Exploring uterine inflammation in postpartum primiparous precocious and conventional and multiparous Bos indicus beef cows.

This study aimed to compare the postpartum uterine dynamics of primiparous precocious (PP), primiparous conventional (PC) and multiparous conventional (MC) Bos indicus beef cows. For this purpose, PP (n = 8), PC (n = 18) and MC (n = 12) cows were enrolled in this study. These cows were evaluated at 20 and 10 days prepartum and weekly from parturition to 42 days postpartum (DPP). During this period, body weight (BW), subcutaneous fat thickness (SFT) and serum concentrations of glucose, ß-hydroxybutyrate, albumin and haptoglobin were measured. Proportion of polymorphonuclear (PMN) cells, and abundance of mRNA transcripts of genes involved in uterine inflammation and uterine health were evaluated. The PP cows had lower (p < .05) BW and SFT than that for PC and MC cows during the study period. The serum concentration of albumin after 35 DPP was lower (p < .05) in PP cows. The PP cows had the highest proportion of PMN on 28 and 35 DPP compared to PC and MC cows. The relative mRNA abundance of IL-1ß and IL-8 increased after 21 DPP in PP cows compared to the other groups. The PC had the highest, MC had an intermediate, and PP cows had the lowest relative abundance of IL10 mRNA. Overall, these findings indicated that uterine inflammation was more pronounced in PP cows. Moreover, based on the proportion of PMN and abundance of transcripts associated with inflammation in the uterus, PP cows may require a longer period to recover their uterine health after calving.

An assessment on the effects of buffers on the productive, behavioral and metabolic parameters of Holstein dairy cows.

This study aimed to evaluate the impact of supplementing sodium bicarbonate or a commercial blend of buffering agents (BBA) comprising calcareous calcitic, magnesium oxide, calcareous algae, and sodium bicarbonate on the productive, behavioral and metabolic parameters of Holstein cows fed starchy diets. Over a 60-day trial period, thirty-six multiparous cows with an average milk yield of 38.84 ± 9.24 kg/day and 63.74 ± 18.63 days in milk (DIM), were randomly divided into two groups. The control group (n = 18) received a supplementation of 1.1% dry matter (DM) of sodium bicarbonate (Raudi®, Totalmix, Brazil), while the BBA group (n = 18) was administered with 0.5% DM of a blend of buffering agents (Equalizer®, Nutron/Cargill, Brazil). The mean values of ruminal pH (control 6.80 ± 0.06 and BBA 6.77 ± 0.06; P > 0.05) and volatile fatty acid (VFA) production (control: acetate 62.63 ± 1.29%, propionate 22.99 ± 1.07%, butyrate 14.30 ± 0.52%; BBA: acetate 63.07 ± 1.32%, propionate 23.47 ± 1.10%, butyrate 13.70 ± 0.57%), were similar (P > 0,05) between the two groups. The value of faecal pH was higher (P < 0.05) in the BBA group (6.25 ± 0.02) than the control group (6.12 ± 0.02). Animals treated with BBA exhibited lower (P < 0,05) dry matter intake (DMI) (24.75 ± 0.64 kg/day), higher feed efficiency (FE) (1.64 ± 0.03), and reduced feeding frequency (52.89 ± 3.73 n°/day) than the control group (DMI, 26.75 ± 0.62 kg/day; FE, 1.50 ± 0.03; feeding frequency, 66.07 ± 3.64 n°/day). Milk production remained similar across both groups (control, 39.11 ± 0.92 kg/day and BBA, 39.87 ± 0.92 kg/day; P > 0.05). Notably, the control group displayed a higher (P < 0,05) concentration of milk protein (1.21 ± 0.05 kg/day) than the BBA (1.18 ± 0.05 kg/day) group. The study concluded that both treatments effectively buffered the rumen and mitigated the risk of ruminal acidosis. Moreover, the higher faecal pH in the BBA-treated group suggests potential intestinal action attributable to the synergistic effects of diverse additives with buffering properties. Despite a reduced DMI, BBA-treated animals exhibited improved FE.

Metabolic profile of transition period in ewes and its influence on passive immunity transference in lambs.

