Sperm cryopreservation of tiete tetra Brycon insignis (Characiformes): effects of cryoprotectants,extenders, thawing temperatures and activating agents on motility features

The aim of this study was to test the effects of cryoprotectants [dimethyl sulphoxide (DMSO) and methylglycol], extenders (0.9% NaCl, 5% glucose, Beltsville Thawing Solution^? and Merck III^?), thawing temperatures (30 and 60 °C) and activating agents (0.29% NaCl and 1% NaHCO₃) on the cryopreservation process of tiete tetra Brycon insignis sperm. Sperm was loaded in 0.5 mL straws, frozen in nitrogen vapour at ?170 °C and stored in liquid nitrogen. Post‐thaw sperm quality was evaluated in terms of subjective motility rate, quality motility score (0=no movement; 5=rapidly swimming spermatozoa), duration of motility and vitality (eosin–nigrosin staining). Post‐thaw sperm motility rate was greater in methylglycol (76–88%), compared with DMSO (23–59%). In general, the highest quality motility scores were observed when sperm was thawed at 30 °C and triggered in 1% NaHCO₃ (3.5–4.3). Duration of motility was longer when triggered in 1% NaHCO₃ (95–120 s) compared with 0.29% NaCl (69–107 s). Sperm vitality was not affected by any of the parameters tested and varied from 51% to 69% intact sperm. Brycon insignis sperm frozen in methylglycol combined with any of the extenders tested and using the methods described above yields motility above 57% and that should last long enough to fertilize oocytes.     

Development of an appropriate procedure for estimation of RUSLE EI30 index and preparation of erosivity maps for Pulau Penang in Peninsular Malaysia

Pluviographic data at 15 min interval from 6 stations in Pulau Penang of Peninsular Malaysia were used to compute rainfall erosivity factor (R) for the revised universal soil loss equation (RUSLE). Three different modelling procedures were applied for the estimation of monthly rainfall erosivity (EI₃₀) values. While storm rainfall (P) and duration (D) data were used in the first approach, the second approach used monthly rainfall for days with rainfall ≥ 10 mm (rain₁₀) and monthly number of days with rainfall ≥ 10 (days₁₀). The third approach however used the Fournier index as the independent variable. Based on the root mean squared error (RMSE) and the percentage error (PE) criteria, models developed using the Fournier index approach was adjudged the best with an average PE value of 0.92 and an average RMSE value of 164.6. Further, this approach was extended to the development of a regional model. Using data from additional sixteen stations and the Fournier index based regional model, EI₃₀ values were computed for each month. ArcView GIS was used to generate monthly maps of EI₃₀ values and also annual rainfall erosivity (R). The rainfall erosivity factor (R) in the region was estimated to vary from 9000 to 14,000 MJ mm ha^− 1 h^− 1 year^− 1.     

鸽毛滴虫外泌体的分离鉴定及蛋白质谱分析

旨在研究鸽毛滴虫是否分泌外泌体及虫源外泌体的蛋白组成和功能,为探索外泌体在鸽毛滴虫寄生过程中的作用奠定基础。采集鸽毛滴虫感染个体口腔及咽部病灶处样本,进行分离培养及纯化,使用显微镜鉴定虫体形态,提取DNA并比对其序列,确定培养物是否为鸽毛滴虫虫株;通过对培养液上清进行超速离心,透射电子显微镜、Western blot技术和纳米颗粒追踪分析（nanoparticle tracking analysis,NTA）鉴定培养液上清中是否含有虫源外泌体;最后通过Label-free技术分析虫源外泌体的蛋白质谱表达。结果显示,虫体具有明显的毛滴虫形态及特征,与鸽毛滴虫ITS1/5.8S/ITS2基因比对率达98%。从虫体培养液中提取的囊泡经透射电子显微镜观察具有明显的茶托样结构;NTA结果显示,粒径集中于125.1 nm,峰值粒径为132.3 nm,占比99.3%;CD63和TSG101等外泌体标记蛋白的条带明显,表明提取物为外泌体。质谱结果显示,烯醇酶、3-磷酸甘油醛-脱氢酶、磷酸甘油酸变位酶和延伸因子为表达量较高的蛋白质。GO功能注释显示外泌体蛋白富集到细胞内和胞浆等细胞组分,发挥GTP连接和GTP酶活性功能,参与小GTPase介导的信号转导与糖酵解等过程。KEGG通路富集表明,虫源外泌体蛋白主要富集于代谢途径、糖酵解/糖异生、抗生素的生物合成和次级代谢产物的生物合成等通路。本研究发现鸽毛滴虫可以分泌外泌体,并对虫源外泌体进行蛋白质谱分析,提示虫源外泌体中的蛋白质在寄生虫的能量代谢、信号转导及生物合成等过程中发挥重要作用。     

Landscape structure shapes carnivore-mediated seed dispersal kernels

Context

Seed dispersal is recognized as having profound effects on the distribution, dynamics and structure of plant populations and communities. However, knowledge of how landscape structure shapes carnivore-mediated seed dispersal patterns is still scarce, thereby limiting our understanding of large-scale plant population processes.

Objectives

We aim to determine how the amount and spatial configuration of forest cover impacted the relative abundance of carnivorous mammals, and how these effects cascaded through the seed dispersal kernels they generated.

