Plasma metabolite changes in anestrous dairy cows with negative energy balance identified using 1H NMR technology / [Alterações do metabólito plasmático em vacas leiteiras com balanço energético negativo identificado usando a tecnologia 1H NMR]

The objective of the present study was to investigate the different plasma metabolites between anestrus and estrus postpartum dairy cows and to provide a theoretical basis for prevention of anestrus in dairy farm cows. In the experiment, one hundred and sixty-seven Holstein dairy cows were selected with similar age and parity. According to the concentration of ß-hydroxybutyric acid, non-esterified fatty acids and glucose in plasma during 14 to 21 days in milk, all dairy cows were determined as having a status of energy balance. According to the results of clinical symptom, rectal and B ultrasound examination at 60 to 90 days postpartum, these cows were divided into twenty estrus and twenty-four anestrus group, other dairy cows were removed. 1H nuclear magnetic resonance technology was utilized to detect the plasma metabolites changes and screen different plasma metabolites between anestrus and estrus cows. Ten different metabolites including alanine, glutamic acid, asparagine, creatine, choline, phosphocholine, glycerophosphocholine, low-density lipoprotein, and very-low-density lipoprotein were significantly decreased in anestrous cows compared with estrous cows. Metabolic pathway analyses indicated that differential metabolites were primarily involved in amino acid and glycerophospholipid metabolism. These metabolites and their enrichment pathways indicate that reduced steroid hormone synthesis precursors result in lower levels of estradiol and progesterone and cause anestrus in negative energy balance. These data provide a better understanding of the changes that may affect estrus of postpartum dairy cows at NEB status and lay the ground for further research.(AU)

O objetivo do presente estudo foi investigar os diferentes metabolitos do plasma entre o cio e o cio pós-parto de vacas leiteiras e fornecer uma base teórica para a prevenção do cio de vacas em fazendas de leite. No experimento, foram selecionadas 127 vacas leiteiras Holstein com idade e paridade similares. De acordo com a concentração de ß- ácido hidroxibutírico, ácidos graxos não esterificados e glicose no plasma entre 14 e 21 dias no leite, todas as vacas leiteiras foram determinadas em estado de equilíbrio energético. De acordo com os resultados dos sintomas clínicos, do exame de ultra-som retal e B aos 60 a 90 dias pós-parto, estas vacas foram divididas em vinte cios e vinte e quatro grupos de cio, outras vacas leiteiras foram removidas. A tecnologia de ressonância magnética nuclear 1H foi utilizada para detectar as alterações dos metabólitos plasmáticos e para triar diferentes metabólitos plasmáticos entre as vacas do cio e do cio. Dez diferentes metabólitos incluindo alanina, ácido glutâmico, asparagina, creatina, colina, fosfocholina, glicerofosfocolina, lipoproteína de baixa densidade e lipoproteína de muito baixa densidade foram significativamente diminuídos nas vacas antróficas em comparação com as vacas estro. As análises da via metabólica indicaram que os metabólitos diferenciais estavam principalmente envolvidos no metabolismo de aminoácidos e glicerofosfolipídios. Estes metabólitos e suas vias de enriquecimento indicam que a redução dos precursores da síntese de hormônios esteróides resulta em níveis mais baixos de estradiol e progesterona e causa anestros no balanço energético negativo. Estes dados fornecem uma melhor compreensão das mudanças que podem afetar o cio das vacas leiteiras pós-parto no estado de NEB e preparam o terreno para mais pesquisas.(AU)

Effects of blood pathological changes before TAI on pregnancy of dairy cows with anestrus and estrus / [Efeitos de mudanças patológicas de plasma antes de (TAI) na gestação de vacas em anestro e estro]

