Publications by authors named "Gabriel Ribas Pereira"

18 Publications

  • Page 1 of 1

Testicular shape, scrotal skin thickness and testicular artery blood flow changes in bulls of different ages.

Reprod Domest Anim 2021 Jul 18;56(7):1034-1039. Epub 2021 May 18.

Deparment of Animal Science, State University of Londrina, Londrina, Paraná, Brazil.

The aim of this study was to compare the biometric testicular characteristics, skin thickness and haemodynamics of the testicular artery of 12- and 24-month-old bulls using Doppler ultrasonography, the study was conducted using 48 indicus-taurus animals. The scrotal circumference (SC) and biometry characteristics of the bulls were measured to calculate the testicular volume. Doppler ultrasonography was used to obtain the haemodynamic values of the testicular artery. The skin thickness and volume were lower (p<.01) in the younger bulls (12 months:4.68 ± 0.68 mm; 168.76 ± 47.96 cm ) versus 24 months (5.05 ± 0.89; 499.73 ± 129.24 cm ) animals (p<.01). During diastole, mean velocity was lower in the 12 months (7.98 ± 3.83) than in the 24 months (11.37 ± 4.15) animals (p <.05). The 12-month-old animals had higher pulsatility and resistivity indices (0.49 ± 0.02; 0.51 ± 0.20) compared to the 24-month-old animals (0.32 ± 0.16; 0.40 ± 0.15) (p < .05). The final testicular end velocity was lower in animals with long/moderate-shaped (L/M) (7.31 ± 2.91) than in those moderate/oval-shaped (M/O) (11.48 ± 3.88) testicles (p < .05). Animals with L/M testes presented higher pulsatility values and resistivity indices (0.51 ± 0.05; 0.55 ± 0.04) compared to animals with M/O shape (0.29 ± 0.20; 0.36 ± 0.15). We showed that the blood flow of the supra testicular artery between the two evaluated ages differed, and that 24-month-old bulls presented better thermoregulation capacity. Animals with a long/moderate testicular format presented a greater vascular resistance, which was imposed on the blood flow due to the anatomical differences in the testicular artery, resulting in lower velocity, and indicating better heat dissipation in this format.
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http://dx.doi.org/10.1111/rda.13947DOI Listing
July 2021

Bovine seminal plasma osteopontin: Structural modelling, recombinant expression and its relationship with semen quality.

Andrologia 2021 Feb 22;53(1):e13905. Epub 2020 Nov 22.

Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.

Osteopontin (OPN) is a multifunctional phosphoprotein that has been linked to fertility in bulls. However, the exact mechanism by which OPN contributes to fertilisation is yet unknown. The biotechnological use of OPN in bovine reproduction is promising but some gaps remain unfilled. The present work aimed: (a) to verify whether the seminal plasma OPN is associated with seminal traits and a standard breeding soundness exam; (b) to predict OPN interactions with integrins, CD44 and glycosaminoglycans through molecular docking; and (c) to develop a protocol for recombinant expression of OPN from vesicular gland cDNA. Ejaculates from top ranked bulls had higher amounts of seminal plasma OPN in comparison with bulls classified as questionable (p < .01). The structural modelling and molecular docking predictions indicated that bovine OPN binds to heparin disaccharide, hyaluronic acid and hyaluronan. In addition, docking studies described the binding complexes of OPN with CD44 and the integrin heterodimers α5β1 and αVβ3. Finally, expression of rOPN-6His was successfully obtained after 3 hr of induction with 0.5 mM IPTG at 37°C and a denaturing purification protocol resulted in efficiently purified recombinant OPN. The present results contribute to the development of biotechnological uses of OPN as a biomarker in bovine reproduction.
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http://dx.doi.org/10.1111/and.13905DOI Listing
February 2021

Effect of scrotal insulation on sperm quality and seminal plasma proteome of Brangus bulls.

Theriogenology 2020 Mar 9;144:194-203. Epub 2020 Jan 9.

