Bacillary hemoglobinuria in a cow

Clinical History: A 436 kg, 2-year-old, pregnant Angus cross cow was presented for necropsy with a history of sudden death during calving season. Two other heifers on the property recently had late term abortions with the calves having to be pulled out from the birth canal. Gross Findings: The subcutaneous tissue, omentum, mesentery, perirenal adipose tissue, and intima of the major vessels were diffusely and markedly yellow (Figure 1). The right liver lobe had a large (15x11x7 cm) area of pale tan discoloration covered by a thin layer of fibrin. On the cut surface, this area was pale red and dry (Figure 2). Throughout the remaining hepatic parenchyma were irregularly shaped, serpiginous, dark green to black tracks. Some of these tracks contained adult Fasciola hepatica flukes in the lumen. The gallbladder was distended and filled with large amounts of dark green, thick bile. The kidneys were diffusely dark red with disseminated pinpoint, dark red foci in the cortex (Figure 3). The urinary bladder contained approximately 300 mL of dark red urine (Figure 4). The brain had a slight yellow discoloration. Approximately 300 mL of red-tinged fluid was in the thorax. Large numbers of fibrous adhesions were present between the visceral pleura, the parietal pleura, and the pericardium. Approximately 200 mL of red-tinged fluid was in the pericardial sac. There was extensive hemorrhage on the epicardium.

Gestational follow-up in ewes - placentome size and fecal progesterone metabolite levels

Background: Transabdominal ultrasonography in sheep allows real-time diagnosis and monitoring of pregnancy as well as measurement of the size of placentomes, which are structures that connect the fetal and maternal portions, that increase as pregnancy progresses. Progesterone is involved in the maintenance of gestation, with high levels noted during pregnancy. In this context, it is hypothesized that measurements of placentome and progesterone could help in monitoring pregnancy in sheep. Therefore, the main objectives of this study included evaluating whether placentome size and fecal progesterone levels are associated with gestational age in ewes and whether measurement of the placentome and progesterone could be used as an estimate of gestational age. Materials, Methods & Results: A total of 63 pregnant adult (between 2 and 5 years of age) Santa Inês ewes were monitored in the city of Boa Vista, Roraima. All ewes were kept in the paddock with Panicum maximun cv. tanzania, mineral salt, and water ad libitum, supplemented with 200 g/animal/day of soybean residue. Ewes were subjected to ultrasonography every 21 days from day 42 of pregancy until term to monitor pregnancy. Ultrasonography was performed using a linear probe to measure the placentome, including length × width in centimeters. Also every 21 days, fecal samples were collected for measurement of progesterone level. Feces were collected directly from the rectal ampulla, stored individually in plastic bags, and frozen at -20ºC until processing. In the laboratory, these feces samples were freeze-dried and subjected to hormonal extraction with 80% methanol. Then, the levels of fecal progesterone metabolites were measured by enzyme immunoassay. Data was tabulated and submitted to statistical analyses, including descriptive stastistic, correlation, KruskalWallis test and Tukey's test. The levels of fecal progesterone metabolites increased significantly between the 3rd and 4th months of pregnancy, with a decrease close to delivery. Placentome size increased significantly between the 2nd and 3rd months of pregnancy. Placentome measurements did not correlate with progesterone levels and showed a weak correlation with gestational age. Therefore, we concluded that the measurements of placentome size and a single evaluation of fecal progesterone metabolites were not considered efficient methods for estimating gestational age. Discussion: Measurement of progesterone level in fecal samples is a non-invasive hormone monitoring method that provides values like that of blood levels and is less stressful for the animals at the time of sample collection. The wide physiological variation makes it difficult or even useless to use fecal progesterone, when is measured only once per animal, as an indicator of the presence of pregnancy. Despite this limitation, serial sampling is useful for monitoring of hormones during the pregnancy period and is an important tool for research purposes. Placentome size measurements correlated weakly with gestational age and did not correlate with progesterone level. Thus, the presence of placentomes is indicative of the presence of pregnancy but was considered inefficient for determining gestational age in sheep.

Understanding the trade-off between the environment and fertility in cows and ewes

The environment contributes to production diseases that in turn badly affect cow performance, fertility and culling. Oestrus intensity is lower in lame cows, and in all cows 26% potential oestrus events are not expressed (to avoid getting pregnant). To understand these trade-offs, we need to know how animals react to their environment and how the environment influences hypothalamus-pituitary-adrenal axis (HPA) interactions with the hypothalamus-pituitary-ovarian axis (HPO). Neurotransmitters control secretion of GnRH into hypophyseal portal blood. GnRH/LH pulse amplitude and frequency drive oestradiol production, culminating in oestrus behaviour and a precisely-timed GnRH/LH surge, all of which are disrupted by poor environments. Responses to peripheral neuronal agents give clues about mechanisms, but do these drugs alter perception of stimuli, or suppress consequent responses? In vitro studies confirm some neuronal interactions between the HPA and HPO; and immuno-histochemistry clarifies the location and sequence of inter-neurone activity within the brain. In both species, exogenous corticoids, ACTH and/or CRH act at the pituitary (reduce LH release by GnRH), and hypothalamus (lower GnRH pulse frequency and delay surge release). This requires inter-neurones as GnRH cells do not have receptors for HPA compounds. There are two (simultaneous, therefore fail-safe?) pathways for CRH suppression of GnRH release via CRH-Receptors: one being the regulation of kisspeptin/dynorphin and other cell types in the hypothalamus, and the other being the direct contact between CRH and GnRH cell terminals in the median eminence. When we domesticate animals, we must provide the best possible environment otherwise animals trade-off with lower production, less intense oestrus behaviour, and impaired fertility. Avoiding life-time peri-parturient problems by managing persistent lactations in cows may be a worthy trade-off on both welfare and economic terms – better than the camouflage use of drugs/hormones/feed additives/intricate technologies? In the long term, getting animals and environment in a more harmonious balance is the ultimate strategy.

Understanding the trade-off between the environment and fertility in cows and ewes

The environment contributes to production diseases that in turn badly affect cow performance, fertility and culling. Oestrus intensity is lower in lame cows, and in all cows 26% potential oestrus events are not expressed (to avoid getting pregnant). To understand these trade-offs, we need to know how animals react to their environment and how the environment influences hypothalamus-pituitary-adrenal axis (HPA) interactions with the hypothalamus-pituitary-ovarian axis (HPO). Neurotransmitters control secretion of GnRH into hypophyseal portal blood. GnRH/LH pulse amplitude and frequency drive oestradiol production, culminating in oestrus behaviour and a precisely-timed GnRH/LH surge, all of which are disrupted by poor environments. Responses to peripheral neuronal agents give clues about mechanisms, but do these drugs alter perception of stimuli, or suppress consequent responses? In vitro studies confirm some neuronal interactions between the HPA and HPO; and immuno-histochemistry clarifies the location and sequence of inter-neurone activity within the brain. In both species, exogenous corticoids, ACTH and/or CRH act at the pituitary (reduce LH release by GnRH), and hypothalamus (lower GnRH pulse frequency and delay surge release). This requires inter-neurones as GnRH cells do not have receptors for HPA compounds. There are two (simultaneous, therefore fail-safe?) pathways for CRH suppression of GnRH release via CRH-Receptors: one being the regulation of kisspeptin/dynorphin and other cell types in the hypothalamus, and the other being the direct contact between CRH and GnRH cell terminals in the median eminence. When we domesticate animals, we must provide the best possible environment otherwise animals trade-off with lower production, less intense oestrus behaviour, and impaired fertility. Avoiding life-time peri-parturient problems by managing persistent lactations in cows may be a worthy trade-off on both welfare and economic terms better than the camouflage use of drugs/hormones/feed additives/intricate technologies? In the long term, getting animals and environment in a more harmonious balance is the ultimate strategy.(AU)

