Efficiency of three boar sperm enrichment techniques

The objective of the study was to investigate the efficiency of three enrichment methods to separate boar spermatozoa. Twenty-four ejaculates from 12 boars (2 ejaculates/boar) were extended (30 × 10⁶ spermatozoa/mL) in commercial Beltsville Thawing Solution. Each semen sample was processed with glass wool column (GW) and glass beads (GB) filtration and with the single-layer centrifugation (SLC) technique. Semen samples before (control; C) and after treatment were evaluated for sperm CASA motility/kinetics and concentration, viability, morphology and chromatin integrity. Data were analysed with mixed models. The concentration of total and motile spermatozoa was significantly decreased after treatment in groups GW and SLC, but not in group GB. Group GW showed increased values of WOB compared with both groups C and GB. Group GB showed greater values of rapid movement spermatozoa and lower values of slow movement spermatozoa compared with group C. In group SLC, higher values of VSL, LIN and STR were observed compared with group C. In conclusion, all techniques under examination enhanced various CASA variables. Based on our results, the GB method is a promising alternative separation technique for boar sperm and deserves further research regarding swine in vitro fertilization.     

An atypical Bacillus anthracis infection in a bull—A potential occupational health hazard

Bacillus anthracis infecting cattle is usually identified based on the typical symptom: sudden death. Bacillus anthracis causing atypical symptoms may remain undiagnosed and represent a potential occupational health hazard for, that is veterinarians and producers, butchers and tanners. In the year 2004, one case of sudden death in a dairy farm in southern Finland was diagnosed as bovine anthrax. Four years later 2008, an atypical case of anthrax was diagnosed in the same holding. The bull was taken to the Production Animal Hospital of the Faculty of Veterinary Medicine, University of Helsinki because of fever, loss of appetite and a symmetrically swollen scrotal sac. Penicillin treatment cured the fever but not the swollen scrotum. Before the intended therapeutic castration, a punctuate consisting of 10 ml fluid collected into a syringe from the scrotal sac was cultivated on blood agar at 37°C. After 24 hr, an almost pure culture of a completely non‐hemolytic Bacillus cereus‐like bacteria was obtained. The strain was identified as B. anthracis using Ba‐specific primers by the Finnish Food Safety Authority (RUOKAVIRASTO). After the diagnosis, the bull was euthanized and destroyed, the personnel were treated with prophylactic antibiotics and the clinic was disinfected. In this particular case, treatment with water, Virkon S and lime seemed to be effective to eliminate endospores and vegetative cells since no relapses of anthrax have occurred in 10 years. This case is the last reported anthrax case in Finland.     

Plastination—A scientific method for teaching and research

Over the last four decades, plastination has been one of the best processes of preservation for organic tissue. In this process, water and lipids in biological tissues are replaced by polymers (silicone, epoxy, polyester) which are hardened, resulting in dry, odourless and durable specimens. Nowadays, after more than 40 years of its development, plastination is applied in more than 400 departments of anatomy, pathology, forensic sciences and biology all over the world. The most known polymers used in plastination are silicone (S10), epoxy (E12) and polyester (P40). The key element in plastination is the impregnation stage, and therefore depending on the polymer that is used, the optical quality of specimens differs. The S10 silicone technique is the most common technique used in plastination. Specimens can be used, especially in teaching, as they are easy to handle and display a realistic topography. Plastinated silicone specimens are used for displaying whole bodies, or body parts for exhibition. Transparent tissue sections, with a thickness between 1 and 4 mm, are usually produced by using epoxy (E12) or polyester (P40) polymer. These sections can be used to study both macroscopic and microscopic structures. Compared with the usual methods of dissection or corrosion, plastinated slices have the advantage of not destroying or altering the spatial relationships of structures. Plastination can be used as a teaching and research tool. Besides the teaching and scientific sector, plastination becomes a common resource for exhibitions, as worldwide more and more exhibitions use plastinated specimens.     

