Physiological and hematological parameters of sheep reared in the tropics and subtropics / [Parâmetros fisiológicos e hematológicos de ovinos criados nos trópicos e nos subtrópicos]

This study aimed to determine the physiological and hematological parameters of sheep in Brazil. Therefore, 5,081 observations were used from previous experiments with animals from different Brazilian regions with the time of day, gender, breed, age, physiological characteristics, hematological and climatic variables. The animals were classified according to their stress level, and only data of non-stressed animals were used to calculate the reference values. Respiratory rate was the parameter that most differed from the normal range, with values ​​above the limits usually reported for the species, being higher in commercial wool sheep. For hematological variables, differences were observed between the present results and the previously reported values in the literature. A higher number of red blood cells (RBC), lower hemoglobin and mean corpuscular hemoglobin concentration (MCHC) were observed in young animals. A higher number of RBC and MCHC were noted in sheep crossed with local breeds. Considering that physiological and hematological parameters are indicators of the animal's pathological or physiological state, for a correct interpretation of the results, the use of reference values based on animals with similar racial characteristics and subjected to similar environmental conditions is recommended.(AU)

O objetivo deste estudo foi determinar parâmetros fisiológicos e hematológicos de ovinos no Brasil. Portanto, foram utilizadas 5.081 observações de experimentos anteriores com animais de diversas regiões do Brasil, com diferentes horário, sexo, raça, idade, características fisiológicas, variáveis ​​hematológicas e climáticas. Os animais foram classificados de acordo com o nível de estresse, e apenas os dados dos animais não estressados ​​foram utilizados para o cálculo dos valores de referência. A frequência respiratória foi o parâmetro que mais diferiu em relação à normalidade, com valores acima dos limites usualmente relatados para a espécie, sendo maior em ovinos de lã comercial. Para as variáveis ​​hematológicas, também foram verificadas diferenças entre os resultados observados e os valores previamente relatados na literatura. Maior número de hemácias, menor hemoglobina e concentração de hemoglobina corpuscular média (CHbCM) foram observados em animais jovens e maior número de hemácias e CHbCM em ovinos cruzados com raças locais. Considerando que os parâmetros fisiológicos e hematológicos são indicadores do estado patológico ou fisiológico do animal, para uma correta interpretação dos resultados recomenda-se a utilização de valores de referência baseados em animais com características raciais semelhantes e submetidos a condições ambientais também semelhantes.(AU)

Sheep Erythrocyte Preparation for Hemolytic Tests Exploring Complement Functional Activities.

Sheep erythrocytes (SE) are commonly used in complement functional tests. Non sensitized SE are useful to study the FH activity of cell protection. Indeed, as the cell surface of sheep erythrocytes is rich in sialic acids, Factor H (FH) is able to bind on it and therefore they represent a model of nonactivating surface. Because of their high capacity of complement regulation SE need to be modified to explore other functionality of the complement pathways, like the Complement hemolytic 50 (CH50) or the AP C3 convertase decay assays. For these tests, SE are sensitized with an anti-sheep red blood cell stroma antibody. In presence of serum or plasma complement components, sensitized SE may initiate complement cascade activation via the classic pathway explored in the CH50 assay. Sensitized SE may also be used to prepare C3b-coated SE that, with the use of buffers favoring AP, are suitable for the C3 Nef hemolytic assay and for the hemolytic assay studying the AP decay activity of FH. In this chapter we describe how to prepare SE for these different hemolytic tests.

Differences in mean grey levels of uterine ultrasonographic images between non-pregnant and pregnant ewes may serve as a tool for early pregnancy diagnosis.

The aim of this study was to determine how physiological and hormonal changes in the uterus during the estrous cycle and early gestational period affect the average grey values of pixels when conducting computer-assisted analysis of uterine ultrasonic images in ewes. For this purpose, 60 ewes on which there had been an estrous synchrony regimen imposed were included in the study. Animals were assigned to two groups with ewes not being mated and assessments occurred during the subsequent estrous cycle (Group 1; n = 25) and ewes being mated and assessments occurring during the subsequent early gestational period (Group 2; n = 35). Ewes were examined utilizing real time ultrasonic procedures and uterine images were obtained. Digital analysis of uterine ultrasonographic images was performed using image J program and mean grey levels (MGL) were determined. Values for progesterone concentrations were consistent with those previously reported in non-pregnant and pregnant ewes. There was a close association between MGL values in ewes of both Group I (P < 0.05) and II (P < 0.05) and days of the estrous cycle. There was also an association between MGL values and day of the gestational period in ewes of Group 2(P < 0.001). In conclusion, there are differences in MGL values between non-pregnant and pregnant ewes with there being changes as days of the estrous cycle and day of gestation period advances, therefore, this procedure could be used as a pregnancy diagnostic criterion during the early period of gestation in ewes.

