First Production of Larvae Using Cryopreserved Sperm: Effects of Preservation Temperature and Cryopreservation on European Eel Sperm Fertilization Capacity

Sperm cryopreservation is a useful tool in captive fish reproduction management, that is to synchronize gamete production, especially in the case of species as the European eel, where the time of female spawning readiness is unpredictable. Several protocols to cryopreserve sperm of this species have been described, but until recently fertilization trials were not feasible. This study evaluated the effect of cold storage of diluted sperm prior to fertilizations and tested whether a previously defined protocol for European eel sperm cryopreservation can be successfully applied in fertilization trials to produce viable offspring. In our experiment, the sperm motility was evaluated after the extraction and the best samples were selected and pooled. Until stripping of eggs and fertilization, diluted sperm samples were maintained at either 4 or 20°C, or cryopreserved, following existing protocols. Fertilization of two egg batches was attempted. Diluted sperm caused a similar percentage of fertilized eggs and a similar number of embryos and larvae, independently of storage temperature (4 or 20°C). The cryopreserved sperm resulted in a lower percentage of fertilized eggs, but embryos developed and a few larvae (‘cryolarvae’) were obtained 55 h after fertilization in one of the two egg batches. This result evidences that the tested cryopreservation protocol is applicable for eel reproduction management, although improvements will be required to enhance fertilization success.     

Effects of Gonadotropins on In Vitro Maturation and of Electrical Stimulation on Parthenogenesis of Canine Oocytes

The objective of this study was to determine the effects of gonadotropins on in vitro maturation (IVM) and electrical stimulation on the parthenogenesis of canine oocytes. In experiment I, cumulus oocyte complexes were collected from ovaries at a random phase of the oestrus cycle and cultured on maturation medium treated with hCG or eCG for 48 or 72 h. There were no significant differences in the effects on the metaphase II (MII) rate between the hCG and eCG treatment groups over 48 h (5.4% vs 5.5%). The MII rate in the co-treatment group of hCG and eCG for 48 h was higher than in each hormone treated group (15.5%, p < 0.05). In experiment 2, the parthenogenetic effect on oocyte development, at various electrical field strengths (1.0, 1.5, 2.0 kV/cm DC) for 60 or 80 μs with a single DC pulse after IVM on the co-treatment of hCG and eCG, was examined. The rate of pronuclear formation (37.1%) in electrical activation at 1.5 kV/60 μs without cytochalasin B (CB) was higher than that of oocytes activated in the other groups (p < 0.05). However, we did not observe the cleavage stages. Also, CB did not influence parthenogenesis of canine oocytes. The results showed that the pronucleus formation rate, indicative of the parthenogenesis start point, could be increased by electrical stimulation. Therefore, these results can provide important data for the parthenogenesis of canine oocytes and suggest the probability of parthenogenesis in canines.     

Cilia and cilia-related disorders

Abstract

Extract

Among the puzzling aspects of the recently described autosomal recessive polycystic kidney disease (ARPKD) of sheep (Johnstone et al 2005 Johnstone, AC, Davidson, BI, Roe, AR, Eccles, MR and Jolly, RD. 2005. Congenital polycystic kidney disease in lambs. New Zealand Veterinary Journal, 53: 307–314. [Taylor &; Francis Online], [Web of Science ®] , [Google Scholar]) is the multiplicity of organ systems affected and the phenotypic variations that occur between the various animal species. It has been shown in studies of other inherited polycystic kidney disease syndromes that the normal gene products of a mutant are likely to be involved in ciliary structure and function. The studies that have led to this recognition can be expected to eventually provide the basis for a better understanding of the functioning of this organelle, and the pathogenesis of lesions in the related diseases.

It is well known that cilia are important in the perception of light, olfactory stimuli and sound and that motile cilia provide cell motility (e.g. sperm) and transport of mucus and other fluids. Recent research has indicated that the involvement of the primary cili-um/basal body complex is of central importance in the detection and cellular response to extracellular movement of fluid, critical phases of embryonic development, cell cycle regulation and maintenance of cell polarity.

