Effects of polyethylene glycol and a synthetic ice blocker during vitrification of immature porcine oocytes on survival and subsequent embryo development

We evaluated the effects of polyethylene glycol (PEG) and Supercool X‐1000 (SC) as supplements during the vitrification of immature cumulus‐enclosed porcine oocytes in a solution based on 17.5% ethylene glycol + 17.5% propylene glycol. After warming, the oocytes were subjected to in vitro maturation, fertilization and embryo culture. In Experiment 1, equilibration and vitrification solutions were supplemented with or without 2% (w/v) PEG (PEG+ and PEG‐, respectively). The survival rate, cleavage and blastocyst development were similar between PEG+ and PEG‐ groups; however, all values were lower than those in the non‐vitrified control. In Experiment 2, vitrification solution was supplemented with or without 1% (v/v) SC (SC+ and SC‐, respectively). The percentages of survival and blastocyst development were similar between SC+ and SC‐ groups but lower than those in the non‐vitrified control. The percentage of cleavage in the SC‐ group was significantly lower than the control and the SC+ groups, which were in turn similar to one another. In both experiments, the cell numbers in blastocysts were not significantly different among the non‐vitrified and vitrified groups. In conclusion, PEG did not improve oocyte survival and embryo development, whereas SC improved the ability of surviving oocytes to cleave but not to develop into blastocysts.     

Secreted phosphatase activities in trypanosomatid parasites of plants modulated by platelet-activating factor

ABSTRACT The secreted phosphatase activities of two trypanosomatid parasites were characterized and compared with supernatants of living cells. The plant parasite Phytomonas fran?ai and the phytophagous hemipteran parasite Herpetomonas sp. hydrolyzed p-nitrophenylphosphate at a rate of 15.54 and 6.51 nmol Pi/mg of protein per min, respectively. Sodium orthovanadate (N(a)VO(3)) and sodium fluoride (NaF) decreased the phosphatase activities. The phosphatase activity of P. fran?ai was drastically diminished (73% inhibition) in the presence of sodium tartrate, whereas the phosphatase activity of Herpetomonas sp. was inhibited by 23%. Cytochemical analysis showed the localization of these enzymes on the external surface and in the flagellar pocket of the two trypanosomatids. Sodium tartrate inhibited this reaction, confirming the biochemical data. Platelet-activating factor modulated the phosphatase activities, inhibiting P. fran?ai activity and stimulating Herpetomonas sp. phosphatase activity.     

Hybrids of <Emphasis Type="Italic">Passiflora</Emphasis>: <Emphasis Type="Italic">P. gardneri</Emphasis> versus <Emphasis Type="Italic">P. gibertii,</Emphasis> confirmation of paternity,morphological and cytogenetic characterization

Two new varieties of interspecific hybrids of Passiflora have been developed from the cross between P. gardneri versus P. gibertii, both registered under the Passiflora Society International. Twelve putative hybrids were analyzed. Hybridization was confirmed using RAPD and SSR markers. Primer UBC11 (5′-CCGGCCTTAC-3′) generated informative bands. Primer SSR Pe75 has amplified species-specific fragments and a heterozygote status was observed with two parent bands 300 and 350 bp. The molecular markers generated have been analyzed for the presence or absence of specific informative bands. Based on the morphological characterization, we have identified two hybrid varieties: P. ‘Gabriela’ and P. ‘Bella’. P. ‘Gabriela’ produced flowers in bluish tones, bluish petals on the adaxial and abaxial faces, light blue sepals on the adaxial and light green on the abaxial faces, corona with the base of filaments in intense lilac color and white apex. P. ‘Bella’ produced flowers in lilac tones, intense lilac petals on the adaxial and abaxial faces, dark lilac sepals with whitish edges on the adaxial and light green on the abaxial faces, corona with the base of filaments in intense lilac color and white apex. The cytogenetic analysis verified that the hybrids have the same chromosomal number as the parents (2n = 18); the formation of bivalents between the homeologous chromosomes (n = 9) was observad, leading to regular meiosis, which allows the sexual reproduction and use of these hybrids in breeding programs.     

Performance of four <Emphasis Type="Italic">Trichogramma</Emphasis> species (Hymenoptera: Trichogrammatidae) as biocontrol agents of <Emphasis Type="Italic">Heliothis virescens</Emphasis> (Lepidoptera: Noctuidae) under various temperature regimes