Spatial distribution of serologically reactive sheep to Leptospira spp. in the northeast region of Brazil

Considering the importance of leptospirosis in sheep farming and public health and the significance of identifying which serogroups circulate in sheep within each region, the objective of this study was to determine the seroprevalence and spatial distribution of the most frequent serogroups causing infection by Leptospira sp. in ovine herds in the Northeast region of Brazil. Blood samples were collected from 4197 sheep from 229 herds in 7 Northeastern States. Sera were analyzed via microscopic agglutination test (MAT). The frequency of seroreactive sheep for Leptospira sp. was 14.06%. The states of Alagoas, Ceará, Paraíba, Piauí, Rio Grande do Norte, and Sergipe, located in the Caatinga biome, had the highest frequencies of serologically reactive sheep, and Maranhão, in the Cerrado biome, had the lowest frequency. The most frequent serogroups were Autumnalis (19.49%), Australis (15.76%) and Serjoe (14.41%). In the states of Ceará, Paraíba, Rio Grande do Norte, and Sergipe, 100% of their municipalities had at least one seroreactive animal. The highest frequencies of seropositive animals were found in the municipalities of União (50%), Passagem (49.06%), Canindé (48.89%), Igaci (28.95%), Gararu (31.2%), Pirapemas (17.5%), and Angicos (16%) located in the states of Piauí, Paraíba, Ceará, Alagoas, Sergipe, Maranhão and Rio Grande do Norte, respectively. The animal-level prevalence (14.06%) obtained in the present study is significant, especially considering the rustic nature of the species and the adverse conditions of the region for the infectious agent. In semi-arid conditions, it has been suggested that perhaps sheep do not seroconvert detectable titers on MAT with a cut-off point of 1:100. It is important to highlight that the ovine population in the Northeast region of Brazil is composed of mixed animals, which have been considered more resistant to infection by Leptospira spp. Also, environmental factors hostile to the survival of the infectious agent in the studied region should be taken into consideration, since they may have influenced the seropositive animal-level prevalence. A noteworthy variation was observed in agglutinin titers, which ranged from 100 to 1,600, where 80.2% of the positive samples had titers ≤ 200. It is important to highlight that more elevated titers (≥ 400) were obtained in all seven states, which may suggest an acute infection caused by a non-adapted serovar, indicating that preventive and control measures focused on possible infection sources for sheep should be adopted. Although some states showed the same serogroups as the most frequent, a variety of serogroups was observed in municipalities, which may indicate different sources of infection, whether interspecies, intraspecies, or via alternative routes of transmission in semi-arid conditions, such as venereal. This indicates that even though sheep are more resistant to infection, they become exposed due to the environment or management conditions. As such, identification, isolation, and treatment of the affected animals are alternative measures recommended for prevention and control of leptospirosis in sheep in the semi-arid region. It is evident that despite the lack of rain observed in the last decade in the Northeast region of Brazil, which prevented the formation of favorable environments for the presence of Leptospira, the infectious agent remains among the sheep, as well as other production and wild animals in the region. Some factors may be contributing to this scenario, such as the fact that sheep farming in the region is = characterized mainly by subsistence systems, where veterinary assistance and adequate sanitary management are absent, thus increasing the possibility of contact with Leptospira. (AU)

Metabolic profile of female lambs on annual ryegrass pasture managed under different grazing intensities and methods / [Perfil metabólico de cordeiros fêmeas em pastagem de azevém manejada sob diferentes intensidades e métodos de pastejo]

The aim of this study was to evaluate the effect of grazing methods and intensities on the metabolic profile of lambs grazing ryegrass. This study was performed in Eldorado do Sul (RS). The treatments consisted of two grazing intensities with rotational and continuous stocking methods: moderate and low. The experimental design was a randomized block arranged in a factorial scheme with four replicates. To evaluate the metabolic and nutritional conditions of the lambs, blood samples were collected to evaluate phosphorus, albumin, glucose, magnesium, globulin, cortisol, urea and total proteins. The highest blood urea value was observed in the month of September in lambs kept in continuous method grazing. There was no statistical difference between the grazing intensities and grazing methods for albumin, glucose, magnesium, globulin, cortisol and total protein levels in profile lambs. The cortisol values suggested that the animals were not subjected to high stress levels. In conclusion, rearing female lambs on ryegrass, irrespective of grazing methods or intensities, allowed the maintenance of body homeostasis and did not cause any metabolic unbalances, nutritional unbalances or stress.(AU)

O objetivo deste estudo foi avaliar o efeito dos métodos e das intensidades de pastejo no perfil metabólico de cordeiros. Este estudo foi realizado em Eldorado do Sul (RS). Os tratamentos consistiram em duas intensidades de pastejo (moderada e baixa), com métodos lotação rotativa e lotação contínua. O delineamento experimental foi em blocos ao acaso, em esquema fatorial com quatro repetições. Para avaliar as condições metabólicas e nutricionais dos cordeiros, foram coletadas amostras de sangue para avaliar fósforo, albumina, glicose, magnésio, globulina, cortisol, ureia e proteínas totais. O maior valor de ureia no sangue foi observado no mês de setembro nos cordeiros mantidos em pastejo contínuo. Não houve diferença estatística entre as intensidades de pastejo e os métodos de pastejo para albumina, glicose, magnésio, globulina, cortisol e níveis de proteína total nos cordeiros perfilados. Os valores de cortisol sugeriram que os animais não foram submetidos a altos níveis de estresse. Concluindo, a criação de cordeiros em azevém, independentemente dos métodos ou das intensidades de pastejo, permitiu a manutenção da homeostase corporal e não causou desequilíbrios metabólicos, desequilíbrios nutricionais ou estresse.(AU)

