Irrigation effects on quality, phenolic composition, and selected volatiles of virgin olive oils cv. Leccino

Field-grown olive trees (Olea europaea L. cv. Leccino) were used over two growing seasons to determine the effect of deficit irrigation regimes on virgin olive oil (VOO) quality. Drip irrigation was managed to maintain a predawn leaf water potential (PLWP): (a) higher than -1.1 MPa (full irrigation: FI); (b) between -1.0 and -3.3 MPa (deficit irrigation: DI); (c) higher than -4.2 MPa (severe deficit irrigation: SI). The fruit yield and oil yield of DI trees were over 90% of those of FI treatments in both years, respectively, whereas yields of SI trees ranged from 61 to 76%. The irrigation regime had minor effects on the free acidity, peroxide value, and fatty acid composition of VOO. The concentrations of phenols and o-diphenols in VOO were negatively correlated with PLWP. The concentrations of the dialdehydic form of decarboxymethyl elenolic acid linked to (3,4-dihydroxyphenyl)ethanol (3,4-DHPEA-EDA), the isomer of the oleuropein aglycon (3,4-DHPEA-EA), and the dialdehydic form of decarboxymethyl elenolic acid linked to (p-hydroxyphenyl)ethanol (p-HPEA-EDA) were lower in FI than in SI treatments. The concentrations of lignans (+)-1-acetoxipinoresinol and (+)-1-pinoresinol were unaffected by the irrigation regime. The tree water status had a marked effect on the concentration of volatile compounds, such as the C(6)-saturated and unsaturated aldehydes, alcohols, and esters.     

Dynamics of public attitudes toward bears and the role of bear hunting in Croatia

Successful carnivore conservation depends on public attitudes and acceptance levels of carnivores, and these are likely to change as circumstances change. Attitude studies repeated in time that can demonstrate such change are rare. Our study surveyed Croatian rural inhabitants in 2002 and in 2008 and analyzed their responses to detect a change in attitudes toward brown bears (Ursus arctos) over time. Important developments occurring in Croatia at the time of our research included a more centralized and more clearly defined bear management strategy, and an increase in the bear population. We constructed models to explain respondent’s value orientations, their level of perceived threat and their acceptance capacity for bears. Findings show that while value orientations and the overall level of perceived threat did not change over time, bear acceptance capacity was reduced. This suggests that the increase in the bear population and perhaps the more centralized bear management reduced respondents’ willingness to accept a larger bear population. We conclude that continuous public involvement in bear management is essential in order to maintain a feeling of control over the bear among the local population. Furthermore we argue that hunting is an important form of public involvement in the region, serving to reinforce existence and bequest values of the bear and increase its public acceptance.     

Evaluation of phenolic compounds in virgin olive oil by direct injection in high-performance liquid chromatography with fluorometric detection

Hydrophilic phenols are the most abundant natural antioxidants of virgin olive oil (VOO), in which tocopherols and carotenes are also present. The prevalent classes of hydrophilic phenols found in VOO are phenyl alcohols, phenolic acids, secoiridoids such as the dialdehydic form of decarboxymethyl elenolic acid linked to (3,4-dihydroxyphenyl)ethanol or (p-hydroxypheny1)ethanol (3,4-DHPEA-EDA or p-HPEA-EDA) and an isomer of the oleuropein aglycon (3,4-DHPEA-EA), lignans such as (+)-1-acetoxypinoresinol and (+)-pinoresinol, and flavonoids. A new method for the analysis of VOO hydrophilic phenols by direct injection in high-performance liquid chromatography (HPLC) with the use of a fluorescence detector (FLD) has been proposed and compared with the traditional liquid-liquid extraction technique followed by the HPLC analysis utilizing a diode array detector (DAD) and a FLD. Results show that the most important classes of phenolic compounds occurring in VOO can be evaluated using HPLC direct injection. The efficiency of the new method, as compared to the liquid-liquid extraction, was higher to quantify phenyl alcohols, lignans, and 3,4-DHPEA-EA and lower for the evaluation of 3,4-DHPEA-EDA and p-HPEA-EDA.     