Trichogramma spp. are major parasitoids of lepidopteran pest eggs, but there is large variation in efficacy toward a given pest among the numerous described Trichogramma species. It is important to select the Trichogramma species that most effectively parasitize and develop in target pest eggs for biological control. In this context, Trichogramma pretiosum, T. exiguum, T. atopovirilia and T. acacioi were studied in Heliothis virescens eggs under different thermal conditions. The parasitoids were reared at constant temperatures of 20, 25 and 30°C and tested at these respective temperatures, while parasitoids reared at 25°C were also tested at 20 and 30°C, for a total of 20 species–temperature combinations. About 30 H. virescens eggs were offered to the parasitoids for 24 h. Among the four species, parasitism rate by T. atopovirilia was highest at all temperature conditions, whereas T. acacioi had the lowest rates of parasitism at 25°C and 25/30°C. Parasitism ranged from 13.8 to 43.8% among all species–temperature combinations. Viability (emerged parasitoids) ranged from 80.8 to 98.4%, and was deemed satisfactory. The emergence rates of T. exiguum and T. acacioi were affected by temperature. Temperature also affected the sex ratio of T. exiguum at 25/30°C, whereas T. pretiosum and T. acacioi produced females predominantly independent of temperature. Overall, the parasitoid T. atopovirilia was the most efficient in parasitizing H. virescens eggs, though the levels of parasitism obtained might not ensure its successful use in biological control programs. The temperature-related differences in biological traits observed in the four Trichogramma species tested hint at the importance of making careful choices regarding climatic conditions where the parasitoid is going to be used when selecting a species for biological control programs.     

A common near infrared—based partial least squares regression model for the prediction of wood density of Pinus pinaster and Larix?×?eurolepis

Wood density is defined as the ratio of mass to volume and therefore in principle it should be possible to calculate a unique partial least squares regression (PLS-R) model for several species. PLS-R models for wood density based on X-ray microdensity data were calculated for each species Pinus pinaster and Larix × eurolepis and for both species together. After cross-validation and test set validation the data sets were combined and final models were calculated. The common model gave a residual prediction deviation (RPD) of 3.1, a range error ratio (RER) of 11.7, and a SEP/SEC of 1.06. The single models for Pinus pinaster and Larix?×?eurolepis gave RPD’s of 3.5 and 3.2, RER’s of 13 and 11, and a SEP/SEC of 1.2. To the best knowledge of the authors all obtained PLS-R models are the first ones that fulfil the requirements according to AACC Method 39-00 (AACC in AACC Method, 39-00:15, 1999) to be used at least for screening (RPD?≥?2.5). Although this method and the defined limits were developed for the analysis of grains they can be used as a rough rule of thumb until limits for wood are available. The improvement of the PLS-R models, compared to published results, might be due to three facts (1) the higher number of scans collected for a single spectrum, (2) that the samples were better represented by the NIR spectra and X-ray microdensity values, and (3) that the sites for the measurement of NIR spectra and X-ray microdensity were coincided as strictly as possibly.     

Immunohistochemical expression of HER - 2 and Ki - 67 in granulosa cell tumor in bitches

Granulosa cell tumour, an ovarian neoplasm of stromal origin, is an important tumour related to oestrogenic dominance syndrome and cystic endometrial hyperplasia–pyometra complex. In order to analyse ovarian tumour´s malignant potential, immunohistochemical markers can be used, such as anti-HER2 and anti-Ki-67. The aim of this study was to evaluate the expression of immunohistochemical markers HER-2 and Ki-67 in granulosa cell tumour from bitches´ ovaries. In HER-2 immunomarker analysis using the HercepTest^® method, most tumours were classified as 2+ (moderate labelling). Concerning Ki-67 immunomarker, only one case was described as having a high proliferative index. An association was found between immunostained cell percentage by anti-HER-2 antibodies and high pleomorphism, represented by the pattern of follicular/trabecular tumour arrangement. There was no correlation between anti-Ki-67 and anti-HER-2 antibody immunostaining intensities, probably due to only one case with a high Ki-67 index. With an effective protocol for HER-2 and Ki-67 immunohistochemical identification in granulosa cell tumours in bitches, it was possible to characterize this neoplasm proliferation profile.     

Establishment techniques affect productivity,nutritive value and atmospheric N2 fixation of two sunn hemp cultivars

Nitrogen fertilization is a common practice for sustaining forage production in forage systems in southeastern United States. Warm-season annual legumes may be an alternative forage to warm-season perennial grasses that do not require N fertilization. Sunn hemp (Crotalaria juncea L.) is a fast-growing, warm-season annual legume native to India and Pakistan. The objective of this 2-year study was to assess the herbage accumulation (HA), atmospheric N₂ fixation (ANF) and nutritive value of sunn hemp. Treatments were the factorial arrangement of two sunn hemp cultivars (“Crescent Sun” and “Blue Leaf”), three seeding rates (17, 28 and 39 kg seed/ha) and seed inoculation (inoculated or non-inoculated seeds), distributed in a randomized complete block design with four replicates. Crescent sun had greater HA (3,218 vs. 1764 kg DM/ha) and ANF (41 vs. 25 kg N/ha). Blue leaf had greater crude protein (CP) (188 vs. 176 g/kg) and in vitro digestible organic matter (IVDOM) concentrations (564 vs. 531 g/kg) than crescent sun. Non-inoculated seed had greater CP than inoculated seed, 188 and 177 g/kg, respectively, and inoculation did not affect HA. Intermediate seeding rate (28 kg/ha) decreased HA (2002 kg DM/ha), while HA from high and low seeding rates (17 and 39 kg/ha, respectively) did not differ (2,863 and 2,615 kg DM/ha respectively). Planting non-inoculated crescent sun at 17 kg/ha seeding rate is a feasible management practice to produce sunn hemp in subtropical regions; however, inoculation should always be recommended for proper establishment.