Assessment of fetal lung maturity using analysis of amniotic fluid from ewes that gave birth prematurely and at term / Avaliação da maturidade pulmonar fetal utilizando a análise de líquido amniótico de ovelhas que deram à luz prematuramente e a termo

The aim of this study was to evaluate the lung maturity of premature and full-term lambs by analyzing amniotic fluid using the following methods: Clements test, Nile blue cytology test, hematoxylin-Shorr stain, lamellar body count, and radiographic tests. The use of these methods is intended to identify high-risk newborns and provide immediate clinical intervention after birth. Altogether, 56 animals (24 ewes and 32 lambs) were included in the study and divided into 3 groups. Group I consisted of 8 ewes that were at approximately 145 days of gestation; this group delivered 10 lambs naturally. Group II consisted of 8 ewes that were at 138 days' gestation; this group delivered 11 lambs by cesarean section. Group III consisted of 8 ewes at 138 days' gestation; this group was administered intramuscular dexamethasone (16mg/animal) 36 hours prior to a cesarean section. Group III delivered11 lambs. Cytological tests were performed using a microscope with a maximum magnification of 1000x, while the Clements test was visually observed by one of the researchers. Amnioticfluid lamellar body counts were measured using transmission electron microscopy. Among the staining methods, hematoxylin-Shorr was reliable, and Group III had a greater number of orangeophilic cells when compared to Group II, probably due to corticoid administration. The Clements test showed pulmonary maturity in approximately 20% of Group I lambs and Group II showed 9.1% of bubbles; however, Group III had the highest pulmonary maturity percentage (36.4%). The lamellar bodies were measured, and all groups had sizes between 0.019 and 0.590μm. Radiographic evaluation revealed that the majority of lambs presented some level of pulmonary radiodensity, indicating an acinar pattern at birth. These results are in line with the expectations of each group. We found that the normal group showed greater pulmonary maturity, whereas Group II presented pulmonary immaturity, which is expected because this group comprised lambs born prematurely and Group III showed pulmonary maturity almost comparable to the normal delivery group (Group I). This is due to the fact that although these animals are premature, the use of dexamethasone helped in pulmonary maturation. Therefore, these pulmonary maturity tests are considered effective when more than one technique is used and can be used routinely in the care of a pregnant ewe in labor, where a simple collection of amniotic fluid can predict a high-risk pregnancy and alert the veterinarian if the newborn needs intensive supportive treatment.(AU)

O objetivo deste estudo foi avaliar a maturidade pulmonar de cordeiros prematuros e a termo por meio da análise do líquido amniótico utilizando os seguintes métodos: teste de Clements, teste de citologia do azul do Nilo, coloração de hematoxilina-Shorr, contagem de corpos lamelares e testes radiográficos. Um desses métodos tem por objetivo identificar recém-nascidos de alto risco e fornecer intervenção clínica imediata após o nascimento. Ao todo, 56 animais (24 ovelhas e 32 cordeiros) foram incluídos no estudo e divididos em 3 grupos. O grupo I foi composto por 8 ovelhas com aproximadamente 145 dias de gestação; este grupo deu à luz 10 cordeiros naturalmente. O Grupo II foi composto por 8 ovelhas com 138 dias de gestação; este grupo deu à luz 11 cordeiros por cesariana. Grupo III consistiu de 8 ovelhas com 138 dias de gestação; este grupo recebeu dexametasona intramuscular (16 mg / animal) 36 horas antes de uma cesariana. O Grupo III entregou 11 cordeiros. Os testes citológicos foram realizados em microscópio com aumento máximo de 1000x, enquanto o teste de Clements foi observado visualmente por um dos pesquisadores. A contagem de corpos lamelares de líquido amniótico foi medida usando microscopia eletrônica de transmissão. Dentre os métodos de coloração, o hematoxilina-Shorr foi confiável, sendo que o Grupo III apresentou maior número de células orangeofílicas quando comparado ao grupo II, provavelmente devido à administração de corticóide. O teste de Clements mostrou maturidade pulmonar em aproximadamente 20% dos cordeiros do Grupo I e o Grupo II apresentou 9,1% de bolhas; entretanto, o Grupo III apresentou o maior percentual de maturidade pulmonar (36,4%). Os corpos lamelares foram medidos e todos os grupos apresentaram tamanhos entre 0,019 e 0,590μm. A avaliação radiográfica revelou que a maioria dos cordeiros apresentava algum grau de radiodensidade pulmonar, indicando padrão acinar ao nascimento. Esses resultados estão alinhados com as expectativas de cada grupo. Verificamos que o grupo normal apresentou maior maturidade pulmonar, enquanto o Grupo II apresentou imaturidade pulmonar, o que é esperado por se tratar de cordeiros nascidos prematuramente e o Grupo III apresentou maturidade pulmonar quase comparável ao grupo de parto normal (Grupo I). Isso se deve ao fato de que, embora esses animais sejam prematuros, o uso da dexametasona auxiliou na maturação pulmonar. Portanto, esses testes de maturidade pulmonar são considerados eficazes quando mais de uma técnica são utilizadas e podem ser usadas rotineiramente no cuidado de uma ovelha gestante em trabalho de parto, onde uma simples coleta de líquido amniótico pode prever uma gravidez de alto risco e alertar o veterinário se o recém-nascido precisa de tratamento de suporte intensivo.(AU)

Assessment of fetal lung maturity using analysis of amniotic fluid from ewes that gave birth prematurely and at term / Avaliação da maturidade pulmonar fetal utilizando a análise de líquido amniótico de ovelhas que deram à luz prematuramente e a termo

The aim of this study was to evaluate the lung maturity of premature and full-term lambs by analyzing amniotic fluid using the following methods: Clements test, Nile blue cytology test, hematoxylin-Shorr stain, lamellar body count, and radiographic tests. The use of these methods is intended to identify high-risk newborns and provide immediate clinical intervention after birth. Altogether, 56 animals (24 ewes and 32 lambs) were included in the study and divided into 3 groups. Group I consisted of 8 ewes that were at approximately 145 days of gestation; this group delivered 10 lambs naturally. Group II consisted of 8 ewes that were at 138 days' gestation; this group delivered 11 lambs by cesarean section. Group III consisted of 8 ewes at 138 days' gestation; this group was administered intramuscular dexamethasone (16mg/animal) 36 hours prior to a cesarean section. Group III delivered11 lambs. Cytological tests were performed using a microscope with a maximum magnification of 1000x, while the Clements test was visually observed by one of the researchers. Amnioticfluid lamellar body counts were measured using transmission electron microscopy. Among the staining methods, hematoxylin-Shorr was reliable, and Group III had a greater number of orangeophilic cells when compared to Group II, probably due to corticoid administration. The Clements test showed pulmonary maturity in approximately 20% of Group I lambs and Group II showed 9.1% of bubbles; however, Group III had the highest pulmonary maturity percentage (36.4%). The lamellar bodies were measured, and all groups had sizes between 0.019 and 0.590μm. Radiographic evaluation revealed that the majority of lambs presented some level of pulmonary radiodensity, indicating an acinar pattern at birth. These results are in line with the expectations of each group. We found that the normal group showed greater pulmonary maturity, whereas Group II presented pulmonary immaturity, which is expected because this group comprised lambs born prematurely and Group III showed pulmonary maturity almost comparable to the normal delivery group (Group I). This is due to the fact that although these animals are premature, the use of dexamethasone helped in pulmonary maturation. Therefore, these pulmonary maturity tests are considered effective when more than one technique is used and can be used routinely in the care of a pregnant ewe in labor, where a simple collection of amniotic fluid can predict a high-risk pregnancy and alert the veterinarian if the newborn needs intensive supportive treatment.(AU)

