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)

Avaliação da lactatemia e das enzimas musculares de equinos Quarto de Milha submetidos à prova de laço em dupla com diferentes tipos de treinamento / Evaluation of lactatemia and muscle enzymes in Quarter Horses subjected to team ropping with different types of training

O objetivo do trabalho foi avaliar a influência de diferentes tipos de treinamento sobre o condicionamento físico de equinos por meio da determinação do lactato sanguíneo e da atividade sérica de creatina quinase, aspartato aminotransferase e lactato desidrogenase após exercício físico de alta intensidade e curta duração. Amostras de sangue venoso foram obtidas de 16 equinos da raça Quarto de Milha, divididos em dois grupos: grupo de treinamento regular (GTR) e grupo de treinamento esporádico (GTE), em sete diferentes momentos: 30 minutos antes do exercício (M0), imediatamente após (M1), 30 minutos (M2), uma (M3), duas (M4), seis (M5) e 24 (M6) horas após o exercício. Para a análise estatística, os dados foram testados quanto à normalidade e homogeneidade de variâncias. Para comparar os grupos e os momentos em cada grupo foram utilizados testes paramétricos (ANOVA) para a análise das atividades séricas das enzimas musculares e não paramétricos (Mann-Whitney e Friedmann) para a análise do lactato sanguíneo (P<0,05). Não houve diferença significativa entre os grupos para nenhuma variável. No entanto, dentro dos grupos experimentais foi possível observar diferenças significativas entre os momentos avaliados, em relação ao lactato e à LDH. No GTE, foram observadas diferenças significativas quanto ao lactato, entre o M0 e o M1, com valores respectivos de 0,90 mmol/L (mín. 0,8 - máx. 1,6) e 3,65mmo/L (mín. 1,0 - máx. 5,7) e quanto à LDH, onde os valores descritos no M6 diferiram significativamente de M0, M1, M2, M3 e M4. No GTR, diferenças significativas entre os momentos experimentais foram observadas em relação à LDH, sendo que os valores observados no M6 foram os menores e diferiram significativamente daqueles encontrados no M1, M2, M3 e M4. Em conclusão, não houve diferença entre o condicionamento físico dos animais treinados regularmente e aqueles treinados esporadicamente. A baixa magnitude das elevações das concentrações de CK, AST e LDH após o exercício e o rápido retorno aos valores basais, inclusive do lactato, observados em ambos os grupos, sugere que todos os animais avaliados estavam condicionados e aptos a realizar tal atividade física.(AU)

The aim of this study was to evaluate the influence of different types of training on physical fitness through the determination of blood lactate and serum creatinine kinase, aspartate aminotransferase, and lactate dehydrogenase activity after high intensity and short duration physical exercise. Venous blood samples were obtained from 16 Quarter Horses, divided into two groups: the regular training group (GTR) and sporadic training group (GTE), in seven different moments: 30 minutes before exercise (M0), immediately after the exercise (M1), 30 minutes (M2), one hour (M3), two hours (M4), six hours (M5) and 24 hours (M6) after the exercise. For statistical analysis, data was tested for normality and homogeneity of variances. To compare the groups and times in each group, parametric tests (ANOVA) were used for muscular enzymes activity and not parametric tests (Mann-Whitney and Friedmann) were used to analyze blood lactato (P<0.05). There was no significant difference between groups for any variable. However, within the experimental groups it was possible to observe significant differences between the evaluated moments, in relation to lactate and LDH. In the GTE, significant differences were observed for lactate between M0 and M1, with respective values ​​of 0.90 mmol/L (min. 0.8, max. 1.6) and 3.65 mmol/L (min. 1.0, max. 5,7) and for LDH, where the values ​​described in M6 differed significantly from M0, M1, M2, M3 and M4. In the GTR, significant differences between the experimental moments were observed in relation to LDH, and the values ​​observed in M6 were the lowest and differed significantly from those found in M1, M2, M3 and M4. In conclusion, there was no difference between the fitness of animals regularly trained and those trained sporadically. The low magnitude of elevations of serum CK, AST and LDH activity after exercise and the quick return to baseline values, including the blood lactate observed in both groups, suggest that all of evaluated animals were conditioned and able to perform such physical activity.(AU)

Avaliação da vitalidade de cordeiros nascidos de partos eutócicos e cesarianas / Evaluation of vitality in lambs born by normal delivery and cesarean section

