Prediction of tenderness in bovine longissimus thoracis et lumborum muscles using Raman spectroscopy.

OBJECTIVE: This study was conducted to evaluate Raman spectroscopy technique as a noninvasive tool to predict meat quality traits on Braford longissimus thoracis et lumborum muscle. METHODS: Thirty samples of muscle from Braford steers were analyzed by classical meat quality techniques and by Raman spectroscopy with 785 nm laser excitation. Water holding capacity (WHC), intramuscular fat content (IMF), cooking loss (CL), and texture profile analysis recording hardness, cohesiveness, and chewiness were determined, along with fiber diameter and sarcomere length by scanning electron microscopy. Warner-Bratzler shear force (WBSF) analysis was used to differentiate tender and tough meat groups. RESULTS: Higher values of cohesiveness and CL, together with lower values of WHC, IMF, and shorter sarcomere were obtained for tender meat samples than for the tougher ones. Raman spectra analysis allows tender and tough sample differentiation. The correlation between the quality attributes predicted by Raman and the physical measurements resulted in values of R2 = 0.69 for hardness and 0,58 for WBSF. Pearson's correlation coefficient of hardness (r = 0.84) and WBSF (r = 0.79) parameters with the phenylalanine Raman signal at 1,003 cm-1, suggests that the content of this amino acid could explain the differences between samples. CONCLUSION: Raman spectroscopy with 785 nm laser excitation is a suitable and accurate technique to identify beef with different quality attributes.

Feeding strategies and ageing time alter calpain system proteins activities and meat quality of Braford steers.

OBJECTIVE: The aim of this study was to evaluate the effect of ageing and feeding strategies on the calpain protease system and meat quality traits in Braford steers. METHODS: Thirty Braford steers were employed; 15 animals were supplemented with corn silage during finishing and 15 were kept only on pasture. Meat quality traits and calpain system protein activity were evaluated in longissimus thoracis et lumborum (LTL) steaks aged for 2, 7, 14, and 21 days. RESULTS: Aged meat showed higher pH and calcium content, while Warner Bratzler shear force (WBSF) decreased to day 21. No interaction between ageing and diet was seen for quality traits. Steers finished with corn silage showed higher values of water holding capacity, WBSF and free calcium, and lower values of pH and cooking loss. Calpain and calpastatin activities decreased with ageing. Finishing steers on pasture produced higher values of calpains and lower values of calpastatin activities. The higher values of calpain 1 activity were observed in muscles aged 2 days from pasture finished animals, and the lower activity of the inhibitor in the 21 days aged samples of the same group. CONCLUSION: These results suggest a diet by ageing interaction in calpains and calpastatin and this interaction impact in Warner Bratzler Shear Force in Braford LTL muscle.

Follicular wave dynamics and Growth factors gene expression in Braford heifers.

The objectives of the present study were (experiment 1) to characterized development and dynamics of the dominant follicles (DF) and the corpus luteum (CL) to determine patterns of two (W2) and three (W3) follicular waves in beef heifers, and (experiment 2) to determine gene expression of growth factors gene expression in follicular cells of W2 and W3 heifer. Twenty-eight Braford heifers were used. Dominant follicular and CL were monitored daily by ultrasonography to identify the development W2 and W3 in heifers. Pre-ovulatory DF were aspirated on day 19 in W2 and on day 22 in W3 heifers. In W2 and W3, follicular cells (FC) of gene expression of growth differentiation factor 9, bone morphogenetic protein 15 (BMP15), fibroblast growth factor basic, transforming growth factor beta receptor 1, bone morphogenetic protein receptor type IB and fibroblast growth factor receptor 2 were evaluated. The regression of the DF of the first follicular wave and the emergency of the DF of the second follicular wave began later in the heifers W2 than in W3 (p = .02 and p < .01). The regression of the CL began earlier in the W2 than in W3 group (p < .01). Gene expression of growth factors and receptors was similar between groups. However, higher relative levels of BMP15 was observed in W2 group (p = .07). Results propose that wave patterns were regulated by the development time of the DF in the first wave and the life of the CL. Furthermore, higher levels of BMP15 could produce shorter life of CL. The present work suggest that ultrasonography associated with molecular assays could be used as an easy and effective tool to characterize follicular wave patterns.

Feeding strategies alter gene expression of the calpain system and meat quality in the longissimus muscle of Braford steers.

OBJECTIVE: The aim of the present study was to determine the effect of supplementing pasture-finished steers with corn silage on the expression level of the calpain system proteins and beef tenderization. METHODS: Thirty Braford steers grazing on summer pasture were used for the study. For 120 days fifteen animals were supplemented with corn silage at 1% of body weight per head per day (Suppl) whereas the remaining 15 steers only received pasture (Contr). Carcass and meat traits were evaluated and compared between groups. Gene expression and activities of proteases (calpain 1 and calpain 2) and inhibitor (calpastatin) were measured using real-time polymerase chain reaction and casein zymography. RESULTS: Carcass and meat traits were significantly different between feeding systems. Supplemented steers showed higher hot carcass weight (p<0.01), fat content (p = 0.02), and Warner-Bratzler shear force (p = 0.03). Furthermore, the control group showed higher protease:inhibitor ratios, at mRNA (p = 0.01) and protein levels (p<0.10). Warner-Bratzler shear force and mRNA calpains:calpastatin ratio were associated in both feeding systems (p<0.01). CONCLUSION: Based on the results obtained in the study, beef tenderness differences among finishing strategies could be modulated through differential expression of the calpain system proteins.