O objetivo deste estudo foi avaliar a maturidade pulmonar de cordeiros prematuros e a termo por meio da análise do líquido amniótico utilizando os seguintes métodos: teste de Clements, teste de citologia do azul do Nilo, coloração de hematoxilina-Shorr, contagem de corpos lamelares e testes radiográficos. Um desses métodos tem por objetivo identificar recém-nascidos de alto risco e fornecer intervenção clínica imediata após o nascimento. Ao todo, 56 animais (24 ovelhas e 32 cordeiros) foram incluídos no estudo e divididos em 3 grupos. O grupo I foi composto por 8 ovelhas com aproximadamente 145 dias de gestação; este grupo deu à luz 10 cordeiros naturalmente. O Grupo II foi composto por 8 ovelhas com 138 dias de gestação; este grupo deu à luz 11 cordeiros por cesariana. Grupo III consistiu de 8 ovelhas com 138 dias de gestação; este grupo recebeu dexametasona intramuscular (16 mg / animal) 36 horas antes de uma cesariana. O Grupo III entregou 11 cordeiros. Os testes citológicos foram realizados em microscópio com aumento máximo de 1000x, enquanto o teste de Clements foi observado visualmente por um dos pesquisadores. A contagem de corpos lamelares de líquido amniótico foi medida usando microscopia eletrônica de transmissão. Dentre os métodos de coloração, o hematoxilina-Shorr foi confiável, sendo que o Grupo III apresentou maior número de células orangeofílicas quando comparado ao grupo II, provavelmente devido à administração de corticóide. O teste de Clements mostrou maturidade pulmonar em aproximadamente 20% dos cordeiros do Grupo I e o Grupo II apresentou 9,1% de bolhas; entretanto, o Grupo III apresentou o maior percentual de maturidade pulmonar (36,4%). Os corpos lamelares foram medidos e todos os grupos apresentaram tamanhos entre 0,019 e 0,590μm. A avaliação radiográfica revelou que a maioria dos cordeiros apresentava algum grau de radiodensidade pulmonar, indicando padrão acinar ao nascimento. Esses resultados estão alinhados com as expectativas de cada grupo. Verificamos que o grupo normal apresentou maior maturidade pulmonar, enquanto o Grupo II apresentou imaturidade pulmonar, o que é esperado por se tratar de cordeiros nascidos prematuramente e o Grupo III apresentou maturidade pulmonar quase comparável ao grupo de parto normal (Grupo I). Isso se deve ao fato de que, embora esses animais sejam prematuros, o uso da dexametasona auxiliou na maturação pulmonar. Portanto, esses testes de maturidade pulmonar são considerados eficazes quando mais de uma técnica são utilizadas e podem ser usadas rotineiramente no cuidado de uma ovelha gestante em trabalho de parto, onde uma simples coleta de líquido amniótico pode prever uma gravidez de alto risco e alertar o veterinário se o recém-nascido precisa de tratamento de suporte intensivo.(AU)

Application of low dose of equine chorionic gonadotropin at acupoint Hou Hai for fixed-time artificial insemination in beef cows / Aplicação de baixa dose de gonadotrofina coriônica equina no acuponto Hou Hai em protocolos de inseminação artificial em tempo fixo para vacas de corte

This study evaluated the efficiency application of low dose of equine chorionic gonadotropin (eCG) using Hou Hai acupoint for fixed-term artificial insemination (FTAI) in beef cows. Seventy cows received intravaginal devices with progesterone, and 3 mg of estradiol benzoate on day zero (D0) of FTAI. On D9, the devices were removed, and 150 μg of prostaglandin F2α was applied; the animals were then randomly distributed to three treatment groups: T1 (n = 23), 300 IU of eCG intramuscularly (IM); T2 (n = 23), 90 IU of eCG at acupoint Hou Hai; and T3 (n = 25), 90 IU of eCG at false acupoint IM. On D10, the animals received 1 mg of gonadotropin-releasing hormone IM, and FTAI was performed, 52 h after the devices were removed. The quantitative variables with normal distribution were assessed using analysis of variance at 5% probability. For variables that did not show normal distribution, Kruskal-Wallis test at 5% probability was used. No significant difference was noted among the groups (P > 0.05) for the interval between device withdrawal at ovulation (58.2 ± 12.2 h), diameter of the largest follicle on D9 (9.9 ± 2.2 mm), diameters of the ovulatory follicle (12.2 ± 3.0 mm) and the second largest follicle (6.7 ± 2.1 mm), follicular growth rate (0.8 ± 0.3 mm/d), ovulation rate (82%), corpus luteum size (2.32 ± 0.35 cm2), and pregnancy rate (58.67%). The protocol cost per animal was US$ 10.67 (T1) and US$ 8.50 (T2 and T3). The use of 90 IU of eCG applied at Hou Hai acupoint or at false acupoint caused satisfactory synchronization of estrus in beef cows. In addition, this procedure was cost-effective.(AU)

Avaliou-se a eficiência da aplicação de subdose gonadotrofina coriônica equina (eCG) utulizando como via de administração o acuponto Hou Hai em protocolos de inseminação artificial em tempo fixo (IATF) para vacas de corte. Setenta vacas receberam no dia zero (D0) do protocolo dispositivos intravaginais com progesterona, e 3mg de benzoato de estradiol. No D9 foram retirados os dispositivos, aplicado 150µg de prostaglandina F2α e distribuídos os animais aleatoriamente em três tratamentos: T1 (n=23) - 300UI de eCG por via intra muscular (IM); T2 (n=23) - 90UI de eCG no acuponto Hou Hai e T3 (n=25) - 90UI de eCG em falso acuponto, IM. No D10 os animais receberam 1mg de GnRH por via IM e foi realizada a IATF, 52 horas após a retirada dos dispositivos. Para as variáveis quantitativas com distribuição normal foi utilizada ANOVA, a 5% de probabilidade. Para as variáveis que não apresentaram distribuição normal, utilizou-se o teste de Kruskal-Wallis, a 5% de probabilidade. Não houve diferença (P >0,05) para o intervalo entre a retirada do dispositivo à ovulação (58,2 ± 12,2h), diâmetro do maior folículo no D9 (9,9 ± 2,2mm), diâmetros do folículo ovulatório (12,2 ± 3,0mm) e do segundo maior folículo (6,7 ± 2,1mm), taxa de crescimento folicular (0,8 ± 0,3mm/d), taxa de ovulação (82%), área de corpo lúteo (2,32 ± 0,35 cm2) e taxa de prenhez (58,67%). O custo do protocolo por animal foi de US$ 10,67 (T1) e US$ 8,50 (T2 e T3). Sugere-se que a utilização de 90UI de eCG aplicada no acuponto Hou Hai ou em falso acuponto foram satisfatórios na sincronização de estro de vacas de corte. Além disso, possibilita a redução do custo do protocolo.(AU)

Application of low dose of equine chorionic gonadotropin at acupoint Hou Hai for fixed-time artificial insemination in beef cows / Aplicação de baixa dose de gonadotrofina coriônica equina no acuponto Hou Hai em protocolos de inseminação artificial em tempo fixo para vacas de corte

This study evaluated the efficiency application of low dose of equine chorionic gonadotropin (eCG) using Hou Hai acupoint for fixed-term artificial insemination (FTAI) in beef cows. Seventy cows received intravaginal devices with progesterone, and 3 mg of estradiol benzoate on day zero (D0) of FTAI. On D9, the devices were removed, and 150 μg of prostaglandin F2α was applied; the animals were then randomly distributed to three treatment groups: T1 (n = 23), 300 IU of eCG intramuscularly (IM); T2 (n = 23), 90 IU of eCG at acupoint Hou Hai; and T3 (n = 25), 90 IU of eCG at false acupoint IM. On D10, the animals received 1 mg of gonadotropin-releasing hormone IM, and FTAI was performed, 52 h after the devices were removed. The quantitative variables with normal distribution were assessed using analysis of variance at 5% probability. For variables that did not show normal distribution, Kruskal-Wallis test at 5% probability was used. No significant difference was noted among the groups (P > 0.05) for the interval between device withdrawal at ovulation (58.2 ± 12.2 h), diameter of the largest follicle on D9 (9.9 ± 2.2 mm), diameters of the ovulatory follicle (12.2 ± 3.0 mm) and the second largest follicle (6.7 ± 2.1 mm), follicular growth rate (0.8 ± 0.3 mm/d), ovulation rate (82%), corpus luteum size (2.32 ± 0.35 cm2), and pregnancy rate (58.67%). The protocol cost per animal was US$ 10.67 (T1) and US$ 8.50 (T2 and T3). The use of 90 IU of eCG applied at Hou Hai acupoint or at false acupoint caused satisfactory synchronization of estrus in beef cows. In addition, this procedure was cost-effective.