Laboratório de Biotecnologia, Universidade do Vale do Taquari- Univates, 95914-014, Lajeado, RS, Brazil. Electronic address:

Seminal plasma (SP) contributes to sperm physiology and metabolism, prevents premature capacitation, and protects sperm against oxidative stress. In order to evaluate the impact of heat stress in the semen of tropically adapted Brangus breed and in their seminal plasma proteome, we studied the effects of scrotal insulation for 72 h. Semen samples from six bulls, between 7 and 8 years of age, were collected prior to scrotal insulation (pre-insulation), and at 4 and 11 wk after insulation. Seminal plasma samples were analyzed by 2D SDS-PAGE and liquid chromatography coupled with mass spectrometry (LC-MS/MS). Insulation caused decrease in vigour, gross and total motility after 4 wk of scrotal insult (P < 0.001). Total defects in sperm were higher after 4 wk compared to pre-insulation and 11 wk after scrotal insulation (P < 0.001). The analysis of the 2D protein profile of the SP resulted in the identification 183 unique protein spots in all gels evaluated. There was no difference in mean number of protein spots amongst time points. Eight protein spots were more abundant in SP after scrotal insulation, returning to the same expression level at 11 wk post-insulation. One spot had higher abundance at 11 wk post-insulation, and one spot had decreased abundance 4 wk after insulation. The ten protein spots with differential abundance amongst time points were identified as Seminal plasma protein PDC-109, Seminal plasma protein A3, Seminal plasma protein BSP-30 kDa, Spermadhesin-1 and Metalloproteinase inhibitor 2. The validation of these five proteins as biomarkers for thermal testicular stress in Brangus breed would allow the development of new biotechnologies that could improve bovine semen analysis in breeding systems in tropical and subtropical conditions. A close association between the identified BSP and Spermadhesin-1 was evidenced in protein-protein interaction analysis. Based on gene ontology analysis, variation in sperm function after insulation could be explained by variation in the expressed proteins in the SP. Further studies are required to verify if these proteins could be used as biomarkers for the identification of bulls with increased seminal resistance to heat stress in Brangus breed.
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http://dx.doi.org/10.1016/j.theriogenology.2020.01.014DOI Listing
March 2020

Effect of breed on testicular blood flow dynamics in bulls.

Theriogenology 2018 Sep 26;118:16-21. Epub 2018 May 26.

REPROA - Animal Reproduction Laboratory, Deparment of Animal Science, State University of Londrina, Rd. Celso Garcia Cid 445, Km 380 Campus Universitário, Londrina, Paraná, 86051-990, Brazil. Electronic address:

The evaluation of testicular hemodynamics can contribute significantly to the understanding of the thermoregulatory mechanisms and oxygen supply of the testis in domestic animals. The present study aimed to characterize circulatory dynamics using the mean velocity (MV), pulsatility index (PI) and resistive index (RI) of the supratesticular artery in bulls. We evaluated 334 bulls of five different breeds (Nelore, Hereford, Aberdeen Angus, Braford and Brangus) by performing a velocimetry analysis using Doppler ultrasonography. Data were compared by Welch's ANOVA, Games-Howell (post-hoc test) and Spearman correlation with a significance level of 5%. The overall MV of 12.14 ± 0.30 cm/s differed among breeds. In addition, we observed that Brangus bulls showed higher (P < 0.05) MV (16.28 ± 1.02 cm/s) compared to Nelore bulls (8.76 ± 0.40 cm/s). The RI had an overall mean of 0.41 ± 0.01 and differed among breeds. We observed higher (P < 0.05) RI values in Hereford (0.44 ± 0.01) compared to Brangus (0.36 ± 0.02) animals. Overall, the PI values (0.33 ± 0.01) did not differ (P > 0.05) among breeds. The correlation between the PI and RI (0.936; P < 0.001) was high and positive; however, the correlations were low and negative between MV and the PI (-0.228; P < 0.001) and between MV and the RI (-0.270; P < 0.001). We concluded that there are differences in the MV and RI of the bulls' supratesticular arteries among the different evaluated breeds. Moreover, the presented values attributed to blood flow dynamics can serve as parameters in future studies and can be used to identify alternative diagnostic tools for infertility or to understand issues of adaptability in bulls.
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http://dx.doi.org/10.1016/j.theriogenology.2018.05.022DOI Listing
September 2018

Infrared thermography relationship between the temperature of the vulvar skin, ovarian activity, and pregnancy rates in Braford cows.

Trop Anim Health Prod 2017 Dec 28;49(8):1787-1791. Epub 2017 Aug 28.

Animal Production Department, College of Veterinary Medicine, University of Northern Paraná - UNOPAR, PR 218, Km 01 Jardim Universitário, Arapongas, PR, 86702-670, Brazil.

