Development of cryopreservation protocols for early stage zebrafish (Danio rerio) ovarian follicles using controlled slow cooling

Cryopreservation of germplasm of aquatic species offers many benefits to the fields of aquaculture, conservation and biomedicine. Although successful fish sperm cryopreservation has been achieved with many species, there has been no report of successful cryopreservation of fish embryos and late stage oocytes which are large, chilling sensitive and have low membrane permeability. In the present study, cryopreservation of early stage zebrafish ovarian follicles was studied for the first time using controlled slow freezing. The effect of cryoprotectant, freezing medium, cooling rate, method for cryoprotectant removal, post-thaw incubation time and ovarian follicle developmental stage were investigated. Stages I and II ovarian follicles were frozen in 4 M methanol and 3 M DMSO in either L-15 medium or KCl buffer. Ovarian follicle viability was assessed using trypan blue, FDA + PI staining and ADP/ATP assay. The results showed that KCl buffer was more beneficial than L-15 medium, methanol was more effective than DMSO, optimum cooling rates were 2-4 °C/min, stepwise removal of cryoprotectant improved ovarian follicle viability significantly and stage I ovarian follicles were more sensitive to freezing. The results also showed that FDA + PI staining and ADP/ATP assay were more sensitive than TB staining. The highest follicle viabilities after post-thaw incubation for 2 h obtained with FDA + PI staining were 50.7 ± 4.0% although ADP/ATP ratios of the cryopreserved follicles were significantly increased indicating increased cell damage. Studies are currently being carried out on in vitro maturation of these cryopreserved ovarian follicles.     

Brood size, sex ratio and egg load of Metaphycus lounsburyi (Hymenoptera: Encyrtidae) when parasitizing adult females of black scale Saissetia oleae (Hemiptera: Coccidae) in the field

Black scale Saissetia oleae (Olivier) (Hemiptera: Coccidae) is a widespread olive pest in California and Europe. Metaphycus lounsburyi (Howard) (Hymenoptera: Encyrtidae) is often one of its main parasitoids. Augmentative releases of M. lounsburyi have been proposed in those areas in which biological control is ineffective. In this paper, we study the relationship between black scale and M. lounsburyi in the field. According to our data, M. lounsburyi parasitizes mainly ovipositing females of black scale. Parasitism rates of ovipositing females reach high levels. In those scales, M. lounsburyi develops as a gregarious parasitoid, with an average of 13 and a maximum of 40 parasitoids developing per scale. The secondary sex ratio is female biased (proportion of males = 0.13) and appears to be brood size dependent. Metaphycus lounsburyi eggs are encyrtiform; however, its egg load is much higher than other Metaphycus that also parasitize black scale. The implications of these results on the behavioral ecology of M. lounsburyi and its use in biological control of black scale are discussed.     

The variances of the distributions of the combinations of the sexes within mammalian litters: Notes to mark the centenary of the problem

It is 100 years since Gini noted that in some samples of litters of mice and rabbits, the variances of the distributions of the combinations of the sexes are sub-binomial. In other words, in contrast with binomial expectation, there are too many litters in which the sexes are equally balanced, and there are too few unisexual litters. In the intervening years, this finding has been replicated in a number of further species, but no explanation has become established. Potential explanations are reviewed here, and it is suggested that the most likely cause is that, at the time of formation of the zygotes, p, the probability that a zygote will be male, varies from one zygote to another within litters, thus constituting an example of Poisson variation. And it is a standard result in probability theory that such variation causes sub-binomial variance.     

