The use of ultrasonography in the reproductive evaluation of boars

The objective was to study the use of ultrasound as a complementary test in the breeding soundness evaluation in male pigs and study the pattern of echogenicity of the testicular parenchyma in boars of different racial groups. Twenty‐six adult boars from four different racial groups were used, 10 from the Piau breed (group 1), four from the commercial and finishing group (group 2), six Pietrain breed (group 3) and six from the Duroc breed (group 4). All animals were evaluated for breeding soundness evaluation and the ultrasound examination of the testicles. The groups of animals that were evaluated showed no difference in the main semen parameters that were evaluated, except for the sperm volume, concentration of the ejaculated sperm and the supravital staining; the lowest figures were for the animals from the Piau breed (group 1). In relation to the testicular biometrics, Duroc animals (group 4) had a greater scrotal width compared to the other groups. But when we assessed the intensity of pixels of the testicles, there was a difference between groups. The groups 2 (finishing animals), 3 (Pietrain) and 4 had no difference between themselves. Group 3 had greater pixel intensity in relation to group 1. Of the 26 animals studied, five showed an abnormality during ultrasound evaluation, like hydrocele, hyperechoic mass in the testicular parenchyma, cyst in the head of the epididymis and the presence of fluid in the head and tail of the epididymis. The various animal groups studied did not differ in the principal reproductive parameters evaluated, showing that despite the great variability of reproductive traits between breeds and within the same breed, the breeding soundness evaluation, the more complete it is, is essential for the selection of breeders and the ultrasonography of the reproductive system becomes an important addition in this examination.     

Finding a needle in the haystack: Regional analysis of suitable Eld’s deer (Cervus eldi) forest in Southeast Asia

For species whose decline preceded the modern era and whose distribution is in the developing world, it is difficult to map suitable habitat across its former range. Eld’s deer (Cervus eldi) is an endangered cervid whose range across Southeast Asia was reduced during the last century to disjoint populations in Myanmar and Cambodia. We used ecological data from the present populations to determine landscape and habitat parameters that would help us predict the occurrence of the species in forests not yet surveyed. The suitable-forest GIS model was created using four readily available datasets for elevation, forest type, canopy closure, and human density. Comparison of the GIS model with 24 verified sightings of Eld’s deer during recent large mammal surveys in Cambodia, found 22 sightings (92%) within predicted suitable forest. Use the suitable-forest GIS model to survey a province in southern Lao People’s Democratic Republic, located a single, previously unreported population from 9 patches surveyed. In a separate analysis, a logistic regression model to predict Eld’s deer habitat in Northern Cambodia found percent tree cover, presence of wetlands, and distance to villages as the best predictors of deer, similar to variables used in the GIS model, with the exception of the importance of wetlands. Using mean annual rainfall to rank suitable-forest patches identified in the GIS model indicated dry dipterocarp forests in Northeastern Cambodia and Northern Myanmar have the highest potential to conserve eld’s deer. Examination of the suitable-forest GIS map and current protected areas indicated only Cambodia, with 11% suitable forest protected, has placed sufficient dry dipterocarp forest under protected status. Other Southeast Asia countries have not recognized dry dipterocarp forest as a significant ecotype worthy of conservation status.     

Oestradiol Induced Inhibition of Neuroendocrine Marker Expression in Leydig Cells of Adult Rats

The objectives of this work were to determine the changes in the expression of neuroendocrine markers in Leydig cell by oestradiol treatment, and to determine whether testosterone is able to recover partially the effects of hormonal suppression induced by oestradiol. Adult male rats were injected daily with either 50 microg of oestradiol or oestradiol plus testosterone propionate (25 mg every 3 days) for 15 days. The animals were sacrificed and testicles were dissected and processed by routine histological protocols. FSH and LH serum levels were determined by radioimmunoassay. The visualization of antigens was achieved by the streptavidin-peroxidase immunohistochemical method. Antibodies against chromogranin A (CrA), S-100 protein (S-100), P substance (PS), synaptofisin (SYN), neurofilament protein (NF), gliofibrillary acidic protein (GFAP) and neuron specific enolase (NSE) were used. The mean LH and FSH serum concentrations were consistently suppressed with hormonal treatments. Intermediate filaments (NF and GFAP) showed no difference in their expression. The expression of S-100, NSE and SYN was significantly lower in both hormone-treated groups. In oestradiol-treated rats, the immunoreactivity of CrA and SP decreased significantly but was restored after testosterone supplementation. Although the nature and functions of many of these substances in Leydig cells remain unknown, these results are consistent with the hypothesis that the expression of some neuroendocrine markers is hormonally controlled.     

