Unveiling the Role of IGF-I in Fertility: Effect of Long-Acting Bovine Somatotropin (bST) on Terminal Follicular Development and Fertility during an Annual Reproductive Cycle in Sheep.

The study aimed to assess the effect of long-acting bST treatment, in a dose that only increases IGF-I plasma concentrations, on ovarian and fertility markers of estrous synchronized ewes that were fed to keep their bodyweight. Three experiments were designed to evaluate this effect: in Experiment 1, 18 ewes were distributed in groups (bST 0, 30, 50 mg) to measure plasma IGF-I and insulin for 15 days; in Experiment 2, 92 ewes (5 replicates) in two groups (0 and 30 mg bST) were synchronized using a 6-day progesterone protocol during the breeding season to assess the effect of bST on follicular and luteal performances, estrous and ovulation, and fertility after mating. In Experiment 3, 50 ewes (3 replicates) were used to repeat the study before but during anestrus. Results indicate that 50 mg bST increased IGF-I and insulin plasma concentrations, but 30 mg bST only increased IGF-I concentrations; and that only during the breeding season did 30 mg bST increase the number of lambs born and the reproductive success of ovulatory-sized follicles compared to controls. This occurred without it affecting any other reproductive marker. In conclusion, 30 mg bST treatment may improve oocyte competence for fertility during the breeding season.

Effect of recombinant bovine somatotropin on the reproductive efficiency of beef cows subjected to differently timed-artificial insemination protocols.

This study aimed to verify the reproductive efficiency of beef cows treated with recombinant bovine somatotropin (rbST). Study 1, Bos indicus cows were distributed (three groups). The control group (CG) was subjected: on day zero (d0), the animals received a CIDR and oestradiol benzoate (EB); on (d8, CIDR was removed, and PGF2α and oestradiol cypionate (EC) were administered; on d10, timed Artificial Insemination (TAI) was performed; on d45, pregnancy diagnosis was made. The rbST on d0 group (bST0G) was subjected to an identical protocol as CG, except for the addition of 250 mg rbST on d0. The rbST on d8 group (bST8G) was subjected to the same protocol as bST0G, except that the rbST was administered on d8 rather. In study 2, the animals followed the same design which was used in Bos taurus cows. The follicular growth rate (FGR) was calculated between d8 and d10. In study 1, pregnancy/artificial insemination (P/AI) did not differ among the treatments. FGR in bST8G was higher than in other groups. In study 2, bST0G showed higher Pregnancy/Artificial Insemination (P/AI) (p < .05) when compared with other groups. bST0G showed a different FGR (p < .0001) than the other groups. In conclusion, rbST (Bos indicus cows) did not increase P/AI, but it did promote follicular growth when administered on d8; the rbST administered on d0 improved P/AI (p < .05) and the FGR in Bos taurus cows.

Use of new recombinant proteins for ovarian stimulation in ruminants.

Currently, gonadotropin products (follicle stimulating hormone, FSH, and luteinizing hormone, LH) used in animal reproduction are produced by extraction and purification from abattoir-derived pituitary glands. This method, relying on animal-derived materials, carries the potential risk of hormone contamination and pathogen transmission. Additionally, chorionic gonadotropins are extracted from the blood of pregnant mares (equine chorionic gonadotropin; eCG) or the urine of pregnant women (human chorionic gonadotropin; hCG). However, recent advancements have introduced recombinant gonadotropins for assisted animal reproduction therapies. The traditional use of FSH for superovulation has limitations, including labor requirements and variability in superovulation response, affecting the success of in vivo (SOV) and in vitro (OPU/IVEP) embryo production. FSH treatment for superstimulation before OPU can promote the growth of a homogenous follicular population and the recovery of competent oocytes suitable for IVEP procedures. At present, a single injection of a preparation of long-acting bovine recombinant FSH (rFSH) produced similar superovulation responses resulting in the production of good-quality in vivo and in vitro embryos. Furthermore, the treatment with eCG at FTAI protocol has demonstrated its efficacy in promoting follicular growth, ovulation, and P/AI, mainly in heifers and anestrous cows. Currently, treatment with recombinant glycoproteins with eCG-like activity (r-eCG) have shown promising results in increasing follicular growth, ovulation, and P/AI in cows submitted to P4/E2 -based protocols. Bovine somatotropin (bST) is a naturally occurring hormone found in cows. Recombinant bovine somatotropin (rbST), produced through genetic engineering techniques, has shown potential in enhancing reproductive outcomes in ruminants. Treatment with rbST has been found to improve P/IA, increase donor embryo production, and enhance P/ET in recipients. The use of recombinant hormones allows to produce non-animal-derived products, offering several advantages in assisted reproductive technologies for ruminants. This advancement opens up new possibilities for improving reproductive efficiency and success rates in the field of animal reproduction.