The intensification of ewe production, focusing on increasing prolificity, results in a higher incidence of metabolic disorders, especially in the transition period, which can lead to production losses with consequences for the health of the animals. The objective of this study was to evaluate the metabolic profile in transition period Lacaune ewes according to the parity and litter size born and its influence on the transference of passive immunity. Nineteen Lacaune ewes, with initial mean body weight of 76.36 ± 10.37 kg (SD), kept in a semi-extensive system and receiving the same diet, were used. Ewes were classified according to parity (primiparous and multiparous) and the litter size born at parturition (1, 2, and 3). On days - 14, - 7, and - 4 of the expected date of parturition and on days 1, 4, 7, and 14 days postpartum, blood sampling, the evaluation of the body condition score (BCS), and the weighing of animals were performed. Blood samples were taken from the lambs 48 h after birth. Colostrum was collected up to 6 h postpartum to determine the brix percentage. Serum concentrations of total plasma proteins (TPP), albumin, aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), beta-hydroxybutyrate (BHB), non-esterified fatty acids (NEFA), paraoxonase (PON1), calcium (Ca), and magnesium (Mg) were evaluated. In lambs, the blood parameters evaluated were TPP, albumin, GGT, and PON1. Lamb plasma was used to estimate colostrum efficiency, brix percentage immunity, and total plasma protein (PPT). Productive data, pre- and postpartum metabolic variables for ewes, lambs, and those related to colostrum brix were analyzed. The BCS had a reduction (p < 0.0001) in the postpartum period, which is evidenced on days 7 and 14 (2.88 and 2.73, respectively). Serum concentration of BHB presented the highest concentrations in the prepartum period for ewes that gave birth to triplets, from day - 14, and for ewes that gave birth to twins on day - 4 (p = 0.0245). Serum concentration of NEFA demonstrated the highest value on day 14 in primiparous (1.07; p = 0.0402). Calcium had the lowest concentrations on day 1 (8.85 mg/dl, p < 0.0001) and magnesium on day 3 (1.79 mg/dl, p < 0.0001) postpartum. The multiparous with two lambs had higher brix percentages (30.5%), and the lowest value was observed in primiparous ewes that gave birth to a single lamb (27.06%; p = 0.0395). Triplet lambs had the lowest weight (3.73 kg, p = 0.0007), and the best brix percentage in plasma was observed in twins (10.29%, p = 0.0174). Regardless of the parity, the ewes that presented the greatest metabolic challenge were those that gave birth to triplets, and these lambs presented the worst immunity and the lowest live weight. The quality of colostrum was influenced by the parity, and multiparous ewes that gave birth to triplets had the lowest brix percentage in the colostrum.

Zootechnical and health performance of Holstein x Gir crossbred calves.

Crossbreeding is used to increase production and disease resistance in adult animals, and there is no research to assess the performance of animals in the early stages. The aim of this study was to evaluate the zootechnical and health performance of Holstein x Gir calves (½ HG: ½ Dutch ½ Gir and ¾ HG: ¾ Dutch » Gir), from birth to 80 days of age, and compare metabolic parameters between groups. In this sense, calves were monitored for zootechnical parameters; epidemiological indexes such as morbidity, mortality, recurrence of diarrhea, pneumonia, and other diseases; as well as serum concentrations of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma-glutamyl transferase, total proteins, cholesterol, triglycerides, albumin, urea, and paraoxonase1 (PON1). ¾ HG calves showed higher morbidity for diarrhea and remained with diarrhea for longer compared to ½ HG calves, and this was reflected in the average daily weight gain until the 42nd day, with ½ HG calves performing better. There were no differences regarding passive immune transfer between groups, as well as no differences in morbidity and mortality from pneumonia. Regarding biochemical analyses, a difference was found only in the concentrations of PON1, which were higher in ¾ HG calves. The findings show that blood degree influences the occurrence and duration of diarrhea, negatively impacting the zootechnical performance of the animals. Crossbreeding bulls with zebu cattle can be an alternative to increase calf resistance and reduce diarrhea, thus lowering economic losses and improving animal performance.

Effects of substituting sweet potato flour for ground corn on performance, feeding behavior, and metabolism of dairy cows.