O trabalho teve como objetivo avaliar a vitalidade de cordeiros nascidos de parto normal e de cesariana, ao longo das primeiras 48 horas de vida, por meio da pontuação no escore Apgar, do seu comportamento e da aferição dos parâmetros vitais. Os animais foram divididos em três grupos, sendo o primeiro constituído por cordeiros nascidos de parto normal (NORMAL) e os outros dois de cesarianas, porém com dois protocolos anestésicos diferentes, ou seja, por meio de anestesias inalatória (INAL) e total intravenosa (ATI). Os animais do grupo NORMAL obtiveram pontuação maior no escore Apgar. Ao nascimento e aos 15 minutos, 93,75% dos cordeiros do grupo NORMAL obtiveram pontuação máxima (7-8), enquanto que nenhum dos animais nascidos de cesarianas atingiu essa pontuação, ao nascimento. Aos 15 minutos, 25,00% dos animais do grupo INAL recebeu a mesma soma de pontos. A frequência cardíaca permaneceu praticamente inalterada para ambos os grupos, com tendência à diminuição dos valores, às 24 horas, para animais do grupo ATI. A frequência respiratória (FR) ao nascimento foi significativamente menor nos animais nascidos de cesarianas (26±25mpm, INAL; 5±5mpm, ATI), encontrando-se, porém, próxima aos valores dos animais do grupo NORMAL, aos 60 minutos. Constatou-se tendência à diminuição nos valores de temperatura até os 15 minutos de vida em todos os grupos, prolongando-se, até os 60 minutos, nos animais pertencentes ao grupo ATI. Os animais nascidos de cesarianas possuem vitalidade inferior aos nascidos de partos normais.(AU)

The aim of this study was to evaluate vitality of lambs born by normal delivery and cesarean section, during 48 hours of life, by means of APGAR score, behavior and vital parameters. The first group was formed with lambs born by normal delivery (NORMAL) and the other two groups were formed with lambs born by cesarean sections, but with two different anesthetic protocols, one used inhalation anesthesia (INAL) and the other total intravenous anesthesia (TIVA). Lambs in the NORMAL group obtained higher pontuation in Apgar score. At birth and 15 minutes, 93.75% of lambs NORMAL group obtained maximum score (7-8), while no lambs born by cesarean section obtained this score at birth. At 15 minutes, 25.00% of the lambs INAL group received the same score. The heart rate remained virtually unchanged for both groups, with a tendency to decrease values, at 24 hours for group TIVA animals. The respiratory rate at birth for the animals delivered by cesarean section (26±25 bpm, INAL; 5±5 bpm, TIVA) was significantly lower than for the ones of the normal delivery. Nevertheless, that value at 60 minutes was found quite close. A tendency for decreasing temperature values was verified until the 15 minutes in all groups, lasting until 60 minutes in animals in the TIVA group. Lambs born by c-section have lower vitality than lambs born by normal delivery.(AU)

Efeito da tosquia na temperatura corpórea e outros parâmetros clínicos em ovinos / Effect of shearing on the skin temperature and other clinical parameters in sheep

Os objetivos deste estudo foram determinar a variação da temperatura após a tosquia de ovinos em condição ambiental quente e seca e comparar as mudanças de temperatura com variações de frequências cardíaca e respiratória, movimentos ruminais e estado de hidratação. Vinte ovelhas Suffolk não tosquiadas foram estudadas. O exame físico foi realizado em todos os animais três vezes ao dia às 7:00, 1:00 PM e 7:00 PM, durante 42 dias (22 dias antes do corte e 20 dias após o corte). A temperatura da superfície corporal foi medida pelo termómetro de infravermelho sobre vários pontos. Os dados foram submetidos à análise de variância, para comparações entre os grupos (tosquiado x não tosquiado) em cada tempo, e a diferença significativa foi avaliada em nível de P<0,05 pelo teste de Tukey. A frequência respiratória foi estatisticamente significativa em todos os horários de cólera (P<0,05). Quando a umidade do ar estava alta, as freqüências respiratórias foram baixas (P= -1). O estresse térmico foi claro em ovinos deste estudo, refletindo mudanças acentuadas nas frequências cardíaca e respiratória e temperatura retal. A frequência respiratória foi o parâmetro mais confiável para estabelecer um quadro de estresse térmico em ovelhas não tosquiadas, com valores em média de três vezes maior do que os relatados na literatura. A correlação entre as temperaturas de superfície corpórea com a temperatura ambiente e umidade do ar foi negativa, explicado pelo efeito de isolamento de lã, ou seja, mesmo com um aumento da temperatura ambiente e umidade, a temperatura do corpo tende a manter um equilíbrio de compensação. Nos animais tosquiados, a correlação entre a temperatura da pele com a temperatura ambiente e umidade do ar mostraram que a temperatura da pele aumenta quando aumenta a temperatura ambiente. O aumento da temperatura ambiente não afeta a temperatura do corpo dos animais não tosquiados devido ao efeito isolante da lã. No entanto, quando a temperatura ambiente sobe, a presença da lã começa a afetar o conforto térmico, já que a presença desta dificulta a perda de calor pela sudação. Neste estudo, os melhores indicadores de estresse térmico foram a frequência respiratória e temperaturas retais e da pele. As temperaturas da pele medidas na face interna da coxa, axilas e períneo foram consideradas as mais confiáveis.