Microsphere Peptide-Based Immunoassay for the Detection of Recombinant Bovine Somatotropin in Injection Preparations.

The use of peptides in immunoassays can be favored over the use of the full protein when more cost effective or less toxic approaches are needed, or when access to the full protein is lacking. Due to restricted access to recombinant bovine somatotropin (rbST), a protein enhancing growth and lactating performances of livestock, which use has been banned in the EU, Canada and Australia (amongst others), we developed a peptide-based biorecognition assay on an imaging planar array analyzer. For this, we identified the rbST epitope that is responsible for binding to the rbST-targeting monoclonal antibody 4H12 (MAb 4H12) to be 115DLEEGILALMR125. This linear peptide was synthesized and coupled to microspheres, after which it was tested in a biorecognition competitive inhibition assay format. We observed IC50 values of approximately 0.11 µg mL-1, which are lower than observed for the full rbST protein (IC50 = 0.20 µg mL-1). Importantly, there was no binding with the scrambled peptide. Preliminary results of directly coupled peptides in a microsphere biorecognition assay for detection of rbST are presented. Real-life applicability for detection of somatotropins (STs) in injection preparations of bovine-, porcine- and equine ST are shown. This newly developed immunoassay strongly supports future developments of peptide-based immunoassays to circumvent the limited access to the full protein.

Exogenous bovine somatotropin and mist-fan cooling synergistically promote the intramammary glucose transport for lactose synthesis in crossbred Holstein cows in the tropics.

BACKGROUND AND AIM: Milk synthesis by the mammary gland is negatively influenced in part by high ambient temperature (AT). This study aimed to clarify the pathway of intramammary glucose utilization involved in mediating lactose synthesis during treatment with somatotropin under housing with misters and fans. MATERIALS AND METHODS: A single subcutaneous injection of 500 mg of recombinant bovine somatotropin (rbST) was administered 3 times once every 14 days to 87.5% crossbred Holstein cattle in early-/mid-/late lactation, under housing in a normal shaded barn and in a shaded barn with a mist-fan cooling system. RESULTS: The milk yields of the cooled cows tended to increase compared with those of uncooled cows and exhibited more potentiated effects in response to rbST treatment, coinciding with increases in mammary plasma flow and glucose uptake, but not in the mammary extraction of glucose. Treatment with rbST in the cooled cows resulted in a greater increase in the milk glucose concentration and a greater decrease in the milk glucose-6-phosphate concentration at all stages of lactation. CONCLUSION: rbST treatment exerted its galactopoietic action more by local intramammary factors than by other extramammary factors at a low AT and the synergistic effect between rbST treatment and low AT increased the availability of intramammary glucose transport in activating the process of lactose synthesis.

Detection of methionine- and alanine-recombinant bovine somatotropins and their induced antibodies in serum and milk of cows suggests blood-milk barrier specificity for these compounds.

The elimination of recombinant bovine somatotropin (rbST) and its induced antibodies through milk of 2 formulations is studied to propose a control strategy for its use or abuse. Two dairy cows were treated with alanine-rbST (Ala-rbST), which is identical to endogenous bovine somatotropin, and ten dairy cows were treated with methionine-rbST (Met-rbST), which differs by 1 amino acid from endogenous bovine somatotropin. We developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method able to measure rbST at a decision limit (CCα) of 0.8 ng/mL and 2.3 ng/mL for serum and milk, respectively. The results show that the administered Ala-rbST is transferred from blood to milk but that this is not the case for Met-rbST. This suggests a blood-milk barrier-related specificity for these compounds. In addition, rbST-induced antibodies were formed in animals treated with Ala-rbST and those treated with Met-rbST. In both treatments, the rbST-induced antibodies were transferred from blood to milk, showing no blood-milk barrier specificity for these antibodies. These elimination patterns show that, for enforcement purposes, the detection of rbST-induced antibodies in tank milk can serve to screen for rbST administration, and subsequent confirmatory serum analysis by LC-MS/MS is needed to identify whether Ala-rbST or Met-rbST has been used.

Effects of administering exogenous bovine somatotropin to beef heifers during the first trimester on conceptus development as well as steroid- and eicosanoid-metabolizing enzymes.