The objective of this study was to evaluate the effect of substituting sweet potato flour for ground corn in rations fed to lactating dairy on milk yield and composition, blood metabolites, and feeding behavior. Twenty lactating Holstein cows from 30 to 60 days postpartum were randomly assigned to one of two groups (n = 10 each) and used in a cross-over design trial with two treatments: a standard concentrate with ground corn as an energy source or experimental concentrate with sweet potato flour (SPF) replacing all of the ground corn. Each of the 35-day periods consisted of 14 days for adaptation to diet and 21 days for data and sample collection. Milk yield, dry matter intake (DMI), and feeding behavior were evaluated daily throughout the trial. Milk samples were collected weekly and blood samples were collected every 3 days. Milk was analyzed for fat, protein, lactose, and total solid constituents. Blood was analyzed for glucose, non-esterified fatty acid (NEFA), gamma-glutamyl transferase (GGT), aspartate aminotransferase (AST), total protein (TP), albumin, and urea concentrations. Milk yield (P = 0.62) and composition (fat: P = 0.71; protein: P = 0.12; lactose: P = 0.82; total solids: P = 0.56) were not affected by dietary treatments. There were no differences between treatments in DMI or meal frequency, but total eating time (P = 0.001), feeding time (P = 0.001), and meal duration (P = 0.001) was higher for control compared with SPF. However, feeding rate (P = 0.001) and serum urea concentration (P = 0.001) were higher for SPF compared with control. No differences were observed in serum metabolites and enzymes measured among treatments. The results of this trial indicate that SPF can be substituted for ground corn without impairing the performance, feeding behavior, and metabolism in dairy cows.

Supplementation with beta-hydroxy-beta-methylbutyrate impacts glucose homeostasis and increases liver size in trained mice.

ß-hydroxy-ß-methyl butyrate (HMB) is a bioactive metabolite derived from the amino acid leucine, usually applied for muscle mass increase during physical training, as well as for muscle mass maintenance in debilitating chronic diseases. The hypothesis of the present study is that HMB is a safe supplement for muscle mass gain by strength training. Based on this, the objective was to measure changes in body composition, glucose homeostasis and hepatic metabolism of HMB supplemented mice during strength training. Two of four groups of male mice (n = 6/group) underwent an 8-week training period session (climbing stairs) with or without HMB supplementation (190 mg/kgBW per day). We observed lower body mass gain (4.9 ± 0.43% versus 1.2 ± 0.43, p < 0.001) and increased liver mass (40.9 ± 0.9 mg/gBW versus 44.8 ± 1.3, p < 0.001) in the supplemented trained group compared with the non-supplemented groups. The supplemented trained group had an increase in relative adipose tissue mass (12.4 ± 0.63 mg/gBW versus 16.1 ± 0.88, P < 0.01) compared to the non-supplemented untrained group, and an increase in fasting blood glucose (111 ± 4.58 mg/dL versus 122 ± 3.70, P < 0.05) and insulin resistance (3.79 ± 0.19 % glucose decay/min versus 2.45 ± 0.28, P < 0.05) comparing with non-supplemented trained group. Adaptive heart hypertrophy was observed only in the non-supplemented trained group (4.82 ± 0.05 mg/gBW versus 5.12 ± 0.13, P < 0.05). There was a higher hepatic insulin-like growth factor-1 expression (P = 0.002) in supplemented untrained comparing with non-supplemented untrained group. Gene expression of gluconeogenesis regulatory factors was increased by training and reduced by HMB supplementation. These results confirm that HMB supplementation associated with intensive training protocol drives changes in glucose homeostasis and liver metabolism in mice.

Early embryonic development of bovine oocytes challenged with LPS in vitro or in vivo.