Immunohistochemical Expression of Growth Factors in the Follicular Wall of Normal and Cystic Ovaries of Sows

The expression of growth factors was evaluated immunohistochemically in normal and cystic ovaries of sows. The immunohistochemically stained area (IHCSA) was quantified by image analysis to analyse the expression of these proteins in the follicular wall of secondary, tertiary and cystic follicles. IGF‐I immunoreactivity was strong in the granulosa cell layer (GC), moderate in the theca interna (TI) and mild in the theca externa (TE) of the normal follicles. There was severe reduction of the labelling to IGF‐I in the GC of the follicular and luteinized cysts. In the normal follicles, the reactivity for IGF‐II was very similar to pattern noted in IGF‐I. There was reduction of the IHCSAs in the GC of the follicular and luteinized cysts, but the decrease was not significant. The staining of the IGF‐II in the TI and TE of the cysts was increased, in comparison with normal follicles. The IHCSAs for VEGF were higher in the GC and TE of the normal follicles in contrast to TI, but this difference was noted only in the tertiary follicle. The VEGF reactivity increased in the GC of the cysts, in relation to normal follicles. The results of the current study show that the formation of ovarian cysts in sows is associated with alterations in the immunohistochemical expression of some growth factors.     

Cryopreservation of Piau‐Breed Wild Boar Sperm: Assessment of Cooling Curves and Centrifugation Regimes

This study aimed to assess the effects of different cooling curves and centrifugation regimes used in cryopreservation protocols on the post‐thaw viability of Piau‐breed wild boar (Sus scrofa) sperm using in vitro assessment tests. Two centrifugations (800  g for 10 min and 2400  g for 3 min) and two cooling curves (conventional cooling using nitrogen vapour – freezing 1 and automated cooling using a programmed freezing machine – freezing 2) were tested. Therefore, the treatments were divided into M3 – centrifugation at 2400  g for 3 min and freezing 2; M10 – centrifugation at 800  g for 10 min and freezing 2; R3 – centrifugation at 2400  g for 3 min and freezing 1; and R10 – centrifugation at 800  g for 10 min and freezing 1. No significant differences (p > 0.05) between treatments occurred post‐thawing regarding the total sperm motility means recorded. The mean values of the different treatments were not different from each other regarding the supravital staining (SV), hypo‐osmotic test (HO), sperm–egg binding assay or sperm morphology. This study showed that both the cooling curve and the centrifugation regime affected the quality of post‐thaw sperm, and centrifugation for shorter times and cooling curves using automated cooling are the most suitable for minimizing sperm injury.     

Factors affecting oat haploid production following oat × maize hybridization

Doubled haploids (DHs) are becoming increasingly important in crop breeding programmes but methods for producing oat DHs remain inefficient. In this study haploid and DH oat plants were produced using the oat × maize hybridization method. Factors influencing the rate of caryopsis and haploid embryo production including genotype, post‐pollination plant growth regulator application and temperature were investigated. The four growth regulators tested showed significant differences in their capacity to induce caryopsis formation with dicamba producing the highest numbers of caryopses, followed by picloram, 2,4‐dichlorophenoxyacetic acid (2,4‐D) and gibberellic acid (GA₃). No significant differences were observed between these growth regulators for their effect on embryo production. The concentration of dicamba was also important and was found to influence caryopsis but not embryo production, with 50 and 100 mg/l dicamba producing significantly more caryopses than 25 or 5 mg/l. Temperature had a significant impact on both caryopsis and embryo production with the magnitude and direction of response depending on genotype. Rates of haploid embryo production observed were between 0.8% and 6.7% of the pollinated florets. The proportion of haploids, which survived and were successfully doubled with colchicine following transfer to soil was between 72% and 81%.     

Interfering with regular meiotic behaviour in Avena sativa as a method of incorporating the gene for mildew resistance from A. barbata

Summary The regular meiotic behaviour of the cultivated oat Avena sativa (2n=6x=42) is genetically controlled. The factors which control the diploid-like meiotic behaviour also restrict the amount of pairing that occurs between alien chromosomes and their homoeologues in A.sativa, and hence increases the difficulties of introducing desirable variation from wild species into the cultivated oat. A genotype of the diploid species A.longiglumis which interferes with the regular meiotic behaviour of A. sativa can be used to induce pairing between alien chromosomes and their corresponding chromosomes in A. sativa. Using this procedure the dominant gene conferring mildew resistance has been transferred from the tetraploid weed species A. barbata into the cultivated oat.     

Volatile emissions of navel oranges as predictors of freeze damage

Volatile emissions of navel orange (Citrus sinensis L. Osbeck cv. Washington) fruit were evaluated as a means for predicting and gauging freeze damage. The fruits were subjected to -5 or -7 degrees C treatments in a laboratory freezer for various time periods of 2-9.5 h and stored at 23 degrees C for 1, 2, or 7 days, after which time the emission of volatiles from the fruit was measured. Following the final day of volatile measurements the fruits were stored at 5 degrees C for an additional 2-3 weeks and then evaluated for fruit quality characteristics. Peel injury in the form of brown lesions, drying of the juice vesicles, a decline in acidity, and a loss of flavor were observed to occur as a result of freezing. Corresponding to the loss in fruit quality were large increases in the emissions of ethanol, ethyl butanoate, methyl hexanoate, and ethyl octanoate. With the exception of methyl hexanoate, for which volatile emissions decreased during storage for 7 days at 23 degrees C, all of the other volatiles were relatively unchanged in amount by storage. Treatment at -7 degrees C caused greater injury, quality loss, and more volatile emanation than did freezing at -5 degrees C. The measurement of volatile emissions appears to be a useful approach to identify freeze-damaged navel oranges.