Interrelationships Between Apoptosis and Fertility in Bull Sperm

Male fertility, the ability of sperm to fertilize and activate the egg and support early embryogenesis, is vital for mammalian reproduction. Despite producing adequate numbers of sperm with normal motility and morphology, some males suffer from low fertility whose molecular mechanisms are not known. The objective was to determine apoptosis in sperm from high and low fertility bulls and its relationship with male fertility. DNA damage, phosphatidylserine (PS) translocation, and expression of pro- and anti-apoptotic proteins (BAX and BCL-2) in the sperm were determined using TUNEL, Annexin V, and immunoblotting approaches, respectively. Amounts of apoptotic spermatozoa were 2.86 (± 1.31) and 3.00 (± 0.96) in high and low fertility bulls, respectively (P=0.548), and were not correlated with fertility. There was a negative correlation between early necrotic spermatozoa and viable spermatozoa (r = –0.99, P<0.0001). Fertility scores were correlated with live spermatozoa detected by eosin-nigrosin test and necrotic spermatozoa determined via flow cytometry (r = –0.49, P<0.006 and r = –0.266, P<0.0113, respectively). BAX level was higher in low fertile group than high fertile group; however, this difference was not statistically significant due to the variations of bull samples (Bull 1–3 vs. Bull 4–5) in low fertile group (P<0.283). BCL-2 was not detectable in any of the sperm samples. The results shed light onto molecular and cellular underpinnings of male fertility.     

Optimization of a culture technique for the isolation of Listeria monocytogenes from faecal samples

A culture technique employing cold enrichment at 4 degrees C followed by selective enrichment and plating at higher temperatures (30 degrees C) was used to isolate Listeria monocytogenes from faecal samples. The samples were held at 4 degrees C for 15 weeks and cultured weekly to assess the sensitivity of the culture after cold storage for different lengths of time. No media, Listeria selective enrichment broth (LSEB), nutrient broth (NB) and saline were used as cold storage medium. Cold storage increased the frequency of Listeria positive samples. The sensitivity of the culture for Listeria spp. and L. monocytogenes was 72 and 94%, and 56 and 61% after third and seventh week of cold storage, respectively. When the results of third and seventh week of cold storage were combined, the sensitivity was 100% for Listeria spp. and 94% for L. monocytogenes. LSEB and NB as storage medium increased Listeria positive samples after the first week of cold storage but did not maintain the increase thereafter while saline had an adverse effect on the growth of the bacteria. However, samples held in no media in a pilot study involving monthly sampling of a herd revealed better results. Detection limit of the culture media was also investigated. The lowest concentration detected by culture media was 3.17 organisms/ml. This was seven organisms/g for known Listeria positive sample. The faecal samples spiked with 10-fold dilutions of L. monocytogenes and held at 4 degrees C revealed that the sample spiked with 3.17 x 10-1 cfu/ml organisms resulted in growth after the second week of cold storage. The results suggest that the culture technique employing cold enrichment followed by selective enrichment and plating is more sensitive, the storage of faecal samples in no media when compared with the samples in storage medium, LSEB, NB and saline, during cold enrichment is a better application and culture of faeces, immediately after collection, at third and seventh week of cold enrichment produce more satisfactory results.     

Anatomical Study of the Gastrointestinal Tract in Free‐living Axis Deer (Axis axis)

The macroscopic anatomy of the stomach and intestines of adult axis deer (Axis axis), a cervid species considered intermediate/mixed feeder, was observed and recorded. Nine adult wild axis deers of both sexes were used and studied by simple dissection. The ruminal papillae were distributed unevenly in the overall area of the inner surface of rumen and primarily were more large and abundant within the atrium. The ruminal pillars had no papillae. There was an additional ruminal pillar located between the right longitudinal and right coronary ventral pillars connected to the caudal pillar. No dorsal coronary pillars were found, and the ventral coronary pillars are connected. The reticulum was the third compartment in size, and the maximum height of the reticular crests was 1.0 mm. The Cellulae reticuli were not divided and rarely contained secondary crests. There were no Papillae unguiculiformes. The omasum was the smallest gastric compartment. The abomasum had about twelve spiral plicae, and a small pyloric torus was present. The intraruminal papillation was similar to those species that are characterized by a higher proportion of grass in their natural diet. The finding of the small reticular crests is typical for browser ruminants and was coincident with data reported for other deer. The comparative ratio of the small intestine to the large intestine was 1.69, in terms of length measurements in axis deer and appears below of the ‘browser range’. We concluded that the gastrointestinal system of axis deer reflected similar morphological characteristics of the both types of ruminants: browser and grazer, and we consider it as an intermediate feeder.     