The aim of this study was to evaluate the effect of exposing bovine oocytes to lipopolysaccharides (LPS) in vivo and in vitro on early embryo development. In experiment 1, cumulus oocyte complexes (COCs, n = 700/group) were challenged with 0, 0.1, 1.0 or 5.0 µg/mL of LPS during in vitro maturation (IVM). Later, in vitro fertilization (IVF) and in vitro culture (IVC) were performed. In experiment 2, COCs (n = 200/group) matured and in vitro fertilized without LPS were subjected to IVC with the same doses of LPS from experiment 1. In experiment 3, heifers received two injections of saline solution (n = 8) or 0.5 µg/kg of LPS (n = 8) 24 h apart, and 3 days later, COCs were recovered and submitted to IVM, IVF, and IVC. In experiments 1 and 3, the expression of TLR4, TNF, AREG and EREG genes in cumulus cells was evaluated. Exposure to 1 and 5 µg/mL of LPS during IVM decreased nuclear maturation (39.4 and 39.6%, respectively) compared with control (63.6%, P < 0.05). Despite that, no effect on cleavage and blastocyst rates were observed. Exposure to LPS during IVC did not affect embryonic development. In vivo exposure to LPS decreased the in vitro cleavage rate (54.3 vs 70.2%, P = 0.032), but cleaved embryos developed normally. Number of cells per embryo and gene expression were not affected by the LPS challenge in any experiment. In conclusion, although in vitro exposure to LPS did not affect early embryo development, in vivo LPS exposure reduced cleavage rate.

Intrafollicular paraoxonase 1 activity and the steroidogenic potential of the first post-partum dominant follicle in dairy cows.

Cows experiencing high levels of inflammation and specific metabolic conditions tend to have slower follicular growth and lower serum and follicular concentrations of oestradiol (E2). Paraoxonase1 (PON1) activity decreases during inflammatory processes. Therefore, the aim of this study was to evaluate the association between serum and intrafollicular (FF) PON1 activity and the serum and intrafollicular levels of E2 and progesterone (P4), as well as the mRNA expression of genes related to steroidogenesis, metabolism and inflammation in the first post-partum dominant follicle of Holstein cows. No correlation was found between PON1 activity, the expression of the analysed genes and levels of follicular E2 and P4, except for a negative correlation between serum E2 and follicular PO1 activity. Also, no correlation was found between serum and follicular PON1 during the first post-partum follicular wave.

Effect of high-density lipoprotein on oocyte maturation and bovine embryo development in vitro.

High-density lipoprotein (HDL) is the main lipoprotein in the follicular fluid, and it has anti-inflammatory, antioxidant and cryoprotectant properties. The anti-inflammatory potential and antioxidant potential are derived from its lipid composition, especially the apolipoprotein AI (ApoAI) and paraoxonase 1 (PON1). The aim of this study was to evaluate the effect of HDL during in vitro maturation (IVM) on oocyte maturation and early bovine embryo development. For this, cumulus-oocyte complexes (COCs) were obtained from bovine ovaries collected at a local slaughterhouse. COCs (n = 2,250) were allocated into three groups (n = 50 COCs/group) according to the addition of HDL protein (HDL-P) during IVM for 22 hr: 0 (control), 50 and 150 mg/dl. After IVM, COCs were inseminated (in vitro fertilization) and cultivated for 7 days. Total cholesterol concentration, total protein, triglycerides and ApoAI concentrations on IVM medium increased proportionally to HDL-P addition. However, PON1 activity was not detected in any treatment. The addition of HDL-P did not affect nuclear maturation rate, endogenous reactive oxygen species and glutathione levels in COCs (p > 0.05). The highest HDL-P concentration (150 mg/dl) decreased cleavage and blastocyst rate (p < 0.05). Moreover, the HDL-P 150 mg/dl group had lower cellular count/blastocyst than the 50 mg/dl group (p < 0.05). However, the addition of HDL-P did not affect relative gene expression of evaluated genes. In conclusion, the complex HDL/ApoAI obtained from human plasma, in the absence of PON1 activity during in vitro oocyte maturation, decreased initial embryo development.

Differential effects of a high-fat diet on serum lipid parameters and ovarian gene expression in young and aged female mice.