Reference values for biochemical analytes in feral sheep from Socorro Island, Revillagigedo Archipelago, Mexico / [Valores de referência para analitos bioquímicos em carneiros selvagens na Ilha Socorro, Arquipélago de Revillagigedo, México]

To establish reference values for biochemical analytes related to freshwater shortage adaptation, a total of 376 blood samples were collected from feral sheep at Socorro Island, Revillagigedo Archipelago. Year-round variation was assessed by sampling at the beginning of each season defined by the March equinox, June solstice, September equinox, and December solstice. The resulting data set was analyzed using Gaussian distribution and descriptive statistics. Confidence intervals of 95% were established. Analysis of variance was used to compare the mean values of each season. Total cholesterol, triglycerides, urea, albumin, total protein, sodium ion, anion gap, creatine kinase, arginine vasopressin, and aldosterone showed concentrations above the reference range for domestic sheep. Triglycerides, urea, albumin, sodium ion, and aldosterone showed concentrations within the reference range for domestic goats. Most biochemical analytes showed differences (P<0.05) between seasons, with the highest values occurring during winter, and the lowest during spring. Results could help improve the accuracy of metabolic profiles used as a tool for evaluating dehydration indicators, and to describe the physiological mechanisms employed by feral sheep to cope with seasonal availability of freshwater.(AU)

Para estabelecer valores de referência para analitos bioquímicos relacionados à adaptação da escassez de água doce, um total de 376 amostras de sangue foram coletadas de carneiros selvagens na ilha de Socorro, no arquipélago de Revillagigedo. A variação durante todo o ano foi avaliada por amostragem no início de cada estação definida pelo equinócio de março, solstício de junho, equinócio de setembro e solstício de dezembro. O conjunto de dados resultante foi analisado usando distribuição Gaussiana e estatística descritiva. Intervalos de confiança de 95% foram estabelecidos. A análise de variância foi usada para comparar os valores médios de cada estação. O colesterol total, triglicerídeos, ureia, albumina, proteína total, íon sódio, hiato aniônico, creatina quinase, arginina vasopressina e aldosterona apresentaram concentrações acima do intervalo de referência para carneiros domésticos. Triglicerídeos, ureia, albumina, íon sódio e aldosterona apresentaram concentrações dentro da faixa de referência para cabras domésticas. A maioria dos analitos bioquímicos apresentou diferenças (P<0,05) entre as estações, com os maiores valores ocorrendo no inverno e os menores na primavera. Os resultados podem ajudar a melhorar a precisão dos perfis metabólicos usados como uma ferramenta para avaliar os indicadores de desidratação e descrever os mecanismos fisiológicos empregados pelas carneiros selvagens para lidar com a disponibilidade sazonal de água doce.(AU)

Uncovering Roles of Streptococcus gordonii SrtA-Processed Proteins in the Biofilm Lifestyle.

Streptococcus gordonii is a commensal oral organism. Harmless in the oral cavity, S. gordonii is an opportunistic pathogen. S. gordonii adheres to body surfaces using surface adhesive proteins (adhesins), which are critical to subsequent formation of biofilm communities. As in most Gram-positive bacteria, S. gordonii surface proteins containing the C-terminal LPXTG motif cleavage sequence are processed by sortase A (SrtA) to become covalently attached to the cell wall. To characterize the functional diversity and redundancy in the family of SrtA-processed proteins, an S. gordonii DL1 markerless deletion mutant library was constructed of each of the 26 putative SrtA-processed proteins. Each library member was evaluated for growth in rich medium, biofilm formation on plastic, saliva and salivary fractions, cell surface hydrophobicity (CSH), hemagglutination, and integration into an ex vivo plaque biofilm community. Library members were compared to the non-SrtA-processed adhesins AbpA and AbpB. While no major growth differences in rich medium were observed, many S. gordonii LPXTG/A proteins impacted biofilm formation on one or more of the substrates. Several mutants showed significant differences in hemagglutination, hydrophobicity, or fitness in the ex vivo plaque model. From the identification of redundant and unique functions in these in vitro and ex vivo systems, functional stratification among the LPXTG/A proteins is apparent.IMPORTANCES. gordonii interactions with its environment depend on the complement of cell wall proteins. A subset of these cell wall proteins requires processing by the enzyme sortase A (SrtA). The identification of SrtA-processed proteins and their functional characterization will help the community to better understand how S. gordonii engages with its surroundings, including other microbes, integrates into the plaque community, adheres to the tooth surface, and hematogenously disseminates to cause blood-borne infections. This study identified 26 putative SrtA-processed proteins through creation of a markerless deletion mutant library. The library was subject to functional screens that were chosen to better understand key aspects of S. gordonii physiology and pathogenesis.