Methods

Camera traps activated by animal movement were used for carnivore sampling. Colour-coded seed mimics embedded in common figs were used to know the exact origin of the dispersed seed mimics later found in carnivore scats. We applied this procedure in two sites differing in landscape structure.

Results

We did not find between-site differences in the relative abundance of the principal carnivore species contributing to seed dispersal patterns, Martes foina. Mean dispersal distance and the probability of long dispersal events were higher in the site with spatially continuous and abundant forest cover, compared to the site with spatially aggregated and scarcer forest cover. Seed deposition closely matched the spatial patterning of forest cover in both study sites, suggesting behaviour-based mechanisms underpinning seed dispersal patterns generated by individual frugivore species.

Conclusions

Our results provide the first empirical evidence of the impact of landscape structure on carnivore-mediated seed dispersal kernels. They also indicate that seed dispersal kernels generated strongly depend on the effect that landscape structure exerts on carnivore populations, particularly on habitat-use preferences.

     

Effect of mixed inoculations with Fusarium oxysporum f. sp. lycopersici and F. oxysporum f. sp. dianthi on the phenols content of tomato plants

Forms ofFusarium oxysporum specific on hosts other than tomato induce in this plant greater initial increases of the phenols content than the pathogenic f. sp.lycopersici. Mixed inoculations of f. sp.lycopersici and f. sp.dianthi are on the contrary no more effective in inducing the phenol accumulation 24 h after the infection than f. sp.lycopersici alone. This observation suggests that the pathogen can suppress the phenolic response that is typical of the incompatible combinations.Samenvatting Vormen vanFusarium oxysporum welke pathogeen zijn voor andere planten dan de tomaat induceren in deze plant aanvankelijk een grotere toename van het fenolgehalte dan de pathogene f. sp.lycopersici. Inoculaties met een gemengd inoculum van de f. sp.lycopersici en f. sp.dianthi hebben daarentegen geen groter effect op de toename van het fenolgehalte 24 uur na infectie dan de inoculaties met f. sp.lycopersici alleen. Verondersteld wordt dat het pathogeen de toename van het fenolgehalte, dat typerend is voor de incompatibele combinatie, kan onderdrukken.     

Reduction and stabilization of hip luxation by the transposition of the ligamentum sacrotuberale in dogs: an in vivo study

OBJECTIVES: To evaluate the effectiveness of transposition of the sacrotuberous ligament (LST) for the treatment of coxofemoral luxation in the dog. STUDY DESIGN: In vivo experimental study. SAMPLE POPULATION: Ten mixed-breed dogs (weighing 12 to 26 kg). METHODS: After general anesthesia, the LST was exposed and released from the sacrum with a sacral bone fragment. Coxofemoral luxation was created by capsulotomy and transection of the ligament of the head of the femur. Suture was passed through 2 small holes in the bone fragment to guide transposition of the LST through a tunnel drilled through the acetabulum and femoral head and neck. The suture material was pulled tight and an interference screw was placed into the femoral tunnel to lock the sacral bone fragment and LST securely in the tunnel. Butorphanol was administered for pain relief and dogs were allowed unrestricted activity. Coxofemoral radiographs were obtained at 15-day intervals. Two dogs were euthanatized for macroscopic and histopathologic examinations at 3 months. RESULTS: Release of the sacral bone was moderately difficult. The transposed ligament and bone fragment were of an appropriate length and were easily and securely fixed by a interference screw method. Visible severe lameness, during the first 7 to 10 days improved to mild lameness within 10 to 21 days. Gait was subjectively normal after 5 weeks. No radiographic abnormalities were observed at any time point. Grossly, the LST was intact and apparently viable and functional. On histopathology, the space in the bone tunnel was filled with new tissue and a hyaline-like layer surrounded the LST. The sacral bone fragment attached to the LST was united with the femoral bone. CONCLUSIONS: Transposition of the LST reduced and stabilized experimentally induced coxofemoral luxation in 10 dogs. Survival of the LST up to 3 months and its adaptation to transposition suggest that this surgical technique could be considered as a permanent treatment option for hip luxations. Transposition and fixation of the LST was easy, but releasing the sacral edge of the LST was difficult because of its deep location and division of the origin into 2 branches in some dogs. CLINICAL RELEVANCE: Results of this experimental study suggest that the technique may be satisfactorily used in dogs with coxofemoral luxation.     

Avipoxvirus infection in peregrine falcons (Falco peregrinus) from a reintroduction programme in Germany

Poxvirus infections are common in domestic birds in Germany, but they are rare in birds of prey. Only species of falconidae imported from Arabian or Asian countries have so far tested positive for poxvirus, and, among these, only raptors kept for falconry. As part of a reintroduction programme in the northern county of Mecklenburg-Western Pomerania, which is adjacent to the Baltic Sea, 21 young peregrine falcons were released into the wild; six of them died and one was examined postmortem, its tissues being examined by light and electron microscopy. In addition, an ELISA for fowlpox, pigeonpox and canarypox was applied. No virus could be isolated and propagation in culture failed, but virus particles were detected by electron microscopy in lesions from its skin and tongue.