Citrantifidiene and citrantifidiol: bioactive metabolites produced by Trichoderma citrinoviride with potential antifeedant activity toward aphids

Two novel metabolites with potential antifeedant activity were isolated from cultures of the fungus Trichoderma citrinoviride strain ITEM 4484 grown in solid-state fermentation on sterile rice kernels. The producing strain was identified at species level by sequence analysis of the internal transcribed spacer regions ITS-1 and ITS-2 of the nuclear rDNA and a fragment of the translation elongation factor gene TEF-1alpha. Fractionation by column chromatography and TLC of the culture organic extract, followed by feeding preference tests on the aphid pest Schizaphis graminum (Rondani), allowed the purification of 5.8 and 8.9 mg/kg of culture of two bioactive metabolites, which were named citrantifidiene and citrantifidiol ( 1 and 2). Citrantifidiene and citrantifidiol, whose structures were determined by spectroscopic methods (NMR and MS) are a symmetrical disubstituted hexa-1,3-dienyl ester of acetic acid and a tetrasubstituted cyclohexane-1,3-diol, respectively. The pure metabolites influenced the feeding preference of S. graminum restraining individuals from feeding on wheat leaves dipped in 5% aqueous methanol solution containing 0.57 mg/mL of citrantifidiene or 0.91 mg/mL of citrantifidiol.     

Genetic diversity in seven populations of Nicaraguan teosinte (<Emphasis Type="Italic">Zea nicaraguensis</Emphasis> Iltis et Benz) as estimated by microsatellite variation

Teosintes are the closest relatives to modern maize, Zea mays L. ssp. mays. They are wild grasses with a native distribution area from Mexico to Nicaragua and represent an important genetic resource. However, the genetic diversity of Nicaraguan teosinte (Zea nicaraguensis Iltis et Benz) has not yet been determined. This teosinte species has decreased in the last 25 years and now must be regarded as an endangered species. An analysis of the genetic diversity of Zea nicaraguensis was carried out in a total of 240 individuals from seven populations. Eleven Simple Sequences Repeat (SSR) primer pairs were used. A total of 42 alleles were found, the range of alleles per locus was 2–5 (mean 3.8) and the numbers of genotypes varied between primers. The primer Bnlg 1538 showed the highest value, with 45 genotypes through all populations. The genetic diversity observed (Ho) between all populations varied from 0.51 to 0.63, with an average of 0.563. One of the populations had as many as 40 alleles. Analysis of molecular variance (AMOVA) revealed that most of the variation was within population, at a significantly high level (P < 0.001). Rare alleles were detected in all populations, but unique alleles were only found in four populations. These results are highly relevant when developing conservation strategies and show that preserving populations in their natural habits is highly important.     

Genetic diversity in farmer grown spring barley material from Kyrgyzstan

Barley (Hordeum vulgare L.) is one of the economically important cereal crops grown in the highlands of the Kyrgyz Republic. In marginal agricultural areas of the country barley is considered as irreplaceable and very suitable fodder crop for livestock. In this study the genetic diversity of currently grown farmers’ spring barley material from Naryn and Issik-Kul provinces is described. In order to capture maximum diversity present in farmer field’s different morphological types were collected. The 22 spring barley accessions collected in 2008 were described morphologically and analysed using fourteen simple sequences repeat (SSR) markers. Most material was two-rowed barley, 78 %, whereas six-rowed barley was only 22 %. The spike colours present were light yellow, dark yellow, brown and violet. The material varied also for such characters as length of spike and number of kernels per spike. For the SSR loci 129 alleles were detected with an average of 9.3 alleles per locus and a genetic diversity with an average of h = 0.721 per locus. Based on analysis of molecular variance the 71.3 % of genetic diversity accounted for within accession variation, whereas 27.4 % for among accession variation. The variation between provinces was not significantly different. A group of two-rowed barley was identified that was more closely related to six-rowed barley than to the rest of the two-rowed barley material. Two-rowed material currently grown in Kyrgyzstan is more diverse than six-rowed barley (h = 0.65, A = 8.8 vs. h = 0.59, A = 3.1). From this study we could conclude that the materials nowadays grown by farmers are mixtures. Since there are no geographic differentiation between these mixtures suggests that materials is frequently inter-exchanged between farmers and are most probably not landrace material. The question concerning the origin of these mixtures remains unsolved and further investigation will be carried out to identify sources of variation in farmers’ field.     