The objective of this study was to compare the presence of ovarian activity and pregnancy rates to temperature variation at the vulvar skin measured by infrared thermography (IRT). In addition, we also aimed to evaluate the IRT as a non-invasive method to evaluate animal breeding from fixed timed artificial insemination (FTAI). The study comprises 150 non-lactating beef Braford cows (5/8 Hereford × 3/8 Nellore) aged between 3 and 10 years. Data were collected along the FTAI protocol period during animal management. Animals were subjected to reproductive ultrasound evaluation and thermal images were performed by an infrared camera. Mean skin vulvar temperature (°C) and ovarian structures data were compared using Tukey's t test used as follow-up test to ANOVA. We observed a statistical difference in the mean vulvar skin temperature between animals that had the presence of ovarian follicles (34.2 ± 1.8) compared to no activity (35.4 ± 1.0; P < 0.05). However, vulvar skin temperature were similar between pregnant (34.5 ± 1.5) compared to non-pregnant (34.3 ± 1.9) animals (P > 0.05). In conclusion, the IRT technique was efficient to detect changes on vulvar skin temperature observed during FTAI protocol in Braford cows. Therefore, the use of IRT as an indirectly diagnostic tool to detect ovarian activity seems promising and further studies are required to validate their potential in beef cattle production.
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http://dx.doi.org/10.1007/s11250-017-1378-5DOI Listing
December 2017

Performance of beef heifers supplemented with sodium lasalocid.

Trop Anim Health Prod 2017 Feb 12;49(2):273-279. Epub 2016 Dec 12.

Faculty of Agronomy, Department of Animal Science, Núcleo de Estudos em Sistemas de Produção de Bovinos de Corte e Cadeia Produtiva - NESPRO, Federal University of Rio Grande do Sul - UFRGS, Ave. Bento Gonçalves 7712, 91540-000, Porto Alegre, RS, Brazil.

This study was conducted on 78 13-month-old crossbred beef heifers that weighed 215 kg in Southern Rio Grande do Sul (RS) State, Brazil. We evaluated the performance of beef heifers that were reared in a pasture system that received a mineral supplement energy-type protein with added sodium lasalocid (LAS). The heifers were randomly and uniformly divided into 2 groups, with 39 animals in each group. One group of animals received a mineral supplement energy-type protein without sodium lasalocid (CON), and the other group received a mineral supplement energy-type protein with added LAS. The mean feed intake, the body weight (BW), the average daily gain (ADG), the body condition score (BCS), and ovarian cyclicity were recorded, and economic parameters were calculated. No differences in supplement intake were observed between the groups, which ensures adequate intake of the other components of the mineral mixture, which are part of the nutritional requirements for the production process. Similarly, no difference in the ADG was observed between treatments. We observed that the heifers in the LAS group had a higher BW gain (51 kg) that the CON heifers (40 kg; P < 0.05). In addition, LAS-supplemented heifers had a higher BCS (3.53) than CON heifers (3.38) at the end of the experiment (P < 0.05). The heifers supplemented with LAS had a higher profitability than the CON heifers, even with the higher cost of the supplement containing LAS; this effect was due to the higher live BW at the end of the study. We concluded that the administration of a mineral supplement energy-type protein with added LAS has beneficial effects on beef heifers in terms of production and economic feasibility.
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http://dx.doi.org/10.1007/s11250-016-1188-1DOI Listing
February 2017

Effects of ambient air temperature, humidity, and wind speed on seminal traits in Braford and Nellore bulls at the Brazilian Pantanal.

Int J Biometeorol 2016 Nov 11;60(11):1787-1794. Epub 2016 Apr 11.

Department of Animal Science, Federal University of Rio Grande do Sul, Av. Bento Gonçalves, n.: 7.712, Porto Alegre, RS, 91540-000, Brazil.

The aim of this study was to evaluate the bioclimatic thermal stress assessed by Equivalent Temperature Index (ETI) and Temperature Humidity Index (THI) on Braford and Nellore bulls sperm quality during the reproductive seasons at the tropical region in the Brazilian Pantanal. We used 20 bulls aged approximately 24 months at the beginning of the study. Five ejaculates per animal were collected using an electroejaculator. Temperature, air humidity, and wind speed data were collected every hour from the automatic weather station at the National Institute of Meteorology. Infrared thermography images data were collected to assess the testicular temperature gradient in each animal. Data were analyzed with ANOVA using MIXED procedure of SAS and means were compared using Tukey's HSD test. The THI and ETI at 12 days (epididymal transit) were higher in January (89.7 and 28.5, respectively) and February (90.0 and 29.0, respectively) compared to other months (P < 0.01). Total seminal defects differ only in Bradford bulls between the months of November and February. Nellore bulls had lower major defects (MaD) and total defects (TD) compared to Braford. Nellore bulls showed correlation between minor defects (MiD) and THI for 30 days (0.90) and 18 days (0.88; P < 0.05). Braford bulls showed correlation for MaD (0.89) in ETI for 12 days (P < 0.05). Infrared thermography showed no difference between animals. Reproductive response to environmental changes is a consequence of Nellore and Braford adaptation to climate stress conditions. Both THI and ETI environmental indexes can be used to evaluate the morphological changes in the seminal parameters in Nellore or Braford bulls; however, more experiments should be performed focusing on larger sample numbers and also in reproductive assessment during the consecutive years to assess fertility potential.
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http://dx.doi.org/10.1007/s00484-016-1167-2DOI Listing
November 2016