The aim of this study was to evaluate the effect of bovine somatotropin (bST) on fetal and placental development during the first third of gestation in beef heifers. Angus heifers (n = 97) were randomly assigned to either receive a 500-mg injection of bST (BST) biweekly on days 0, 15, 29, 43, and 57 of gestation or not receive bST (CTL) throughout the experiment. Body weight (BW) was assessed on days -9, -3, 0, 15, 22, 29, 43, 50, 57, 64, and 77, while blood samples were collected on days 0, 22, 50, and 64. Pregnancy status was determined via transrectal ultrasonography on days 29 and 64. A subset of pregnant heifers (BST, n = 7; CTL, n = 5) were harvested on day 84, and complete gravid reproductive tracts and liver tissue were collected for analysis. Cytochrome P450 1A (CYP1A), 2C (CYP2C), 3A (CYP3A), and uridine 5'-diphospho-glucuronosyltransferase (UGT) activities were determined. Mean change in BW and average daily gain of heifers between fixed-time artificial insemination (day 0) and day 77 did not differ between treatments (P ≥ 0.05). Mean concentrations of insulin-like growth factor 1 (IGF-1) were greater (P < 0.001) in BST (347 ± 27.7 ng/mL) compared with CTL (135 ± 32.8 ng/mL) heifers. Mean placental weight, fetal membrane weight, uterine weight, and ovarian and corpus luteum (CL) weights, as well as fetal morphometric data, did not differ (P ≥ 0.05) between treatments. However, BST heifers had greater (P = 0.03) quantities of combined fetal fluid compared with CTL (521.6 ± 22.9 vs. 429.6 ± 27.14 g, respectively). Tendencies were observed for BST heifers to have reproductive tracts with fewer placentomes (P = 0.08) and fetuses with greater umbilical diameters (P = 0.09) compared with CTL. The activity of CYP1A did not differ (P ≥ 0.05) within the maternal and fetal liver, caruncle, cotyledon, or CL tissue samples between treatments. Furthermore, CYP3A activity was only observed in maternal liver samples and was not different between treatments (P ≥ 0.05). Interestingly, CYP2C activity was greater (P = 0.01) in the liver of BST vs. CTL heifers, and UGT activity was greater (P = 0.02) in the CL from BST heifers compared with CTL. In conclusion, the administration of bST during the first third of gestation increased plasma concentrations of IGF-1, which resulted in an increase in fetal fluid, decrease in placentome number, and greater umbilical diameter, but failed to alter fetal development.

Impact of recombinant bovine somatotropin, progesterone, and estradiol benzoate on ovarian follicular dynamics in Bos taurus taurus cows using a protocol for estrus and ovulation synchronization.

This study aimed to evaluate the effect of recombinant bovine somatotropin (bST) in combination with progesterone (P4) and estradiol benzoate (EB) on ovarian follicular dynamics using a protocol for estrus and ovulation synchronization in crossbred Bos taurus taurus cows. Twenty-four non-lactating multiparous cows were randomly assigned to two groups: the recombinant bovine somatotropin group (GbST; n = 11) received an intravaginal P4 device (1.5 g), estradiol benzoate (EB = 1.0 mg IM), bST (500 mg SC), and an ovarian ultrasonography (US) on day zero (d0 = beginning of the study); d-cloprostenol (150 µg, IM), US, and P4 removal on d8; 1.0 mg of EB (IM) on d9; and US on d10 and d15. On the other hand, to the control group (GC; n = 13), the same protocol as the GbST was applied, except for the non-receipt of bST on d0. The follicles were measured and evaluated on d0, d8, and d10, as were the corpora lutea (CL) on d15 (using ultrasonography). The effect of the two treatments (GbST vs. GC) on the follicle size, CL (F-test), and ovulation rate (logistic regression) were evaluated. The GbST showed a greater follicle diameter on d10 (14.5 mm) than the GC (12.1 mm; P < 0.03), as well as a greater diameter of CL on d15 (19.7 vs. 16.9 mm, P < 0.01). In addition, in the former, the ovulation rate (90.9 vs. 69.2%, P = 0.09) was observed to be greater. It was concluded that the combination of bST, P4, and EB in synchronization for estrus and ovulation protocols significantly increased the diameter of the preovulatory follicle, produced a higher follicular growth rate, and a greater diameter of the corpus luteum. Additionally, there was a higher percentage of cows with ovulation compared to the group that did not receive bST.

Impact of recombinant bovine somatotropin, progesterone, and estradiol benzoate on ovarian follicular dynamics in Bos taurus taurus cows using a protocol for estrus and ovulation synchronization.

