sperm quality (SQ; morphofunctional features) and sperm molecular components. Due to the fact thatsperm quality is not always related with fertility, the effect of many molecular factors on fertility has been intensively studied. miRNAs, which have post transcriptional action, are present on sperm cells and are important to spermatogenesis, sperm maturation and embryo development. However, the mechanisms by which they regulate male fertility are not completely understood. Thus, the objective of the present studies was to investigate the role of miRNAs on fertility regulation. On the first study, frozen-thawed semen batches of high (54.3±1.0%; HF; n=3) and low fertility (41.5±2.3%; LF; n=3) Aberdeen Angus (Bos taurus) bulls were analyzed in concern to SQ; were used in embryo in vitro production to produce and collect embryos of one cell, two cells and blastocysts; and were analyzed to the profile of 380 sperm miRNAs. The target genes of the miRNAs that presented different abundance levels on sperm cells and embryos of HF and LF bulls were investigated. Blastocysts diameter, cell number and proliferation index were also evaluated. Besides, in this study, we validated a new model to prove that some miRNAs are delivered by sperm cell to embryo. Data were analyzed by analyzes of variance (ANOVA) and Chi-Square. Signicante level was 5% unless otherwise stated. Among miRNAs evaluated, miR- 216b presented less abundance on HF sperm cells (P=0.08) and HF zygotes (P<0.05). miR- 216b target gene, KRAS, that is related with cell proliferation, presented high abundance (P<0.05) on HF two cells embryos such as the first clivage rate and the number of cells on blastocysts that were greater on this group. We also showed that miR-216b is delivered by sperm cells to embryos. On the second study, we used six Nelore (Bos indicus) bulls. Three bulls were submitted to testicular heat stress (HS group). Semen was collected three days before and 21 days after heat stress. Samples were analyzed in concern to SQ and to the profile of 380 miRNAs on sperm cells and on exossomes. Data were analyzed by ANOVA. Signicante level was 5%. HS presented 21 sperm miRNAs diferently abundant, but miRNAs from exossomes were not different between the HS and control groups. miR-126-5p and -146a presented less abundance on HS sperm cells and on BF bulls sperm cells of the first study. miR-216b was not different in the second study. Thus, we can conclude that 1) miRNAs are important to fertility; 2) there are miRNAs that are important to fertility and can be altered by spermatogenesis injuries; and 3) miR-216b is delivered by sperm cells to embryos and is important to initial embryo development regulating HF and LF embryos differently; showing that sperm miRNAs are important molecules for regulation and establishment of fertility. The studies here presented generate knowledge upon sperm miRNAs importance and present novel results that confirm that bull sperm cells contribute with more than DNA to embryo development

Treatment of testicular degeneration in rams supplemented with vitamin A or low level laser therapy