Avaliação da atividade funcional dos neutrófilos sanguíneos em ovelhas primíparas e pluríparas da raça Lacaune / [Functional evaluation of blood neutrophils in primiparous and pluriparous Lacaune ewes]

This study aimed to evaluate the neutrophil oxidative metabolism and phagocytosis of Zymonsan particles of primiparous and pluriparous Lacaune ewes during the first 30 days after lambing. A total of 20 ewes were evaluated, 10 primiparous (GPR) and 10 pluriparous (GPL). Evaluation of basal oxidative metabolism was performed using the nitroblue tetrazolium (NBT) technique, stimulating neutrophil phagocytosis with Zymosan particles. Blood samples were collected at parturition day (M1) and 1, 3, 7, 15 and 30 days after parturition, corresponding to M2, M3, M4, M5 and M6, respectively. In relation to the groups, GPR presented lower oxidative basal metabolism neutrophils in M1 compared to M4, in M3 with M1, M2, M4 and M5. In M4 and M5 differences were found at all times and in M6 with M4 and M5. Higher percentage of neutrophils than phagocytes were found in M4, M5 and M6 than in M1 and M2 in GPL animals. At all times GPR presented a lower percentage of phagocytosis than GPL. Thus it is concluded that the immune response in pluriparous sheep was more efficient than in primiparous sheep.(AU)

Desenvolvimento ponderal e parâmetros hematológicos e bioquímicos de cordeiros Pantaneiros submetidos a diferentes manejos de amamentação / [Ponderal development and hematological and biochemical parameters of Pantaneiro lambs subjected to different suckling regimens]

Este estudo teve por objetivo avaliar o desenvolvimento ponderal e a dinâmica dos parâmetros hematológicos e bioquímicos de cordeiros Pantaneiros submetidos a diferentes manejos de amamentação dos 15 aos 43 dias. Foram separados 30 cordeiros em três diferentes grupos (n=10). Os grupos foram caracterizados conforme o tempo de permanência das ovelhas com suas crias em diferentes sistemas de amamentação: MAM24 - ovelhas e cordeiros 24 horas em conjunto; MAM12 - ovelhas e cordeiros 12 horas em conjunto durante a noite; MAM2x30 - ovelhas e cordeiros 30 minutos de manhã e 30 minutos à tarde em conjunto. As coletas de amostras sanguíneas e as pesagens ocorriam a cada sete dias. Houve aumento significativo no peso com o avanço da idade dos cordeiros nos três manejos, mas os tratamentos não diferiram entre si para o ganho de peso. Houve diferenças entre os três tratamentos para a variável hematológica CHGM e para as variáveis bioquímicas AST, glicose, ureia e proteína total. As variáveis bioquímicas foram influenciadas pela faixa etária dos animais. Os diferentes manejos de amamentação não influenciaram o desenvolvimento ponderal dos cordeiros. Não ocorreram alterações patológicas. Alguns parâmetros hematológicos e bioquímicos podem ser influenciados pelo desenvolvimento etário dos cordeiros.(AU)

This study aimed to evaluate the weight development and the dynamics of hematological and biochemical parameters of Pantaneiro lambs submitted to different breastfeeding management from 15 to 43 days. First, 30 lambs were separated into three different groups (n = 10). The groups were characterized according to the length of stay of ewes with their young in different breastfeeding systems: MAM24 - ewes and lambs 24 hours together. MAM12- Sheep and lambs 12 hours together at night. MAM2x30 - sheep and lambs 30 minutes in the morning and 30 minutes in the afternoon together. Blood samples were collected and weighed every seven days. Significant increase in weight was observed with the age of the lambs in the three managements, but the treatments did not differ for weight gain. Differences were identified between the three treatments for the hematological variable CHGM and for the biochemical variables AST, glucose, urea and total protein. Correlating the age of the lambs with the biochemical variables, there was variation influenced by the age of the animals. The different management of breastfeeding did not influence the weight development of the lambs. Hematological and biochemical variations did not represent pathological changes. Some hematological and biochemical parameters may be influenced by the age development of lambs.(AU)

Characterization of immunological, biochemical and inflammatory response of clinical and subclinical endometritis in ewes in the subtropics.

Pluriparus Ossimi (n = 50) ewes were used to investigate the immune profile of the affected ewes to accurately diagnose clinical and subclinical endometritis and associations with biochemical variables. Ewes were slaughtered and animals were classified into control (no fertility problems), subclinical endometritis (SCE) and clinical endometritis (CE) groups based on pre-slaughter determinations of conception failure. Serum was collected from ewes to estimate concentrations of pro-inflammatory cytokines including interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8) as well as nitric oxide (NO) concentration. The results from immunological evaluations indicated there were greater (P < 0.001) serum concentrations of IL-6, IL-8, TNF-α and NO in ewes classified with SCE and CE as compared to ewes of the control group. Furthermore, values for concentrations of TNF-α were positively correlated with IL-6 and IL-8 concentrations in ewes of the SCE and CE groups. In ewes classified with CE and SCE there were greater (P < 0.01) concentrations of blood glucose, ALT, AST, urea and creatinine than in ewes of the control group. It is concluded that serum pro-inflammatory cytokines IL-6, IL-8, and TNF-α are diagnostic markers for CE and SCE in ewes and serve as a criterion for different inflammatory complications in ewes classified as having CE or SCE.

Greater concentrations of IGF-1 are associated with increasing pregnancy rate in melatonin implanted anestrous Barki ewes.

