Biochemical profile and oocyte quality of primiparous Bos indicus cows submitted to a timed artificial insemination protocol.

The objective of this study was to verify the causes of the lower response of primiparous Bos indicus cows to the ovulation synchronization protocol. Two experiments were performed to evaluate the biochemical profile, oocyte and follicular cell quality (Experiment 1) and pregnancy (Experiment 2). In Experiment 1, suckled primiparous (n = 24) and multiparous cows (n = 24) were submitted to ovum pick up (OPU). On Day 0 (D0), all cows received 2 mg of estradiol benzoate (EB) and an intravaginal progesterone insert (P4). Five days (D5) after the first hormonal administration (EB + P4), all follicles larger than 3 mm were counted on each ovary, and ovum pick-up (OPU) was performed. On day 12 (D12), the intravaginal progesterone insert was removed, and measurement and aspiration of the largest follicle were performed. Blood samples were collected on D5 and D12 to evaluate the concentrations of glucose, cholesterol, NEFA, IGF-1 and insulin. In Experiment 2, suckled primiparous (n = 50) and multiparous (n = 50) cows were subjected to an ovulation synchronization protocol based on E2/P4 (D0: 2 mg EB plus P4 intravaginal insert; D8: 500 µg of cloprostenol, 1 mg cypionate estradiol and 300UI of eCG; D10: artificial insemination). In addition, blood samples were collected on D10 for evaluation of the same hormones and metabolites described in Experiment 1. In all studies, calves remained with the cows during the experimental period. In experiment 1, the number of oocytes grade 1 (P = 0.83), grade 2 (P = 0.23) and grade 3 (P = 0.51), total number of retrieved oocytes (P = 0.14), oocyte quality index (P = 0.93) and total viable oocytes (P = 0.38) did not differ between primiparous and multiparous cows. The number of follicles at the time of follicular aspiration (20.7 ± 1.5 vs. 18.0 ± 1.9; P = 0.05) and the diameter of the largest follicle on D12 (13.5 ± 0.6 vs. 11.4 ± 0.6; P = 0.04) were greater in multiparous cows, and the number of degenerated oocytes was greater in primiparous cows (1.9 ± 0.7 vs. 1.2 ± 0.3; P = 0.05). On D5, the concentrations of IGF-1 (P = 0.47), insulin (P = 0.08), total cholesterol (P = 0.47), NEFA (P = 0.77) and glucose (P = 0.55) in the blood and IGF-1 (P = 0.97) and insulin (P = 0.11) in the follicular fluid did not differ between parity groups. On D12, there was no difference in the concentrations of IGF-1 (P = 0.26), total cholesterol (P = 0.32), NEFAs (P = 0.31) and glucose (P = 0.93) in the blood between primiparous and multiparous cows, however, the serum insulin concentration (P = 0.04) was higher in primiparous cows. There was no correlation between serum and follicular fluid insulin concentrations (r = 0.17; P = 0.31), however, there was a low correlation between serum and follicular fluid IGF-1 concentrations (r = 0.47; P = 0.002). Quantification of transcripts did not differ between parity groups. In experiment 2, concentrations of NEFA (P = 0.12) and insulin (P = 0.16) in the blood and P/AI (P = 0.93) did not differ between parity [60 % (30/50) primiparous vs. 60 % (30/50) multiparous]. In contrast, blood concentrations of IGF-1 (P = 0.0001), total cholesterol (P = 0.005) and glucose (P = 0.01) were greater in primiparous cows. It was concluded that the oocyte quality and expression of the genes evaluated in the granulosa cells were not different between primiparous and multiparous cows. Unexpectedly, the pregnancy rate did not differ between parity. Nevertheless, the blood concentrations of IGF-1, total cholesterol and glucose were greater in primiparous cows.

