Effect of pregnancy on the genetic evaluation of dairy cattle

We investigated the effect of stage of pregnancy on estimates of breeding values for milk yield and milk persistency in Gyr and Holstein dairy cattle in Brazil. Test-day milk yield records were analyzed using random regression models with or without the effect of pregnancy. Models were compared using residual variances, heritabilities, rank correlations of estimated breeding values of bulls and cows, and number of nonpregnant cows in the top 200 for milk yield and milk persistency. The estimates of residual variance and heritabilities obtained with the models with or without the effect of pregnancy were similar for the two breeds. Inclusion of the effect of pregnancy in genetic evaluation models for these populations did not affect the ranking of cows and sires based on their predicted breeding values for 305-day cumulative milk yield. In contrast, when we examined persistency of milk yield, lack of adjustment for the effect of pregnancy overestimated breeding values of nonpregnant cows and cows with a long days open period and underestimated breeding values of cows with a short days open period. We recommend that models include the effect of days of pregnancy for estimation of adjustment factors for the effect of pregnancy in genetic evaluations of Dairy Gyr and Holstein cattle.     

Measurement of cooling and warming rates in vitrification‐based plant cryopreservation protocols

Cryopreservation protocols include the use of additives and pretreatments aimed to reduce the probability of ice nucleation at all temperatures, mainly through micro‐viscosity increase. Still, there is a risk of ice formation in the temperature region comprised between the equilibrium freezing (T_f) and the glass transition (T_G) temperatures. Consequently, fast cooling and warming, especially in this region, is a must to avoid ice‐derived damage. Vitrification and droplet‐vitrification techniques, frequently used cryopreservation protocols based in fast cooling, were studied, alongside with the corresponding warming procedures. A very fast data acquisition system, able to read very low temperatures, down to that of liquid nitrogen, was employed. Cooling rates, measured between ?20°C and ?120°C, ranged from ca. 5°C s^?1 to 400°C s^?1, while warming rates spanned from ca. 2°C s^?1 to 280°C s^?1, for the different protocols and conditions studied. A wider measuring window (0°C to ?150°C) produced lower rates for all cases. The cooling and warming rates were also related to the survival observed after the different procedures. Those protocols with the faster rates yielded the highest survival percentages. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:1177–1184, 2014     

The Pht1;9 and Pht1;8 transporters mediate inorganic phosphate acquisition by the Arabidopsis thaliana root during phosphorus starvation

? The activation of high-affinity root transport systems is the best-conserved strategy employed by plants to cope with low inorganic phosphate (Pi) availability, a role traditionally assigned to Pi transporters of the Pht1 family, whose respective contributions to Pi acquisition remain unclear. ? To characterize the Arabidopsis thaliana Pht1;9 transporter, we combined heterologous functional expression in yeast with expression/subcellular localization studies and reverse genetics approaches in planta. Double Pht1;9/Pht1;8 silencing lines were also generated to gain insight into the role of the closest Pht1;9 homolog. ? Pht1;9 encodes a functional plasma membrane-localized transporter that mediates high-affinity Pi/H? symport activity in yeast and is highly induced in Pi-starved Arabidopsis roots. Null pht1;9 alleles exhibit exacerbated responses to prolonged Pi limitation and enhanced tolerance to arsenate exposure, whereas Pht1;9 overexpression induces the opposite phenotypes. Strikingly, Pht1;9/Pht1;8 silencing lines display more pronounced defects than the pht1;9 mutants. ? Pi and arsenic plant content analyses confirmed a role of Pht1;9 in Pi acquisition during Pi starvation and arsenate uptake at the root-soil interface. Although not affecting plant internal Pi repartition, Pht1;9 activity influences the overall Arabidopsis Pi status. Finally, our results indicate that both the Pht1;9 and Pht1;8 transporters function in sustaining plant Pi supply on environmental Pi depletion.     

Alternative stable equilibria and critical thresholds created by fire regimes and plant responses in a fire‐prone community

Wildfire is a dominant disturbance in many ecosystems, and fire frequency and intensity are being altered as climates change. Through effects on mortality and regeneration, fire affects plant community composition, species richness, and carbon cycling. In some regions, changes to fire regimes could result in critical, non‐reversible transitions from forest to non‐forested states. For example, the Klamath ecoregion (northwest United States) supports extensive conifer forests that are initially replaced by hardwood chaparral following high‐severity fire, but eventually return to conifer forest during the fire‐free periods. Climate change alters both the fire regime and post‐fire recovery dynamics, potentially causing shrubland to persist as a stable (i.e. self‐renewing) vegetation stage, rather than an ephemeral stage. Here, we present a theoretical investigation of how changes in plant traits and fire regimes can alter the stability of communities in forest‐shrub systems such as the Klamath. Our model captures the key characteristics of the system, including life‐stage‐specific responses to disturbance and asymmetrical competitive interactions. We assess vegetation stability via invasion analysis, and conclude that portions of the landscape that are currently forested also can be stable as shrubland. We identify parameter thresholds where community equilibria change from stable to unstable, and show how these thresholds may shift in response to changes in life‐history or environmental parameters. For instance, conifer maturation rates are expected to decrease as aridity increases under climate change, and our model shows that this reduction decreases the fire frequencies at which forests become unstable. Increases in fire activity sufficient to destabilize forest communities are likely to occur in more arid future climates. If widespread, this would result in reduced carbon stocks and a positive feedback to climate change. Changes in stability may be altered by management practices.     