The uses of infrared thermography to evaluate the effects of climatic variables in bull's reproduction.

Int J Biometeorol 2016 Jan 7;60(1):151-7. Epub 2015 Jun 7.

Department of Animal Science, Federal University of Rio Grande do Sul, 91540-000, Porto Alegre, RS, Brazil.

The objective of this study was to evaluate the seasonal effects of the environment on sperm quality in subtropical region determined by temperature and humidity index (THI). We used 20 Brangus bulls (5/8 Angus × 3/8 Nellore) aged approximately 24 months at the beginning of the study. Semen evaluations were performed twice per season during 1 year. Climate THI data were collected from an automatic weather station from the National Institute of Meteorology. Infrared thermography images were used to determine the temperature of the proximal and distal poles of the testis to assess the testicular temperature gradient (TG). The seasonal effects on seminal and climatic variables were analyzed with ANOVA using MIXED procedure of SAS. Sperm motility in spring (60.1%), summer (57.6%), and autumn (64.5%) showed difference compared to winter (73.0%; P < 0.01). TG was negatively correlated with THI at 18 days (spermiogenesis) (-0.76; P < 0.05) and at 12 days (epididymal transit) (-0.85; P < 0.01). Ocular temperature (OcT) had a positive correlation with THI at 18 days (0.78; P < 0.05) and at 12 days (0.84; P < 0.01). Motility showed a negative correlation with THI only at 18 days (-0.79; P < 0.05). During spermiogenesis, the TG had higher negative correlation compared to OcT (-0.97; P < 0.01) and rectal temperature (-0.72; P < 0.05). Spermatozoa with distal midpiece reflex were correlated with THI during transit epididymis (0.72; P < 0.05). Seminal parameters are not affected when THI reaches 93.0 (spermiogenesis) and 88.0 (epididymal transit). We concluded that infrared thermography can be adopted as an indirect method in order to assess the effect of environmental changes in TG and OcT of Brangus bulls.
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http://dx.doi.org/10.1007/s00484-015-1013-yDOI Listing
January 2016

Technologies that affect the weaning rate in beef cattle production systems.

Trop Anim Health Prod 2015 Oct 6;47(7):1255-60. Epub 2015 Jun 6.

Center for Studies and Research in Agribusiness - CEPAN. Núcleo de Estudos em Sistemas de Produção de Bovinos de Corte e Cadeia Produtiva - NESPRO, Federal University of Rio Grande do Sul - UFRGS, Ave. Bento Gonçalves 7712, 91540-000, Porto Alegre, RS, Brazil.

We investigated the differences between weaning rates and technologies adopted by farmers in cow-calf production systems in Rio Grande do Sul State, Brazil. Interviews were carried out with 73 farmers about 48 technologies that could affect reproductive performance. Data were analyzed by multivariate analysis using a non-hierarchical cluster method. The level of significance was set at P < 0.05. Three distinct clusters of farmers were created (R (2) = 0.90), named as low (LWR), intermediate (IWR), and high (HWR) weaning rate, with 100, 91, and 96 % of the farmers identified within their respective groups and average weaning rates of 59, 72, and 83 %, respectively. IWR and HWR farmers used more improved natural pasture, fixed-time artificial insemination, selection for birth weight, and proteinated salt compared to LWR. HWR farmers used more stocking rate control, and IWR farmers used more ultrasound to evaluate reproductive performance compared to the LWR group. IWR and HWR adopted more technologies related to nutrition and reproductive aspects of the herd in comparison to LWR. We concluded that farmers with higher technology use on farm had higher weaning rates which could be used to benefit less efficient farmers.
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http://dx.doi.org/10.1007/s11250-015-0856-xDOI Listing
October 2015

A specific adenylyl cyclase inhibitor (DDA) and a cyclic AMP-dependent protein kinase inhibitor (H-89) block the action of equine growth hormone on in vitro maturation of equine oocytes.