Aspectos que influenciam a contagem bacteriana total e a contagem de células somáticas do leite de propriedades rurais do interior do estado de São Paulo, Brasil / Aspects that influence milk total bacterial count and somatic cell count from farms in interior of São Paulo State, Brazil / Aspectos que influyen el contaje bacteriano total y el contaje de células somáticas de la leche de propiedades rurales del interior del estado de São Paulo, Brasil

Este estudo objetivou avaliar aspectos que exercem influência sobre a contagem bacteriana total (CBT) e a contagem de células somáticas (CCS) do leite e avaliar a adequação das amostras do produto à Instrução Normativa nº 76, de 26 de novembro de 2018, do Ministério da Agricultura e Pecuária do Brasil (IN 76). A pesquisa foi realizada em propriedades rurais da microrregião de Birigui, São Paulo, Brasil, e se baseou em coleta de dados, por entrevista estruturada, e coletas de leite nas propriedades. Foi realizada amostragem por conveniência, sendo escolhidos 15 produtores de cada estrato (de acordo com a produção diária de leite: pequeno - até 100 L; médio - 101 a 300 L; grande - mais de 300 L), totalizando 45 produtores de leite. Participaram da pesquisa somente propriedades com base da alimentação volumosa em sistema de pastejo ou em semiconfinamento. Para avaliar a influência de diferentes fatores sobre a contagem bacteriana total (CBT) e a contagem de células somáticas (CCS), separadamente, realizou-se regressão multivariada. Ainda, foi determinado o coeficiente de correlação entre variáveis desta pesquisa. As variáveis resposta foram transformadas em logaritmo para normalização dos dados. Foi feita comparação entre os resultados da pesquisa e os parâmetros da IN 76 para verificar o cumprimento das normas pelos produtores. Foram encontrados valores médios de 6.986.977,818.961.790,7 UFC/mL para CBT e de 608.911,1 ± 414.802,9 CS/mL para CCS. A utilização de tanque de resfriamento individual mostrou-se favorável à baixa CBT, em comparação com não uso de tanque ou uso de tanque comunitário, conforme o esperado, pois quanto mais tempo leva para resfriar o leite, maior é a proliferação de bactérias no produto. Foi verificado que o leite de produtores que realizam o California Mastitis Test (CMT) com maior frequência apresentou CCS mais elevada, o que não era esperado. As análises físico-químicas médias foram densidade 1,031 g/mL a 15°C, índice crioscópico -0,538 °H, teor de proteína 3,58%, teor de gordura 4,06%, extrato seco total 12,75%, extrato seco desengordurado 8,69%, acidez titulável 16,2 °D e estabilidade ao álcool (Alizarol 72%) 100%. Em 13,3% (6/45) das amostras foi detectada presença de antibióticos. Considerando a IN 76, o leite de somente 8,9% (4/45) dos produtores apresentou-se adequado simultaneamente para todos os parâmetros analisados (CBT, CCS, densidade, índice crioscópico, proteína, gordura, extrato seco total, extrato seco desengordurado, acidez titulável, estabilidade ao alizarol e pesquisa de antibióticos). A utilização de tanque de resfriamento individual e a realização de CMT favoreceram menor CBT e maior CCS, respectivamente.