The aim of this study was to determine the variation of the temperature after shearing in sheep under dry and hot environment conditions and to compare the temperature changes with variation in cardiac and respiratory frequencies, ruminal movements and hydration status. Twenty Suffolk unshorn ewes were studied. Physical examination was performed in all animals three times a day at 7:00 AM, 1:00 PM and 7:00 PM, during 42 days (22 days before shearing and 20 days after shearing). The skin temperature was measured by infrared thermometer over several surfaces of the body. Data were submitted to analysis of variance, for comparisons between groups (shorn versus unshorn) at each time, and the significant difference was evaluated at level of P<0.05 by Tukey test. The respiratory frequency was statistically significant at all times. When air humidity was high, the respiratory frequencies were low. The thermal stress was clear in sheep of this study, reflecting marked changes in cardiac and respiratory frequencies and rectal temperature. The respiratory frequency was the parameter more reliable to establish a framework of thermal stress in the unshorn sheep, with values on average three times higher than those reported in the literature. The heart rate monitors the thermal variation of the environment and is also an indicator of heat stress. This variation shows the Suffolk breed is well adapted to hot climates. The correlation between the body surface temperatures with environment temperature and air humidity was negative, as explained by the effect of wool insulation, i.e. even with an increase in environment temperature and humidity, the body temperature tends to maintain a compensating balance. In the shorn animals, the correlation between skin temperature with environment temperature and air humidity showed that the skin temperature increases when the environment temperature increases. The increase in the environment temperature does not affect the body temperature of unshorn animals due the insulating effect of the wool. However, when environment temperature rises, the presence of the wool starts to affect the thermal comfort as the heat absorption is larger than the capacity of heat loss. In this study, the best thermal stress indicators were the respiratory frequency and rectal and skin temperatures. The temperatures of the skin measured at the perineum, axillae and inner thigh were considered the most reliable.

Molecular characterization of Cryptosporidium spp. in dairy calves from the state of São Paulo, Brazil.

Cryptosporidiosis is a common protozoan disease observed in a wide range of vertebrate hosts, including ruminants. Cattle can be a potential reservoir of Cryptosporidium spp., leading to environmental contamination with oocysts of zoonotic species. The molecular characterization of Cryptosporidium spp. isolated from cattle from the state of São Paulo, Brazil, was accomplished using nested polymerase chain reaction for amplification of fragments of the 18S rRNA gene and the glycoprotein GP60 gene, following sequencing of amplified fragments. Positivity for Cryptosporidium was found in 10.7% (21/196) of the samples. Four species of Cryptosporidium were identified: C. andersoni, C. bovis, C. parvum subtype IIaA15G2R1, and C. ryanae. To the best of our knowledge, this is the first report of infection by C. ryanae and C. parvum IIaA15G2R1 in cattle from Brazil.

Evaluation of a portable clinical analyzer for the determination of blood gas partial pressures, electrolyte concentrations, and hematocrit in venous blood samples collected from cattle, horses, and sheep.

OBJECTIVE: To compare results reported for blood gas partial pressures, electrolyte concentrations, and Hct in venous blood samples collected from cattle, horses, and sheep and analyzed by use of a portable clinical analyzer (PCA) and reference analyzer (RA). ANIMALS: Clinically normal animals (24 cattle, 22 horses, and 22 sheep). PROCEDURES: pH; Pco(2); Po(2); total carbon dioxide concentration; oxygen saturation; base excess; concentrations of HCO(3)(-), Na(+), K(+), and ionized calcium; Hct; and hemoglobin concentration were determined with a PCA. Results were compared with those obtained for the same blood sample with an RA. Bias (mean difference) and variability (95% confidence interval) were determined for all data reported. Data were also subjected to analyses by Deming regression and Pearson correlation. RESULTS: Analysis of Bland-Altman plots revealed good agreement between results obtained with the PCA and those obtained with the RA for pH and total carbon dioxide concentration in cattle, K(+) concentration in horses and sheep, and base excess in horses. Except for Na(+) concentration and Hct in horses and sheep, correlation was good or excellent for most variables reported. CONCLUSIONS AND CLINICAL RELEVANCE: Data from blood gas and electrolyte analyses obtained by use of the PCA can be used to evaluate the health status of cattle, horses, and sheep. Furthermore, the handheld PCA device may have a great advantage over the RA device as a result of the ability to analyze blood samples on farms that may be located far from urban centers.