Zygote 2015 Dec 26;23(6):795-801. Epub 2014 Sep 26.

Department of Population Health and Reproduction,School of Veterinary Medicine,University of California,Davis,CA,USA.

The objectives of this study were firstly to determine whether the stimulatory function of equine growth hormone (eGH) on equine oocyte maturation in vitro is mediated via cyclic adenosine monophosphate (cAMP); and secondly if the addition of eGH in vitro influences oocyte nuclear maturation and if this effect is removed when GH inhibitors are added to the culture. Cumulus-oocyte complexes (COCs) were recovered from follicles <25 mm in diameter and randomly allocated as follows: (i) control (no additives); and (ii) 400 ng/ml of eGH. A specific inhibitor against cyclic AMP-dependent protein kinase (H-89; 10-9, 10-11 or 10-15 M concentration) and a specific adenylate cyclase inhibitor, 2',3'-dideoxyadenosine (DDA; 10-8, 10-10 or 10-14 M concentration) were used to observe whether they could block the eGH effect. After 30 h of in vitro maturation at 38.5°C with 5% CO2 in air, oocytes were stained with 10 μg/ml of Hoechst to evaluate nuclear status. More mature oocytes (P < 0.05) were detected when COCs were incubated with eGH (29 of 84; 34.5%) than in the control group (18 of 82; 21.9%). The H-89 inhibitor used at a concentration of 10-9 M (4 of 29; 13.8%) decreased (P < 0.05) the number of oocytes reaching nuclear maturation when compared with eGH (11 of 29; 38%). The DDA inhibitor at a concentration of 10-8 M (2 of 27; 7.4%) also reduced (P < 0.05) the number of oocytes reaching maturity when compared with the eGH group (9 of 30; 30%). Results from the present study show that H-89 and DDA can be used in vitro to block the eGH effect on equine oocyte maturation.
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http://dx.doi.org/10.1017/S0967199414000434DOI Listing
December 2015

Vitrification of equine oocytes with a polyvinyl alcohol after in vitro maturation with equine growth hormone and insulin-like growth factor-I.

Cryo Letters 2014 Mar-Apr;35(2):90-4

Animal Pathology Department, Veterinary School, Federal University of Pelotas, Brazil.

Background: In vitro fertilization (IVF) procedures are limited by the inability to mature equine oocytes on in vitro methods.

Objective: The aim of this study was to evaluate structural integrity of equine oocytes subjected to vitrification with a synthetic polymer (PVA).

Methods: The effect of eGH and its relationship with IGF-I on in vitro maturation (IVM) were evaluated. Compact cumulus oocytes complexes (n=122) were cultured in TCM-199 with eGH, IGF-I or eGH+IGF-I for 30h at 38.5C in air with 5 % CO2. Oocytes were fixed after IVM or subjected to the vitrification protocol. Cryopreserved oocytes were exposed to 1.4M DMSO+1.8M EG+1 percent PVA for 3min, and then transfer to 2.8M DMSO+3.6M EG+0.6M sucrose+1 % PVA for 1min. After rewarming, oocytes were evaluated by confocal microscopy.

Results: Maturation rates of oocytes were not significant different among groups (P > 0.05), however eGH+IGF-I group can develop the assessment of resumption of meiosis (MI+MII = 86.7 %).

Conclusion: The oocyte did not show morphological alterations. The use of PVA-copolymer may represent a potential alternative for vitrification of equine oocytes after IVM.
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June 2014

Scrotal infrared digital thermography as a predictor of seasonal effects on sperm traits in Braford bulls.

Int J Biometeorol 2015 Mar 22;59(3):357-64. Epub 2014 May 22.

Department of Animal Science, Federal University of Rio Grande do Sul, Av. Bento Gonçalves, no 7.712, 91540-000, Porto Alegre, RS, Brazil.