The aim of this study was to compare serum lipid profiles and ovarian gene expression between aged and younger female mice fed a control or a high-fat diet for 2 months. For this 16 female mice (C57BL/6) of 4 months (Young, n = 8) or 13 months (Old, n = 8) of age were used. The females were divided into four groups: (i) young females fed a normal diet; (ii) young females fed a high-fat diet; (iii) old females fed a normal diet; and (iv) old females fed a high-fat diet. Food intake was reduced (P < 0.05) in mice fed with a high-fat (2.9 ± 0.1 g) diet in comparison with control mice (3.9 ± 0.1 g). Body weight was higher for old females on the high-fat diet (35.1 ± 0.3 g) than for young females on the same diet (23.3 ± 0.4 g; P < 0.05). PON1 activity was lower in the high-fat than control diet group (114.3 ± 5.8 vs. 78.1 ± 6.0 kU/L, respectively) and was higher in older than younger females (85.9 ± 6.4 vs. 106.5 ± 5.3; P < 0.05, respectively). Females fed a high-fat diet had lower expression of Igf1 mRNA (P = 0.04). There was an interaction between age and diet for the expression of Gdf9 and Survivin, with lower expression in older females in both diets and young females that received the high-fat diet (P < 0.05). Concluding, the high-fat diet reduced the expression of ovarian Igf1 mRNA, and Gdf9 and Survivin mRNA in younger females, which can indicate lower fertility rates. High-density lipoprotein concentration and PON1 activity were higher in aged female mice.

Rumen-protected methionine modulates body temperature and reduces the incidence of heat stress temperatures during the hottest hours of the day of grazing heat-stressed Bos indicus beef cows.

This study evaluated the effects of supplementation of rumen-protected methionine (RPM) on body thermoregulation and conception rate of Nelore cows exposed to high temperature-humidity index (THI). On -31 days before the artificial insemination protocol, 562 lactating, multiparous cows were assigned to receive (MG) or not (CG) RPM supplementation (3 g/cow mixed into 100 g of mineral supplement). Both groups remained in tropical pastures and received supplementation for 77 days. A subset of cows (n = 142) remained with an intravaginal thermometer collecting intravaginal temperature (IT). The respective minimum, average, and maximum environmental THI were 72.8, 78.0, and 83.3. Effects of treatment × hour of the day were detected (P < 0.0001) for IT. From 1330 to 1730 h and 1830 to 1900 h, IT was higher (P < 0.05) for CG versus MG cows when exposed to moderate and high THI. The supplementation with RPM did not affect conception rate (CG = 64.4% vs. MG = 58.2%; P > 0.05). In conclusion, 3 g of RPM supplementation lowered internal body temperature and possibly altered critical THI threshold in Nelore cows with no impact on reproduction.

Predictive value of prepartum serum metabolites for incidence of clinical and subclinical mastitis in grazing primiparous Holstein cows.

The aim of this study was to measure changes in biochemical markers in the peripartum period of primiparous Holstein cows diagnosed with subclinical and clinical mastitis. In this study, 37 dairy cows were monitored daily during milking until 60 days postpartum and were categorized according to the occurrence of clinical mastitis (group mastitis (GM), n = 9) or subclinical mastitis (group subclinical mastitis (GSUB), n = 10) or absence of symptoms (control group (CG), n = 18). Blood samples were collected weekly from -30 to 60 days from calving. Samples were grouped for prepartum (-30 to 0 days from calving), early postpartum (0 to 30 days from calving), and late postpartum (30 to 60 days from calving) periods. Prepartum serum non-esterified fatty acid (NEFA) concentration was higher in GM than in CG (P < 0.01). In addition, CG had higher prepartum serum glucose concentration than GM (P = 0.03). In the early postpartum period, aspartate aminotransferase (AST) activity was lower in CG than in GSUB (P < 0.05), and in the late postpartum period, AST activity was lower in CG than GSUB and GM (P = 0.01). Somatic cell count was higher during the early and late postpartum periods for GM and GSUB when compared to CG (P < 0.01). In this study, primiparous cows with low glucose and higher NEFA in the prepartum were more susceptible for mastitis in the early postpartum, probably due to low immune function associated to a more negative energy balance. In sum, increased prepartum serum NEFA concentration and decreased glucose in primiparous cows were associated with clinical mastitis incidence in the postpartum period.

Association between chronic Anaplasma marginale and Babesia spp. infection and hematological parameters of taurine heifers.