The objective of this study was to investigate the influence of plasma pathological changes before timed artificial insemination (TAI) on pregnancy of cows. The contents of estrogen (E2), progesterone (P4), glucose (Glu), selenium (Se), brain-derived neurotrophic factor (BDNF), and histamine (HIS) in plasma of 48 Holstein cows were measured before TAI. According to the estrus detection, the cows were divided into estrus (E) and anestrus (A) groups. After pregnancy testing at 28 d after TAI, two groups of E and A were divided into positive pregnancy of E group (EP+), negative pregnancy of E group (EP-), positive pregnancy of A group (AP+), and negative pregnancy of A group (AP-). The contents of E2, P4, Glu, Se, BDNF and hIS significantly differed among the four groups (P<0.01). The ROC analysis was used to determine the risk of negative pregnancy test (-) after TAI was increased when plasma E2 was less than 46.45 pmol/L in cows before TAI. The changes in E2, P4,hIS, Glu, and BDNF in the blood of natural estrus and natural anestrus cows affected the pregnancy after TAI. the level of E2 in plasma may be used to assess the risk of negative pregnancy after TAI.(AU)

O objetivo do presente estudo foi investigar a influência de mudanças patológicas de plasma antes de inseminação artificial (TAI) na gestação de vacas. O conteúdo de estrogênio (E2), progesterona (P4), glucose (Glu), selênio (Se), fator neurotrófico derivado do cérebro (BDNF), e histamina (HIS) no plasma de 48 vacas Holstein foi medido antes de TAI. De acordo com a detecção de estro, as vacas foram divididas em dois grupos: estro (E) e anestro (A). Após teste de gestação 28 d após TAI, dois grupos de E e A foram formados em gestação positiva do grupo E (EP+), gestação negativa do grupo E (EP-), gestação positiva do grupo A (AP+), e gestação negativa do grupo A (AP-). Os valores de E2, P4, Glu, Se, BDNF e hIS foram significativamente diferentes entre os quatro grupos (P<0,01). A análise ROC foi utilizada para determinar o risco de teste de gestação negativo (-) após aumento de TAI quando plasma E2 estava abaixo de 46,45 pmol/L em vacas antes de TAI. Alterações em E2, P4,hIS, Glu e BDNF no sangue de estro natural e anestro natural em vacas afetou a gestação após TAI. O nível de E2 no plasma pode ser usado para avaliar o risco de gestação negativa após TAI.(AU)

Profile of plasmatic progesterone on pregnancy, and the postpartum estrus of Dasyprocta prymnolopha (Rodentia: Dasyproctidae) / Perfil de la progesterona plasmática durante durante la preñez y celo posparto del Dasyprocta prymnolopha (Rodentia: Dasyproctidae)

Abstract:The agouti (Dasyprocta sp.) is a hystricomorph rodent found in some regions of the Americas. It is an important cynegetic species, which indicates that the overhunting is a threat to their conservation. Very little is known about this wildlife in relation to what already has been studied in domestic animals. Thus, the knowledge on reproduction of wildlife becomes necessary and essential for the management and conservation of these natural resources. Specifically, studies regarding hormonal monitoring are important as a basic tool for research in modern reproductive biotechnology, and currently, there is no information on the progesterone changes during pregnancy of Dasyprocta sp., compared to other hystricomorphs. The aim of this study was to describe the profile of plasmatic progesterone during pregnancy, and report the restart of ovarian cycle in agouti after parturition. For this purpose, 18 black-rumped agoutis (Dasyprocta prymnolopha) born in captivity were used, and one or more consecutive pregnancies were observed. Copulation was confirmed by the presence of spermatozoa observed in the colpocytological examination. Blood samples were collected two times per week, and concentrations of progesterone, assessed in ten agoutis, were determined by radioimmunoassay. The onset of ovarian activity in six females was observed daily by colpocytological examination, starting on the seventh day postpartum. The gestational period observed in this study was 104.04 days (SD = 1.31) (101-106 days) (Number of cases, N = 26), and the interval between births was 126.03 days (SD = 18.40) (109-184 days). The plasmatic profile of progesterone during pregnancy showed a progressive increase from the 1st to the 5th week. The higher progesterone levels over this period (6.88 ng / mL, SD = 3.01) were detected in the 5th week. This value was similar (One-Way ANOVA, p > 0.05) to that observed in the 4th and 6th weeks, but was statistically different (One-Way ANOVA, p < 0.05) when compared to the other weeks. After the 6th week there was a progressive decrease in plasmatic progesterone levels. The animals showed a postpartum estrus of 12.04 days (SD= 4.29) (7-24 days) (N= 23). It was observed that 80.95 % (N= 19) of copulations during this period were fertile. This work contributed to understanding the dynamic changes in the progesterone levels during the pregnancy in agouti. Nevertheless, more studies are needed for a better appreciation of other endocrine and biological changes, in the mother and feto-placental unit of the agouti. Rev. Biol. Trop. 64 (4): 1519-1526. Epub 2016 December 01.