The aim of this study was to assess the seasonal effects of the environment on semen quality in bulls, using infrared thermography. Sperm motility (M), mass motion (MM), and vigor (VIG) were evaluated in sperm samples from 17 Bradford bulls aged approximately 24 months at the beginning of the study. Infrared thermography images and data were collected using an infrared FLIR T 300 camera and Quick Report 1.2 SP2 software to determine the temperature of the proximal and distal poles of the testis and to assess the testicular temperature gradient. The seasonal effects on physiological, seminal, and climatic variables were analyzed by the GLM ANOVA and CORR procedures using SAS®. The microclimatic factors were recorded in hourly intervals, and the daily mean temperature and mean relative humidity were calculated to determine the daily temperature-humidity index (THI) every day for 1 year. The temperature gradient (TG) variations of the testes were significantly higher in the autumn (4.5 °C), winter (4.0 °C), and spring (2.9 °C) compared to summer (0.9 °C) (P < 0.05). Ocular globe temperatures were lower in the winter (27.6 °C) and autumn (26.8 °C) compared to summer (33.9 °C) and spring (31.1 °C) (P < 0.05). The average MM (2.58), M (52.64), and VIG (2.70) of the semen decreased in the summer compared to other seasons (P < 0.01). The TG was negatively correlated with THI (-0.44; P < 0.05). For the seminal variables, MaD (-0.45; P < 0.05) and TD (-0.50; P < 0.01) presented a negative correlation with TG. The TG had a positive correlation between M and VIG, which had values of 0.36 and 0.35, respectively (P < 0.05). We have concluded that infrared thermography can be used to assess the testicular temperature gradient and its consequences on physical and quantitative aspects of sperm.
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http://dx.doi.org/10.1007/s00484-014-0847-zDOI Listing
March 2015

Higher feeding diets effects on age and liveweight gain at puberty in crossbred Nelore × Hereford heifers.

Trop Anim Health Prod 2014 Aug 23;46(6):953-60. Epub 2014 Apr 23.

Department of Animal Science, Núcleo de Estudos em Sistemas de Produção de Bovinos de Corte e Cadeia Produtiva - NESPRO, Federal University of Rio Grande do Sul, UFRGS, Ave. Bento Gonçalves 7712, 91540-000, Porto Alegre, RS, Brazil.

This study was conducted to determine the age and liveweight at puberty of 120 crossbred beef heifers submitted to four diets to achieve predetermined weight gains (kg/day): 0.5 (G500; n = 32), 0.75 (G750; n = 32), 1.0 (G1000; n = 29), and 1.25 (G1250; n = 27). Animals were classified depending on their level of crossing between Nelore (N) and Hereford (H): 25%N-75%H, 37.5%N-63.5%H, 43.7%N-56.7%H, 50%N-50%H, and 75%N-25%H. Reproductive evaluation was performed at the beginning and at the end of the experimental period by ultrasonography and hormone analysis. The average age and liveweight at puberty were 388.0 ± 1.9 days and 331.4 ± 1.3 kg, respectively. Animals from the 25%N-75%H group reached puberty earlier than heifers from other genetic groups supplemented with G1250 diet (P < 0.05). Heifers with higher degree of Nelore (75%N-25%H) fed with G1000 diet showed estrus 42 days prior to mating, but only 57% reached puberty at mating (P < 0.05). Heifers with follicles of higher diameter reached puberty (P < 0.05) earlier. Higher average daily weight gain showed a positive effect on follicular diameter and IGF-I level at puberty (P < 0.01). Concentrations of GH were lower in heifers fed G1250 compared to G1000 diet (P < 0.05). There was a significant interaction between nutritional level and insulin levels at puberty (P < 0.01). We demonstrated the relationship between IGF-I and average daily gain on the onset of puberty in heifers. In conclusion, heifers submitted to the higher feeding level showed a higher follicular diameter and were younger at puberty.
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http://dx.doi.org/10.1007/s11250-014-0593-6DOI Listing
August 2014

Neospora caninum DNA detection by TaqMan real-time PCR assay in experimentally infected pregnant heifers.

Vet Parasitol 2014 Jan 30;199(3-4):129-35. Epub 2013 Oct 30.

Laboratório de Biotecnologia e Reprodução Animal - BioRep, Departamento de Clínica de Grandes Animais (DCGA), Centro de Ciências Rurais, Universidade Federal de Santa Maria, 97105-900 Santa Maria, Brazil.