This study aimed to evaluate the effect of recombinant bovine somatotropin (bST) in combination with progesterone (P4) and estradiol benzoate (EB) on ovarian follicular dynamics using a protocol for estrus and ovulation synchronization in crossbred Bos taurus taurus cows. Twenty-four non-lactating multiparous cows were randomly assigned to two groups: the recombinant bovine somatotropin group (GbST; n = 11) received an intravaginal P4 device (1.5 g), estradiol benzoate (EB = 1.0 mg IM), bST (500 mg SC), and an ovarian ultrasonography (US) on day zero (d0 = beginning of the study); d-cloprostenol (150 µg, IM), US, and P4 removal on d8; 1.0 mg of EB (IM) on d9; and US on d10 and d15. On the other hand, to the control group (GC; n = 13), the same protocol as the GbST was applied, except for the non-receipt of bST on d0. The follicles were measured and evaluated on d0, d8, and d10, as were the corpora lutea (CL) on d15 (using ultrasonography). The effect of the two treatments (GbST vs. GC) on the follicle size, CL (F-test), and ovulation rate (logistic regression) were evaluated. The GbST showed a greater follicle diameter on d10 (14.5 mm) than the GC (12.1 mm; P < 0.03), as well as a greater diameter of CL on d15 (19.7 vs. 16.9 mm, P < 0.01). In addition, in the former, the ovulation rate (90.9 vs. 69.2%, P = 0.09) was observed to be greater. It was concluded that the combination of bST, P4, and EB in synchronization for estrus and ovulation protocols significantly increased the diameter of the preovulatory follicle, produced a higher follicular growth rate, and a greater diameter of the corpus luteum. Additionally, there was a higher percentage of cows with ovulation compared to the group that did not receive bST.

Development of a confirmatory method for detecting recombinant bovine somatotropin in plasma by immunomagnetic precipitation followed by ultra-high performance liquid chromatography coupled to tandem mass spectrometry.

Recombinant bovine somatotropin (rbST), a synthetic growth hormone, is used to stimulate growth and enhance milk production in dairy cows. Both its use and the sale of dairy products from treated animals are prohibited in the European Union, as well as in Australia, Canada, Japan, and New Zealand, but authorised in several countries (e.g. Brazil, USA). Screening methods involve detecting anti-rbST antibodies (biomarkers) in treated cows. Confirmatory methods are required to prove rbST abuse. The major challenges in determining rbST are its potentially low levels, its high similarity to native bST, and matrix interferences. To overcome these obstacles, we have developed a method involving immunomagnetic precipitation followed by UHPLC-MS/MS for rbST detection. Briefly, protein G magnetic beads pre-coated with an in-house produced monoclonal antibody were added to plasma. Incubation at room temperature allowed rbST present in the sample to bind to the magnetic beads. After that, magnetic beads were isolated by centrifugation and thoroughly washed (PBS, PBS + 0.2% Tween 20). Finally, rbST was released by alkalinisation and the samples were trypsin digested prior to UHPLC-MS/MS analysis in the MRM mode. Validation was done in accordance with European Commission Decision 2002/657/CE. Matrix-matched calibration with internal standards was used. The decision limit (CCα) reached with this approach was 0.11 µg l-1.

Quantitative Risk Assessment of Antimicrobial-Resistant Foodborne Infections in Humans Due to Recombinant Bovine Somatotropin Usage in Dairy Cows.

Recombinant bovine somatotropin (rbST) is a production-enhancing technology that allows the dairy industry to produce milk more efficiently. Concern has been raised that cows supplemented with rbST are at an increased risk of developing clinical mastitis, which would potentially increase the use of antimicrobial agents and increase human illnesses associated with antimicrobial-resistant bacterial pathogens delivered through the dairy beef supply. The purpose of this study was to conduct a quantitative risk assessment to estimate the potential increased risk of human infection with antimicrobial-resistant bacteria and subsequent adverse health outcomes as a result of rbST usage in dairy cattle. The quantitative risk assessment included the following steps: (i) release of antimicrobial-resistant organisms from the farm, (ii) exposure of humans via consumption of contaminated beef products, and (iii) consequence of the antimicrobial-resistant infection. The model focused on ceftiofur (parenteral and intramammary) and oxytetracycline (parenteral) treatment of clinical mastitis in dairy cattle and tracked the bacteria Campylobacter spp., Salmonella enterica subsp. enterica, and Escherichia coli in the gastrointestinal tract of the cow. Parameter estimates were developed to be maximum risk to overestimate the risk to humans. The excess number of cows in the U.S. dairy herd that were predicted to carry resistant bacteria at slaughter due to rbST administration was negligible. The total number of excess human illnesses caused by resistant bacteria due to rbST administration was also predicted to be negligible with all risks considerably less than one event per 1 billion people at risk per year for all bacteria. The results indicate a high probability that the use of rbST according to label instructions presents a negligible risk for increasing the number of human illnesses and subsequent adverse outcomes associated with antimicrobial-resistant Campylobacter, Salmonella, or E. coli .