A degeneração testicular (DT) possui grande relevância dentre os distúrbios da reprodução e pode ser causada pelo aumento da temperatura testicular. Este provoca aumento do metabolismo celular, levando ao estresse oxidativo (EO) e apoptose. O tratamento usual consiste na retirada do agente causador e administração de antioxidantes; entretanto, pode não ser eficiente. Dessa forma, o presente estudo preconizou o tratamento da DT por meio de agentes com poder proliferativo: vitamina A e laserterapia de baixa intensidade (LTBI). Foram realizados três experimentos; o experimento 1 objetivou definir a dose de energia da LTBI necessária para a bioestimulação testicular. Foram utilizados seis carneiros distribuídos em três grupos: GC) insulação escrotal (IE) e sem tratamento (n=2); G28) IE e tratado com LTBI com 808 nm de comprimento de onda, 30 mW de potência e 28 J/cm² de densidade de energia por 15 dias a cada 48 horas (n=2); G56) IE e tratado com LTBI com 808 nm, 30 mW e 56 J/cm² por 15 dias a cada 48 horas (n=2). Foram feitas análises clínicas, reprodutivas e histopatológicas. Os dados foram submetidos à análise de variância (ANOVA) e teste de Tukey. Apesar da LTBI diminuir as taxas de espermatozoides com membrana acrossomal íntegra, esta foi eficiente em aumentar a população celular dos túbulos seminíferos no G28. Portanto, a LBTI provocou efeito bioestimulatório em testículos degenerados de carneiros. O experimento 2 objetivou validar a técnica de avaliação do EO espermático por meio da sonda fluorescente CellROX Deep Red®. Foram realizados dois experimentos; o primeiro utilizou ejaculados de três carneiros tratados em T0 (ejaculado não submetido à indução de EO), T50 (50% não induzido e 50% induzido ao EO) e T100 (submetido ao EO). Os dados foram submetidos à regressão linear. No segundo experimento foram utilizados 16 carneiros submetidos à IE. Foram feitas avaliações do EO antes e após a IE. Os dados foram submetidos à ANOVA e teste LSD de Fisher. O coeficiente de determinação foi de 0,728 e houve aumento do EO após a IE. Assim, a sonda CellROX® foi capaz de detectar o EO espermático. No experimento 3 foi proposto tratamento para a DT baseado na suplementação vitamínica ou LTBI. Foram utilizados 33 carneiros distribuídos em seis grupos: CC) sem IE e sem tratamento (n=5); CA) sem IE e tratado com vitamina A IM 120.000 UI/animal, duas vezes por semana durante três semanas (n=6); CL) sem IE e tratado com LTBI protocolo G28 (experimento 1) (n=5); IC) IE e sem tratamento (n = 5); IA) IE e tratado com vitamina A IM 120.000 UI/animal, duas vezes por semana durante três semanas (n=6); IL) IE e tratado com LTBI protocolo G28 (n=6). Foram realizadas análises clínicas, reprodutivas, hormonais e histopatológicas. Os dados foram analisados usando o procedimento de modelos mistos e os efeitos dos tratamentos foram avaliados utilizando contrastes ortogonais. Não houve efeito benéfico dos tratamentos para as características ultrassonográficas, qualidade espermática, concentração de testosterona e aspectos histopatológicos. Assim, os tratamentos não foram eficientes para melhorar a qualidade espermática nem promover a proliferação celular.The testicular degeneration (TD) has great significance among the reproductive disorders and one of the main causes is the increase in testicular temperature. High testicular temperature results in increase cellular metabolism, leading to oxidative stress and apoptosis. The treatment consists in removing the causative agent and administration of antioxidants; however, it could be not efficient. The objective of this study is to recommend the treatment of TD by administering agents with proliferative action: vitamin A and low level laser therapy (LLLT). For this, three experiments were conducted. In experiment 1 the objective was to define the dose of energy for LLLT testicular biostimulation; it was used six rams distributed in three groups: GC) scrotal insulation (SI) and untreated (n=2); G28) SI and treated with LLLT with 808 nm, 30 mW and 28 J/cm ² of power density for 15 days every 48 hours (n=2); G56) SI and treated with LLLT with 808 nm, 30 mW and 56 J/cm² for 15 days every 48 hours (n=2). Clinics, reproductive and histopathological analyzes were done. Data were subjected to analysis of variance and Tukey test. The rates of sperm with intact acrosome membrane were decreased by LLLT, but the LLLT was effective in increasing the cell population of the seminiferous tubules in the G28. Thus, LLLT was able of causing stimulatory effect in degenerate testis of rams. The objective of experiment 2 was to assess the technique of evaluation of sperm oxidative stress (OS). This study was divided in two experiments; in experiment 1 was used ejaculates of three rams treated in T0 (ejaculate that was not submitted to OS induction), T50 (50% without OS and 50% inducted to OS) and T100 (entire submitted to OS induction). Data obtained were evaluated by linear regression analysis. In experiment 2, sixteen rams were submitted to SI. Analyses of OS were done before and after the SI. Data obtained were evaluated by analysis of variance and Fisher\'s LSD test. The determination coefficient was of 0.728 and there were increase in sperm showing OS after SI period. Thus, CellROX® fluorescent probe was able to detect sperm OS. The objective of experiment 3 was establish a treatment for TD based on vitamin A supplementation or LLLT; 33 rams were distributed in six groups: CC) no SI and non-treated (n=5); CA) no SI and treated with 120,000 IU/animal of IM vitamin A, twice a week for three weeks (n=6); CL) no SI and treated with LLLT G28 protocol (experiment 1) (n=5); IC) SI and untreated (n=5); IA) SI and treated with 120,000 IU/animal of IM vitamin A, twice a week for three weeks (n=6); IL) SI and treated with LLLT G28 protocol (n=6). Clinics, reproductive, hormonal and histopathological analyzes were performed. Data were analyzed using the mixed models procedure and treatments effects were evaluated using orthogonal contrasts. There was no beneficial effect of treatments for ultrasonographic characteristics, sperm quality, testosterone concentration and histopathological aspects. Thus, the treatments were not effective for improving sperm quality or promoting cell proliferation