Resumen:El agouti (Dasyprocta sp.) es un roedor histricomorfo encontrado en algunas regiones de América. Es una especie cinegética importante, lo que indica que la caza excesiva es una amenaza para su conservación. Muy poco es lo que se conoce acerca de la biología de los animales silvestres, en relación al conocimiento acumulado sobre los domésticos. Por lo tanto, el estudio sobre la reproducción de los animales silvestres se hace necesario e imprescindible para el manejo y conservación de la especie como recurso natural. En concreto, los estudios relativos a la monitorización hormonal son una herramienta básica para la investigación en biotecnología reproductiva moderna. No hay información sobre los cambios de progesterona durante la preñez de Dasyprocta sp. en comparación con otros histricomorfos. El objetivo de este estudio fue describir el perfil de la progesterona plasmática durante la preñez, y reportar el reinicio del ciclo ovárico después del parto. Para ello, se utilizaron 18 agoutis de rabo negro (Dasyprocta prymnolopha) nacidos en cautiverio. Fueron estudiadas una o más preñeces consecutivas. La copulación fue confirmada por la presencia de espermatozoides observados en examen colpocitológico. Las muestras de sangre se recogieron dos veces por semana, y las concentraciones de progesterona se determinaron en 10 individuos mediante radioinmunoensayo. Se observó el inicio de la actividad ovárica diaria por examen colpocitológico, comenzando en el día séptimo posparto. El período gestacional observado en este estudio fue de 104.04 días (SD= 1.31, rango entre 101-106 días, N= 26), y el intervalo entre los nacimientos fue 126.03 días (SD= 18.40, rango entre 109-184 días). El perfil plasmático de la progesterona durante el embarazo mostró un aumento progresivo de la 1ª a la 5ª semana. Los mayores niveles de progesterona durante este período (6.88 ng / mL, SD= 3.01) se detectaron en la quinta semana. Este valor fue similar (ANOVA de um factor, p > 0.05) al observado en las semanas cuatro y seis, pero fue estadísticamente diferente (ANOVA de um factor, p<0.05) en comparación con las otras semanas. Después de la sexta semana hubo una disminución progresiva en los niveles plasmáticos de progesterona. Los animales mostraron un estro posparto de 12.04 días (SD= 4.29, rango entre 7-24 días, N= 23). Se observó que el 80.95 % (N= 19) de cópulas durante este período fueron fértiles. Este trabajo contribuyó a la comprensión de los cambios dinámicos en los niveles de progesterona durante la preñez del agouti. Sin embargo, se necesitan más estudios para una mejor comprensión de otros cambios biológicos y endocrinos, y en la unidad materna y feto-placentaria del agouti.

Efeito do nível de uréia na dieta sobre o desempenho, a qualidade e o estádio de desenvolvimento embrionário em cabras Alpinas / Effect of the dietary urea level on performance, quality and embryonic development stage on Alpine goats