The present study was conducted to determine if increases in IGF-1 concentration, associated with treatment of ewes with melatonin, has beneficial effects on pregnancy rates when there is induction of estrus in anestrous ewes. A total of 120 multiparous lactating ewes were assigned to three groups (n = 10/group). Ewes of Group 1 were treated with a melatonin implant for 42 days followed by insertion of a controlled internal drug release (CIDR) device for 14 days with administration of equine chorionic gonadotropin (eCG) at day of CIDR removal. The ewes of Group 2 were treated with a CIDR and eCG at the same times as ewes of Group 1 and ewes of Group 3 were assigned to an be untreated control group. Melatonin implantation resulted in an increase in IGF-1 concentrations and lesser estradiol (E2) and triiodothyronine (T3) concentrations. Ewes of Groups 1 and 2 had the greatest progesterone (P4) concentration compared ewes of Group 3. The E2:P4 ratio was less in ewes of Group 1 compared Group 3. Melatonin implantation of ewes resulted in a greater pregnancy rate compared to treatment with the CIDR and eCG which, in turn, had a greater rate than ewes of the control group. In conclusion, melatonin implantation modulates the hormonal milieu including P4, E2, T3 and IGF-1 in seasonally anestrous ewes. Increased IGF-1concentrations, as a result of melatonin treatment, are associated with a greater percentage pregnancy rate when there is treatment of anestrous ewes to induce onset of estrus.

Synthesis and biological evaluation of 12 novel (-)-camphene-based 1,3,4-thiadiazoles against Mycobacterium tuberculosis.

Aim: To evaluate the activity, cytotoxicity and efflux pumps inhibition of a series of 12 novels (-)-camphene-based 1,3,4-thiadiazoles (TDZs) against Mycobacterium tuberculosis (Mtb). Materials & methods: The minimum inhibitory concentration (MIC), cytotoxicity for three cell lines, ethidium bromide accumulation and checkerboard methods were carried out. Results: Compounds (6a, 6b, 6c, 6g, 6h and 6j) showed significant anti-Mtb activity (MIC 3.9-7.8 µg/ml) and no antagonism with anti-TB drugs already used in the TB treatment. Selectivity index (SI) was also determined, with values reaching 42.9 for H37Rv strain and 97.1 for clinical isolate. Five compounds also showed bacterial efflux pumps inhibition and one showed modulator effect with three drugs. Conclusion: These six TDZs should be considered as new scaffolds to develop anti-TB drugs.

Avaliação hematológica e hepática de ovinos sob tratamento com torta de neem (Azadirachta indica) / Hematological and liver evaluation of sheep under treatment with neem pie (Azadirachta indica)

Devido aos altos índices de resistência dos parasitas aos diferentes princípios ativos comerciais, novas alternativas de controle vêm sendo estudadas, entre elas a fitoterapia. Essas medidas visam a busca de métodos auxiliares no controle das parasitoses, entretanto, muitos produtos estão disponíveis no mercado e não têm comprovação científica de sua eficácia ou de possíveis efeitos colaterais. O objetivo deste estudo foi avaliar os efeitos hematológicos e hepáticos após a administração de torta de Neem (Azadirachta indica) em ovinos. Foram testadas três dosagens da torta de Neem adicionada ao sal mineral (1, 2 e 4%), administradas por 126 dias para 32 ovinos da raça Lacaune, divididos em quatro grupos sendo três grupos para os diferentes tratamentos e um controle, o qual recebeu somente sal mineral. Amostras de sangue foram colhidas a cada 21 dias para realização do hemograma completo, dosagem de proteína plasmática total e fibrinogênio e avaliação da bioquímica clínica hepática. Nestas mesmas ocasiões, amostras de fezes foram coletadas para a quantificação de ovos por grama de fezes (OPG). Foram observadas diferenças estatísticas entre momentos e grupos para diversas variáveis, porém sem estarem relacionadas à administração de torta de Neem. Os resultados obtidos de hemograma completo, dosagem de proteína plasmática total, fibrinogênio e de bioquímica clínica hepática indicaram que a administração de torta de Neem nas concentrações de 1, 2 e 4%, não interfere nos valores hematológicos, nem sobre a integridade e função hepática de ovinos da raça Lacaune.

Due to high levels of parasite resistance to different commercial active ingredients, new control alternatives are being studied, including the phytotherapy. These measures aim to search for auxiliary methods in the control of parasitic diseases. However, there are many products available in the market and there are no scientific proof of its efficacy. The objective of this study was to evaluate the hematological and hepatic effects following administration of Neem pie (Azadirachta indica) in sheep. Three concentrations of Neem cake was added to the mineral salt (1, 2 and 4%) and administered during 126 days to 32 Lacaune breed sheeps, divided into four groups: three groups for different treatments and a and a control were tested, the ladder receiving only mineral salt. Blood samples were taken every 21 days to perform the complete blood count, serum total plasma protein and fibrinogen and liver biochemical evaluation. In those same times, fecal samples were collected for quantification of eggs per gram of feces (EPG). Statistical differences between times and groups for several variables were observed, but without being related to the administration of Neem pie. The results of complete blood count, measurement of total plasma protein, fibrinogen and hepatic clinical biochemistry indicated that administration of Neem pie at concentrations of 1, 2 and 4%, does not interfere in hematological values, or on the integrity and liver function Lacaune sheep breed.

Short communication: Characterizing ovine serum stress biomarkers during endotoxemia.