The aim of this study was to investigate the association between chronic Anaplasma marginale and Babesia spp. infection and hematological parameters of pregnant and non-pregnant taurine heifers. Blood samples from 94 females were collected on the first day (D-10) of timed artificial insemination (TAI) protocol and on pregnancy diagnosis (D+34). Hematological parameters were determined and compared between pregnant (PG) and non-pregnant (NPG) heifers, and within group at different sampling days. Real-time PCR (qPCR) was used to determine A. marginale and Babesia bovis infection, and for absolute quantification of Babesia spp. between PG and NPG groups. Correlation analysis was performed between the number of gDNA copies (CN) of Babesia spp. and hematological parameters. On D-10, mean hemoglobin concentration was higher for NPG, and hematocrit and total plasma protein were higher on D+34 for both groups. There was no difference in Babesia spp. CN between groups. In the first qPCR, all heifers were positive for A. marginale and B. bovis. Significant correlations were found between hemoglobin and erythrocyte and between hemoglobin and hematocrit (r = 0.8082 and r = 0.3009, respectively). Low levels of A. marginale and Babesia spp. did not affect hematological parameters of chronically infected pregnant and non-pregnant taurine heifers.

Feed intake, milk production and metabolism of Holstein, Gyr and Girolando-F1 heifers with high body condition score during the transition period.

The transition period in taurine dairy cows is widely reported in the literature. However, little is known about the metabolism of zebu animals and their crossbreeding with taurine breeds during this phase. Considering the importance of these breeds in tropical and subtropical regions, this study aimed to evaluate the feed intake, milk production and hepatic metabolism in Holstein, Gyr and Girolando-F1 (½ Holstein × ½ Gyr) heifers presenting high body condition score (BCS) during the transition period (prepartum weeks -2 and -1 and postpartum weeks +1 and + 3). Twelve heifers of each genetic group were used, totaling 36 animals. Variables considered were blood metabolites related to liver function, dry matter intake (DMI), body condition score (BCS), body weight (BW), milk yield (MY), and fat and protein concentrations in milk. Gyr heifers had the lowest concentrations of paraoxonase in weeks -2 (43.13 U/mL), -1 (62.10 U/mL) and + 3 (77.89 U/mL), albumin in week -1 (3.07 g/dL), and the highest concentration of non-esterified fatty acids (NEFA) in weeks -2 (1.35 mmol/L) and -1 (1.19 mmol/L). ß-hydroxybutyrate (BHB) and NEFA values were negatively correlated with prepartum DMI (-0.82 and -0.57, respectively), while paraoxonase was positively correlated to DMI (0.54). Gyr cows were more susceptible to inflammation despite having intermediate BCS and lower milk yield. Girolando-F1 animals showed the highest BCS among groups, although their hepatic metabolism had better results than Gyr cows. Holstein animals had lower lipomobilization and higher DMI and MY between breeds. These results suggest that the metabolism of zebu and crossbred cows does not react equally to the metabolism of Holstein cows concerning stress factors such as transition period and obesity. Therefore, the present study addresses an emerging theme that highlights the need for differentiated management during the transition period between the different breeds studied in order to ensure the maximum health and welfare of these animals.

Butaphosphan Effects on Glucose Metabolism Involve Insulin Signaling and Depends on Nutritional Plan.

Butaphosphan is an organic phosphorus compound used in several species for the prevention of rapid catabolic states, however, the mechanism of action remains unclear. This study aimed at determining the effects of butaphosphan on energy metabolism of mice receiving a normal or hypercaloric diet (HCD) and submitted or not to food restriction. Two experiments were conducted: (1) during nine weeks, animals were fed with HCD (n = 28) ad libitum, and at the 10th week, were submitted to food restriction and received butaphosphan (n = 14) or saline injections (n = 14) (twice a day, for seven days) and; (2) during nine weeks, animals were fed with a control diet (n = 14) or HCD (n = 14) ad libitum, and at the 10th week, all animals were submitted to food restriction and received butaphosphan or saline injections (twice a day, for seven days). In food restriction, butaphosphan preserved epididymal white adipose tissue (WAT) mass, increased glucose, NEFA, and the HOMA index. In mice fed HCD and submitted to food restriction, the butaphosphan preserved epididymal WAT mass. Control diet influences on PI3K, GCK, and Irs1 mRNA expression. In conclusion, butaphosphan increased blood glucose and reduced fat mobilization in overweight mice submitted to caloric restriction, and these effects are influenced by diet.

Association of polymorphisms in the IGF-I, GHR and STAT5A genes with serum IGF-I concentration and reproductive performance of Holstein dairy cows.