Utilizaram-se 22 cabras da raça Alpina, distribuídas aleatoriamente em quatro tratamentos (T): as cabras do T1 (n=5) formaram o grupo-controle; as do T2 (n=7) receberam 0,73 por cento de uréia na matéria seca da dieta; as do T3 (n=4) receberam 1,46 por cento de uréia; e as do T4 (n=6), 2,24 por cento de uréia. As cabras foram superovuladas e os embriões, coletados entre sete e oito dias após a primeira monta, foram avaliados quanto à qualidade e ao estádio de desenvolvimento. Amostras de sangue para dosagem dos teores de uréia e glicose foram coletadas nos dias do estro e da coleta de embriões. Houve efeito linear crescente do nível de uréia nas dietas sobre o consumo de MS (kg/dia) e de proteína bruta (kg/dia). O peso das cabras não diferiu (P>0,05) entre os tratamentos nem entre as semanas experimentais. Dezoito cabras (81,8 por cento) manifestaram estro após a sincronização. A duração do estro e o intervalo da remoção da esponja ao início do estro não foram influenciados (P>0,05) pelos tratamentos. Quatorze cabras (77,8 por cento) responderam à superovulação. O número de estruturas e de embriões coletados não diferiu (P>0,05) entre os tratamentos. O número (Y= 10,90 - 11,64NS U + 4,93§U²; R² = 0,67; P<0,10) e a percentagem (Y= 94,08 - 39,59NS U + 18,16ªU²; R² = 0,94; P<0,07) de embriões viáveis, e o número (Y= 10,83- 12,18NS U + 5,02mU²; R² = 0,78; P<0,08) e a percentagem (Y= 94,83- 52,31NS U + 21,56*U²; R² = 0,90; P<0,05) de embriões excelentes e bons apresentaram comportamento quadrático em função do nível de uréia nas dietas. Os teores de uréia e glicose no plasma não foram influenciados (P>0,05) pelos tratamentos. A uréia pode ser fornecida no nível de 2,24 por cento na MS da dieta de cabras não lactantes

Twenty-two Alpine goats were allocated at random into four treatments: 0.0 percent (T1 - control, n=5); 0.73 percent (T2, n=7); 1.46 percent (T3, n=4) or 2.24 percent (T4, n=6) of urea in the dry matter (DM) of the diet. Embryos collected from 7 to 8 days after mating of superovulated goat were evaluated by quality and development stage. Blood samples for urea and glucose analyses were collected at estrus and at embryos collection day. The DM (kg/day) and crude protein (kg/day) intake increased linearly in function of dietary urea level. Goat body weights were not affected by treatments out experimental weeks (P>0.05). Eighteen goats (81.8 percent) came in estrus after the synchronization. The estrus length and the interval from sponge removal to the beginning of estrus were not affected (P>0.05) by treatments. Fourteen goats (77.8 percent) were responsive to superovulation protocol. The levels of urea (treatments) did not affect structures and embryo numbers (P>0.05). Number (Y= 10.90 - 11.64NS U + 4.93§U²; R²= 0.67; P<0.10) and percentage (Y= 94.08 - 39.59NS U + 18.16ªU²; R²= 0.94; P<0.07) of viable embryos, and number (Y= 10.83- 12.18NS U + 5.02mU²; R²= 0.78; P<0.08) and the percentage (Y= 94.83- 52.31NS U + 21.56*U²; R²= 0.90; P<0.05) of excellent and good embryos were quadraticaly effected by urea dietary level. The treatments did not affect plasma urea and glucose levels (P>0.05). Diets for no nursing goats can be supplied by urea at 2.24 percent of DM

Isolation, identification and characterization of secretory proteins of IVMFC embryos and blood circulation of estrus and early pregnant goat.

The aim of the present study was to isolate, identify and characterize the secretory proteins of IVM oocytes and IVMFC embryos to evaluate its immunogenecity. and identify of such proteins if any, in blood circulation of estrus and early pregnant goats. Oocytes were matured in TCM-199 with 1 microg/ml, estradiol-17beta; 0.5 microg/ml, FSH; 100 IU/ml, LH and 10% FCS on granulosa cell monolayer. After 18 hr of maturation, oocytes were further cultured in maturation medium containing 3 mg/ml polyvinyl alcohol (PVA) without serum and BSA for 12 hr and medium was collected. The IVF embryos of 4-8 cell stage were cultured in medium containing PVA without serum and BSA. Embryo culture medium was collected after 24 hr of culture and was pooled. The proteins were analyzed on SDS-PAGE (12.5%). Four secretory proteins of oocytes with approximately molecular weight of 45, 55, 65 and 95 kDa and three secretory proteins of embryos 45, 55 and 65 kDa were obtained on SDS-PAGE in silver staining. The protein profile of midluteal, estrus and early pregnant goat serum was similar and no variation was observed among the proteins on SDS-PAGE. Two secretory proteins of 55 and 65 kDa of both IVM oocytes and IVMFC embryos were observed on Western analysis. None of such proteins was observed in midluteal, estrus and early pregnant goat serum on western blotting. It can be concluded that IVM oocytes and IVMFC embryos secrete proteins in medium and two of them can develop antibody. The proteins secreted from embryos till morula stage was similar to that of oocytes. None of these oocyte/embryo released proteins were observed in blood circulation of estrus and early pregnant goats.

