Culture system and long-term storage of culture media in the in vitro production of bovine embryos

The optimum culture system for in vitro matured and fertilised oocytes still remains to be clarified. Culture media (CM) for mammalian embryos are routinely prepared fresh for use and preserved under refrigeration during one or two weeks. The purposes of this work were (1) to compare the efficiency of a synthetic oviduct fluid (SOF) with two different bovine serum albumin (BSA) concentrations (3 and 8 g/L) for the in vitro production of bovine blastocysts, (2) to test the effect of timing on adding fetal calf serum (FCS) to the SOF, and (3) to evaluate the effects on bovine embryo development of freezing and lyophilisation as procedures for preserving the SOF. Supplementation of SOF with 3 g/L BSA increased Day-7 blastocyst expansion rates (18.3 ± 1.6 vs. 14.4 ± 0.7; P < 0.05), although no differences in hatching rates were found. Addition of FCS to SOFaa (SOF with amino acids) medium supplemented with sodium citrate (SOFaaci) at 48 and at 72 h post-insemination (PI) allowed obtaining higher Day-6 embryo development rates than when FCS was added at 18 or 96 h PI (Day-6 morulae + blastocyst rate: 30.0 ± 1.1, 40.8 ± 1.1, 43.9 ± 2.3 and 39.3 ± 0.5 for FCS addition at 18, 48, 72 and 96 h, respectively). Hatching rates were significantly improved when serum was added at 72 h PI. Finally, both refrigeration and lyophilisation appeared as useful cryopreservation procedures for SOFaaci, although a significant loss of its ability to support embryo development, compared to the control fresh culture medium, was observed.     

Quantification of pulmonary to systemic flow ratio by a Doppler echocardiographic method in the normal dog: Repeatability, reproducibility, and reference ranges

Objective

To determine if the pulmonary to systemic flow ratio (Qp/Qs) could be assessed in healthy awake dogs using Doppler echocardiography.

Background

Qp/Qs could provide reliable information in assessing the severity of intracardiac shunts (ICS) by quantifying pulmonary overcirculation. Qp/Qs has been validated against electromagnetic flowmeter methods in experimental canine models. However, its clinical applicability in awake dogs has never been assessed.

Animals, materials and methods

Six healthy dogs were used to determine the repeatability and reproducibility of the technique (Study 1); Qp/Qs was then prospectively assessed in 50 healthy dogs (Study 2). In both studies Qp/Qs was calculated in awake animals using a standardized Doppler echocardiographic method.

Results

Within- and between-day coefficients of variation for Qp/Qs were <10% (Study 1). For Study 2, a relatively wide range of Qp/Qs was found (reference range = 0.71-1.29; mean ± SD = 1.00 ± 0.15).

Conclusions

Qp/Qs can be assessed with good repeatability and reproducibility in healthy dogs. However, the wide range of Qp/Qs obtained in the healthy population may suggest a limited usefulness of this variable for accurately assessing ICS severity in diseased animals. This needs to be assessed in further prospective and longitudinal studies including a large number of animals with ICS of various grades.     

Long-term effects of good handling practices during the pre-weaning period of crossbred dairy heifer calves

The aim of this study was to determine whether applying good practices of handling during the pre-weaning period have long-term effects on behavioral and physiological indicators, health status, and average daily gain (ADG) of crossbred Bos taurus × Bos indicus heifer calves. During the pre-weaning period, 98 crossbred of Holstein × Gir heifer calves were allotted into three treatments: (1) good practices of handling + brushing (GPB; n?=?25), (2) good practices of handling (GP; n?=?25), and (3) control (n?=?48). Every 2 months, four evaluation periods (EV₁ to EV₄) were conducted to record data. Behavioral indicators comprised time to drive (TD), flight speed (FS), flight distance (FD), and composite reactivity score (CRS). Physiological indicators of acute stress during handling comprised respiratory and heart rates. Health status comprised data regarding occurrence of most common diseases (i.e., pneumonia and anaplasmosis). Collected data were analyzed by using a linear mixed model for repeated measures, Tukey’s test, and chi-squared procedures. Treatment influenced (P?<?0.05) TD, FS, and FD but not CRS (P?=?0.78). From EV₁ to EV₃, the control calves had the lowest TD. The GPB group had lower FS than the control but did not differ from GP. The GPB group had lower FD means than the other two groups in EV₂, EV₃, and EV₄. No differences (P?>?0.05) due to treatment were observed on heart and respiratory rates, ADG, or occurrence of pneumonia and anaplasmosis. It was concluded that adoption of good practices of handling during pre-weaning period may lead to long-term positive effects.     