Avaliou-se a eficiência da aplicação de subdose gonadotrofina coriônica equina (eCG) utulizando como via de administração o acuponto Hou Hai em protocolos de inseminação artificial em tempo fixo (IATF) para vacas de corte. Setenta vacas receberam no dia zero (D0) do protocolo dispositivos intravaginais com progesterona, e 3mg de benzoato de estradiol. No D9 foram retirados os dispositivos, aplicado 150µg de prostaglandina F2α e distribuídos os animais aleatoriamente em três tratamentos: T1 (n=23) - 300UI de eCG por via intra muscular (IM); T2 (n=23) - 90UI de eCG no acuponto Hou Hai e T3 (n=25) - 90UI de eCG em falso acuponto, IM. No D10 os animais receberam 1mg de GnRH por via IM e foi realizada a IATF, 52 horas após a retirada dos dispositivos. Para as variáveis quantitativas com distribuição normal foi utilizada ANOVA, a 5% de probabilidade. Para as variáveis que não apresentaram distribuição normal, utilizou-se o teste de Kruskal-Wallis, a 5% de probabilidade. Não houve diferença (P >0,05) para o intervalo entre a retirada do dispositivo à ovulação (58,2 ± 12,2h), diâmetro do maior folículo no D9 (9,9 ± 2,2mm), diâmetros do folículo ovulatório (12,2 ± 3,0mm) e do segundo maior folículo (6,7 ± 2,1mm), taxa de crescimento folicular (0,8 ± 0,3mm/d), taxa de ovulação (82%), área de corpo lúteo (2,32 ± 0,35 cm2) e taxa de prenhez (58,67%). O custo do protocolo por animal foi de US$ 10,67 (T1) e US$ 8,50 (T2 e T3). Sugere-se que a utilização de 90UI de eCG aplicada no acuponto Hou Hai ou em falso acuponto foram satisfatórios na sincronização de estro de vacas de corte. Além disso, possibilita a redução do custo do protocolo.

Subplacental development in Galea spixii / Desenvolvimento subplacentário do preá (Galea spixii)

Animal models are essential to understand healthy human placentation. Guinea pig related rodents became on focus for such purposes. In particular, processes of trophoblast invasion are similar. The latter is associated with a specialized area, the subplacenta. Since previous results showed differences between the guinea pig and its close relative Galea spixii, we aimed to study subplacental development with more detail. We investigated 16 pregnant females of 14 to 55 days of gestation by means of histology, morphometrics, immunohistochemistry and electron microscopy. The overlap between the fetomaternal blood systems resulted as intimate, suggesting some exchange processes. Proliferation was revealed by three independent methods, being most active in early and mid-gestation, which was in accordance to former results. Though degeneration of tissues took place, the subplacenta was maintained towards term with access to the fetal vascularization, supporting a hypothesis about the release of substances to the fetal unit in advanced gestation. In contrast to other species, the extraplacental trophoblast showed a shift from syncytial streamers to giant cells during mid-gestation. Views on placentation in caviomorphs were influenced by the guinea pig, but our data supported recent studies that the subplacenta had a much greater placidity. In regard to subplacental grow, degeneration and likely also exchange processes, Galea and other species showed a more basal pattern of caviomorphs than the guinea pig. Such differences should be considered, when choosing most adequate animal models for special purposes in comparison to human placentation.(AU)

Modelos animais são essenciais para entender a placenta humana sadia. Neste sentido os roedores relacionados ao porquinho da índia tornaram-se foco para tal entendimento. Em particular, os processos de invasão trofoblástica são semelhantes. O último está associado a uma área especializada, a subplacenta. Uma vez que os resultados anteriores mostraram diferenças entre o porquinho da índia e seu relativo o preá, buscamos estudar o desenvolvimento subplacentário com mais detalhes. Pesquisamos 16 fêmeas gestantes de 14 a 55 dias de gestação por meio de histologia, morfometria, imuno-histoquímica e microscopia eletrônica. A sobreposição entre os sistemas sanguíneos materno e fetal apresentou-se com íntima relação, sugerindo alguns processos de troca. A proliferação foi revelada por três métodos independentes, sendo mais ativos no início e metade da gestação, o que corroborou com os resultados anteriores. Embora a degeneração dos tecidos tenha ocorrido, a subplacenta foi mantida até o termo gestacional com acesso à vascularização fetal, apoiando uma hipótese sobre a liberação de substâncias para a unidade fetal em gestação avançada. Em contraste com outras espécies, o trofoblasto extraplacentário mostrou uma mudança de flâmulas sinciciais para células gigantes durante a metade da gestação. As visualizações sobre a placentação em caviomorfos foram influenciadas pelo porquinho da índia, mas nossos dados apoiaram estudos recentes de que a subplacenta apresentava uma plasticidade muito maior. Em relação ao crescimento subplacentário, a degeneração e provavelmente também os processos de troca, o preá e outras espécies apresentaram um padrão mais basal de caviomorfos do que o porquinho da índia. Tais diferenças devem ser consideradas, ao escolher os modelos animais mais adequados para fins especiais em comparação com a placentação humana.(AU)

Subplacental development in Galea spixii / Desenvolvimento subplacentário do preá (Galea spixii)

Animal models are essential to understand healthy human placentation. Guinea pig related rodents became on focus for such purposes. In particular, processes of trophoblast invasion are similar. The latter is associated with a specialized area, the subplacenta. Since previous results showed differences between the guinea pig and its close relative Galea spixii, we aimed to study subplacental development with more detail. We investigated 16 pregnant females of 14 to 55 days of gestation by means of histology, morphometrics, immunohistochemistry and electron microscopy. The overlap between the fetomaternal blood systems resulted as intimate, suggesting some exchange processes. Proliferation was revealed by three independent methods, being most active in early and mid-gestation, which was in accordance to former results. Though degeneration of tissues took place, the subplacenta was maintained towards term with access to the fetal vascularization, supporting a hypothesis about the release of substances to the fetal unit in advanced gestation. In contrast to other species, the extraplacental trophoblast showed a shift from syncytial streamers to giant cells during mid-gestation. Views on placentation in caviomorphs were influenced by the guinea pig, but our data supported recent studies that the subplacenta had a much greater placidity. In regard to subplacental grow, degeneration and likely also exchange processes, Galea and other species showed a more basal pattern of caviomorphs than the guinea pig. Such differences should be considered, when choosing most adequate animal models for special purposes in comparison to human placentation.(AU)