Breeding stress-resilient livestock is a potential strategy to help mitigate the negative effect of environmental and pathogenic stressors. The hypothalamic-pituitary-adrenal axis and immune system are activated during stress events and release mediators into the circulation that help restore physiological homeostasis. The purpose of this study was to assess a comprehensive set of circulatory mediators released in response to an acute immune stress challenge to identify candidate biomarkers that can be used for the selection of stress-resilient animals. Fifteen female lambs were stress challenged with an intravenous bolus of lipopolysaccharide (LPS; 400 ng/kg), and blood was collected from the jugular vein at 0, 2, 4, and 6 h after LPS challenge to identify and monitor candidate stress biomarkers; temperature was also recorded over time. Biomarker responses were evaluated with a repeated-measures model to compare time points with baseline values. As expected, all sheep had a monophasic febrile response to LPS challenge, and cortisol increased and returned to baseline by 6 h. The cytokines tumor necrosis factor-α, IL-6, IFN-γ (proinflammatory), and IL-10 (anti-inflammatory) increased, but only tumor necrosis factor-α returned to baseline during the monitoring period. The cytokines IL-1α, IL-1ß, IL-17α (proinflammatory), and IL-4 (anti-inflammatory) did not respond to LPS challenge. All chemokines (CCL2, CCL3, CCL4, CXCL10, and IL-8) responded to LPS challenge; however, only CCL2, CCL3, CCL4, and CXCL10 increased over time, and only CCL3, CCL4, and CXCL10 returned to baseline during the monitoring period. MicroRNA (miR-145, miR-233, and miR-1246) also increased and remained elevated during the study. In summary, the LPS challenge induced a strong stress response in Rideau-Dorset sheep that could be monitored with a distinct profile of circulatory biomarkers.

Dietary supplementation with ovine serum immunoglobulin modulates correlations between mucin, microbiota and immunity proteins in the growing rat.

The objective of this study was to evaluate the relationship among the number of bacteria, number of goblet cells, gut mucin gene expression, mucin protein and immunity protein levels of rats fed a diet containing freeze-dried ovine Ig (FD). Sprague Dawley male rats were used in a 21-days study and were fed a basal control diet (BD; no Ig) and a test diet containing freeze-dried ovine Ig (FD). Diets were isocaloric and contained the same amount of the first limiting amino acids, methionine plus cysteine. Pearson's correlation analysis was conducted on the data (stomach, ileum and colon) obtained from individual rats (n = 10) fed either casein-based diet (BD) or ovine serum Ig (FD) to evaluate the relationship between number of bacteria, number of goblet cells, gut mucin gene expression and gut mucin protein levels. Pearson's correlation analysis was then conducted with the data from the FD fed rats to evaluate the relationship among the above said variables. In the stomach content, a significant (p < .05) correlation was found between the Muc5Ac gene expression and mucosal mucin protein. In the ileum and colon, a significant (p < .05) correlation was observed among the mRNA levels of mucin (Muc2 and Muc4) genes. There was also evidence of a strong relationship (p < .05) between digesta mucin and mucosal mucin protein concentrations. A negative correlation of mucosal IgA protein concentration with total Lactobacillus (in ileum and colon) and total bacteria (in the ileum) was not evident with FD fed rats when compared to the results obtained using both BD and FD fed rats. In conclusion, this study suggests that feeding freeze-dried ovine Ig in growing rats results in a strong correlation between the number of bacteria, mucin and immunity proteins.

Effect of supplementation of pelleted hazel (Corylus avellana) leaves on blood antioxidant activity, cellular immune response, and heart beat parameters in sheep1.

Hazel leaves (Corylus avellana) fed to sheep resulted in decreased methane emissions without negatively affecting feed intake and were found to have antioxidant properties in vitro. The objective of this study was to evaluate effects of hazel leaves, rich in tannins, on blood antioxidant activity, cellular immune response, and heart beat parameters in sheep. Four experimental pellets were produced by mixing alfalfa and hazel leaves in different proportions, including alfalfa alone as a control, 30% and 60% of hazel leaves, the latter also with 3.8% polyethylene glycol (PEG). Six adult, nonpregnant, nonlactating female sheep (71 ± 5.7 kg of body weight) were allocated to 4 treatments in a 6 × 4 crossover design with four 18-d periods. The diet consisted of experimental pellets and ryegrass-dominated hay (ratio 80% to 20% in dry matter), resulting in hazel leaf proportions of approximately 0%, 25%, and 50% in the total diet. Blood samples were collected at the end of each period to determine plasma total phenol concentration and markers of oxidative status as well as peripheral blood mononuclear cells (PBMC) activation and proliferation response in vitro. Heart rate (HR) and HR variability parameters were measured for 2 consecutive days in each period, during different activities (i.e., eating pellets or hay, or lying). Treatments were compared with multiple comparisons and contrast analysis was used to test for linear and quadratic relations. Compared with control, feeding a high dosage of hazel leaves enhanced (P = 0.006) the plasma total antioxidant capacity, which linearly (P = 0.016) increased with increasing level of hazel leaves in the diet. The total phenol concentration and activities of the antioxidant enzymes superoxide dismutase, catalase, and glutathione reductase in the plasma were not different (P ≥ 0.23) among the treatments; however, the latter slightly increased linearly (P = 0.047) with increasing hazel leaves proportion. No differences were observed in the activation and proliferation of PBMC among treatments. The HR decreased linearly (P ≤ 0.009) during pellet eating and lying and the root mean square of successive differences of interbeat intervals (RMSSD) increased linearly (P = 0.037) when lying with increasing level of hazel leaves in the diet. In conclusion, our findings indicate that hazel leaves are a promising supplement to improve oxidative status with no effect on cellular immune response and cardiac stress level of sheep.