This study aimed to evaluate aspects that exerts influence on the total bacterial count (TBC) and somatic cell count (SCC) of milk and evaluate the conformity of the samples to the Normative Instruction no. 76, from November 26, 2018, of the Brazilian Agriculture and Livestock Ministry (NI 76). The research was done in farms around Birigui micro-region, São Paulo State, Brazil, and was based on data collection obtained by structured interviews and milk collection in the farms. Convenience sampling was carried out from 15 producers of each stratum (according to daily milk production: small - up to 100 L; medium - 101 to 300 L; large - more than 300 L), totalizing 45 milk producers. Only farms with grazing or semi-feedlot system for roughage feeding base were included. To evaluate the influence of different factors on Total Bacterial Count (TBC) and Somatic Cell Count (SCC), separately, a multivariate regression was done. Moreover, the correlation coefficient among variables was determined. The response variables were transformed into logarithms for the normalization of the data. A comparison between results and NI 76 parameters was done to verify the standard compliance by producers. Mean TBC was 6.986.977,8 ± 18.961.790,7 CFU/mL and mean SCC was 608.911,1414.802,9 cells/mL. The use of individual cooler tank was favorable to lower TBC, compared with the non-use of the tank or with the use of the communitarian tank, as expected, since the longer it takes to cool the milk, the higher the proliferation of bacteria in the product. It was verified that milk from farmers that more frequently perform California Mastitis Test (CMT) showed higher SCC, which was not expected. The average physicochemical analyzes were density 1.031 g/mL at 15°C, cryoscopic index -0.538 °H, protein content 3.58%, fat content 4.06%, total dry extract 12.75%, dry extract defatted 8.69%, titratable acidity 16.2 °D and alcohol stability (Alizarol 72%) 100%. The presence of antibiotics was detected in 13.3% (6/45) of the samples. Considering NI 76, milk from only 8.9% (4/45) farmers were proper simultaneously for all the parameters analyzed (TBC, SCC, density, cryoscopic index, protein, fat, total dry extract, defatted dry extract, titratable acidity, alizarol stability and antibiotics). The use of individual cooling tank and the CMT performance support lower TBC and higher SCC respectively.

Este estudio tuvo como objetivo evaluar los aspectos que influyen en el contaje bacteriano total (CBT) y el contaje de células somáticas (CCS) de la leche y evaluar la adecuación de las muestras del producto a la Instrucción Normativa nº 76, de 26 de noviembre de 2018, del Ministerio de Agricultura y Ganadería en Brasil (IN 76). La investigación se llevó a cabo en propiedades rurales en la microrregión de Birigui, São Paulo, Brasil, y se basó en la recolección de datos, a través de entrevistas estructuradas, y colectas de leche en las propiedades. Se realizó un muestreo por conveniencia, escogiendo 15 productores de cada estrato (de acuerdo con la producción diaria de leche: pequeño - hasta 100 L; medio - 101 hasta 300 L; grande ­ más de 300 L), totalizando 45 productores de leche. Participaron de la investigación únicamente propiedades basadas en alimentación voluminosa en sistema de pastoreo o, a lo sumo, en semiconfinamiento. Para evaluar la influencia de diferentes factores sobre el contaje bacteriano total (CBT) y el contaje de células somáticas (CCS), por separado, se realizó una regresión multivariada. Asimismo, se determinó el coeficiente de correlación entre las variables de esta investigación. Las variables de respuesta se transformaron en logaritmos para la normalización de datos. Se realizó una comparación entre los resultados de la investigación y los parámetros de la IN 76 para verificar el cumplimiento de las normas por parte de los productores. Se encontraron valores medios de 6.986.977,818.961.790,7 UFC/mL para CBT y de 608.911,1 +414.802,9 CS/mL para CCS. El uso de un tanque de enfriamiento individual se ha mostrado favorable a la baja CBT, en comparación con no usar un tanque o usar un tanque comunitario, conforme lo esperado, ya que cuanto más se tarda en enfriar la leche, mayor es la proliferación de bacterias en el producto. Se verificó que la leche de los productores que realizan el California Mastitis Test (CMT) con mayor frecuencia presentó CCS más alto, lo cual no se esperaba. Los análisis fisicoquímicos promedio fueron densidad 1,031 g/mL a 15°C, índice crioscópico -0,538 °H, contenido de proteína 3,58%, contenido de grasa 4,06%, extracto seco total 12,75%, extracto seco desgrasado 8,69%, acidez titulable 16,2°D y estabilidad del alcohol (Alizarol 72%) 100%. En el 13,3% (6/45) de las muestras se detectó la presencia de antibióticos. Considerando la IN 76, la leche de solo el 8,9% (4/45) de los productores se presentó adecuado simultáneamente para todos los parámetros analizados (CBT, CCS, densidad, índice crioscópico, proteína, grasa, extracto seco total, extracto seco desgrasado, acidez titulable, estabilidad al alizarol e investigación de antibióticos). El uso de un tanque de enfriamiento individual y la realización de CMT favorecieron un CBT más bajo y un CCS más alto respectivamente.