Modelos animais são essenciais para entender a placenta humana sadia. Neste sentido os roedores relacionados ao porquinho da índia tornaram-se foco para tal entendimento. Em particular, os processos de invasão trofoblástica são semelhantes. O último está associado a uma área especializada, a subplacenta. Uma vez que os resultados anteriores mostraram diferenças entre o porquinho da índia e seu relativo o preá, buscamos estudar o desenvolvimento subplacentário com mais detalhes. Pesquisamos 16 fêmeas gestantes de 14 a 55 dias de gestação por meio de histologia, morfometria, imuno-histoquímica e microscopia eletrônica. A sobreposição entre os sistemas sanguíneos materno e fetal apresentou-se com íntima relação, sugerindo alguns processos de troca. A proliferação foi revelada por três métodos independentes, sendo mais ativos no início e metade da gestação, o que corroborou com os resultados anteriores. Embora a degeneração dos tecidos tenha ocorrido, a subplacenta foi mantida até o termo gestacional com acesso à vascularização fetal, apoiando uma hipótese sobre a liberação de substâncias para a unidade fetal em gestação avançada. Em contraste com outras espécies, o trofoblasto extraplacentário mostrou uma mudança de flâmulas sinciciais para células gigantes durante a metade da gestação. As visualizações sobre a placentação em caviomorfos foram influenciadas pelo porquinho da índia, mas nossos dados apoiaram estudos recentes de que a subplacenta apresentava uma plasticidade muito maior. Em relação ao crescimento subplacentário, a degeneração e provavelmente também os processos de troca, o preá e outras espécies apresentaram um padrão mais basal de caviomorfos do que o porquinho da índia. Tais diferenças devem ser consideradas, ao escolher os modelos animais mais adequados para fins especiais em comparação com a placentação humana.(AU)

Effects of maternal high fat intake during pregnancy and lactation on total cholesterol and adipose tissue in neonatal rats / Efeitos do consumo materno de dieta hiperlipídica durante a gestação e lactação no colesterol total e tecido adiposo de ratos neonatos

AimObesity during pregnancy is one of the most established risk factors for negative long-term programming. The aim of the present study was to investigate the effects of maternal consumption of a high-fat diet during pregnancy and lactation on the weight gain, visceral adipose tissue and cholesterolemia in neonatal rats.MethodsWistar rats were divided into two groups according to the mother's diet during pregnancy and lactation: Control group (CG, n = 12) were the offspring of rats fed a standard diet (4% lipid) and the Test group (TG, n = 12) were pups rats fed on a high fat diet (23% lipid). The weight of the animals was measured on alternate days until the 22nd day of life, when collected visceral adipose tissue and blood were collected for biochemical analysis. For statistical analysis the Student t test, Sidak´s teste and two way ANOVA was used, with p 0.05.Resultsthe test group showed differences in weight gain, visceral adipose tissue and higher cholesterol. ConclusionA maternal exposure to a high-fat diet during pregnancy and lactation can promote changes in weight gain, hypercholesterolemia and an increase in adipose tissue in neonatal rats.(AU)

ObjetivoA obesidade durante a gestação é um dos fatores de risco mais estabelecidos para uma programação negativa em longo prazo. O objetivo do presente estudo foi investigar os efeitos do consumo materno de uma dieta hiperlipídica durante a gestação e lactação no aumento do peso, do tecido adiposo visceral e colesterolemia em ratos neonatos. MétodosRatos Wistar foram divididos em dois grupos de acordo com a dieta da mãe durante a gestação e lactação: grupo controle (GC, n=12) composto por filhotes de ratas alimentadas com uma dieta padrão (lipídios 4%) e o grupo teste (GT, n=12) composto de filhotes de ratas alimentadas com dieta hiperlipídica (lipídios 23%). O peso dos animais foi aferido em dias alternados até o 22° dia de vida, quando foi coletado sangue para análises bioquímicas. Para a análise estatística utilizou-se os seguintes testes: two way ANOVA, teste de Sidak e teste t de Student, com p 0,05.ResultadosO grupo teste mostrou diferença no ganho de peso, no tecido adiposo visceral e nos níveis de colesterol.ConclusãoUma exposição materna a uma dieta hiperlipídica durante a gestação e lactação pode promover maior ganho ponderal, hipercolesterolemia e um aumento do tecido adiposo em ratos neonatos.(AU)

Maternal plasma levels of vascular endothelial growth factor and interleukin-6 in the term-pregnant and non-pregnant rats

Female reproductive functions are associated with changes in the level of systemic cytokines which play pivotal roles during ovulation, development of corpus luteum, embryo implantation, and labor. Although many studies reported elevated level of VEGF during early pregnancy and IL-6 at the onset of labors, there is still incomplete elucidation of the level of these systemic cytokines in the very-late pregnancy or in the non-pregnant states. The purpose of this study was to determine the maternal plasma level of vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) in the term-pregnant and non-pregnant rats to see if their levels are changed at the very end of pregnancy. Blood samples were collected from twelve term-pregnant rats and compared by aged-matched seven non-pregnant rats. Plasma was separated carefully from all samples and the concentrations of VEGF and IL-6 were determined by ELISA techniques. The plasma concentration of VEGF was significantly higher in the non-pregnant rats (0.237 pg/ ml) than in the term-pregnant (0.2 pg/ ml; P < 0.001). The plasma concentration of IL-6 was significantly higher in the term-pregnant rats (50.12 pg/ ml) than in the non-pregnant ones (40.19 pg/ ml; P < 0.05). These findings suggest that the level of VEGF is decreased few days before labor possibly to decrease the ovarian blood flow and to cause luteolysis while the level of IL-6 is increased at the end of pregnancy possibly to cause a local inflammation. Together, these changes are necessary to stimulate the onset of labor.

Maternal plasma levels of vascular endothelial growth factor and interleukin-6 in the term-pregnant and non-pregnant rats

Female reproductive functions are associated with changes in the level of systemic cytokines which play pivotal roles during ovulation, development of corpus luteum, embryo implantation, and labor. Although many studies reported elevated level of VEGF during early pregnancy and IL-6 at the onset of labors, there is still incomplete elucidation of the level of these systemic cytokines in the very-late pregnancy or in the non-pregnant states. The purpose of this study was to determine the maternal plasma level of vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) in the term-pregnant and non-pregnant rats to see if their levels are changed at the very end of pregnancy. Blood samples were collected from twelve term-pregnant rats and compared by aged-matched seven non-pregnant rats. Plasma was separated carefully from all samples and the concentrations of VEGF and IL-6 were determined by ELISA techniques. The plasma concentration of VEGF was significantly higher in the non-pregnant rats (0.237 pg/ ml) than in the term-pregnant (0.2 pg/ ml; P < 0.001). The plasma concentration of IL-6 was significantly higher in the term-pregnant rats (50.12 pg/ ml) than in the non-pregnant ones (40.19 pg/ ml; P < 0.05). These findings suggest that the level of VEGF is decreased few days before labor possibly to decrease the ovarian blood flow and to cause luteolysis while the level of IL-6 is increased at the end of pregnancy possibly to cause a local inflammation. Together, these changes are necessary to stimulate the onset of labor.(AU)

Efeito da indução da atividade ovariana e da ovulação, com gonadotropinas exógenas (eCG, hCG), na recuperação, viabilidade e congelabilidade de embriões de gatos domésticos / Effect of induction of ovarian activity and ovulation, with exogenous gonadotropins (eCG, hCG), on the recovery, viability and freezing of domestic cat embryos

Objetivou-se a avaliação da efetividade do protocolo de aplicação, em dose única, de gonadotrofinas exógenas (150 UI eCG associado a 100 UI hCG), na indução da atividade ovariana e da ovulação em gatas domésticas, bem como a competência desenvolvimental in vitro e in vivo de embriões felinos produzidos por monta natural, e ainda o efeito da congelação padronizada no desempenho pós-descongelamento de embriões felinos congelados em diferentes fases de desenvolvimento, como modelo experimental para potencial uso em espécies felídeas não domésticas. Foram utilizados três machos e 22 gatas para indução reprodutiva e duas gatas com manifestação natural do cio. Em todas as gatas foram computadas, na superfície ovariana, a presença de corpos lúteos e folículos anovulatórios. Dos animais induzidos e inseminados naturalmente, foram coletados 189 embriões, dos quais: em 27 foi testada a viabilidade de cultivo a fresco; 24 foram transferidos a quatro gatas receptoras, sincronizadas com o mesmo método; e em 21 embriões foi testada a viabilidade de desenvolvimento in vitro, após congelamento com meio (10% glicerol, 0,1mol L-1 sacarose). Conclui-se que o protocolo foi efetivo na produção de embriões felinos, embora haja indícios sugerindo a diminuição da dose de eCG e aumento da dose de hCG. Os embriões produzidos foram viáveis para uso em transferência interespecífica, sendo que 75% (3/4) das fêmeas receptoras ficaram gestantes e 50% levaram a gestação a termo. O protocolo de congelamento reduziu em 51% a taxa de desenvolvimento (8/21), in vitro de embriões felinos e reduziu significativamente (P = 0,01) o número de blastômeros (139,1/161) após 24 horas de cultivo em meio TCM 199 modificado.(AU)