Hepatic mRNA expression for genes related to somatotropic axis, glucose and lipid metabolisms, and inflammatory response of periparturient dairy cows treated with recombinant bovine somatotropin.

Objectives of this experiment were to evaluate the effects of recombinant bovine somatotropin (rbST) treatment of periparturient dairy cows on hepatic mRNA expression for genes related to the somatotropic axis, insulin, glucose, and lipid metabolism, inflammation, and oxidative stress. Holstein cows were enrolled in the experiment at 253 ± 3 d of gestation and assigned to 1 of 3 treatments: untreated control (n = 53), 87.5 mg of rbST (n = 56; rbST87.5), and 125 mg of rbST (n = 57; rbST125). Cows in the rbST87.5 and rbST125 treatments received weekly injections of rbST from -21 to 28 d relative to calving. A subsample of cows (control = 20, rbST87.5 = 20, rbST125 = 20) was randomly selected for collection of liver samples according to expected calving date, BCS, and previous lactation 305-d mature equivalent milk yield. Only cows that had liver sampled at -21 ± 3, -7 ± 3, and 7 ± 3 d relative to calving were used in the current experiment. Blood, sampled weekly from -28 to 21 d relative to calving, was used to determine the concentrations of growth hormone, insulin-like growth factor 1, insulin, cortisol, fatty acids, ß-hydroxybutyrate, glucose, haptoglobin, and tumor necrosis factor-α. Liver samples were used to determine hepatic mRNA expression of 50 genes. Treatment with rbST increased growth hormone concentrations during the postpartum period (control = 9.0 ± 0.7, rbST87.5 = 15.3 ± 1.0, rbST125 = 18.5 ± 1.3 ng/mL) and increased insulin-like growth factor 1 concentrations during the prepartum period (control = 107.4 ± 7.2, rbST87.5 = 126.9 ± 6.6, rbST125 = 139.4 ± 6.9 ng/mL). Control cows had greater postpartum concentrations of ß-hydroxybutyrate (control = 776.4 ± 64.0, rbST87.5 = 628.4 ± 59.7, rbST125 = 595.4 ± 60.9 µmol/L) than rbST cows. The rbST87.5 and rbST125 treatments upregulated the hepatic mRNA expression for somatotropic axis genes (GHR, GHR1A, IGF1, IGFBP3, and SOCS2) on d -7 relative to calving and upregulated the mRNA expression for SOCS2 on d 7. On d -7, rbST87.5 and rbST125 treatments increased mRNA expression for genes involved in hepatic lipid transport (ANGPTL4, APOA5, APOB100, and SCARB1) and downregulated mRNA expression for PPARD, which is involved in lipid storage. On d 7, rbST tended to upregulate the mRNA expression for genes involved in gluconeogenesis (PCK1) and fatty acid ß-oxidation (ACOX1), and downregulated the mRNA expression for genes involved in inflammation (TNFRSF1A, ICAM1, CXCL1, MYD88, HIF1A, IL1RN, NFKBIA, and SOCS3) and oxidative stress (XBP1). Administration of rbST during the periparturient period may improve liver function and health by increasing hepatic capacity for gluconeogenesis and lipid transport and by reducing inflammation and oxidative stress.

Serum responsiveness to recombinant bovine somatotropin in buffalo: a three-month lactation study using an acid-stripping ELISA for screening.

The misuse of recombinant bovine somatotropin (rbST) to increase milk yield involves buffalo not just cows. Screening methods to identify rbST-treated cattle have already been proposed. However, there have been no studies on prolonged periods with a high number of animals. In this study, we developed a new enzyme-linked immunosorbent assay (ELISA) to measure the serum responsiveness towards rbST, based on an acid-stripping procedure and relatively simple integral calculation dilution curves. We also applied the analysis to 640 serum and 96 milk samples collected from 16 buffalo treated with rbST and 16 controls, over a period of approximately three months. Its suitability as a screening method, in compliance with EU law, was also assessed. A bi-factorial approach was also evaluated, including the measurement of insulin-like growth factor 1 concentration in serum. Results showed that our ELISA could be used on its own for screening purposes, without the need to assess other biomarkers. Copyright © 2016 John Wiley & Sons, Ltd.

A LC-MS-MS method to detect recombinant bovine somatotropin misuse in buffalos.

