Preharvest applications of ethephon reduce superficial scald of ‘Fuji’ and ‘Granny Smith’ apples in storage

In 1989 and 1990, preharvest applications of 2-chloroethyl phosphonic acid (ethephon) at dosages from 50–400 mg 1’1 were applied to ‘Fuji’ and ‘Granny Smith’ apples. In 1989, the greatest reduction in scald after storage on both ‘Fuji’ and ‘Granny Smith’, relative to the untreated control resulted from applying 400 mg 1^?1 ethephon five weeks before harvest and was 45% and 55% of controls, respectively. On ‘Granny Smith’ 400 mg 1^? was also effective when applied three weeks before harvest. In 1990, preharvest applications of ethephon at either 200 or 400 mg 1^?1 reduced scald in both cultivars relative to the controls. Ethephon applied six weeks before harvest had no dosage effect on scald reduction after storage. There was no consistent effect on fruit maturity at harvest from any treatment in either year, and no effect on quality of fruit out of storage.     

Ultrastructure of epicuticular wax aggregates during fruit development in apple (Malus domestica Borkh.)

Summary

Development of wax platelets on the surface of ‘Delicious’ (Malus domestica Borkh.) apple fruit was investigated throughout the growing season using field emission scanning electron microscopy. At 5,000 and greater, wax crystalline structures appeared to be composed of microtubules (MT), aggregates of individual MT to form single platelets, and of one or more platelets. The thickness of a single wax platelet ranged from 116–128 nm, approximately the diameter of a single MT; whereas multiple-platelet aggregates ranged in thickness from 307–428 nm. Individual platelets, multiple platelet aggregates and MT were visible throughout fruit development, as well as on different apple cultivars. Examination of the cuticle from young fruitlets (receptacle diameter = 3 mm) of ‘Chinese Crabapple’ (M. hupehensis Rehd.), which develops without trichomes, best demonstrated early platelet formation. A model for wax platelet and cuticle development on apple is proposed based on these data.     

Effects of Hormonal Supplementation on Nuclear Maturation and Cortical Granules Distribution of Canine Oocytes During Various Reproductive Stages

The aim of this study was to evaluate the effects of hCG, progesterone and oestradiol supplementation on nuclear and cytoplasmic maturation of canine oocytes cultured for 24, 48, 72 and 96 h. Oocytes obtained from 18 healthy bitches were divided into three groups according to their reproductive status (follicular, luteal and anoestrus stages) and cultured in TCM 199 + 25 UI/ml of hCG + 1 μg/ml of progesterone + 1 μg/ml of 17‐β oestradiol or without hormonal supplementation (control) for different periods. Then, they were stained with FITC‐LCA‐Hoescht for chromatin configuration and cortical granules distribution and evaluated under an epifluorescence microscope. Culture time and the influence of different stages of the oestrous cycle were also evaluated. The present study demonstrated that there was no significant difference among the reproductive stages. With regards to culture medium, only oocytes from the supplemented medium were able to complete meiosis; however, significant difference was only noticed in the percentage of MI stage oocytes (p < 0.05) in the follicular and luteal group at 72 h of culture. Most oocytes in germinal vesicle, germinal vesicle breakdown and metaphase I stage had cortical granules distributed throughout the cytoplasm (immature pattern), irrespective of the culture period (p < 0.05). Cortical granules distributed immediately beneath the plasma membrane (mature) was only observed in metaphase II stage oocytes, but not all of them presented matured cytoplasm. Our results reveal that cortical granules distribution in canine oocytes matured in vitro did not progressed in correspondence with nuclear stage changes and are in accordance with those from other species.     

Hypoosmotic Swelling Test in Alpaca (Vicugna pacos) Epididymal Spermatozoa

The present study intended to develop the hypoosmotic swelling (HOS) test in alpaca for its use in epididymal spermatozoa, to evaluate the membrane functional integrity and determine an appropriate hypoosmotic solution and whether the incubation time of 15 or 60 min is sufficient for the execution of the test. Hypoosmotic solutions (HS) with the following concentrations were used: 50, 100, 150, 200 and 275 mOsm/kg of sodium citrate tribasic dihydrate and d ‐fructose. Ten microlitres of epididymal sperm sample was mixed in 150 μL of the respective HS and incubated for 15 or 60 min at 38°C. From the proportion of reacted (swollen) spermatozoa, the 150 mOsm/kg HS was the most sensitive (p < 0.05). The exposure times (15 and 60 min) did not have significant differences (p > 0.05) in the proportion of both strong‐ and total‐coiled sperm tails. In conclusion, 150 mOsm/kg HS and 15 min exposure time are optimal to evaluate the plasma membrane functional integrity through the HOS test in alpaca epididymal spermatozoa.     