The early impact of diets enriched with saturated and unsaturated fatty acids on intestinal inflammation and tight junctions.

The gut has been suggested as the first organ to be affected by unbalanced diets contributing to the obesogenic process. This study aimed to test a short time-course exposition model to a known pro- or anti-inflammatory enriched fatty diet to understand the early gut alterations. Male mice were exposed to the chow diet (CT), high-fat (HF) diet, or a high-fat diet partially replaced on flaxseed oil (FS), rich in omega-3 (ω3), for 14 days. HF and FS increased the total body weight mass compared with the CT group, but FS reduced the epididymal fat depot compared to HF. The bioinformatics from mice and human databases showed the Zo1-Ocln-Cldn7 tight junctions as the main protein-triad. In the ileum, the HF diet has increased IL1ß transcript and IL1ß, TNFα, and CD11b proteins, but reduced the tight junctions (Zo1, Ocln, and Cld7) compared to the CT group. Despite the FS diet being partially efficient in protecting the ileum against inflammation, the tight junctions were increased, compared to the HF group. The GPR120 and GPR40 receptors were unaffected by diets, but GPR120 was colocalized on the surface of ileum macrophages. The short period of a high-fat diet was enough to start the obesogenic process, ileum inflammation, and reduce the tight junctions. Flaxseed oil did not protect efficiently against dysmetabolism. Still, it increased the tight junctions, even without alteration on inflammatory parameters, suggesting the protection against gut permeability during early obesity development.

Plasma Estradiol Profile After Administration of Different Types of Estradiol Esters in Acyclic Mares.

Although the use of different estradiol esters has been extensively studied in hormonal protocols in cows, such information is lacking in mares. The present study aimed to assess the effects of treatment with the same doses and administration frequency of estradiol cypionate, estradiol benzoate and 17ß estradiol on plasma estradiol (E2) concentrations of acyclic mares and correlate the E2 profile to the endometrial edema score. Sixteen treatments were performed in 14 mares randomly divided into three groups: EB (n = 5), EC (n = 5), and 17ß (n = 6), receiving 10 mg on day 0 (D0), 6 mg on D1, and 4 mg on D2 of estradiol benzoate, estradiol cypionate, and estradiol 17ß, respectively. Blood samples, rectal palpations, and ultrasound evaluations were performed once daily, starting before the first estradiol treatment (D0) until edema disappearance or D8. Moderate to high edema was observed in all groups 24 hours after the first estradiol administration. Edema persisted above score 2 until D7, D5, and D4 in groups EC, EB, and 17ß, respectively. Higher edema was found on D2 in EB group compared to 17ß, on D6 and D7 in EC compared to 17ß, and on D8 in EC compared to EB (p ≤ .05). Maximum E2 concentrations were detected on D1 in groups: EB, showing a sharp decrease from D2 to D3 (p < .0001); and 17ß, where no differences were observed between treatment days (p ≥ .05). In the EC group, maximum concentrations were observed on D2, which remained high from D2 to D4 compared to the other days (p < .0001). Plasma concentration of E2 was higher in EB than the other groups on D0 and D1, and EB>EC>17ß on D2. Plasma E2 concentrations and edema score were positively correlated, being moderate in groups EB and EC and weak in the 17ß group. In conclusion, the most pronounced plasma E2 concentration was observed in EB group 24 hours after the administration of the first dose. Estradiol concentrations peaked 48 hours after EC administration, while a distinct peak was not observed in a 24h interval evaluation in the 17ß group. Moreover, high edema does not necessarily reflect on high E2 concentrations in acyclic mares.

An older diabetes-induced mice model for studying skin wound healing.