Neosporosis has been considered the main cause of abortion between the first and the second trimester of pregnancy in cattle. Therefore, the objective of this study was to identify the presence of Neospora caninum DNA obtained from experimental models based on the evaluation of different areas of the fetal nervous system and organs from heifers previously inoculated with NC-1 after or before insemination. This study was performed with Hereford × Nelore (n=29) heifers and all animals were considered free of diseases at the beginning of the experiment. All animals were bred by fixed-time artificial insemination (TAI) and allocated as follows: (a) seronegative heifers subjected to TAI (TAI, n=9), (b) heifers infected with N. caninun 60 days prior to TAI (NC-1+TAI, n=9), and (c) heifers submitted to TAI and infected with N. caninum 60 days later (TAI+NC-1, n=11). The pregnancy was confirmed by transrectal ultrasonography 35 days after TAI and evaluated every 30 days until the end of gestation. Fetuses were collected surgically at 170 days of gestation, and immediately necropsied to remove tissues aseptically. Samples of the central nervous system (CNS), heart, kidney, lung, liver, skeletal muscle and caruncle were collected for DNA extraction. Days of gestation at abortion and interval from abortion to first insemination were examined by Student's t-test. At 35 days of gestation the pregnancy rates in the group NC-1+TAI (4/9, 44.4%) was lower than in the control group (8/9, 88.8%, P<0.05). At 60 days, the pregnancy rates in the NC-1+TAI group (0/4, 0%) was lower compared to TAI+NC-1 (5/7, 71.4%) and control (6/8, 75.0%) groups (P<0.05). Animals from the group NC-1+TAI were re-inseminated 60 days after the first TAI. After pregnancy losses throughout the study, 5 animals (TAI), 3 animals (NC-1+TAI) and 5 animals (TAI+NC-1) maintained pregnancy until 170 days of gestation. TaqMan RT-PCR demonstrated the presence of N. caninum DNA in the medulla and right posterior cortex in 3 out of 5 fetuses from the TAI+NC-1 group. We concluded that heifers infected after TAI had a higher incidence of the parasite at the fetus CNS. Identification of N. caninum by TaqMan RT-PCR would assist in the investigation of infection and in the evaluation of vaccines or therapeutic drugs to control neosporosis in cattle.
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http://dx.doi.org/10.1016/j.vetpar.2013.10.018DOI Listing
January 2014

The components of the angiotensin-(1-7) system are differentially expressed during follicular wave in cattle.

J Renin Angiotensin Aldosterone Syst 2015 Jun 13;16(2):275-83. Epub 2013 Jun 13.

Laboratório de Biotecnologia e Reprodução Animal-BioRep, Universidade Federal de Santa Maria, Brazil.

Introduction: This study was based on the hypothesis that some components of the angiotensin-(1-7) (Ang-(1-7)) system are differentially expressed during follicular development and can be involved in the follicular health/atresia transition in bovine.

Material And Methods: The largest (F1) and second largest follicles (F2) were collected from cows before (Day 2), during (Day 3), or after (Day 4) the expected moment of follicular deviation. In the second experiment, F1 was induced to atresia through intrafollicular injection of fulvestrant (estrogen receptor-antagonist) and, in both experiments, mRNA expression of the Mas receptor, ACE2, NEP, and PEP was evaluated in the granulosa and theca cells.

Results: The mRNA expression of Mas receptor was upregulated in the granulosa cells of F2 after the establishment of follicular deviation, while PEP mRNA increased during and after the deviation process. The mRNA expression of ACE2 was upregulated in the granulosa cells of F1 during and after the follicular deviation. The mRNA expression of NEP was not regulated in F1 and F2. Mas receptor expression increased in the F1 induced to atresia.

Conclusions: mRNA for Mas receptor, ACE2, and PEP are differentially expressed in granulosa cells throughout follicular development and the Mas receptor can be involved with the establishment of follicular dominance.
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http://dx.doi.org/10.1177/1470320313491996DOI Listing
June 2015

Growth factor receptor-bound protein 14: a potential new gene associated with oocyte competence.

Zygote 2014 May 17;22(2):103-9. Epub 2013 May 17.

Laboratory of Biotechnology and Animal Reproduction- BioRep, Federal University of Santa Maria, Av. Roraima #1000, CEP 97105-900, Santa Maria, RS 97105-900, Brazil.