Confocal microscopic studies of renal cyst epithelium in ovine ARPKD have shown that only 30% of cells have a cilium and that these are often truncated (McGlashan et al 2005 McGlashan, SR, Poole, CA, Stayner, C, Johnstone, AC, Eccles, MR and Jensen, CG. 2005. “Primary cilia in fibrosis associated with two models of polycystic kidney disease”. In Proceedings of the 45th Annual Conference of the American Society for Cell Biology December 10–14 [Google Scholar]). The observation, although preliminary, strongly supports the argument that the mutant gene in this disease of sheep normally encodes for a protein essential for primary ciliary function.     

Stability of Coniferous Tree Stems in Relation to Damage by Snow

In this work both conical and paraboloidal conifer stems aretaken to bend along curves calculated to produce uniform strainin the outer wood. Such curvature has been observed in windtunnel tests. From these curves for various displacements theresistive bending moment produced by the elasticity of the stemmay be calculated. The maximum value of this is related to thebreaking stress of the timber. The bending moments applied bythe weights of stem, crown and snowload are calculated by obtainingthe appropriate lever arms from the stem bending curves. Appliedand resistive bending moments are compared for 20m-high treesof various types. Densely grown trees of low taper are leaststable. Open grown trees with conical stems and high taper aremost stable. The effects on tree stability of wind, a leaningstem, an asymmetrical crown and basal bowing are also investigated.     

The effects of heavy thinning on stem quality and timber properties of silver birch (Betula pendula Roth)

A study was carried out into the effects of heavy thinning onstem quality and timber properties of silver birch (Betula pendulaRoth). Comparisons were made with lightly thinned and unthinnedbirch stands. Birch stands from the UK were also compared withsimilarly managed stands from Germany and Denmark. In general,trees from lightly thinned and unthinned stands had similarcharacteristics. Heavy thinning enhanced diameter incrementthrough greater ring width in comparison with the other treatments.The timber properties of basic density, grain angle and shrinkagecoefficients in the radial, tangential and longitudinal directions,however, were largely unaffected. More and bigger branches andgreater stem taper were found on trees from heavily thinnedstands, but these stems still met the current Finnish veneerbirch quality and dimension standards. Comparisons between birchstands, managed by heavy and light thinning, from the UK andGermany and Denmark revealed no meaningful differences in timberproperties. The results suggest that veneer dimension and qualitylogs could be produced in the UK in rotations as short as 40years on good sites if heavy thinning is adopted.     

Comparison of the efficacy of macrocyclic lactone anthelmintics,either singly or in combination with other anthelmintic(s), in nine beef herds in southern NSW

Anthelmintic resistance (AR) is a well-recognized challenge in farmed ruminants. The use of anthelmintics in combination is one of the strategies recommended to slow the rate of AR development. Two studies were undertaken in 2017 and 2019 to assess the efficacy of single-dose macrocyclic lactone (ML) anthelmintic and ML combination drenches. In total, 11 Faecal Egg Count Reduction Trials (FECRTs) were set up in 10 different beef herds, with results available from 10 of those FECRTs (9 herds). AR to a single ML anthelmintic was detected in all 9 herds, with resistance to Cooperia and Haemonchus spp on 9 farms, and resistance to Ostertagia and Trichostrongylus spp on 2 farms. In contrast, for the ML combination anthelmintics, all FECRTs resulted in efficacies of 99%–100%. The results suggest that cattle producers should strongly consider using combination drenches in their herds in preference to single actives.     

Molecular Basis of Gene-for-Gene Specificity in Bacterial Speck Disease of Tomato

Transient expression of the Pseudomonas syringae avirulence gene avrPto in plant cells resulted in a Pto-dependent necrosis. The AvrPto avirulence protein was observed to interact directly with the Pto resistance protein in the yeast two-hybrid system. Mutations in the Pto and avrPto genes which reduce in vivo activity had parallel effects on association in the two-hybrid assay. These data suggest that during infection the pathogen delivers AvrPto into the plant host cell and that resistance is specified by direct interaction of Pto with AvrPto.