Peripheral plasma FSH concentration in relation to expression of estrus in Sahiwal cattle (Bos indicus).

An experiment was conducted to investigate the changes in peripheral plasma FSH concentrations in relation to expression of estrus in Sahiwal cows. Out of total five estrus, three were accompanied by overt signs whereas rest two were silent estrus. In cows with overt estrus, plasma FSH concentrations during periestrus, early luteal, midluteal and late luteal phase were 1.65+/-0.34, 1.67+/-0.21, 1.58+/-0.18 and 1.69+/-0.31 ng/ml, respectively and the corresponding values in cows with silent estrus being 0.68+/-0.39, 0.50+/-0.12, 0.75+/-0.13 and 0.46+/-0.12 ng/ml, respectively. The overall plasma FSH levels in cows that exhibited overt estrus was 1.63+/-0.82 ng/ml as against 0.64+/-0.55 ng/ml in silent estrus. It was concluded that FSH levels were higher (P<0.01) in cows that exhibited overt estrus compared to silent estrus.

Peripheral plasma progesterone concentration in relation to estrus expression in Sahiwal cows.

The present study investigated the changes in peripheral plasma progesterone levels in relation to expression of estrus in Sahiwal cows. Out of a total of five estrus, three were accompanied by overt signs whereas the remaining two were silent estrus. In cows with overt estrus, plasma progesterone concentrations during periestrus, early luteal, midluteal and late luteal phase were 0.40 +/- 0.02, 0.74 +/- 0.10, 1.94 +/- 0.22 and 0.63 +/- 0.16 ng/ml, respectively and the corresponding values in cows with silent estrus being 0.47 +/- 0.03, 0.94 +/- 0.08, 1.39 +/- 0.13 and 0.95 +/- 0.19 ng/ml, respectively. The overall plasma progesterone levels in cows that exhibited overt estrus was 1.23 +/- 0.99 ng/ml as against 1.08 +/- 0.09 ng/ml in silent estrus. It was concluded that progesterone levels were lower (P > 0.05) in cows that exhibited silent estrus compared to overt estrus.

Apoptosis in endometrium of mouse during estrous cycle.

The present study was carried out to evaluate apoptosis in endometrium and to correlate these changes with the circulating levels of estradiol and progesterone in the mouse. Apoptosis was observed in various compartments of mouse uterus i.e. stroma, glandular epithelium and luminal epithelium depending on the stage of cycle. Stromal cell apoptosis was observed during various stages of cyclicity except on estrus day. Luminal epithelial cells showed apoptotic changes during all stages of cyclicity except on diestrus day. During metestrus, apoptosis was observed in glandular and luminal epithelia as well as stromal cells. Steroid antagonists such as tamoxifen and onapristone altered the apoptotic changes in the uterus. The results suggest that epithelial cell apoptosis is regulated by estrogen while stromal cell apoptosis is under the control of progesterone.

Natural estrous cycle in normal and diabetic bitches. II) serum nonesterified fatty acids and serum free glycerol levels during glucose and insulin tests