Assessing the risk to nontarget organisms from Bt corn resistant to corn rootworms (Coleoptera: Chrysomelidae): Tier-I testing with Orius insidiosus (Heteroptera: Anthocoridae)

A 14-d continuous dietary exposure bioassay using nymphs of the insidious flower bug, Orius insidiosus (Say) (Heteroptera: Anthocoridae), was conducted to assess nontarget impacts of genetically modified corn event MON 863 expressing the Cry3Bb1 protein for management of corn rootworms, Diabrotica spp. (Coleoptera: Chrysomelidae). Nymphs of O. insidiosus were continuously fed a bee pollen diet inoculated with a maximum hazard exposure dose (930 microg/g of diet) of the Cry3Bb1 protein for 14 d. The Cry3Bb1 protein at a concentration of 930 microg/g of diet had no adverse effect on the survival and development (to adults) of O. insidiosus nymphs. In contrast, when O. insidiosus nymphs were fed bee pollen diet treated with a hazard dose of the protease inhibitor E64 (53 microg/g of diet) or the stomach poison potassium arsenate (8.9 microg/g of diet), all nymphs died before developing to adults. Furthermore, statistical power analysis indicated that at levels of 80% power and a 5% type I error rate, the study design would have been able to detect a minimum 30% reduction in survival of test nymphs and a 20% reduction in nymphal development to the adults relative to the buffer control groups. Based on the maximum level (93 microg/g) of the Cry3Bb1 protein expressed in MON 863 corn tissues including leaves, roots, and pollen, findings from this study indicate that corn hybrids containing the MON 863 event have a minimum 10 times safety factor for nymphs of O. insidiosus and thus pose minimal risk to this beneficial insect.     

Evaluation of characteristics of Pediococcus spp. to be used as a vaginal probiotic

Aims

The aim of our research was to select, identify and characterize an isolate of lactic acid bacteria to be considered as a vaginal probiotic.

Methods and Results

Thirty‐five isolates of Pediococcus spp. showed bacteriocinogenic activity against Listeria monocytogenes and the ability to survive in simulated vaginal fluid (SVF) at pH 4·2. One isolate of Pediococcus spp. was selected and characterized to evaluate its safety before the use as vaginal probiotic. Pediococcus pentosaceus SB83 did not show the presence of virulence factors such as the production of gelatinase, lipase and DNase, haemolytic activity, nor the presence of virulence genes (genes esp, agg, gelE, efaAfm, efaAfs, cylA, cylB and cylM). Pediococcus pentosaceus SB83 was considered sensitive to chloramphenicol, gentamicin, streptomycin, kanamycin, erythromycin and ampicillin. This strain was considered resistant to tetracycline and vancomycin. Pediococcus pentosaceus SB83 was a biofilm producer at different pH values (4·2, 5·5 and 6·5) in SVF and in de Man, Rogosa and Sharpe medium.

Conclusions

The in vitro results provide a basis for the use of P. pentosaceus SB83 as a vaginal probiotic, to prevent colonization by L. monocytogenes in pregnant women.

Significance and Impact of Study

The application of vaginal probiotics could have the potential for preventing vaginal infections and consequently reduce abortion and neonatal infections.     

Non-Polar Chemical Constituents of Atemoya and Evaluation of the Cytotoxic and Antimicrobial Activity

This study aimed to identify the chemical composition of essential oil from fruits (EOAF) and the hexanic crude extract from aerial parts (At-Hex) of atemoya (Annona cherimola x Annona squamosa), a hybrid belonging to the Annonaceae family. Cytotoxic and antimicrobial activity was also evaluated. OEAF was obtained by hydrodistillation using a Clevenger apparatus, and their composition was analyzed by gas chromatography-mass spectrometry (GC-MS) analyses. Cytotoxicity was tested against human tumor cell lines HCT-116 (colon carcinoma), SF-295 (glioblastoma), OVCAR-8 (ovarian carcinoma) and HL60 (leukemia) using 3-(4,5-dimethyl-2-thiazolyl)-2,5- diphenyl-2H-tetrazolium bromide (MTT) assay, while antimicrobial activity was conducted by bioauthography method against eleven microorganisms strains. Twenty-four compounds were identified in the EOAF and twenty-nine in At-Hex. The monoterpenes linalool (25.70%), α-pinene (10.38%), β-pinene (9.12%), transocimene (7.43%), and the sesquiterpene bicyclogermacrene (12.58%) were the major constituents of EOAF, whereas the sesquiterpene spathulenol (13.91%) was the main compound of At-Hex. At-Hex showed a high cytotoxicity against SF-295 (glioblastoma). These findings show an important chemotaxonomic contribution for the Annonaceae family, mainly for the Annona genus. Atemoya proved to be a promising source of substances with potential cytotoxic activity.     