This study was conducted to evaluate associations of polymorphisms in the genes for the growth hormone receptor (GHR), insulin-like growth factor I (IGF-I) and signal transducer and activator of transcription 5A (STAT5A) with serum concentrations of IGF-I, reproductive performance and milk production of postpartum Holstein dairy cows. Days from calving to first ovulation (DTO) and calving to conception interval (CCI) were evaluated in 95 Holstein cows. Serum concentrations of IGF-I and ß-hydroxybutyrate (BHBA) were quantified in samples collected in sequential blood collections. Genotyping of the IGF-I and STAT5A genes was performed. The IGF-I polymorphism distribution was 35.9% CC, 46.1% CT and 18% TT. The IGF-I concentrations in circulation were greater in cows of the TT compared with both the CT and CC groups (P < 0.05). Genotype had a linear association (P < 0.05) with DTO and CCI, which were less for cows of the TT group. There was no association of STAT5A BstEII on serum IGF-I or reproductive variables (P> 0.05). When combining the GHR AluI T allele, obtained in a previous study, and the IGF-I SnaBI T allele from the current study, for the same cows, there were additive associations of both with serum IGF-I, BHBA, number of services per conception, DTO and CCI (P < 0.05). Thus, the IGF-1 SnaBI TT appears to be associated with fewer DTO and lesser CCI of lactating dairy cows and had an additive association with the GHR AluI T allele on indicators for improvement of fertility.

Effect of recombinant bovine somatotropin (rbST) treatment on follicular population and development in non-lactating dairy cows.

The aim of this study was to evaluate the long-term effects of recombinant bovine somatotropin (rbST) on follicle population and ovulatory follicle development in non-lactating dairy cows. Twenty-one Jersey cows were allocated in rbST (n=11) or control (n=10) groups. On day -60, cows in rbST group received 500 mg of somatotropin (s.c. Lactotropin®, Elanco). On day 0, control and rbST cows received an intravaginal progesterone-releasing device (1.9 g, CIDR®, Zoetis) and GnRH (100 mg, IM, Factrel®, Zoetis). On day 8, cows received PGF2α (25 mg, IM, Lutalyse®, Zoetis) and the CIDR® was removed. Twelve hours after device removal (D8), serum, follicular fluid and granulosa cells samples were collected. Serum and follicular concentration of estradiol (E2) and progesterone (P4) were analyzed. Total RNA was extracted from granulosa cells to measure gene expression of LHCGR, STAR, HSD-3B1, CYP11A1, CYP19A1, CYP17A1, IGFR and PAPPA by real-time PCR. Ultrasonography was performed on days -60, -53, -46, -14, -7, 0 and 8 for antral follicle count. Results were analyzed by repeated measures ANOVA and t-test. There was no effect of rbST treatment on the number of follicles during the 60 days period, as well as no effect on serum and follicular fluid E2 and follicular fluid P4 at the moment of follicle aspiration. There was a reduction in PAPPA (P = 0.006), CYP11A1 (P = 0.04) and CYP19A1 (P = 0.002) mRNA levels in granulosa cells of the pre-ovulatory follicle of rbST treated cows. In conclusion, a single dose of rbST did not have long-term effects on antral follicle population, serum and follicular E2/P4 concentrations in non-lactating dairy cows. Despite that, rbST injection decreased granulosa cell expression of genes related to steroidogenesis in the pre-ovulatory follicle.

Insulin treatment does not affect follicular development but alters granulosa cell gene expression in dairy cows.