An in vitro comparative study of follicle stimulating hormone (FSH) and activin A effects on the maturation of preantral follicle-enclosed oocytes from immature Syrian mice

Objectives: It was aimed to investigate the effects of different doses of follicle stimulating hormone (FSH) and activin A on the growth and maturation of preantral mouse follicles during the in vitro culture. Methods: Preantral follicles (90-100 mum in diameter) were harvested from 6-8 week-old Syrian mice and cultured in TCM199 culture medium for 6 days to see the effect of FSH and Activin A. Activin A concentrations in the range of 10-200 ng/ml were used, while 10, 50, 100, 150 and 200 mIU/ml FSH were used in the experiment. Results: Activin A concentration of 100 ng/ml resulted in a significant increase in follicle diameter (170 mum) with the survival rate of 73% as compared to the control (100 mum and 25%, P<0.05). The number of oocytes matured and the percentage of germinal vesicle breakdown (GVBD) was 61 and 70%, respectively as compared to the control (20 and 29%, P<0.05). Follicle diameter (190 mum) and survival rate (85%) increased significantly in the presence of 100 mIU/ml of FSH as compared to the control (P<0.05). But, the administration of activin A+FSH increased the effect of both factors on follicular diameter (205 mum as compared to 100 mum in control, P<0.01). Follicle survival, oocyte maturation and GVBD rates were 91, 81 and 89%, respectively (P<0.01). Conclusion: These results have suggested that exposure to FSH and activin A before the formation of antral cavity had positive effect on follicle survival and oocyte robustness.     

Glyphosate effects on photosynthesis,nutrient accumulation,and nodulation in glyphosate‐resistant soybean

Previous greenhouse studies have demonstrated that photosynthesis in some cultivars of first‐ (GR1) and second‐generation (GR2) glyphosate‐resistant soybean was reduced by glyphosate. The reduction in photosynthesis that resulted from glyphosate might affect nutrient uptake and lead to lower plant biomass production and ultimately reduced grain yield. Therefore, a field study was conducted to determine if glyphosate‐induced damage to soybean (Glycine max L. Merr. cv. Asgrow AG3539) plants observed under controlled greenhouse conditions might occur in the field environment. The present study evaluated photosynthetic rate, nutrient accumulation, nodulation, and biomass production of GR2 soybean receiving different rates of glyphosate (0, 800, 1200, 2400 g a.e. ha^–1) applied at V2, V4, and V6 growth stages. In general, plant damage observed in the field study was similar to that in previous greenhouse studies. Increasing glyphosate rates and applications at later growth stages decreased nutrient accumulation, nodulation, leaf area, and shoot biomass production. Thus, to reduce potential undesirable effects of glyphosate on plant growth, application of the lowest glyphosate rate for weed‐control efficacy at early growth stages (V2 to V4) is suggested as an advantageous practice within current weed control in GR soybean for optimal crop productivity.     

Extracellular alpha-galactosidase from Debaryomyces hansenii UFV-1 and its use in the hydrolysis of raffinose oligosaccharides

Raffinose oligosaccharides (RO) are the factors primarily responsible for flatulence upon ingestion of soybean-derived products. ROs are hydrolyzed by alpha-galactosidases that cleave alpha-1,6-linkages of alpha-galactoside residues. The objectives of this study were the purification and characterization of extracellular alpha-galactosidase from Debaryomyces hansenii UFV-1. The enzyme purified by gel filtration and anion exchange chromatographies presented an Mr value of 60 kDa and the N-terminal amino acid sequence YENGLNLVPQMGWN. The Km values for hydrolysis of pNP alphaGal, melibiose, stachyose, and raffinose were 0.30, 2.01, 9.66, and 16 mM, respectively. The alpha-galactosidase presented absolute specificity for galactose in the alpha-position, hydrolyzing pNPGal, stachyose, raffinose, melibiose, and polymers. The enzyme was noncompetitively inhibited by galactose (Ki = 2.7 mM) and melibiose (Ki = 1.2 mM). Enzyme treatments of soy milk for 4 h at 60 degrees C reduced the amounts of stachyose and raffinose by 100%.     

The path from β-carotene to carlactone, a strigolactone-like plant hormone

Strigolactones, phytohormones with diverse signaling activities, have a common structure consisting of two lactones connected by an enol-ether bridge. Strigolactones derive from carotenoids via a pathway involving the carotenoid cleavage dioxygenases 7 and 8 (CCD7 and CCD8) and the iron-binding protein D27. We show that D27 is a β-carotene isomerase that converts all-trans-β-carotene into 9-cis-β-carotene, which is cleaved by CCD7 into a 9-cis-configured aldehyde. CCD8 incorporates three oxygens into 9-cis-β-apo-10'-carotenal and performs molecular rearrangement, linking carotenoids with strigolactones and producing carlactone, a compound with strigolactone-like biological activities. Knowledge of the structure of carlactone will be crucial for understanding the biology of strigolactones and may have applications in combating parasitic weeds.     