The effects of dietary nitrate on plasma glucose and insulin sensitivity in sheep.

Nitrate (NO3 ¯ ) is an effective non-protein nitrogen source for gut microbes and reduces enteric methane (CH4 ) production in ruminants. Nitrate is reduced to ammonia by rumen bacteria with nitrite (NO2 ¯ ) produced as an intermediate. The absorption of NO2 ¯ can cause methaemoglobinaemia in ruminants. Metabolism of NO3 ¯ and NO2 ¯ in blood and animal tissues forms nitric oxide (NO) which has profound physiological effects in ruminants and has been shown to increase glucose uptake and insulin secretion in rodents and humans. We hypothesized that absorption of small quantities of NO2 ¯ resulting from a low-risk dose of dietary NO3 ¯ will increase insulin sensitivity (SI ) and glucose uptake in sheep. We evaluated the effect of feeding sheep with a diet supplemented with 18 g NO3 ¯ /kg DM or urea (Ur) isonitrogenously to NO3 ¯ , on insulin and glucose dynamics. A glucose tolerance test using an intravenous bolus of 1 ml/kg LW of 24% (w/v) glucose was conducted in twenty sheep, with 10 sheep receiving 1.8% supplementary NO3 ¯ and 10 receiving supplementary urea isonitrogenously to NO3 ¯ . The MINMOD model used plasma glucose and insulin concentrations to estimate basal plasma insulin (Ib ) and basal glucose concentration (Gb ), insulin sensitivity (SI ), glucose effectiveness (SG ), acute insulin response (AIRg) and disposition index (DI). Nitrate supplementation had no effect on Ib (p > .05). The decrease in blood glucose occurred at the same rate in both dietary treatments (SG ; p = .60), and there was no effect of NO3 ¯ on either Gb , SI , AIRg or DI. This experiment found that the insulin dynamics assessed using the MINMOD model were not affected by NO3 ¯ administered to fasted sheep at a low dose of 1.8% NO3 ¯ in the diet.

Comparação dos parâmetros bioquímicos entre gestação única e gemelar de ovelhas da raça Dorper durante gestação, parto e pós-parto / Comparison of biochemical parameters between single and twin pregnancies of Dorper breed ewes during gestation, lambing and postpartum

Nas ovelhas, a demanda de nutrientes aumenta durante a gestação, especialmente nas últimas seis semanas, quando ocorre maior desenvolvimento do feto. Os objetivos deste trabalho foram estudar o perfil bioquímico de ovelhas durante a gestação e no periparto imediato, comparando-se gestação única com gemelar. Foram utilizadas 60 ovelhas, Dorper, divididas em dois grupos experimentais com base no diagnóstico de gestação ultrassonográfico: grupo 1: 30 ovelhas com feto único; grupo 2: 30 ovelhas com gestação gemelar. Em nove momentos de coleta, foram mensurados no soro sanguíneo: ureia, creatinina, proteína total (PT), albumina, globulina, aspartato aminotransferase (AST), gamaglutamiltransferase, fosfatase alcalina, colesterol e triglicérides. O peso vivo das ovelhas do G2 foi maior que do G1 em todos os momentos, inclusive quando estavam vazias. Houve diferença entre as ovelhas do G1 e do G2 nos valores de PT, albumina, AST e triglicérides. A partir de 120 dias de gestação, o perfil proteico se modificou, com menores níveis de ureia, PT, albumina e globulina, mostrando que a demanda metabólica se intensificou no último mês da gestação. Os níveis de colesterol e triglicérides diminuíram de 140 dias de gestação para o parto. Este trabalho mostrou que a gestação proporcionou mudanças significativas no metabolismo da ovelha, principalmente no terço final da gestação, quando há maior necessidade de acompanhamento para evitar a incidência de doenças metabólicas.(AU)

In sheep, nutrient demand increases during gestation, particularly in the last six weeks, when the fetuses develop. The objectives of this study were to evaluate the biochemical profile of ewes during pregnancy and in the immediate peripartum comparing single with twin pregnancies. Sixty ewes of Dorper breed were divided into two groups by pregnancy ultrasonographic diagnosis: 1 - 30 ewes with single fetus gestation; 2 - 30 ewes with twin pregnancy. In nine collection moments the following parameters were measured: urea, creatinine, total protein (TP), albumin, globulin, aspartate aminotransferase, gamma glutamyltransferase, alkaline phosphatase, cholesterol and triglycerides. The weight of G2 sheep was higher than G1 since they were empty. There were differences in G1 and G2 sheep metabolism in TP, Albumin, AST and Triglycerides. From 120 days of pregnancy, the protein profile changed, with lower levels of urea, TP, albumin and globulin, showing that the metabolic demand intensified in the last month of pregnancy. The levels of cholesterol and triglycerides decreased from 140 days of pregnancy to lambing day. This study showed that gestation provided significant changes in the ewe's metabolism, especially in the final third of gestation, when there is a greater need to follow the animals to avoid the incidence of metabolic diseases.(AU)