NEOTROPICAL CARNIVORES: a data set on carnivore distribution in the Neotropics

Mammalian carnivores are considered a key group in maintaining ecological health and can indicate potential ecological integrity in landscapes where they occur. Carnivores also hold high conservation value and their habitat requirements can guide management and conservation plans. The order Carnivora has 84 species from 8 families in the Neotropical region: Canidae; Felidae; Mephitidae; Mustelidae; Otariidae; Phocidae; Procyonidae; and Ursidae. Herein, we include published and unpublished data on native terrestrial Neotropical carnivores (Canidae; Felidae; Mephitidae; Mustelidae; Procyonidae; and Ursidae). NEOTROPICAL CARNIVORES is a publicly available data set that includes 99,605 data entries from 35,511 unique georeferenced coordinates. Detection/non-detection and quantitative data were obtained from 1818 to 2018 by researchers, governmental agencies, non-governmental organizations, and private consultants. Data were collected using several methods including camera trapping, museum collections, roadkill, line transect, and opportunistic records. Literature (peer-reviewed and grey literature) from Portuguese, Spanish and English were incorporated in this compilation. Most of the data set consists of detection data entries (n = 79,343; 79.7%) but also includes non-detection data (n = 20,262; 20.3%). Of those, 43.3% also include count data (n = 43,151). The information available in NEOTROPICAL CARNIVORES will contribute to macroecological, ecological, and conservation questions in multiple spatio-temporal perspectives. As carnivores play key roles in trophic interactions, a better understanding of their distribution and habitat requirements are essential to establish conservation management plans and safeguard the future ecological health of Neotropical ecosystems. Our data paper, combined with other large-scale data sets, has great potential to clarify species distribution and related ecological processes within the Neotropics. There are no copyright restrictions and no restriction for using data from this data paper, as long as the data paper is cited as the source of the information used. We also request that users inform us of how they intend to use the data

NEOTROPICAL XENARTHRANS: a data set of occurrence of xenarthran species in the Neotropics

Xenarthrans—anteaters, sloths, and armadillos—have essential functions for ecosystem maintenance, such as insect control and nutrient cycling, playing key roles as ecosystem engineers. Because of habitat loss and fragmentation, hunting pressure, and conflicts with domestic dogs, these species have been threatened locally, regionally, or even across their full distribution ranges. The Neotropics harbor 21 species of armadillos, 10 anteaters, and 6 sloths. Our data set includes the families Chlamyphoridae (13), Dasypodidae (7), Myrmecophagidae (3), Bradypodidae (4), and Megalonychidae (2). We have no occurrence data on Dasypus pilosus (Dasypodidae). Regarding Cyclopedidae, until recently, only one species was recognized, but new genetic studies have revealed that the group is represented by seven species. In this data paper, we compiled a total of 42,528 records of 31 species, represented by occurrence and quantitative data, totaling 24,847 unique georeferenced records. The geographic range is from the southern United States, Mexico, and Caribbean countries at the northern portion of the Neotropics, to the austral distribution in Argentina, Paraguay, Chile, and Uruguay. Regarding anteaters, Myrmecophaga tridactyla has the most records (n = 5,941), and Cyclopes sp. have the fewest (n = 240). The armadillo species with the most data is Dasypus novemcinctus (n = 11,588), and the fewest data are recorded for Calyptophractus retusus (n = 33). With regard to sloth species, Bradypus variegatus has the most records (n = 962), and Bradypus pygmaeus has the fewest (n = 12). Our main objective with Neotropical Xenarthrans is to make occurrence and quantitative data available to facilitate more ecological research, particularly if we integrate the xenarthran data with other data sets of Neotropical Series that will become available very soon (i.e., Neotropical Carnivores, Neotropical Invasive Mammals, and Neotropical Hunters and Dogs). Therefore, studies on trophic cascades, hunting pressure, habitat loss, fragmentation effects, species invasion, and climate change effects will be possible with the Neotropical Xenarthrans data set. Please cite this data paper when using its data in publications. We also request that researchers and teachers inform us of how they are using these data