Advances in wound treatment depend on the availability of animal models that reflect key aspects of human wound healing physiology. To this date, the accepted mouse models do not reflect defects in the healing process for chronic wounds that are associated with type two diabetic skin ulcers. The long term, systemic physiologic stress that occurs in middle aged or older Type 2 diabetes patients is difficult to simulate in preclinical animal model. We have strived to incorporate the essential elements of this stress in a manageable mouse model: long term metabolic stress from obesity to include the effects of middle age and thereafter onset of diabetes. At six-weeks age, male C57BL/6 mice were separated into groups fed a chow and High-Fat Diet for 0.5, 3, and 6 months. Treatment groups included long term, obesity stressed mice with induction of diabetes by streptozotocin at 5 months, and further physiologic evaluation at 8 months old. We show that this model results in a severe metabolic phenotype with insulin resistance and glucose intolerance associated with obesity and, more importantly, skin changes. The phenotype of this older age mouse model included a transcriptional signature of gene expression in skin that overlapped that observed with elderly patients who develop diabetic foot ulcers. We believe this unique old age phenotype contrasts with current mice models with induced diabetes.

Brown fat triglyceride content is associated with cardiovascular risk markers in adults from a tropical region.

Brown adipose tissue (BAT) is regarded as an interesting potential target for the treatment of obesity, diabetes, and cardiovascular diseases, and the detailed characterization of its structural and functional phenotype could enable an advance in these fields. Most studies evaluating BAT structure and function were performed in temperate climate regions, and we are yet to know how these findings apply to the 40% of the world's population living in tropical areas. Here, we used 18F-fluorodeoxyglucose positron emission tomography - magnetic resonance imaging to evaluate BAT in 45 lean, overweight, and obese volunteers living in a tropical area in Southeast Brazil. We aimed at investigating the associations between BAT activity, volume, metabolic activity, and BAT content of triglycerides with adiposity and cardiovascular risk markers in a sample of adults living in a tropical area and we showed that BAT glucose uptake is not correlated with leanness; instead, BAT triglyceride content is correlated with visceral adiposity and markers of cardiovascular risk. This study expands knowledge regarding the structure and function of BAT in people living in tropical areas. In addition, we provide evidence that BAT triglyceride content could be an interesting marker of cardiovascular risk.

Cyclodextrins Initiated Ring-Opening Polymerization of Lactide Using 4-Dimethylaminopyridine (DMAP) as Catalyst: Study of DMAP/ß-CD Inclusion Complex and Access to New Structures.

Cyclodextrins (CDs) are cyclic oligosaccharides used in many fields. Grafting polymers onto CDs enables new structures and applications to be obtained. Polylactide (PLA) is a biobased, biocompatible aliphatic polyester that can be grafted onto CDs by -OH-initiated ring-opening polymerization. Using 4-dimethylaminopyridine (DMAP) as an organocatalyst, a quantitative functionalization is reached on native α-, ß-, γ- and 2,3-dimethyl- ß-cyclodextrins. Narrow molecular weight distributions are obtained with the native CDs (dispersity < 1.1). The DMAP/ß-CD combination is used as a case study, and the formation of an inclusion complex (1/1) is shown for the first time in the literature, which is fully characterized by NMR. The inclusion of DMAP into the cavity occurs via the secondary rim of the ß-CD and the association constant (Ka) is estimated to be 88.2 M-1. Its use as an initiator for ring-opening polymerization leads to a partial functionalization efficiency, and thus a more hydrophilic ß-CD-PLA conjugate than that obtained starting from native ß-CD. Polymerization results including also the use of the adamantane/ß-CD inclusion complex as an initiator suggest that inclusion of the DMAP catalyst into the CD may not occur during polymerization reactions. Rac-lactide does not form an inclusion complex with ß-CD.

Arcuate Nucleus Overexpression of NHLH2 Reduces Body Mass and Attenuates Obesity-Associated Anxiety/Depression-like Behavior.