Recombinant bovine somatotropin (rbST) is a peptide hormone used to increase milk yield in cows and buffalos. In Europe, its use has been banned. However, rbST is sometimes illegally included in zootechnical practices for profit purposes, undermining the fair trade and the law prescriptions. For this reason, efficient and reliable analytical techniques are required to contrast rbST misuse. A few LC-MS-MS methods have been developed to detect, in cow serum, methyonil-rbST, one of the two main rbST forms available on the market. The other form, which is widespread, is identical to the most abundant variant of bovine somatotropin (bST) and differs from the buffalo somatotropin for one amino acid in the N-terminus. For this reason, it is technically possible to distinguish both rbST forms in serum of buffalos. In this work, we describe a novel LC-MS-MS-based method, capable to quantify, with a high sensitivity and selectivity, the methyonil-rbST and the other bST-identical recombinant form in buffalo serum, previously purified using a solid-phase extraction procedure. The method was internally validated and used to analyse 152 serum samples, collected from eight buffalos administered with rbST for a period of 3 months, according to conventional protocols. The obtained results confirmed the suitability of the method in the detection of illegal hormonal treatments. Graphical abstract ᅟ.

The effect of bovine somatotropin on the cost of producing milk: Estimates using propensity scores.

Annual farm-level data from New York dairy farms from the years 1994 through 2013 were used to estimate the cost effect from bovine somatotropin (bST) using propensity score matching. Cost of production was computed using the whole-farm method, which subtracts sales of crops and animals from total costs under the assumption that the cost of producing those products is equal to their sales values. For a farm to be included in this data set, milk receipts on that farm must have comprised 85% or more of total receipts, indicating that these farms are primarily milk producers. Farm use of bST, where 25% or more of the herd was treated, ranged annually from 25 to 47% of the farms. The average cost effect from the use of bST was estimated to be a reduction of $2.67 per 100 kg of milk produced in 2013 dollars, although annual cost reduction estimates ranged from statistical zero to $3.42 in nominal dollars. Nearest neighbor matching techniques generated a similar estimate of $2.78 in 2013 dollars. These cost reductions estimated from the use of bST represented a cost savings of 5.5% per kilogram of milk produced. Herd-level production increase per cow from the use of bST over 20 yr averaged 1,160 kg.

Effect of administration of recombinant bovine somatotropin on health and performance of lactating dairy cows diagnosed with hyperketonemia.

The effect of administering recombinant bovine somatotropin (rbST) to cows with hyperketonemia during the early postpartum period on health, metabolic parameters, milk production, and early reproductive performance was evaluated in a double-blinded clinical trial. Cows from 8 dairy herds in New York State were tested weekly between 3 and 16d in milk for elevated serum ß-hydroxybutyrate. Cows were enrolled in the study when blood ß-hydroxybutyrate was ≥1.3mmol/L for the first time. Enrolled cows were randomly assigned to a treatment (n=273) or placebo control (n=270) group. Treated cows were given 325mg of rbST subcutaneously on the day of enrollment and again 14d later. Control cows received the same regimen except the syringe contained only the carrier without somatotropin. After enrollment, blood samples were collected weekly for 4wk and submitted to the laboratory to be analyzed for selected metabolites. Risk ratios for clinical diseases subsequent to treatment were calculated using Poisson regression. Continuous data were analyzed using linear mixed models. Time to first insemination was assessed with survival analysis. In the 42d following the first administration of rbST, incidence risks of displaced abomasum, clinical ketosis, metritis, clinical mastitis, and lameness were not different between treatment groups. Cows treated with rbST had a slightly lower body condition score 28d after enrollment compared with control cows. In the 4wk following enrollment, serum nonesterified fatty acids and aspartate amino-transferase were slightly higher for treated than control cows, respectively. Serum glucose, calcium, haptoglobin, and ß-hydroxybutyrate were similar between groups. Treatment had no effect on resolution of hyperketonemia in any of the 4wk after enrollment. Milk production in either of the 2-wk periods after each treatment was not different between treated and control cows. Furthermore, milk production was not different between groups from enrollment to 98d in milk (42.6±0.6 and 42.1±0.7kg/d for treatment and control groups, respectively). Treatment had no effect on time to first insemination (83 and 74d in milk for treatment and control groups, respectively; hazard ratio=0.72) or first insemination pregnancy risk (27 and 29% for treatment and control groups, respectively; risk ratio=0.92). Based on the current results, it is not recommended to use a low dose of rbST as therapy for cows with hyperketonemia.

A carbon nanofiber-based label free immunosensor for high sensitive detection of recombinant bovine somatotropin.