Antioxidative role of selenium against the toxic effect of heavy metals (Cd<Superscript>+2</Superscript>, Cr<Superscript>+3</Superscript>) on liver of rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> Walbaum 1792)

The main purpose of this study is to discuss the effect of Cd⁺², Cr⁺³ and Se metals on biochemical parameters in liver tissue of Oncorhynchus mykiss. The rainbow trout were exposed to heavy metal stress (Cd⁺², Cr⁺³) at 2 ppm dosage. The present study was undertaken to determine the protective effect of selenium treatment at the same dosage (2 ppm) on some biochemical parameters. The activity of catalase (CAT), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and the changes in levels of malondialdehyde (MDA) from biochemical parameters were determined in liver tissue of the fish groups exposed to heavy metals, especially for the selenium-applied groups. Results of this study showed that the activities of CAT, GSH-Px and SOD in the tissues of fish exposed to the stress of Cd⁺² and Cr⁺³ were significantly lower than the control groups (P < 0.05). Meanwhile, the closer values to the control groups were obtained in selenium-added groups (Cr⁺³ + Se⁺⁴, Cd⁺² + Se⁺⁴). For the level of MDA, the last production of lipid peroxidation showed increases (P < 0.05) in the groups exposed to the metal stress, whereas significant decreases were obtained in selenium-applied groups. The result of the statistical evaluation showed that the negative effects occurring in the biochemical parameters of the applied groups exposed to the toxicity of heavy metal were significantly eliminated (P < 0.05) as a result of selenium treatment.     

Potential for using the World Reference Base for Soil Resources to identify less favoured areas

The European Commission (EC) has adopted a resolution seeking to test a proposed set of eight soil and climate criteria that objectively assesses and delimits agricultural areas suffering from natural handicaps. In areas designated as ‘Less Favoured Areas’ (LFA) agricultural production is hampered by natural handicaps, for example difficult climatic conditions, steep slopes in mountain areas or poor soil productivity. To delineate LFAs, a set of factsheets were provided by the EC Joint Research Center for mapping the proposed common soil, climate and terrain criteria. The present study was performed to determine the suitability of the World Reference Base for Soil Resources, 2006 (WRB) soil classification system in terms of estimating LFA thresholds. It was concluded that the WRB soil categories are directly applicable for delineating areas affected by the LFA criteria for soil constraints Organic Soil and Vertic Properties. The WRB categories correctly defined areas constrained by Salinity and Sodicity, but since they are stricter, than the LFA criteria, further analysis is needed to delineate the larger areas corresponding to LFA criteria. The climate and terrain criteria of LFA are only broadly indicated by WRB categories.     

Potential and limitations of isozymes for chromosomal localization of resistance genes against barley mild mosaic virus (BaMMV)

Summary To assess the possibilities offered by isozymes to locate resistance genes against barley mild mosaic virus (BaMMV), the isozyme patterns of 19 barley (Hordeum vulgare L.) genotypes carrying genes different from ym4 were determined. Of the 15 isozyme systems tested, only three were polymorphic, namely aconitate hydratase, esterase, phosphogluconate dehydrogenase, providing markers on four of the seven barley chromosomes. Studies of F₂ progenies derived from three crosses between resistant genotypes and susceptible varieties failed to reveal linkage between resistance genes and isozymes. Another goal of the experiment was to study the linkage relationships between ym4 and the esterase locus (Est1-Est2-Est4). Our estimates of the recombination rate between these two loci (3.41 and 8.32%) were in the range of those reported between these esterases and one of the resistance genes of the Chinese variety Mokusekko 3.