Nescient helix-loop-helix 2 (NHLH2) is a hypothalamic transcription factor that controls the expression of prohormone convertase 1/3, therefore having an impact on the processing of proopiomelanocortin and thus on energy homeostasis. Studies have shown that KO of Nhlh2 results in increased body mass, reduced physical activity, and hypogonadism. In humans, a polymorphism of the NHLH2 gene is associated with obesity; and in Prader-Willi syndrome, a condition characterized by obesity, hypogonadism and behavioral abnormalities, the expression of NHLH2 is reduced. Despite clinical and experimental evidence suggesting that NHLH2 could be a good target for the treatment of obesity, no previous study has evaluated the impact of NHLH2 overexpression in obesity. Here, in mice fed a high-fat diet introduced right after the arcuate nucleus intracerebroventricular injection of a lentivirus that promoted 40% increase in NHLH2, there was prevention of the development of obesity by a mechanism dependent on the reduction of caloric intake. When hypothalamic overexpression of NHLH2 was induced in previously obese mice, the beneficial impact on obesity-associated phenotype was even greater; thus, there was an 80% attenuation in body mass gain, reduced whole-body adiposity, increased brown adipose tissue temperature, reduced hypothalamic inflammation, and reduced liver steatosis. In this setting, the beneficial impact of hypothalamic overexpression of NHLH2 was a result of combined effects on caloric intake, energy expenditure, and physical activity. Moreover, the hypothalamic overexpression of NHLH2 reduced obesity-associated anxiety/depression behavior. Thus, we provide an experimental proof of concept supporting that hypothalamic NHLH2 is a good target for the treatment of obesity.SIGNIFICANCE STATEMENT Obesity is a highly prevalent medical condition that lacks an effective treatment. The main advance provided by this study is the demonstration of the beneficial metabolic and behavioral outcomes resulting from the overexpression of NHLH2 in the hypothalamus. When NHLH2 was overexpressed simultaneously with the introduction of a high-fat diet, there was prevention of obesity by a mechanism dependent on reduced caloric intake. Conversely, when NHLH2 was overexpressed in previously obese mice, there was reduction of the obese phenotype because of a combination of reduced caloric intake, increased physical activity, and increased thermogenesis. In addition, the overexpression of NHLH2 reduced anxiety/depression-like behavior. Thus, NHLH2 emerges as a potential target for the combined treatment of obesity and its associated anxiety/depression-like behavior.

A Smartphone App for Individual Xylazine/Ketamine Calculation Decreased Anesthesia-Related Mortality in Mice.

Currently, experimental animals are widely used in biological and medical research. However, the scientific community has raised several bioethical concerns, such as the number of animals required to achieve reproducible and statistically relevant results. These concerns involve aspects related to pain, discomfort, and unwanted animal loss. Retrospectively, we compare two different approaches for anesthesia dosage: a mobile app for dose calculation and a standard dose calculation. A total of 939 C57BL/6J and Swiss mice were analyzed. We collected data on intraoperative and anesthesia-related mortality as described in electronic or physical handwritten records. Our results showed that the mobile app approach significantly reduces anesthetic-related deaths upon using doses of ketamine and xylazine. The results suggest that anesthesia-related mortality can be minimized even more using information technology approaches, helping to solve an old but transversal challenge for researchers working with experimental mice. The mobile app is a free and open code which could be implemented worldwide as an essential requirement for all anesthetic procedures in mice using xylazine and ketamine combination. As an open code app, the Labinsane initiative could also represent the starting point to unify and validate other anesthetic procedures in different species and strains.

Peri-estrus ovarian, uterine, and hormonal variables determine the uterine luminal fluid metabolome in beef heifers.

In cattle, uterine luminal fluid (ULF) is the main source of molecules that support embryo development and survival during the peri-implantation period. Our overarching hypothesis is that peri-estrus changes in uterine function, including ULF accumulation and absorption, are uneven among individuals, and affect ULF composition and fertility. Our objectives were (1) to characterize temporal and spatial changes in ULF volume, endometrial and luteal blood perfusion, endometrial and luteal size, and circulating progesterone concentrations during the peri-estrus period in beef heifers and (2) to associate such changes with the metabolite composition in the ULF, 4 days after estrus (d 0). Fourteen Bos indicus heifer that presented a PGF2α responsive CL received 500 µg PGF2α analog i.m. and were examined daily by rectal B-mode and pulse-wave color-Doppler ultrasonography until the fifth day after estrus (d 5). The composition of the ULF was analyzed by targeted mass spectrometry on d 4. Multivariate analyses clustered heifers according to ovarian, uterine, and hormonal variables in clusters A (n = 5) and B (n = 8 heifers). Concentrations of Pro, Ala, Leu, Gly, Val, Lys, Ile, Phe, Asp, Orn, Tyr, Arg, Trp, Suc, Cit, ADMA, the sum of essential Amino Acids (AA), sum of nonessential AA, sum of aromatic AA, and total AA were greater in cluster A (FDR ≤ 0.05). ULF volume dynamics and uterine, ovarian, and hormonal variables during the peri-estrus period presented a concerted variation among heifers within clusters, which was associated with the ULF composition 4 days after estrus.