A carbon nanofiber-based label free electrochemical immunosensor for sensitive detection of recombinant bovine somatotropin (rbST) was developed. In this immunosensor design, a mild site-directed antibody immobilization via interaction of boronic acid and oligosaccharide moiety found on Fc region of an antibody was performed to preserve the biological activity of antibody and improve the sensor's sensitivity. Electrochemical characterization of the immunosensor fabrication was carried out by differential pulse voltammetry (DPV) in Fe(CN)6(3-)/Fe(CN)6(4-) probe. A comparison study between different transducer platforms showed carbon nanofiber gave higher current signal response than single-walled carbon nanotube. In this work, calibration curve was obtained from the decrease of DPV peak current of Fe(CN)6(3-)/Fe(CN)6(4-) after immunocomplexed was formed. A linear relationship between DPV current change signal response and rbST concentrations from 1 pg/mL to 10 ng/mL (correlation coefficient of 0.9721) was achieved with detection limit of 1 pg/mL. Our developed immunosensor demonstrated high selectivity in cross-reactivity studies and a good percentage recovery in spiked bovine serum sample.

Influência do Hormônio do Crescimento na concentração de testosterona plasmática e nas características seminais de touros jovens e adultos da raça Nelore / Influence of Growth Hormone on plasma testosterone concentration and seminal characteristics of young and adult Nellore bulls

Estudou-se o efeito do Hormônio do Crescimento bovino (bGH) sobre os parâmetros seminais e a concentração sérica de testosterona de touros da raça Nelore. Dezesseis touros foram distribuídos em um delineamento fatorial 2 x 2 (duas idades e dois níveis de r-bST), com quatro animais em cada tratamento. As aplicações de r-bST foram realizadas a cada 14 dias, totalizando nove aplicações por animal, em um período experimental de 120 dias. As coletas de sêmen foram realizadas a cada 15 dias e, para determinar as concentrações de testosterona, foram realizadas coletas de sangue a cada quatro horas, num total de 24 horas, e a cada 30 dias a partir da primeira aplicação de r-bST. O sêmen dos touros adultos tratados com r-bST apresentaram maior motilidade e vigor (P<0,05) em relação aos animais controles (79,70±11,56% vs 64,06±23,65%) e (3,80±0,88 vs 2,92±1,25), respectivamente; entretanto, as demais características seminais não foram influenciadas (P>0,05) pelo tratamento com r-bST. Os aspectos físicos do sêmen dos touros jovens não foram afetados (P>0,05) pelo tratamento com r-bST. Entretanto o tratamento com r-bST diminuiu os defeitos morfológicos menores (delgado, gigante normal, pequeno normal, globoso, cabeça isolada normal, abaxial, cauda dobrada e enrolada levemente e gota distal) em relação aos jovens não tratados (P<0,05). As concentrações séricas de testosterona foram influenciadas (P<0,05) pelo tratamento, aos 120 dias após a primeira aplicação do r-bST, e apresentaram tendência de três picos no período de 24 horas.

The aim of this trial was to study the effect of Growth Hormone (GH) on the seminal parameters and testosterone profile from Nellore bulls. Sixteen bulls were allocated in a 2 x 2 factorial arrangement (two ages and two r-bST levels), with four animals per treatment. The r-bST was injected every 14 days, with a total of nine injections per animal, during the 120 days of the experimental period. The semen collections were done every 15 days and blood samples were collected in four hour intervals for 24 hours, and in 30 day intervals from the first r-bST injection for testosterone profile. The semen of adult bulls treated with r-bST showed greater MOT and VIG (P<0.05) in relation to the control (79.70±11.56% vs 64.06±23.65%) and (3.80±0.88 vs 2.92±1.25) respectively; however, the other evaluated seminal parameters were not affected (P>0.05) by the r-bST treatment. The physical measurements of the semen of young animals was also unaffected (P>0.05) by r-bST injection. However, the treatment with r-bST decreased the number of minor morphologic defects (narrow head, giant head, small normal head, round head, free normal head, abaxial implantation, simple bent tail and distal droplet) compared to untreated young bulls (P<0.05). The serum testosterone concentration was influenced (P<0.05) by treatment, at 120 days after the first injection of r-bST and they showed a tendency of three picks in a 24 hour period of sampling.

Effect of prepartum administration of recombinant bovine somatotropin on health and performance of lactating dairy cows.