The Effect of Glycerol Concentrations on the Post‐thaw In Vitro Characteristics of Cryopreserved Sex‐sorted Boar Spermatozoa

The objective of this study was to optimize protocols for the cryopreservation of sex‐sorted boar spermatozoa. In the experiment 1, we evaluated the effects of a standard boar sperm cryopreservation procedure (3% final glycerol concentration) on the in vitro characteristics of sex‐sorted sperm frozen at low sperm concentrations (20 × 10⁶ sperm/ml; S20 group). Non‐sorted spermatozoa frozen at 1000 × 10⁶ (C1000 group) and 20 × 10⁶ (C20 group) sperm/ml were used as the freezing control groups. In experiment 2, the effects of different final glycerol concentrations (0.16%, 0.5%, 1.0%, 2.0% and 3.0%) on post‐thaw quality of the S20 and C20 groups were evaluated. In both experiments, the samples were evaluated prior to freezing (5°C) and at 30, 90 and 150 min after thawing. Experiment 1 indicated that freezing sperm at low concentrations decreased (p < 0.05) the total motility (TM) and progressive motility (PM) at 90 and 150 min after thawing regardless of whether the sperm were sorted or not. However, the sperm membrane integrity was not affected at any evaluation step. Inexperiment 2, significant effects on the TM and PM because of increased glycerol concentrations in the S20 and C20 groups were observed only at 90 and 150 min after thawing. The samples frozen in 3% glycerol showed lower (p < 0.05) TM and PM values when compared to those frozen in the presence of 0.5% and 1% glycerol. In both experiments, non‐sorted control samples displayed higher percentages of spermatozoa with damaged DNA than sorted spermatozoa. In conclusion, the optimization of cryopreservation conditions by decreasing the glycerol concentrations can improve post‐thaw motility of sex‐sorted spermatozoa frozen at low concentrations.     

Transfer of Bovine Blastocysts Derived from Short-term In Vitro Culture of Low Quality Morulae Produced In Vivo

The aim of this study was to evaluate if blastocysts arising from in vitro culture of Grade 3 bovine morulae produced in vivo can promote acceptable pregnancy rates when transferred into recipients. Embryos of different stages and qualities were recovered from superovulated Bos taurus and B. indicus donors. Grade 3 morulae were cultured in either Holding Plus® or TCM‐199 (supplemented with 10% bovine fetal serum) media for 24 h at 38.5°C. After this culture period, the resulting blastocysts were morphologically classified (Grades 1, 2 and 3) and transferred into recipients previously synchronized with the donors. Non‐cultured Grades 1 and 3 morulae were used as control. Pregnancy diagnosis was carried out 60 days after embryo transfer and the data were analysed by logistic regression, considering variables, such as embryo quality (Grade), donor breed, culture medium, donor‐recipient synchrony and seasonality. Embryo quality was the only variable, showing significant effect on the pregnancy rate. Pregnancy rates for non‐cultured Grade 1 and 3 morulae, and blastocysts arising from cultured Grade 3 morulae were 58.1% (n = 31), 17.1% (n = 35) and 51.1% (n = 47), respectively (p < 0.05). There were no statistical differences between non‐cultured Grade 1 morulae and cultured blastocysts. Pregnancy rates for Grades 1 (65.0%) and 2 (60.0%) were higher than Grade 3 (29.4%) cultured blastocysts (p < 0.05). It was concluded that short‐term in vitro culture is a very convenient method of identifying morphologically low quality morulae with higher chances of continuing development after the transfer into recipients.     

Prestorage ultraviolet-white light irradiation alters apple peel metabolome

Global metabolic profiling of 'Granny Smith' apple peel was employed for evaluating metabolomic alterations resulting from prestorage UV-white light irradiation. Apples were bagged midseason to restrict sunlight, harvested at the preclimacteric stage prior to bag removal, treated with fluorescent UV-white light for 0-48.5 h, and stored for 6 months at 0 degrees C. Trimethylsilyl (oxime) derivatized or underivatized aliquots of methanolic extracts from peel samples collected immediately after irradiation or following cold storage were evaluated using GC-MS and LC-UV/vis-MS, respectively. The profile, including more than 200 components, 78 of which were identified, revealed changes in the metabolome provoked by UV-white light irradiation and cold storage. Analyses of individual components selected using principal component analysis (PCA) models showed distinct temporal changes, before and after cold storage, related to prestorage irradiation in a diverse set of primary and secondary metabolic pathways. The results demonstrate metabolic pathways associated with ethylene synthesis, acid metabolism, flavonoid pigment synthesis, and fruit texture, are altered by prestorage irradiation, and many of the alterations are detectable after 6 months of cold storage in air.