The objective of this study was to evaluate the effectiveness of the single-dose protocol of exogenous gonadotropins (150 IU eCG associated with 100 IU hCG), to induce ovarian activity and ovulation in domestic cats, as well as developmental competence in vitro and in vivo of feline embryos produced by natural mating. The effect of standardized freezing on the post-thawing performance of frozen feline embryos at different stages of development also was evaluated as an experimental model for potential use in non-domestic felid species. For reproductive induction, 3 males and 22 females were used and 2 females with natural estrus. In all females, the presence of luteal bodies and anovulatory follicles were counted on the ovarian surface. Of induced and naturally inseminated animals, 189 embryos were collected, of which: the viability of fresh culture was tested in 27; 24 were transferred to four receiving cats, synchronized with the same method; and in 21 embryos the viability of in vitro development was tested after freezing with medium (10% glycerol, 0.1 mol L-1 sucrose). It was concluded that the protocol was effective in the production of feline embryos, although there are suggestions to decrease the eCG dose and increase the hCG dose. The embryos produced were viable for use in interspecific transfers, with 75% (3/4) of the recipient females becoming pregnant and 50% leading to full term pregnancy. The freezing protocol reduced the rate of in vitro development (8/21), of feline embryos by 51% and significantly reduced the number of blastomers (P = 0.01), after 24 hours of culture in modified TCM 199 medium (139,1/161).(AU)

Efeito da indução da atividade ovariana e da ovulação, com gonadotropinas exógenas (eCG, hCG), na recuperação, viabilidade e congelabilidade de embriões de gatos domésticos / Effect of induction of ovarian activity and ovulation, with exogenous gonadotropins (eCG, hCG), on the recovery, viability and freezing of domestic cat embryos

Objetivou-se a avaliação da efetividade do protocolo de aplicação, em dose única, de gonadotrofinas exógenas (150 UI eCG associado a 100 UI hCG), na indução da atividade ovariana e da ovulação em gatas domésticas, bem como a competência desenvolvimental in vitro e in vivo de embriões felinos produzidos por monta natural, e ainda o efeito da congelação padronizada no desempenho pós-descongelamento de embriões felinos congelados em diferentes fases de desenvolvimento, como modelo experimental para potencial uso em espécies felídeas não domésticas. Foram utilizados três machos e 22 gatas para indução reprodutiva e duas gatas com manifestação natural do cio. Em todas as gatas foram computadas, na superfície ovariana, a presença de corpos lúteos e folículos anovulatórios. Dos animais induzidos e inseminados naturalmente, foram coletados 189 embriões, dos quais: em 27 foi testada a viabilidade de cultivo a fresco; 24 foram transferidos a quatro gatas receptoras, sincronizadas com o mesmo método; e em 21 embriões foi testada a viabilidade de desenvolvimento in vitro, após congelamento com meio (10% glicerol, 0,1mol L-1 sacarose). Conclui-se que o protocolo foi efetivo na produção de embriões felinos, embora haja indícios sugerindo a diminuição da dose de eCG e aumento da dose de hCG. Os embriões produzidos foram viáveis para uso em transferência interespecífica, sendo que 75% (3/4) das fêmeas receptoras ficaram gestantes e 50% levaram a gestação a termo. O protocolo de congelamento reduziu em 51% a taxa de desenvolvimento (8/21), in vitro de embriões felinos e reduziu significativamente (P = 0,01) o número de blastômeros (139,1/161) após 24 horas de cultivo em meio TCM 199 modificado.

The objective of this study was to evaluate the effectiveness of the single-dose protocol of exogenous gonadotropins (150 IU eCG associated with 100 IU hCG), to induce ovarian activity and ovulation in domestic cats, as well as developmental competence in vitro and in vivo of feline embryos produced by natural mating. The effect of standardized freezing on the post-thawing performance of frozen feline embryos at different stages of development also was evaluated as an experimental model for potential use in non-domestic felid species. For reproductive induction, 3 males and 22 females were used and 2 females with natural estrus. In all females, the presence of luteal bodies and anovulatory follicles were counted on the ovarian surface. Of induced and naturally inseminated animals, 189 embryos were collected, of which: the viability of fresh culture was tested in 27; 24 were transferred to four receiving cats, synchronized with the same method; and in 21 embryos the viability of in vitro development was tested after freezing with medium (10% glycerol, 0.1 mol L-1 sucrose). It was concluded that the protocol was effective in the production of feline embryos, although there are suggestions to decrease the eCG dose and increase the hCG dose. The embryos produced were viable for use in interspecific transfers, with 75% (3/4) of the recipient females becoming pregnant and 50% leading to full term pregnancy. The freezing protocol reduced the rate of in vitro development (8/21), of feline embryos by 51% and significantly reduced the number of blastomers (P = 0.01), after 24 hours of culture in modified TCM 199 medium (139,1/161).

Contamination index. A novel parameter for metal and pesticide analyses in maternal blood and umbilical cord

PURPOSE:To evaluate the contamination index of metals and pesticides in pregnant women, and to relate this to perinatal outcomes.METHODS:Descriptive, retrospective, exploratory study, developed from existing secondary data analyses at Level III maternity center. A total of 40 mothers with their newborns (NB), living in a rural area in Botucatu- Brazil and surrounding region. Blood samples from mothers and newborn were collected to determine the total contamination index for metals and pesticides. The concentrations of each metal and each pesticide were determined in blood samples of mothers and their newborns by Rudge's results. After obtaining these concentrations, the total contamination index in mother and NB was calculated, along with its correlation with clinical parameters of NB.RESULTS:There was no correlation (p> 0.05) between maternal contamination index with NB clinical parameters, and NB contamination index versus NB clinical parameters.CONCLUSION:The maternal contamination index of metals and pesticides was not related to perinatal outcomes, but it could be used as baseline parameter in future toxicological studies, regarding to long-term toxic characteristics as persistent organic pollutants, its long half-lives, bioacumulative, and expected to impose serious health effects on humans.(AU)