A double-blind, randomized clinical trial was conducted in 5 commercial dairy herds in southern Ontario with 1,362 cows enrolled to evaluate the effect of prepartum administration of recombinant bovine somatotropin (rbST) on health and performance. Cows were randomly assigned to receive either 325 mg of sometribove zinc suspension (n=680) or a placebo injection (n=682; control) subcutaneously every 14 d until calving. Treatments started 28 to 22 d before expected calving, with a maximum of 3 treatments per cow. Serum samples taken at the time of enrollment, 1 wk before calving, and weekly for 3 wk after calving were analyzed for nonesterified fatty acids (NEFA), ß-hydroxybutyrate (BHBA), glucose, aspartate aminotransferase, calcium, and haptoglobin. Diseases were recorded by farm staff. Incidences of clinical ketosis, clinical mastitis, displaced abomasum, metritis, retained placenta, milk fever, and lameness were similar between groups. Body condition score was lower for treated than for control cows at 3 wk after calving (3.13 and 3.17, respectively). Serum NEFA tended to be higher for treated than for control cows by 0.01 mmol/L. Overall BHBA was not different between groups, but BHBA for treated cows was higher in wk 1 after calving (750 and 698 µmol/L, respectively) and tended to be higher in wk 2 after calving (779 and 735 µmol/L, respectively). Incidence of hyperketonemia was similar between groups. Treated cows had higher serum glucose compared with control cows (2.8 and 2.7 mmol/L, respectively). We detected no differences in serum aspartate aminotransferase, calcium, or haptoglobin between groups. Milk yield was recorded daily for each cow for 63 d, and did not differ between groups (37.1 ± 0.5 kg and 36.7 ± 0.5 kg, respectively) but we detected a tendency for treated cows to produce 0.8 kg/d more milk than control cows in wk 1 after calving. We observed no difference between groups in the time from calving to first insemination or the probability of pregnancy at the first insemination. Groups did not differ in the proportion of anovular cows at 53 ± 3 d in milk based on serum progesterone measured from a subset of cows (38.0 and 34.3%, respectively, for treated and control groups). We found no difference between groups in dry matter intake from 21 d before calving to 63 d after calving in a subset of cows (17.4 ± 0.4 and 17.5 ± 0.4 kg/d, respectively). Based on results of the current study, biweekly (every 14 d) administration of rbST before calving to prevent disease and enhance performance is not recommended.

Efeito da somatotropina recombinante bovina (rbST) sobre os níveis de marcadores metabólicos de touros semiconfinados das raças Aberdeen Angus e Brangus / Effect of recombinant bovine somatotropin (rbST) on the concentrations of metabolic markers in semi-confined sires Aberdeen Angus and Brangus

O objetivo deste estudo foi avaliar o efeito da somatotropina recombinante bovina (rbST) sobre os marcadores do metabolismoenergético, proteico, mineral e hepático de touros mantidos em semiconfinamento. Foram utilizados 20 touros de raçasAberdeen Angus e Brangus, sendo divididos aleatoriamente em dois grupos (Placebo e rbST), para que no Placebo fosseadministrado via subcutânea duas doses de NaCl, 0,9% e no rbST fosse administrado o rbST (Lactotropin®). Foram realizadascinco coletas de sangue, com intervalo de 14 dias, para avaliação da concentração de triacilgliceróis e colesterol comomarcadores energéticos, ureia, como indicador proteico, fosfatase alcalina e fósforo como marcadores minerais e asenzimas aspartato aminotransferase (AST) e a gama glutamiltransferase (GGT) como indicadoras da função hepática. Aaplicação de rbST não influenciou os marcadores energéticos, minerais, e proteico, exceto a enzima hepática GGT (P 0,05).Estes resultados indicam que a utilização do rbST em touros não alterou o metabolismo, pois os parâmetros avaliadosmantiveram-se dentro dos padrões considerados fisiológicos para a espécie.

The aim of this study was to evaluate the effect of recombinant bovine somatotropin (rbST) on metabolic markers for energy, protein, mineral and hepatic metabolism of sires kept grazing and receiving concentrate. Twenty sires Aberdeen Angus and Brangus used for this experiment were divided in two groups (Placebo and rbST). The group Placebo received subcutaneous administration of NaCl, 0.9% and rbST receive rbST twice. Five blood collections with 14 days of interval were made for evaluation of triglyceride and cholesterol as energetic metabolism markers, urea, as proteic marker, alkaline phosphatase and phosphorus as minerals markers and aspartate aminotransferase (AST) and gamma glutamyltransferase (GGT) as indicators of the hepatic function. The rbST did not influence the energetic, mineral and proteic markers, except the hepatic enzyme GGT (P<0,05). These results indicated that the use of rbST in sires did not affect metabolism, therefore evaluated parameters remaining in the physiological range for the specie.