Transcriptional changes associated with recurrent airway obstruction in affected and unaffected horses.

OBJECTIVE: To identify differentially expressed genes in pulmonary tissues of horses affected with summer pasture-associated obstructive pulmonary disease (SPAOPD), which is a form of recurrent airway obstruction (RAO), compared with those of unaffected horses. ANIMALS: 6 horses with SPAOPD-RAO and 6 unaffected (healthy) horses. PROCEDURES: Horses were assigned to 2 groups on the basis of medical history, clinical score, and transpleural pressure. Total RNA from each of the 5 lung lobes of each of the 6 SPAOPD-RAO-affected horses was extracted and pooled. Similarly, total RNA from unaffected horses was pooled. Differential display (DD) PCR assay was performed, and differentially expressed bands were purified and cloned into a plasmid vector. Plasmids were extracted from recombinant colonies, and purified DNA was sequenced. Genes of interest for RAO pathogenesis were identified. Real-time PCR assay was performed to confirm findings for the DD PCR assay. RESULTS: 18 differentially expressed genes (17 upregulated and 1 downregulated) were identified. Three genes of particular interest were found to be altered (2 upregulated and 1 downregulated) in horses with SPAOPD-RAO by use of real-time PCR assay, and these findings matched the differential expression found by use of the DD PCR assay. CONCLUSIONS AND CLINICAL RELEVANCE: SPAOPD-RAO in horses is a multifactorial, complex disease involving several genes. Upregulated genes, particularly beta2-microglobulin, and the downregulated secretoglobin gene can serve as marker genes that may help to identify SPAOPD-RAO at an early age.

Importância de Cryptosporidium spp. como causa de diarréia em bezerros / Importance of Cryptosporidium spp. as a cause of diarrhea in calves

Avaliou-se a presença de oocistos de Cryptosporidium spp. em amostras de fezes de 14 bezerros e de suas mães até a oitava semana pós parição. A maior taxa de excreção de oocistos foi verificada em bezerros com sete dias de idade. Das vacas, 42,8 por cento foram positivas para Cryptosporidium no período pós-parto. Em outra etapa deste estudo, foram acompanhados 57 bezerros positivos para Cryptosporidium, com até 30 dias de idade, provenientes de 32 propriedades leiteiras, e estudouse o grau de eliminação dos oocistos com a possível ocorrência de diarréia. Em todos os animais positivos para Cryptosporidium foi pesquisada a presença de bactérias enteropatogênicas, vírus (Rotavirus e Coronavirus) e protozoários (Eimeria spp.).(AU)

The aim of this research was to evaluate the shedding of Cryptosporidium spp. oocysts in fecal samples from 14 calves from one dairy farm, from birth until 60 days old and from cows until eight weeks after parturition. The higher percentage of oocysts excreted was observed in 7-day-old calves. In the post-partum period 43.7 percent of cows were positive for Cryptosporidium oocysts. Further analyses were accomplished in 57 calves from another 32 milk farms, previously known as positive for Cryptosporidium, through oocysts fecal screening and clinical signs analyses until calves were 30 days old. Fecal samples from all animals that presented diarrhea were screened for the presence of bacteria, virus (Rotavirus and Coronavirus ) and protozoa (Eimeria spp.).(AU)

Effect of age and abomasal puncture on peritoneal fluid, hematology, and serum biochemical analyses in young calves.

The goals of this study were to evaluate techniques for collection of peritoneal fluid from calves, establish reference ranges for fibrinogen in peritoneal fluid during the 1st month of life, and determine if abomasal puncture would alter peritoneal fluid or hematologic variables. Twenty-two healthy Holstein calves underwent 3 peritoneal fluid collections on day 1, day 15, and day 30 of age. Fibrinogen concentration in peritoneal fluid was 0.20 g/dL and 0.10 g/dL (P < .05) for day 1 and day 30, respectively, and 0.10 at day 15 (P > .05) for calves without abomasal puncture. Plasma fibrinogen concentration was 0.60 g/dL and 0.70 g/ dL (P < .05) for days 15 and 30, respectively, in calves without abomasal puncture. There were no significant differences (P < or = .05) in peritoneal fluid and peripheral blood total protein and fibrinogen concentrations, specific gravity, total and differential cell count, or erythrocyte counts between calves with or without abomasal puncture. We concluded that the reference ranges established for fibrinogen and total protein concentration are important for accurate evaluation of peritoneal fluid in calves for further comparison with similar-aged animals with gastrointestinal-tract or abdominal-cavity disease. Additionally, accidental abomasal puncture does not alter values of fibrinogen, total protein, and nucleated cell count in peritoneal fluid and does not cause apparent clinical abnormalities.