F. M. Monteiro

Evidence of negative relationship between female fertility and feed efficiency in Nellore cattle1,2

The objective of this study was to evaluate phenotypic and genetic relationships between fertility traits and feed efficiency in male and female Nellore cattle. Data from 320 females born between 2004 and 2011 were used for phenotypic evaluation. These animals were evaluated for postweaning residual feed intake (RFI) and classified as negative (RFI < 0, mean = -0.294 ± 0.017 kg DM/d) or positive RFI (RFI > 0, mean = 0.305 ± 0.0189 kg DM/d). Of these, 118 prepuberal heifers were submitted to ultrasonography of the uterus and ovaries for monitoring the presence (or absence) of a corpus luteum and for the measurement of endometrial thickness. The following fertility traits were evaluated in all females: age at first calving, days to calving, first calving interval, calving success, stayability, and longevity. The variance components were estimated by the average information restricted maximum likelihood method under an animal model in 5-trait analysis of backfat and rump fat thickness, scrotal circumference, days to calving, and RFI. The total number of animals with records was 6,718, including 927 males with records of scrotal circumference and RFI and 264 females with records of days to calving and RFI. Negative RFI females consumed 12.5% less DM daily than positive RFI females and had a lower rump fat thickness when evaluated postweaning. Among the fertility traits studied, only first calving interval differed (P = 0.0858) between RFI classes, with the interval of negative RFI females being 45 d shorter than that of positive RFI animals. The heritability estimates were 0.29, 0.34, 0.50, 0.12, and 0.16 for backfat and rump fat thickness, scrotal circumference, days to calving, and RFI, respectively. The genetic correlations between RFI and the other traits studied were unfavorable for selection and were of moderate magnitude with backfat thickness, rump fat thickness, and days to calving (0.53, 0.37, and -0.49, respectively) and close to zero with scrotal circumference (0.07). Scrotal circumference (0.17 and 0.15) and days to calving (-0.10 and -0.22) were weakly and favorably correlated with backfat and rump fat thickness. There is evidence of moderate genetic antagonism between female fertility and feed efficiency, but with no evidence of a genetic correlation between male fertility and feed efficiency. There is also evidence of low genetic synergism between fat thickness and fertility.

Evaluation of cooling and freezing systems of bovine semen

Semen cryopreservation comprises different steps, among them are the cooling and freezing rates which significantly influence the quality of thawed sperm. Different systems with variable freezing rates are used for freezing bull semen in the field, with a consequence of variable success rates. The objective of this study was to compare different systems for freezing bull semen in the field. Five cooling methods of semen and two methods for the subsequent freezing phase (5 × 2 factorial scheme) were used. Two to four ejaculates were collected from 12 bulls with an electroejaculator. The ejaculates were diluted in BotuBov® to a concentration of 50 × 106 spermatozoa/mL in 0.5-mL straws. After dilution, the straws were cooled to 5 °C in five cooling systems: TK 4000® at a cooling rate of -0.25 °C/min (R1); TK 4000® at a rate of -0.5 °C/min (R2); Minitube® refrigerator at a rate of -2.8 °C/min (R3); Botutainer® at a rate of -0.65 °C (R4), and domestic refrigerator at a rate of -2.0 °C/min (R5). After stabilization at 5 °C for 4 h, these straws were then submitted to two freezing systems: TK 4000® at a freezing rate of -15 °C/min (C1) and Styrofoam box with liquid nitrogen at a rate of -19 °C/min (C2). Sperm kinetics were evaluated by computer-assisted sperm analysis at four time points: in fresh semen, after cooling, post-thawing, and after the rapid thermal resistance test (TRT). In addition, plasma and acrosomal membrane integrity, mitochondrial potential and intracellular H2O2 were analyzed after thawing by flow cytometry. The R1, R2 and R4 cooling systems were the most efficient in preserving sperm viability, membrane integrity and intracellular H2O2. Samples frozen in the C1 system exhibited better post-thaw and post-TRT kinetics than C2 samples. In conclusion, slower cooling curves in conjunction with a constant freezing rate obtained with the programmable unit were more efficient for freezing bull semen in the field.

Chemically induced enucleation of activated bovine oocytes: chromatin and microtubule organization and production of viable cytoplasts

As the standard enucleation method in mammalian nuclear transfer is invasive and damaging to cytoplast spatial organization, alternative procedures have been developed over recent years. Among these techniques, chemically induced enucleation (IE) is especially interesting because it does not employ ultraviolet light and reduces the amount of cytoplasm eliminated during the procedure. The objective of this study was to optimize the culture conditions with demecolcine of pre-activated bovine oocytes for chemically IE, and to evaluate nuclear and microtubule organization in cytoplasts obtained by this technique and their viability. In the first experiment, a negative effect on oocyte activation was verified when demecolcine was added at the beginning of the process, reducing activation rates by approximately 30%. This effect was not observed when demecolcine was added to the medium after 1.5 h of activation. In the second experiment, although a reduction in the number of microtubules was observed in most oocytes, these structures did not disappear completely during assessment. Approximately 50% of treated oocytes presented microtubule reduction at the end of the evaluation period, while 23% of oocytes were observed to exhibit the complete disappearance of these structures and 28% exhibited visible microtubules. These findings indicated the lack of immediate microtubule repolymerization after culture in demecolcine-free medium, a fact that may negatively influence embryonic development. However, cleavage rates of 63.6-70.0% and blastocyst yield of 15.5-24.2% were obtained in the final experiment, without significant differences between techniques, indicating that chemically induced enucleation produces normal embryos.

Comparison of Brucella agar, CITA and Farrell media for selective isolation of Brucella abortus from semen of bovine bulls

Brucellosis remains as a public health concern worldwide. In domestic animals, the disease is characterized by reproductive disorders in male and female. Besides extensive use of serological tests and recent development of molecular biology techniques, microbiological culture of Brucella species is yet considered a “gold standard” method for diagnosis. Here, semen of 335 bovine bulls was subjected simultaneously to microbiological culture in Brucella agar, Farrell media, and CITA media to evaluate comparatively the best selective media for isolation of Brucella sp. Among all 335 samples, B. abortus B19 strain was isolated from semen of five (1.49%) bulls using the three selective media. However, Farrell media was considered the best selective media for microbiological diagnosis, because of allowed isolation of B. abortus B19 strain from bull semen without bacterial commensal or fungal contamination of plates. Key words: Brucella agar, Farrell media, CITA media, bovine bulls, semen.