人工红树林湿地系统净化污水研究进展

咸淡水交接处这种特殊的生境造就了红树植物“抗污”和“降污”的功能,但能否将自然红树林湿地作为污水处理场所目前仍有争议。按照构建的人工红树林湿地的类型,分为三大类：模拟潮汐湿地系统、人工红树林污水处理系统以及红树林种植-养殖生态耦合系统。第一类系统模拟了自然海滩上的潮涨潮落的情形,第二类系统与一般人工湿地相类似,而第三类系统是指在养殖塘种植红树植物以去除有机物和营养盐。不同的类型反映了该领域研究工作的不断深入,展现了红树林湿地净化污水从理论探索向实际应用不断靠近的过程。应用人工红树林湿地净化污水具有较大的可行性,前景可观。     

Quantifying the impact of current and future tropospheric ozone on tree biomass, growth, physiology and biochemistry: a quantitative meta-analysis

The northern hemisphere temperate and boreal forests currently provide an important carbon sink; however, current tropospheric ozone concentrations ([O₃]) and [O₃] projected for later this century are damaging to trees and have the potential to reduce the carbon sink strength of these forests. This meta‐analysis estimated the magnitude of the impacts of current [O₃] and future [O₃] on the biomass, growth, physiology and biochemistry of trees representative of northern hemisphere forests. Current ambient [O₃] (40 ppb on average) significantly reduced the total biomass of trees by 7% compared with trees grown in charcoal‐filtered (CF) controls, which approximate preindustrial [O₃]. Above‐ and belowground productivity were equally affected by ambient [O₃] in these studies. Elevated [O₃] of 64 ppb reduced total biomass by 11% compared with trees grown at ambient [O₃] while elevated [O₃] of 97 ppb reduced total biomass of trees by 17% compared with CF controls. The root‐to‐shoot ratio was significantly reduced by elevated [O₃] indicating greater sensitivity of root biomass to [O₃]. At elevated [O₃], trees had significant reductions in leaf area, Rubisco content and chlorophyll content which may underlie significant reductions in photosynthetic capacity. Trees also had lower transpiration rates, and were shorter in height and had reduced diameter when grown at elevated [O₃]. Further, at elevated [O₃], gymnosperms were significantly less sensitive than angiosperms. There were too few observations of the interaction of [O₃] with elevated [CO₂] and drought to conclusively project how these climate change factors will alter tree responses to [O₃]. Taken together, these results demonstrate that the carbon‐sink strength of northern hemisphere forests is likely reduced by current [O₃] and will be further reduced in future if [O₃] rises. This implies that a key carbon sink currently offsetting a significant portion of global fossil fuel CO₂ emissions could be diminished or lost in the future.     

Differential turnover of the photosystem II reaction centre D1 protein in mesophyll and bundle sheath chloroplasts of maize

Photoinhibition is caused by an imbalance between the rates of the damage and repair cycle of photosystem II D1 protein in thylakoid membranes. The PSII repair processes include (i) disassembly of damaged PSII-LHCII supercomplexes and PSII core dimers into monomers, (ii) migration of the PSII monomers to the stroma regions of thylakoid membranes, (iii) dephosphorylation of the CP43, D1 and D2 subunits, (iv) degradation of damaged D1 protein, and (v) co-translational insertion of the newly synthesized D1 polypeptide and reassembly of functional PSII complex. Here, we studied the D1 turnover cycle in maize mesophyll and bundle sheath chloroplasts using a protein synthesis inhibitor, lincomycin. In both types of maize chloroplasts, PSII was found as the PSII-LHCII supercomplex, dimer and monomer. The PSII core and the LHCII proteins were phosphorylated in both types of chloroplasts in a light-dependent manner. The rate constants for photoinhibition measured for lincomycin-treated leaves were comparable to those reported for C3 plants, suggesting that the kinetics of the PSII photodamage is similar in C3 and C4 species. During the photoinhibitory treatment the D1 protein was dephosphorylated in both types of chloroplasts but it was rapidly degraded only in the bundle sheath chloroplasts. In mesophyll chloroplasts, PSII monomers accumulated and little degradation of D1 protein was observed. We postulate that the low content of the Deg1 enzyme observed in mesophyll chloroplasts isolated from moderate light grown maize may retard the D1 repair processes in this type of plastids.