Hormones and monensin use to improve pregnancy rates in grazing lactating beef cows in the semiarid region of Argentina.

The objectives of this study were 1) to determine the effect of monensin treatment, alone or combined with a hormonal estrus synchronization treatment, on the pregnancy rate of lactating beef cows, and 2) to evaluate the effect of monensin capsule administration on ruminal metabolism in steers. In experiment 1, ninety-four cows were selected from a 300 cow herd. The experimental design used was a 2 x 2 factorial with the administration of monensin capsule as first factor (M1 = with monensin vs. M0 = without monensin) and hormonal treatment as second factor (H1 = with hormonal treatment vs. H0 = no hormonal treatment). Thirty-eight days before the beginning of the breeding season, cows were randomly assigned to the first factor, and thirty days later to the second factor, resulting in four treatments: M1H0, M1H1, M0H0 and M0H1. Cow were exposed to bull (bull/cow ratio 1:20) from day 0 (day 0 = start of the breeding season and 38 days after monensin capsule administration) to day 50. Pregnancy diagnosis was performed at 30, 60 and 80 days after start breeding season by ultrasonography. In experiment 2, eight ruminally cannulated crossbred beef steers were randomly assigned to two treatments (M1 and M0). To determine proportion of volatile fatty acids (VFA), ruminal fluid samples were taken on days 0, 40 and 77 of the experimental period, at 0, 4 and 12 h after grazing. In experiment 1, treatments whit monensin did not improve pregnancy rate (P = 0.95), however, hormonal treatment resulted in grater pregnancy rates (P = 0.03). In experiment 2, the proportion of VFA in ruminal fluid of steers was significantly different between treatments. The highest proportion of propionate was found in ruminal fluid from M1 treatment at 12 h after grazing (P = 0.04). In conclusion, the treatment with monensin increased the proportion of propionate. The result might suggest that energy balance was improved in steers, without improvement in cow´s pregnancy rates. Treatment with monensin alone did not improve pregnancy rate, nor did treatment with monensin enhance the pregnancy rate when a hormonal synchronisation treatment was given. Nevertheless, the use of a hormonal treatment increased pregnancy rate, suggesting that it could be used as a suitable tool to enhance the productivity in cows with marginal body condition score.

Expression and localization of nodal in bovine oviduct and uterus during different functional stages of oestrus cycle and pregnancy.

Members of TGF-ß superfamily play a major role in the endometrial changes involved in the establishment and maintenance of pregnancy. Their deregulated expression and action could lead to absolute or partial failure of embryo implantation. Nonetheless, the precise function and mechanism of many of these cytokines remain unclear. Nodal, a transforming growth factor beta (TGF-ß) superfamily member, was characterized in the human and rodent uterus and implicated in the tissue remodeling events during menstruation and embryo implantation. In order to study its possible role in the cattle reproductive process, we have analyzed Nodal expression pattern and localization in the oviduct and uterine horn during the oestrus cycle and early pregnancy (day 20). Nodal was detected both in oviduct and uterus during either the oestrus cycle or pregnancy; however, it shows a differential expression profile in the uterine horn at dioestrus and pregnancy, decreasing 1.5 and 1.4 folds in comparison with oestrus. Nodal immunostaining intensity was observed in stromal and in epithelial cells of the surface and the glandular epithelium. The staining pattern correlates with the RT-qPCR expression profile. This work is the first to evidence the presence of Nodal in the bovine reproductive tract; our data suggest that Nodal is a novel cytokine that would be involved in the remodelling occurring in the endometrium of cattle during the oestrus cycle and in the embryo implantation. The identification of new molecules that participate in endometrium cycling and/or pregnancy may be useful for predicting the ability of the uterine tissue to establish and maintain pregnancy or for detecting the infertility processes. These results highlight Nodal as a possible novel marker of the fertility process, nevertheless further studies should be done to determine its role in the reproductive system.

Biology and biotechnology of follicle development.

Growth and development of ovarian follicles require a series of coordinated events that induce morphological and functional changes within the follicle, leading to cell differentiation and oocyte development. The preantral early antral follicle transition is the stage of follicular development during which gonadotropin dependence is obtained and the progression into growing or atresia of the follicle is made. Follicular growth during this period is tightly regulated by oocyte-granulosatheca cell interactions. A cluster of early expressed genes is required for normal folliculogenesis. Granulosa cell factors stimulate the recruitment of theca cells from cortical stromal cells. Thecal factors promote granulosa cell proliferation and suppress granulosa cell apoptosis. Cell-cell and cell-extracellular matrix interactions influence the production of growth factors in the different follicular compartments (oocyte, granulosa, and theca cells). Several autocrine and paracrine factors are involved in follicular growth and differentiation; their activity is present even at the time of ovulation, decreasing the gap junction communication, and stimulating the theca cell proliferation. In addition, the identification of the factors that promote follicular growth from the preantral stage to the small antral stage may provide important information for the identification for assisted reproduction techniques.