Methylome Patterns of Cattle Adaptation to Heat Stress.

Heat stress has a detrimental impact on cattle health, welfare and productivity by affecting gene expression, metabolism and immune response, but little is known on the epigenetic mechanisms mediating the effect of temperature at the cellular and organism level. In this study, we investigated genome-wide DNA methylation in blood samples collected from 5 bulls of the heat stress resilient Nellore breed and 5 bulls of the Angus that are more heat stress susceptible, exposed to the sun and high temperature-high humidity during the summer season of the Brazilian South-East region. The methylomes were analyzed during and after the exposure by Reduced Representation Bisulfite Sequencing, which provided genome-wide single-base resolution methylation profiles. Significant methylation changes between stressful and recovery periods were observed in 819 genes. Among these, 351 were only seen in Angus, 366 were specific to Nellore, and 102 showed significant changes in methylation patterns in both breeds. KEGG and Gene Ontology (GO) enrichment analyses showed that responses were breed-specific. Interestingly, in Nellore significant genes and pathways were mainly involved in stress responses and cellular defense and were under methylated during heat stress, whereas in Angus the response was less focused. These preliminary results suggest that heat challenge induces changes in methylation patterns in specific loci, which should be further scrutinized to assess their role in heat tolerance.

Functions of the GABAergic system on serum LH concentrations in pre-pubertal Nellore heifers.

This study was conducted to evaluate the luteinizing hormone (LH) secretion pattern after gamma-aminobutyric acid (GABAA) antagonist to determine the effects of the GABAergic system on LH secretion during reproductive maturation in pre-pubertal Nellore heifers. Nellore heifers (n = 10) were administered a picrotoxin injection of 0.18 mg/kg, i.v. Blood samples were collected every 15 min for 3 h at different developmental stages (8, 10, 14 and 17 mo of age). Plasma concentrations of LH were quantified using an RIA (sensitivity of 0.04 ng/mL and CV of 15 %). There was an interaction between treatment and age (P = 0.034). Picrotoxin-treated heifers had lesser (P ≤  0.05) LH mean concentrations during a 3 h period at 10 and 17 mo of age compared to control heifers (P ≤  0.05). Comparing the period before and after Picrotoxin injection in the same animals, there was a 33 % decrease in LH concentration at 8 mo of age (P = 0.0165). These results indicate the GABAergic system has a stimulatory function in inducing LH secretion in pre-pubertal Nellore heifers. These findings corroborate previous results that GABA increases GnRH/LH secretion in other species during the pre-pubertal period.

Early resynchronization of follicular wave emergence among Nelore cattle using injectable and intravaginal progesterone for three timed artificial inseminations.