The Grb14 protein is a member of the Grb7 protein family. This protein family acts by binding to tyrosine kinase receptors, promoting cell proliferation and differentiation. There is evidence of the involvement of tyrosine kinase factors in the bovine oocyte maturation process. However, Grb14 has not been studied for bovine cumulus-oocyte complexes (COCs). The aim of the present study was to characterize Grb14 mRNA expression in bovine COCs during follicular development. Furthermore, we demonstrated that the expression of Grb14 mRNA is not regulated by estradiol. mRNA expression of Grb14 was assessed in 480 COCs from follicles of different sizes (1-3, 4-6, 6-8 or >8 mm) by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Grb14 mRNA expression decreased in COCs throughout follicular growth (P < 0.05). The role of estradiol in the expression of Grb14 mRNA in COCs was studied. Grb14 mRNA abundance did not differ in COCs cultured in the presence or absence of 17β-estradiol or fulvestrant. In conclusion, we showed that Grb14 mRNA is downregulated in COCs during antral follicle development, a finding that suggests a role for Grb14 in oocyte competence.
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http://dx.doi.org/10.1017/S0967199413000221DOI Listing
May 2014

The effect of growth hormone (GH) and insulin-like growth factor-I (IGF-I) on in vitro maturation of equine oocytes.

Zygote 2012 Nov 28;20(4):353-60. Epub 2011 Jul 28.

Laboratory of Biotechnology and Animal Reproduction-BioRep, Federal University of Santa Maria, Santa Maria, RS, Brazil.

The objective of this study was to test the hypothesis that equine growth hormone (eGH), in combination with insulin growth factor-I (IGF-I), influences positively in vitro nuclear and cytoplasmic maturation of equine oocytes. Cumulus-oocyte complexes were recovered from follicles that were < 25 mm in diameter, characterized by morphology and were allocated randomly as follow: (a) control (no additives); (b) 400 ng/ml eGH; (c) 200 ng/ml IGF-I; (d) eGH + IGF-I; and (e) eGH + IGF-I + 400 ng/ml anti-IGF-I antibody. Oocytes were matured for 30 h at 38.5°C in air with 5% CO2 and then stained with 10 μg/ml propidium iodide (PI) to evaluate nuclear status and 10 μg/ml Lens culinaris agglutinin-fluorescein complex (FITC-LCA) to assess cortical granule migration by confocal microscopy. The proportion of immature oocytes that developed to the metaphase II (MII) stage in the eGH + IGF-I group (15 of 45) was greater than in the groups that were treated only with IGF-I (7 of 36, p = 0.03). Oocytes that reached MII in the control group (20 of 56; 35.7%) showed a tendency to be different when compared with eGH + IGF-I group (15 of 45; 33.3%, p = 0.08). The treated group that contained anti-IGF-I (15 of 33; 45.4%) decreased the number of oocytes reaching any stage of development when compared with eGH (47 of 72; 65.3%) and eGH + IGF-I (33 of 45; 73.3%) groups (p = 0.05) when data from MI and MII were combined. We concluded that the addition of eGH to in vitro maturation (IVM) medium influenced the in vitro nuclear and cytoplasmic maturation of equine oocytes. The use of GH and IGF-I in vitro may represent a potential alternative for IVM of equine oocytes.
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http://dx.doi.org/10.1017/S0967199411000335DOI Listing
November 2012

Leukemia inhibitory factor stimulates the transition of primordial to primary follicle and supports the goat primordial follicle viability in vitro.

Zygote 2012 Feb 18;20(1):73-8. Epub 2011 Mar 18.

Laboratory of Biotechnology and Animal Reproduction - BioRep, Federal University of Santa Maria, CEP 97105-900, Santa Maria, RS, Brazil.

The aim of this study was to evaluate the effect of leukemia inhibitory factor (LIF) on the activation and survival of preantral follicles cultured in vitro enclosed in ovarian fragments (in situ). Goat ovarian cortex was divided into fragments to be used in this study. One fragment was immediately fixed (fresh control - FC) and the remaining fragments were cultured in supplemented minimum essential medium (MEM) without (cultured control - CC) or with different concentrations of LIF (1, 10, 50, 100 or 200 ng/ml) for 1 or 7 days, at 39°C in air with 5% CO2. Fresh control, CC and treated ovarian fragments were processed for histological and fluorescence analysis. The percentage of histological normal preantral follicles cultured for 7 days with 1 ng/ml (49.3%), 10 ng/ml (58.6%) and 50 ng/ml (58%) of LIF was higher than in the CC (32.6%; p < 0.05). After 7 days of culture, the percentage of primordial follicles in situ cultured with LIF decreased and primary follicles increased in all LIF concentrations compared with FC and CC (p < 0.05). In conclusion, LIF induced primordial follicle activation and supported preantral follicle viability of goat ovarian tissues cultured for 7 days.
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http://dx.doi.org/10.1017/S0967199411000074DOI Listing
February 2012
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