Mitochondrial import and degradation of amyloid-β peptide

Mitochondrial dysfunctions associated with amyloid-β peptide (Aβ) accumulation in mitochondria have been observed in Alzheimer's disease (AD) patients' brains and in AD mice models. Aβ is produced by sequential action of β- and γ-secretases cleaving the amyloid precursor protein (APP). The γ-secretase complex was found in mitochondria-associated endoplasmic reticulum membranes (MAM) suggesting that this could be a potential site of Aβ production, from which Aβ is further transported into the mitochondria. In vitro, Aβ was shown to be imported into the mitochondria through the translocase of the outer membrane (TOM) complex. The mitochondrial presequence protease (PreP) is responsible for Aβ degradation reducing toxic effects of Aβ on mitochondrial functions. The proteolytic activity of PreP is, however, lower in AD brain temporal lobe mitochondria and in AD transgenic mice models, possibly due to an increased reactive oxygen species (ROS) production. Here, we review the intracellular mechanisms of Aβ production, its mitochondrial import and the intra-mitochondrial degradation. We also discuss the implications of a reduced efficiency of mitochondrial Aβ clearance for AD. Understanding the underlying mechanisms may provide new insights into mitochondria related pathogenesis of AD and development of drug therapy against AD. This article is part of a Special Issue entitled: 18th European Bioenergetic Conference.     

Scent glands in legume flowers

Scent glands, or osmophores, are predominantly floral secretory structures that secrete volatile substances during anthesis, and therefore act in interactions with pollinators. The Leguminosae family, despite being the third largest angiosperm family, with a wide geographical distribution and diversity of habits, morphology and pollinators, has been ignored with respect to these glands. Thus, we localised and characterised the sites of fragrance production and release in flowers of legumes, in which scent plays an important role in pollination, and also tested whether there are relationships between the structure of the scent gland and the pollinator habit: diurnal or nocturnal. Flowers in pre‐anthesis and anthesis of 12 legume species were collected and analysed using immersion in neutral red, olfactory tests and anatomical studies (light and scanning electron microscopy). The main production site of floral scent is the perianth, especially the petals. The scent glands are distributed in a restricted way in Caesalpinia pulcherrima, Anadenanthera peregrina, Inga edulis and Parkia pendula, constituting mesophilic osmophores, and in a diffuse way in Bauhinia rufa, Hymenaea courbaril, Erythrostemon gilliesii, Poincianella pluviosa, Pterodon pubescens, Platycyamus regnellii, Mucuna urens and Tipuana tipu. The glands are comprised of cells of the epidermis and mesophyll that secrete mainly terpenes, nitrogen compounds and phenols. Relationships between the presence of osmophores and type of anthesis (diurnal and nocturnal) and the pollinator were not found. Our data on scent glands in Leguminosae are original and detail the type of diffuse release, which has been very poorly studied.     

Response of in vitro strawberry to antibiotics

By identifying antibiotics that had the least phytotoxic effects on explants during genetic transformation, we evaluated the effect of various antibiotics on callus induction and morphogenesis from leaf explants and in vitro growth of Fragaria × ananassa Duch. cv. Toyonaka. Results showed that kanamycin (Kan) significantly inhibited callus induction, bud differentiation and root morphogenesis while carbenicillin (Carb), cefotaxime (Cef) and an equal concentration of Cef and Carb up to 500 mg L⁻¹ had no significant effects on callus induction and shoot growth. Kan, even at 2.5 mg L⁻¹, significantly inhibited callus induction, shoot regeneration and root formation, while no shoots regenerated at concentrations above 15 mg L⁻¹. Rooting was completely inhibited in the presence of 50 mg L⁻¹ Kan. Cef had negative effects on shoot regeneration from leaf explants and in vitro growth of strawberry. Compared to Cef, Carb at ≤300 mg L⁻¹ significantly promoted shoot and root organogenesis. However, an equal concentration of Carb plus Cef could alleviate the negative effect of Cef on strawberry. Results from relative electrolyte leakage, root and antioxidant activities, O₂^·− production rate, H₂O₂, proline and MDA contents showed that Kan, Cef and Carb caused electrolyte leakage and triggered active enzymatic processes and metabolism. This offers a possible mechanism for the inhibition or stimulation of strawberry growth caused by these antibiotics.