Neoplasias em equinos, asininos e muares

Os equinos, asininos e muares têm importante participação na economia e as neoplasias nestas espécies são importantes causas de perdas econômicas. As neoplasias em equídeos são diagnosticadas através de necropsias e biópsias, sendo o principal sistema afetado o tegumentar, principalmente em decorrência do sarcoide. No entanto é importante incluir neoplasias de outros sistemas, inclusive em órgãos internos como importantes diagnósticos diferenciais que causam debilidade e podem até levar a morte. Esta dissertação foi elaborada em dois capítulos, compostos por dois artigos originais. O primeiro a ser submetido à revista Pesquisa Veterinária Brasileira relata a frequência de tumores diagnosticados em nove sistemas primários de equídeos, descrevendo aspectos epidemiológicos, clínicos e patológicos. Foram diagnosticadas 186 neoplasias em 167 equídeos. O sistema mais afetado foi o tegumentar seguindo do reprodutor masculino e feminino, ocular/periocular, respiratório, digestório, hepático, nervoso e hematopoiético. A neoplasia mais diagnosticada foi o sarcoide, seguida do carcinoma de células escamosas e melanoma. Metástases ocorreram em 14 casos e os sinais clínicos foram variáveis de acordo com a localização anatômica. O segundo artigo será submetido à Journal of Equine Veterinary Science e descreve-se três casos de tumor das células da granulosa diagnosticados em éguas, abordando os aspectos epidemiológicos, clínicos e patológicos. As éguas apresentavam alterações comportamentais e reprodutivos como infertilidade, repetição de cio (ninfomania), comportamento de garanhão (virilidade), indiferença ao garanhão e ausência de sinais de estro. As neoplasias foram visualizados através da ultrassonografia e removidos cirurgicamente pela ovariectomia e enviados para exames histopatológicos. Macroscopicamente as massas variavam de 9 a 20 cm de diâmetro, eram firmes e bem delimitadas com superfícies lisas e ao corte observou-se múltiplas cavidades císticas circundadas por trabéculas de tecido amarelo-esbranquiçado e firmes. Microscopicamente observou-se massas multilobuladas, bem delimitadas, compostas por células dispostas em ninhos semelhantes a folículos abortados ou formando cistos, apoiadas em moderado estroma fibrocolagenoso. Pseudorosetas foram observadas. Na neoplasia da égua 2, observou-se formações semelhantes a túbulos de sertoli e células semelhantes a células de Leydig. Na imuno-histoquímica, houve marcação pela inibina e vimetina, de células da granulosa maduras e neoplásicas que compunham os ninhos e revestiam cistos. Após a cirurgia, a égua 1 não exibiu mais as alterações comportamentais, apresentou cio duas vezes, ficou prenhe e abortou. A égua 3 também ficou prenhe e pariu a termo. Não há informações referentes a éguas 2 após a cirurgia. Foi possível constatar com esta dissertação que embora a maioria das neoplasias diagnosticadas em equídeos no Laboratório de Patologia Animal do Hospital Veterinário Universitário da UFCG, envolvam órgãos relacionados ao tegumento, neoplasias que acometem órgãos internos ocorrem esporadicamente e é importante incluí-las como diagnósticos diferenciais.

Horses, donkeys and mules play an important role in the economy and neoplasms in these species are important causes of economic losses. Neoplasms in equines are diagnosed through necropsies and biopsies, with the integumentary system being the main affected system, mainly due to the sarcoid. However, it is important to include neoplasms from other systems, including internal organs as important differential diagnoses that cause debility and can even lead to death. This dissertation was elaborated in two chapters, composed by two original articles. The first to be submitted to the journal Pesquisa Veterinária Brasileira reports the frequency of tumors diagnosed in nine primary equine systems, describing epidemiological, clinical and pathological aspects. 186 neoplasms were diagnosed in 167 horses. The most affected system was the integumentary, followed by the male and female, ocular/periocular, respiratory, digestive, hepatic, nervous and hematopoietic systems. The most diagnosed neoplasm was sarcoid, followed by squamous cell carcinoma and melanoma. Metastases occurred in 14 cases and clinical signs varied according to anatomical location. The second article will be submitted to the Journal of Equine Veterinary Science and describes three cases of granulosa cell tumor diagnosed in mares, addressing the epidemiological, clinical and pathological aspects. The mares showed behavioral and reproductive changes such as infertility, repetition of heat (nymphomania), stallion behavior (virility), indifference to the stallion and absence of signs of estrus. The neoplasms were visualized through ultrasonography and surgically removed by ovariectomy and sent for histopathological exams. Macroscopically, as masses ranged from 9 to 20 cm in diameter, they were firm and well delimited with smooth surfaces and, when cut, multiple cystic cavities surrounded by firm yellow-white tissue trabeculae were observed. Microscopically, well-delimited, multilobulated masses were observed, composed of cells arranged in nests similar to aborted follicles or forming cysts, supported by a moderate fibrocollagenous stroma. Pseudorosettes were observed. In mare 2 neoplasm, formations similar to sertoli tubules and cells similar to Leydig cells were observed. In immunohistochemistry, there was inhibin and vimetin marking of mature and neoplastic granulosa cells that made up the nests and lined the cysts. After surgery, mare 1 no longer exhibited behavioral changes, presented heat twice, became pregnant and miscarried. Mare 3 also became pregnant and gave birth to term. There is no information regarding mares 2 after surgery. It was possible to verify with this dissertation that although most neoplasms diagnosed in horses in the Animal Pathology Laboratory of the University Veterinary Hospital of UFCG involve organs related to the integument, neoplasms that affect internal organs occur sporadically and it is important to include them as differential diagnoses.

HEMODINÂMICA PLACENTÁRIA DE ÉGUAS: ULTRASSONOGRAFIA DOPPLER ESPECTRAL, ANÁTOMO-HISTOPATOLOGIA E IMUNOEXPRESSÃO DE EZRIN

Este estudo teve como objetivo avaliar a hemodinâmica placentária de éguas com gestações avançadas por meio da ultrassonografia Doppler espectral das artérias uterinas, mensuração da junção útero-placentária com ultrassonografia modo B, e análise morfológica e imunoexpressão de Ezrin da placenta pós-parto. Foram utilizadas 32 éguas da raça Mangalarga Marchador a partir de cinco meses de gestação, a fim de medir mensalmente a espessura da junção útero-placentária (JUP) e os índices espectrais com ultrassonografia Doppler das artérias uterinas direita e esquerda. Após o nascimento dos potros, as placentas coletadas foram analisadas quanto à morfologia. Foram coletados oito fragmentos de diferentes regiões da placenta e acondicionados em formol tamponado para avaliação microscópica e histomorfométrica dos fragmentos placentários e posteriormente a realiazação da técnica de imunohistoquímica para imunoexpressão do anticorpo anti-ezrin. Durante a gestação, o índice de pulsatilidade (IP) e o índice de resistência (IR) das artérias uterinas foram obtidos por ultrassonografia Doppler espectral, a JUP foi obtida mensalmente por ultrassonografia modo B. Na microscopia foram avaliadas as regiões dos cornos gravídicos e não gravídicos quanto à vascularização dos vasos fetais do alantocório e dos microcotilédones e foram contados os números de vasos em cinco campos de cada corno. Para histomorfometria foram contados cinco campos de vasos fetais da lâmina basal do corioalantoide e mensurada luz dos vasos em comprimento e largura em escala de micrômetro. Para classificação da imunoexpressão do anticorpo anti-ezrin nas amostras placentárias foram utilizados os parâmentros de intensidade de cor marrom nos microcotilédones, células trofoblásticas, lâmina basal e vasos do corioalantóide, bem como a distribuição focal ou difusa das marcações e região dos microcotilédones proximal, medial e distal da lâmina basal do alantocório. Os resultados foram separados por capítulos. A espessura uteroplacentária combinada correlacionou-se com o tempo gestacional de até 13 anos de idade, sendo a diferença significativa observada a partir do sexto mês. A JUP ficou dentro das medidas ideais para esta raça e para este período gestacional. Não houve correlação do JUP com o IP, porém foi observada uma correlação negativa e significativa do IR. O índice de resistência diferiu significativamente entre as faixas etárias, e o IR da artéria uterina esquerda tendeu a diminuir em todas as faixas etárias. Além disso, apenas o IR diferiu significativamente entre as medianas da idade gestacional. O PI esquerdo caiu nas éguas mais velhas. Na avaliação morfológica da placenta, não foram encontradas alterações significativas na macroscopia. Na análise microscópica da região do corno gravídico (CG) foi possível observar que o arranjo microcotiledonário se apresentou mais intenso ou equiparado ao corno não gravídico (CNG), com excessão de um caso. A intensidade de vascularização nos microcotilédones e no alantocório teve uma grande variação entre os cornos. Cinco casos apresentaram congestão nos microcotilédones e vasos fetais. A expressão do anticorpo anti-ezrin nos cornos placentários, foi mais intensa na região proximal dos microcotilédones em relação à lâmina basal do alantocório e apresentou-se difusa por toda rede de microcotilédones. O CG apresentou mais expressão nas células trofoblásticas em comparação ao CNG e entre os cornos a intensidade de cor ficou equiparada na maioria dos casos, apresentando coloração moderada e intensa. Na microscopia dos cornos não foi possível observar alterações significativas relacionadas a alguma patologia como necrose, infiltrados inflamatórios e hipoplasia. Não foram encontradas diferença significativa entre os dois cornos relacionados a número de vasos e intensidade de vascularização dos microcotilédones e alantocório. As medidas da luz dos vasos apresentaram igualdade estatística (p 0,05) entre os cornos. Assim, ocorreu um aumento da perfusão sanguínea nas artérias uterinas de éguas com gestações avançadas e entre diferentes categorias de idade durante o desenvolvimento fisiológico da placenta e do feto. Foi possível obter dados das características vasculares da unidade feto-placentária equina que não apresentaram alterações morfológicas. A expressão de ezrin foi bem signficativa nas células trofoblásticas localizadas nos microcotilédones de placentas equinas a termo que tem um papel importante na vascularização e consequentemente na troca materno-fetal. A aplicabilidade acompanhamento ultrassonográfico gestacional em equinos tem como principal objetivo prevenir a perda gestacional de alto valor comercial, com a mensuração do fluxo sanguíneo uterino em éguas predispostas a doenças vasculares e com história de aborto espontâneo e parto prematuro. Para um diagnóstico confiável na espécie, foi essencial obter dados de um padrão de normalidade de placentas saudáveis para comparar esses dados em futuras éguas com prenhez de alto risco, principalmente vascular, e escolher a intervenção clínica necessária.