Soluble carbohydrates content in tomato pollen and its variations along and between blooming periods

The soluble carbohydrates content in the mature (starchless) pollen of the tomato (Solanum lycopersicum L.) cv. Platense was studied at several moments of the blooming period in two consecutive years. The aim of the analysis was to evaluate if the content of soluble carbohydrates is relatively constant or if it can fluctuate along the blooming period. No significant variations in pollen viability were recorded along each season. The soluble carbohydrates found and their concentrations can change significantly among samples, but the fluctuations observed did not follow a strongly definite pattern in any season. Reducing sugars predominated; small quantities of a phosphorylated sugar, UDP-glucose, and maltosaccharides were also recorded. The constant presence of maltosaccharides is a novel record for pollen. Sucrose was absent in one season, but present in the other, in low percentages in contrast to reducing sugars. Changes in the soluble carbohydrates content have been usually related with alterations in pollen fertility. However, there may be some flexibility in the metabolism of the pollen studied this time, at least within a certain range, which may allow constant adjustments to maintain acceptable levels of viability despite the variations in the carbohydrates concentrations.     

Genetic parameters for fertility traits in Nellore bulls

Nellore is the main cattle breed used in Brazil, being the largest commercial herd in the world. Beyond the importance of male reproductive efficiency for farm profit, the use of reproductive techniques, mainly artificial insemination, turns the evaluation of male reproductive traits even more important. Estimation of genetic parameters increases the knowledge on traits variances and allows envisaging the possibility of the inclusion of new traits as selection criterion. Genetic parameters for fifteen traits that can be classified as testicular biometry or physical and morphological semen traits were estimated for a Nellore bull population ranging from 18 to 36 months. Single-trait and bi-trait animal models were used for (co)variance components estimation. The contemporary group was considered as fixed effect and age at measurement as covariable. Scrotal circumference presented heritability of 0.47 ± 0.12. This value is similar to the heritabilities found for all testicular biometry traits (0.34–0.48). Sperm progressive motility, which has a direct effect on bull fertility, presented low heritability (0.07 ± 0.08). Major and total sperm defects presented moderate to high heritabilities (0.49 ± 0.18 and 0.39 ± 0.15, respectively), indicating that great genetic gain can be obtained through selection against sperm defects. High and positive genetic correlations were observed among testicular biometry traits, which also presented favourable genetic correlations with physical and morphological traits of the semen with magnitude ranging from high to low. Scrotal circumference presented moderate to high and favourable genetic correlations with sperm progressive motility, sperm turbulence, major sperm defects and total sperm defects. Thus, the selection for scrotal circumference results in favourable correlated genetic response for semen quality. The results show that the use of scrotal circumference as reference trait for bull fertility is appropriate, since it presents high heritability and favourable genetic correlation with semen quality.     

Evaluation of reproductive parameters in male Neotropical bats during dry and rainy months in a specific area of the Cerrado biome

The aim of this study was to analyse the reproductive aspects of male bats of three common species of the Phyllostomidae family: Artibeus lituratus, Platyrrhinus lineatus and Sturnira lilium, during dry and rainy months in a specific area of the Cerrado biome. Body weight was significantly higher during the dry months for S. lilium. The gonadosomatic index (GSI) and testicular weight were not significantly different between dry and rainy periods. The tubular parameters were significantly bigger in A. lituratus than in the other two species during both periods. No difference in the tubular/interstitial ratio was observed in any of the species during both periods. In both periods, all sperm cells and germ cell developmental stages were visible on seminiferous tubules whereas sperm cells were observed in epididymides of all sampled animals. The percentage of morphologically normal sperm was low (35%–60%), with no difference between periods. Spermatozoa from A. lituratus presented a leaf-shaped head, while the head was round-shaped in the other two species. In conclusion, our data suggest that males from the three studied species did not present reproductive latency during the most critical weather periods (dry and rainy months) in the metropolitan region of Brasilia, Brazil.     

Radiographic anatomy of the cockatiel (Nymphicus hollandicus) axial and appendicular skeleton

Cockatiels are popular pets. Still, despite medical and surgical relevance, the radiographic anatomy of the cockatiel (Nymphicus hollandicus) skeleton, like that of different wild and exotic bird species, has seldom been described. This study set out to describe the radiographic anatomy of the cockatiel skeleton. Twelve adult male and nine adult female specimens were radiographed using a digital X-ray system and different views. The radiographic anatomy of these birds was similar to that of other Psittacidae. However, some particularities inherent to the target species were detected, such as the presence of four flexion zones in the skull (craniofacial, nasal, jugal arch and palatine), complete bony orbit comprising a suborbital arch, 34–38 vertebrae (10 or 11 cervical, 8 or 9 thoracic, 9 or 10 lumbosacral, 5 or 6 caudal vertebrae and a pygostyle comprising 2 fused vertebrae), eight or nine pairs of ribs and a notarium made up of fused T2–T6 vertebrae. Poor radiopacity of the notarium, ribs and respective uncinate processes, and synsacral vertebrae made demarcation of these structures difficult. The appendicular skeleton of the cockatiel was very similar to that of other Psittacidae, and there were no gender-related differences.