Actions and interactions of spontaneous diabetes mellitus (DM) and natural estrous cycles (sex seasons) on the regulation of serum monesterified fatty acids (NEFAs) and free glycerol (FG) levels in bitches in the fasting condition and during i.v. glucose (IVGTT) and insulin (ITT) tolerance tests, were studied. DM increased serum NEFAs concentration both in the basal condition and during IVGTT; it provoked a fall response to glucose load which is absent in normal controls. Estrous cycles did not modify these observations. Serum NEFAs levels during ITT were unresponsive in normal and diabetic bitches at every sex stage; flat, overlapped serum NEFAs profiles were then observed except for the diabetic group at A, which showed an early abrupt fall response of this variable from its high base line. DM increased also serum FG concentration in the fasting condition and during IVGTT. In the normal controls, serum FG base line was not affected by sex status; similary shaped, increasing, overlapped curves during the test were observed. In the diabetic bitches "in season" (either phase), serum FG basal value was hardly above in respect to anestrous, but during IVGTT their flat profiles coincided. DM increased serum FG concentration in the basal condition and during ITT, and modified the profiles of this variable. In normal dogs in the basal condition, serum FG concentration remained unaffected by sex status; this variable hard, transiently increased during ITT, which was not influenced by "sex seasons"; therefore, similarly shaped, overlapped serum FG profiles were then observed. In the normal and diabetic bitches, serum FG base line was not changed by "sex seasons". During ITT, serum FG mean profile in the diabetic bitches at EP was modestly above that observed in those at LP; differences for any other comparisions in normals or diabetic bitches were nonsignificant. As reported by us elsewhere, impaired glucose metabolism and absolute insulin dificiency induced ketose-prone, acidotic, insulin-dependent diabetic chryses in certain normal and diabetic beaches "in season" studied here. The unability of these animals for hydrolizingglyceride-glycerol via lipoproteinlipase (IVGTT) or via hormone sensitive fractions of lipase (ITT) and the abolished serum NEFAs suppressibility during modest hiperinsulinemia (ITT) appear to contribute to the production of such chryses...

Natural estrous cycle in normal and diabetic bitches in relation to glucose and insulin tests

The influence of spontaneous "sex seasons" on blood sugar (BS) and serum insulin levels was studied in bitches with natural diabetes mellitus (DM) and normal controls, in the basal condition and during glucose and insulin tests, was studied. DM increased basal BS, reduced glucose tolerance, distribution space (DS) and clearance from blood, and induced resistance to insulin hypoglycemic action. In normals, occurrence of "seasons", inconsistently modified basal BS, increased glucose tolerance and DS; during estrogenic phase (EP), these variables were above those during luteal phase (LP). In diabetics at LP, BS found in fasting condition and during glucose test were higher than in diabetic biches at EP (respective values at anestrous (A) in between) and glucose DS was smaller. Rate of glucose clearance from blood remained unaffected by "seasons" in both dog groups. Basal serum IRI was not modified by DM or "seasons". In normals, serum IRI response to glucose load was nonsignificant during A and increased during the "seasons"; either insulin DS or the rate of insulin clearance from blood stream remained unchanged under the circumstances the increase being mediated by insulin secretion. During EP, the increase was particulary intense and mean insulinogenic index (MII) rose. During LP, MII returned to A value, whereby diabetic states might be manifest. Serum IRI profiles during insulin test were not modified by "seasons" in normal bitches; such response in diabetic bitches was intense during A, then decreased (EP) or was later abolished (LP). Either in normal or diabetic bitches, the sensitivity to exogenous insulin hypoglycemic action remained unchanged in spite of "seasons". In diabetic bitches at A, serum IRI after glucose challenge peaked higher than in respective normal controls (insulin clearance and insulin DS were similar): They exhibited relative insulin shortage and resistance to insulin hypolgycemic action partly compensated by prometed insulin secretion. Along with "season", abolished serum IRI response to glucose load in diabetics was observed. During EP, extrapancreatic factors regulating serum IRI concentration and MII did not change in respect to A, whereby abolishment appears mediated by depressed insulin secretion. During LP, insulin anatogonism in conjunction with 1) absolute insulin deficiency and 2) intense decrease in MII appears as a powerful factor exposing diabetic bitches to a severe or fatal derangement in diabetic disease.

Changes in cortisol levels during estrous cycle and peripartum period of native Egyptian cattle

Cortisol levels were determined [RIA] in the plasma of five mature cyclic Egyptian heifer cows during green and dry seasons of the year as well as in the peripartum period of five cows till the first postpartum estrus. Cortisol level increased [P <0.01] at the day of estrus with no obvious changes between the green and dry seasons. The slope of cortisol increased [P <0.01] with the approach of calving and insignificantly fluctuated during the postpartum period