The use of strategies to stimulate follicular growth are important, especially for use in timed artificial insemination (TAI) protocols, aiming to increase dairy cow's fertility. The aim of this study was to investigate the effect of insulin on follicular growth, steroid production and expression of genes related to follicular development. For this, cows were submitted to a progesterone (P4) and estradiol (E2) based synchronization protocol. In study 1, eleven primiparous lactating Holstein cows, received a single s.c. application of 0.25 IU/kg human insulin or no treatment (control) on D8 of the protocol. Blood samples were collected, and the dominant follicle diameter was assessed daily via transrectal ultrasonography, from D8 to D12. In study 2, eight multiparous non-pregnant and non-lactating Jersey cows, received a single s.c. application of 0.25 IU/kg human insulin, whereas cows from the control group received a single s.c. injection (1 mL) of saline solution (NaCl 0.9%). Blood samples were collected, and the dominant follicle diameter was assessed daily via transrectal ultrasonography from D6 to D9 of the protocol. Sixteen hours after insulin injection, follicular aspiration was performed. In study 1, insulin treatment decreased systemic glucose levels, but did not affect follicular growth. In study 2, the glucose decrease induced by insulin treatment was accompanied by a tendency of decreased progesterone levels in follicular fluid, along with a decrease in steroidogenic acute regulatory protein (STAR) and insulin like growth factor binding protein 2 (IGFBP2) mRNA abundance in granulosa cells. In conclusion, insulin treatment does not increase follicle growth and estradiol secretion in dairy cows, but decreases IGFBP2 and tends to increase pappalysin (PAPPA) mRNA abundance in granulosa cells, suggesting a positive effect on follicle development.

Effect of bovine somatotropin injection in late pregnant Holstein heifers on metabolic parameters and steroidogenic potential of the first postpartum dominant follicle.

The aim of this study was to determine the effect of pre-partum injections of bovine somatotropin (bST) in dairy heifers on metabolic markers and the steroidogenic potential of the first postpartum dominant follicle. Heifers were assigned to two groups: bST (ST; n = 29), that received two doses of bST (500 mg/dose) at -28 and -14 days relative to calving; and control (CTL; n = 30), that did not received bST. Follicular development was monitored via ultrasound every 3 days starting at 8 days in milk (DIM) in a subset of 20 heifers until the day the first large follicle reached a diameter of 16 mm. From these cows follicular fluid was aspirated and the follicular cells recovered (ST; n = 8 and CTL; n = 10). Blood samples were collected weekly for all heifers. Follicular fluid IGF-I concentrations of the first postpartum dominant follicle was higher (P = 0.05) in ST (87.1 ± 7.7 mg/mL) than CTL cows (64.3 ± 6.8 mg/mL). Also, E2 concentration in the follicular fluid was higher (P = 0.02) for ST (199.7 ± 55.9 ng/mL) than CTL cows (74.5 ± 37.7 ng/mL). The expression of LHCGR and STAR mRNA in follicular cells was higher (P < 0.05) in ST than CTL cows. Nonetheless, HSD3B, P450scc, P450c17, IGFr and CYP19A1 mRNA expression was not different between groups (P > 0.05). Serum IGF-I concentration was higher in ST treated heifers during the pre-partum period (P = 0.01) and no difference was observed in the postpartum period (P = 0.19). In conclusion, pre-partum bST treatment in dairy heifers increased intrafollicular IGF-I and expression of LHCGR and STAR mRNA in follicular cells of the first postpartum dominant follicle. These changes were associated to increased intrafollicular and serum E2 concentration, which can potentially increase the chance of ovulation of the first follicular wave.

Association between polymorphisms in somatotropic axis genes and fertility of Holstein dairy cows.

The aim of this study was to determine the effect of growth hormone receptor (GHR) AluI, insulin-like growth factor type 1 (IGF-I) SnaBI, and signal transducer and activator 5A (STAT5A) BstEII polymorphisms in the reproductive performance of Holstein dairy cows and the frequency of this genotypes in cows managed in different systems. This work studied 381 and 506 Holstein cows from semiextensive and intensive systems, respectively. The frequency of genotypes GHRAluI (+/-), IGF-ISnaBI (-/-) and (+/-), and STAT5ABstEII (-/-) was higher in animals from semiextensive system, whereas the frequency of the genotypes GHRAluI (+/+) and IGF-ISnaBI (+/+) was higher in animals from intensive system (P < 0.05). In the intensive system, cows from the STAT5ABstEII (-/-) genotype had a longer calving-first heat interval (P = 0.03). In conclusion, there was no association between the genotypes of GHRAluI and IGF-ISnaBI and fertility of Holstein cows raised in semiextensive or intensive regimes, although the genotype frequencies of the evaluated polymorphisms were different between the studied systems. The STAT5ABstEII polymorphism was associated with calving-first heat interval in Holstein cows raised in the intensive system, indicating that this gene could be a molecular marker for genetic selection to improve reproductive performance.