Follicular dynamics and pregnancy per AI (P/AI) of cattle submitted to resynchronization 13 d after TAI using a progesterone (P4) insert and supplementary injectable progesterone (iP4) were determined. There was synchronization of ovulation timing imposed (Day 0 = expected estrus; Experiment 1). On Day 13, animals were assigned to: control (only P4 insert; 15 cows and 13 heifers) and iP4 (P4 insert+100 mg iP4; 13 cows and 12 heifers) groups, and submitted to daily ovarian ultrasonography and blood collection (Day 13 to Day 22). In Experiment 2, 760 suckled cows and 498 heifers were submitted to a TAI on Day 0 and assigned into control and iP4 groups on Day 13. In animals with luteolysis on Day 22, there was a second TAI on Day 24, and on Day 37 were subjected to the opposite treatments as imposed in first resynchronization procedure. On Day 37, there was pregnancy diagnosis in animals with functional CL. The third TAI was performed on Day 48. Day of follicular wave emergence did not differ between groups and parities. Dominant follicles were larger in cows than heifers, and in animals of the control group on Day 24. Greater P4 concentrations were detected on Day 14 and Day 15 in the iP4-treated animals. Luteolysis occurred earlier in cows than heifers. Overall P/AI percentage as a result of second and third TAIs, regardless of parity, was greater in iP4-treated animals. In conclusion, females treated with a supplementary iP4 had a greater plasma P4 concentration and P/AI, but there was no effect of iP4 treatment on synchrony of timing of follicular wave.

Serum Progesterone and Conception Rates in Acyclic Embryo Recipient Mares Using a Bovine Progesterone-Releasing Intravaginal Device.

The objective of this study was to quantify serum progesterone levels, uterine features, and pregnancy rates in acyclic, embryo recipient mares using a bovine progesterone-releasing intravaginal device in a commercial embryo transfer (ET) program. The study included 73 recipient mares of unknown breed, aged 3-10 years, weighing 350-500 kg, and kept under an intensive management system on Tifton 85 (Cynodon spp.) pastures with water and mineral salt ad libitum. The horses were divided into two groups: a group with a progesterone-releasing intravaginal device (1 g progesterone, G-IVP4, n = 24) and a control group (G-iP4, n = 49) receiving an injection of 1,500 mg long-acting progesterone. Jugular blood was collected for the G-IVP4 group for subsequent progesterone measurement by radioimmunoassay on three occasions: Day 0 (D0), intravaginal device was placed; Day 5 (D5), day of the ET; and Day 9 (D9), day of pregnancy diagnosis. There was an increase (P < .0001) in serum progesterone levels on D5 and D9 compared with D0 (4.09 ± 0.81 and 6.45 ± 1.03 ng/mL vs. 0.71 ± 0.14 ng/mL). There were no differences among groups in the pregnancy rate (P > .05), with rates of 83.33% and 73.46% for G-IVP4 and G-iP4, respectively. In conclusion, the intravaginal route for absorption of 1 g of progesterone device increased the serum level of progesterone sufficiently to prepare the uterus of acyclic recipient mares for ET, and the conception rate was similar to the standard protocol using long-acting injectable progesterone.

Short Dietary Intervention with Olive Oil Increases Brown Adipose Tissue Activity in Lean but not Overweight Subjects.

BACKGROUND: The brown adipose tissue (BAT) is a potential target for the treatment of obesity and metabolic disorders. Its activation by cold exposure or adrenergic drugs can increase systemic insulin sensitivity and improve lipid metabolism; however, little is known about the effects of specific dietary components on BAT activity. OBJECTIVES: We asked if a short-term (4 weeks) dietary intervention with olive oil could modify BAT activity in lean and overweight/obese volunteers. DESIGN: This was a 4-week open clinical trial in which all participants underwent a dietary intervention with extra-virgin olive oil supplementation. As the initial intake of olive oil was controlled all the participants were controls of themselves. RESULTS: The intervention resulted in significant increase in blood monounsaturated fatty acid levels, which was accompanied by increased BAT activity in lean but not in overweight/obese volunteers. In the lean group, an increase in leptin was detected after the intervention, and low leptin values at the beginning of the study were predictive of greater BAT activity after intervention. In addition, increase in leptin concentration was associated with increased BAT activity. Three known endogenous mediators of BAT activity, secretin, fibroblast growth factor 21 (FGF21), and 12,13-dihydroxy-9Z-octadecenoic acid (12,13-diHOME) were increased by intervention in lean, whereas only secretin and FGF21 were increased in subjects with excessive weight. CONCLUSION: This study provides clinical evidence for the impact of monounsaturated fatty acids on BAT activity and an advance in the understanding of the beneficial health effects of olive oil.