This study aimed to evaluate the placental hemodynamics of mares with advanced pregnancies by means of spectral Doppler ultrasonography of the uterine arteries, measurement of the uteroplacental junction with B-mode ultrasound, and morphological analysis and Ezrin immunoexpression of the postpartum placenta. Thirty-two Mangalarga Marchador mares from five months of gestation were used in order to measure monthly the combined uteroplacental thickness (CUPT) and the spectral indices with Doppler ultrasound of the right and left uterine arteries. After the foals were born, the collected placentas were analyzed for morphology. Eight fragments from different regions of the placenta were collected and placed in buffered formalin for microscopic and histomorphometric evaluation of the placental fragments and later the realization of the immunohistochemical technique for immunoexpression of the anti-ezrin antibody. During pregnancy, the pulsatility index (PI) and resistance index (RI) of the uterine arteries were obtained by spectral Doppler ultrasonography, the CUPT was obtained monthly by B-mode ultrasonography. Under microscopy, the regions of the pregnant and non-pregnant horns were evaluated as to the vascularization of the fetal vessels of the allantochorium and microcotyledons, and the number of vessels in five fields of each horn were counted. For histomorphometry, five fields of fetal vessels from the basal lamina of the chorioallantois were counted, and the vessel lumen were measured in length and width on a micrometer scale. To classify the immunoexpression of anti-ezrin antibody in placental samples, brown color intensity parameters were used in microcotyledons, trophoblast cells, basal lamina and chorioallantoic vessels, as well as focal or diffuse distribution of markings and proximal, medial microcotyledon region and distal from the basal lamina of the allantochorium. The results were separated by chapters. The CUPT was correlated with gestational time up to 13 years of age, with a significant difference observed from the sixth month onwards. CUPT was within the ideal measurements for this breed and for this gestational period. There was no correlation of CUPT with IP, but a negative and significant correlation of RI was observed. The resistance index differed significantly between age groups, and the left uterine artery RI tended to decrease in all age groups. Furthermore, only the RI differed significantly between the medians of gestational age. Left PI fell in older mares. In the morphological evaluation of the placenta, no significant changes were found in the macroscopy. In the microscopic analysis of the gravid horn region it was possible to observe that the microcotyledonary arrangement was more intense or similar to the non-pregnant horn, with the exception of one case. The intensity of vascularization in microcotyledons and allantochorion varied widely between horns. Five cases presented congestion in the microcotyledons and fetal vessels. In the expression of anti-ezrin antibody in placental horns, it was more intense in the proximal region of the microcotyledons in relation to the basal lamina of the allantochorion and it was diffused throughout the network of microcotyledons. Pregnant horn showed more expression in synciotrophoblasts compared to nonpregnant horn and among horns the color intensity was similar in most cases, with moderate and intense coloration. In the microscopy of the horns it was not possible to observe significant alterations related to any pathology such as necrosis, inflammatory infiltrates and hypoplasia. No significant difference was found between the two horns related to the number of vessels and vascularization intensity of microcotyledons and allantochorium. Vessel lumen measurements showed statistical equality (p 0.05) between the horns. Thus, there was an increase in blood perfusion in the uterine arteries of mares with advanced pregnancies and between different age categories during the physiological development of the placenta and fetus. It was possible to obtain data on the vascular characteristics of the equine fetus-placental unit that did not show morphological changes. The expression of ezrin was very significant in trophoblastic cells located in the microcotyledons of term equine placentas, which play an important role in vascularization and consequently in maternal-fetal exchange. The applicability of gestational ultrasound monitoring in horses aims to prevent pregnancy loss of high commercial value, with the measurement of uterine blood flow in mares predisposed to vascular diseases and with a history of miscarriage and premature birth. For a reliable diagnosis in the species, it was essential to obtain data from a normality pattern of healthy placentas to compare these data in future mares with high risk pregnancy, mainly vascular, and to choose the necessary clinical intervention.

Blood flow indices in single and multiple bearing does

The aim of present study was to examine the changes in blood flow indices throughout pregnancy in singleton and multiple pregnant does. Doppler scanning was done to assess resistance index (RI), pulsatility index (PI), time-averaged maximum velocity (TAMV) and blood flow volume (Vol). The placentomes were investigated for blood indices on day 45 as an echogenic structure on the surface of the endometrium. Transrectal ultrasonography was performed every 30 days during the overage of the pregnancy. Diameters of largest six placentomes from each doe were measured using an ultrasonography device, and the mean values of placentome diameters were calculated. Values of RI and PI exhibited a steady decrease toward term and declined more rapidly and earlier in multiple than in single pregnancies. Values of RI and PI were negatively correlated with placetome size; however, values of TAMV and Vol were positively correlated with placentome size. In singleton and multiple pregnancies, the placentome size showed an increase toward term, but there is no difference in placentome size between single and multiple bearing does. In conclusion, the values of TAMV and Vol for multiple pregnancies were significantly higher than for singleton at any gestational month to satisfy the high foetal demand of nutrients and dissolved oxygen for foetal progression. Knowing the relationship between the foetal number and blood flow parameters could be used in the clinical management of such pregnancies and the early detection or prediction adverse pregnancy outcome.

Blood flow indices in single and multiple bearing does

The aim of present study was to examine the changes in blood flow indices throughout pregnancy in singleton and multiple pregnant does. Doppler scanning was done to assess resistance index (RI), pulsatility index (PI), time-averaged maximum velocity (TAMV) and blood flow volume (Vol). The placentomes were investigated for blood indices on day 45 as an echogenic structure on the surface of the endometrium. Transrectal ultrasonography was performed every 30 days during the overage of the pregnancy. Diameters of largest six placentomes from each doe were measured using an ultrasonography device, and the mean values of placentome diameters were calculated. Values of RI and PI exhibited a steady decrease toward term and declined more rapidly and earlier in multiple than in single pregnancies. Values of RI and PI were negatively correlated with placetome size; however, values of TAMV and Vol were positively correlated with placentome size. In singleton and multiple pregnancies, the placentome size showed an increase toward term, but there is no difference in placentome size between single and multiple bearing does. In conclusion, the values of TAMV and Vol for multiple pregnancies were significantly higher than for singleton at any gestational month to satisfy the high foetal demand of nutrients and dissolved oxygen for foetal progression. Knowing the relationship between the foetal number and blood flow parameters could be used in the clinical management of such pregnancies and the early detection or prediction adverse pregnancy outcome.(AU)