Effect of phosphate‐based seed priming on strigolactone production and Striga hermonthica infection in cereals

Strigolactones, plant‐secreted underground signalling molecules, play an important role in agricultural ecosystems, because they mediate the interaction of crops with symbiotic AM fungi and parasitic weeds like Striga hermonthica. Cereal host plants secret these signalling molecules particularly under nutrient‐deficient conditions and especially when phosphate (P) is limiting. The objective of the present study was to see the potential of P seed priming for S. hermonthica management in cereals in relation to strigolactone production. It has been demonstrated that P fertiliser application down‐regulates the production of these signalling molecules in the rhizosphere, which results in lower S. hermonthica infection of cereals. The laboratory study showed maximum production of strigolactones from dry and water‐soaked seeds, while seed soaking in P solution reduced their production. Similarly, maximum S. hermonthica infection was observed under control treatments with dry sowing or water soaking, while P seed soaking decreased S. hermonthica germination, emergence and dry biomass in all cereal crops. Our study shows that P seed priming resulted in lower exudation of strigolactones, which induced less S. hermonthica seeds germination and hence may lead to lower S. hermonthica infection. P‐based seed priming could prove to be an effective and affordable strategy to reduce S. hermonthica infection in cereals. Further research for practical field application is needed.     

Genetic and pathogenic diversity within Ascochyta rabiei(Pass.) Lab. populations in Pakistan causing blight of chickpea (Cicer arietinum L.)

Pathogenic and genetic diversity in Ascochyta rabiei populations in Pakistan were evaluated. Biological pathotyping of 130 A. rabiei isolates (obtained from hierarchically collected samples) was conducted on a set of three chickpea differentials, i.e. ILC 1929 (susceptible), ILC 482 (tolerant) and ILC 3279 (resistant), under controlled conditions. Disease severity data were recorded 12 days after inoculation. Statistical analysis grouped the isolates into three pathotype classes. Four isolates belonged to pathotype I (least aggressive), 79 isolates to pathotype II (medium aggressive) and 47 isolates to pathotype-III (highly aggressive).Genetic analysis was performed using RAPDs and oligonucleotide fingerprinting, where Hinf I-digested DNA was hybridized to the³²P-endlabeled oligonucleotide probes (CAA)₅, (GAA)₅, (GA)₈, (CA)₈and (GATA)₄. Dendrograms produced by cluster analysis discriminated 46 genotypes in the A. rabiei population of Pakistan. Genetic distances and relatedness between isolates were calculated. At a genetic distance of 0.3, genotypes were divided into six distinct genotype groups A, B, C, D, E and F containing 16, 11, 2, 5, 5 and 7 isolates, respectively. Most of the genotypes were area specific or predominated in certain areas but did not belong to a distinct pathotype, while most of the aggressive isolates (pathotype III) occurred in Northern Punjab and in the North Western Frontier Province.     

History and development of plastination techniques

Plastination was a game‐changing invention for macroscopic anatomical preparation. The method yielded dry, odourless, tangible and durable specimens which allowed new exhibition and teaching set‐ups and paved the way for sophisticated preparations and spectacular positioning of specimens. Despite the impact of the new method, there have been similar techniques in place before. Exsiccation techniques, polymer embeddings and specimen impregnation with hardening substances were earlier methods which already included the main concepts that were later combined and refined in plastination. S10 silicone plastination, the technique most commonly known and applied, was followed by plastination methods suitable for research and sectional anatomy teaching. Numerous variations of sheet plastination techniques allow research applications and new ways of presenting topographic relations and mesoscopic insights. Besides the development of plastination techniques in sensu stricto, related techniques had a renaissance with new applications and developments, including corrosion casting and diaphonization methods. This brief review shall provide a historical context of plastination including some anecdotal spotlights on the ideas and innovations that lead to nowadays plastination techniques.