Transferrina como marcador de maturação pulmonar em cordeiros nascidos a termo ou prematuros / Transferrin as a marker of pulmonary maturation in lambs born at term or premature

O objetivo do estudo foi procurar proteínas de fase aguda que possam indicar sinais de maturação no neonato prematuro, por meio da quantificação sérica delas. Identificou-se a imunoglobulina A, a ceruloplasmina, a haptoglobina, a glicoproteína ácida, a transferrina, a albumina e as imunoglobulinas G de cadeias leve e pesada, pela comparação do perfil dos proteinogramas de cordeiros nascidos a termo com os prematuros submetidos a diferentes protocolos terapêuticos, a fim de estimular a atividade respiratória. Constituíram-se seis grupos: PN (n= 9): nascidos de parto normal; CN (n= 7): nascidos de cesariana em tempo normal de gestação; CP (n= 6): nascidos de cesariana prematura sem nenhum tipo de tratamento; DEX (n= 9): prematuros cujas mães receberam dexametasona pré-parto; SURF (n= 6): prematuros tratados com surfactante; e DEXSURF (n= 6): prematuros tratados com surfactante cujas mães receberam dexametasona pré-parto. As avaliações foram realizadas nos momentos imediatamente após o nascimento (M0), após 24 (M24) e após 48 horas (M48). As amostras foram processadas por meio de eletroforese em gel de poliacrilamida contendo dodecil sulfato de sódio (SDS-PAGE). A albumina, as imunoglobulinas e a proteína total dos cordeiros tiveram elevação após a ingestão de colostro. Maiores valores séricos de transferrina são referentes a maior período gestacional, podendo essa proteína ser utilizada como marcador de maturação neonatal.(AU)

The aim of this study was to search for acute phase proteins that could indicate signs of maturation in the premature neonate by quantifying them in serum. Immunoglobulin A, ceruloplasmin, haptoglobin, acid glycoprotein, tranferrin, albumin, light and heavy chain immunoglobulin G were quantified, comparing the profile of proteinograms from term to preterm lambs submitted to different protocols that stimulate respiratory activity. Six groups were used: PN (n= 9): born from normal birth; CN (n= 7): born from caesarean section at normal time of gestation; CP (n= 6): born from premature cesarean without any type of treatment; DEX (n= 9) preterm whose mothers received prepartum dexamethasone; SURF (n= 6) preterm treated with surfactant; DEXSURF (n= 6): preterm treated with surfactant whose mothers received prepartum dexamethasone. The evaluations were performed immediately after birth (M 0), after 24 and 48 hours (M 24 and M 48). Samples were processed with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Albumin, immunoglobulins, and serum total protein of the lambs were elevated, after colostrum ingestion. Higher serum transferrin values refer to a longer gestational period, and this protein may be used as a marker of neonatal maturation.(AU)

Comparação de métodos para avaliação de beta-hidroxibutirato em ovelhas / Comparison of methods for evaluation of beta-hydroxybutyrate in ewes

Atualmente o uso de sensores portáteis para mensuração de corpos cetônicos está padronizado e difundido na rotina clínica, contudo estudos em ovinos são escassos. Assim, a presente pesquisa objetivou avaliar a acurácia dos sensores portáteis de uso humano e de uso veterinário para a determinação de beta-hidroxibutirato (BHB) em ovelhas no final da gestação e no pós-parto recente. Foram utilizadas 37 amostras de sangue provenientes de nove ovelhas mestiças Corriedale. A determinação bioquímica de BHB no soro, considerada como o padrão-ouro, foi realizada utilizando-se metodologia enzimática colorimétrica. A média obtida na bioquímica sérica foi de 0,497mmol/L; no sensor de uso humano, a média foi igual a 0,537mmol/L, enquanto no sensor de uso veterinário foi de 0,751mmol/L. Foi verificada alta correlação entre o dosímetro de uso humano e o padrão-ouro (r=0,93, P<0,001). A média do aparelho de uso veterinário diferiu das demais (51%; P<0,05), superestimando os resultados em ovelhas. As medições obtidas no aparelho veterinário também apresentaram menor precisão e veracidade. Concluiu-se que o sensor portátil de uso humano é mais acurado e mais preciso no diagnóstico precoce de toxemia da gestação em ovelhas.

Currently the use of portable sensors for measuring ketone bodies is standardized and diffused in the clinical routine, however, studies in sheep are scarce. Therefore, the present study aimed to evaluate the accuracy of the human portable sensor and the veterinary portable sensor for the determination of beta-hydroxybutyrate (BHB) in sheep at the end of gestation and postpartum. We used 37 samples of blood from nine crossbred Corriedale sheep. Biochemical determination of serum BHB, considered gold standard, was performed using colorimetric enzymatic methodology. The mean serum biochemistry was 0.497mmol/L, in the human sensor the mean was 0.537mmol/L, while in the veterinary sensor it was 0.751mmol/L. A high correlation was verified between the dosimeter for human use and the gold standard (r= 0.93, P< 0.001). The mean of the veterinary apparatus differed from the others, being 51% (P< 0,05), higher than the standard, that is, it was less accurate and had lower veracity, overestimating the results in sheep. It was concluded that the portable sensor for human use is more accurate and accurate in the early diagnosis of toxemia of pregnancy in sheep.