Retrospective Biomolecular Investigation of Coxiella burnetii and Leptospira spp. DNA in Cases of Abortion,Stillbirth and Neonatal Mortality in Dogs and Cats

Abortion and neonatal mortality are events that can occur in breeding bitches and queens. It has been reported that up to 55% and 33% of these cases remain without a known cause, respectively, in canine and feline pregnancies. Unusual abortigenic and potentially zoonotic agents, including Coxiella burnetii and Leptospira spp., may be involved in these cases. C. burnetii is able to cause reproductive disorders in cattle, sheep and goats, and cases of abortion have been observed in dogs and cats. Moreover, several outbreaks of C. burnetii infection in humans have been caused by delivering bitches and queens, and some of these animals experienced abortion. Leptospira interrogans sensu lato is able to cause abortion or stillbirth in several animal species and its abortigenic role has occasionally been described in bitches and queens. The aim of this study was to search for C. burnetii and Leptospira spp. DNA in a retrospective series of 103 cases of canine and feline abortion, stillbirth, and neonatal mortality submitted for the identification of possible infectious agents. One hundred and fifty-one specimens were tested using PCR assays and found negative for C. burnetii and Leptospira DNA. However, in 49 samples (47.6%) other infectious causes of abortion, stillbirth, and neonatal mortality were identified. These results showed that C. burnetii and Leptospira spp. are probably not common abortigenic agents or causes of neonatal deaths in dogs. However, given the potential abortigentic and zoonotic role of these agents, surveillance of canine and feline abortion, stillbirth, and neonatal mortality could be advisable for a systematic investigation of these events.     

Prioritizing marine invasive alien species in the European Union through horizon scanning

1. The disproportionately low presence of marine species in the list of invasive alien species (IAS) of Union concern of the European Union (EU) Regulation 1143/2014 does not fully acknowledge the threat they pose to the EU marine environment.
2. In this study, the first EU‐scale Horizon Scanning (HS) focusing on marine alien species was performed, aiming to deliver a ranked list of species that should be of high priority for risk assessment (Article 5 of the EU IAS Regulation).
3. Species absent from or with a limited distribution in EU marine waters were targeted. In total, 363 alien species were initially screened for HS by a panel of experts, including a broad range of taxonomic groups. Species were scored for their likelihood of arrival, establishment, spread, and impact in EU waters.
4. A consensus workshop ranked 267 species, including a subset of 26 prioritized species. These species are considered to be mainly introduced by shipping (fouling and ballast water), via the Suez Canal, and aquaculture activities. The 26 priority species were also scrutinized in terms of feasibility of their management; 18 of them were suggested for performing risk assessments on the basis of the EU IAS Regulation.
5. Since biological invasions are dynamic and connected with accelerated globalization and diversified human activities, we recommend HS to be repeated periodically to review the species already listed and assess new ones.

     

Genetic diversity and gene flow in six accessions of Meso-America teosintes

The genus Zea (teosinte) has a native distribution from Mexico to Nicaragua (Meso-America region) and studies on teosinte species may help improve our understanding of maize genetics, population genetics, genome evolution and crop evolution. Moreover, since F1 hybrids between teosinte and maize are fertile, all teosinte species can potentially contribute to maize breeding in the form of traits such as resistance or tolerance to heavy metals in the soil and other adverse environmental conditions. Gene flow between maize and teosinte occurs in Mexico and Central America. This study analysed the genetic diversity of teosintes by using the microsatellite technique on a total of 120 individuals from six different species and 21 Simple Sequence Repeats (SSR). A total of 109 alleles were found and the number of alleles per locus ranged from 2 to 7. The genetic diversity varied between all species from 0.514 to 0.572, with a mean value of 0.548 (Nei index). Rare and unique alleles were detected in all species, with the highest frequencies found in Zea diploperennis Iltis, Doebley et Guzman, Zea perennis (Hitchc.) Reeves et Mangelsdorf and Zea nicaraguensis Iltis et Benz, which had more than seven rare alleles each. Analysis of molecular variance revealed that 19.87% of the variation between species was statistically significant (P < 0.001). When the different species were compared with the Wright (Fst) value, genetic variation between species was confirmed. The data also revealed a moderate rate gene flow between the species. Genetic diversity was found to be somewhat higher in Central America than in Mexico.     

Feasible application of a portable NIR-AOTF tool for on-field prediction of phenolic compounds during the ripening of olives for oil production

Olive fruits of three different cultivars (Moraiolo, Dolce di Andria, and Nocellara Etnea) were monitored during ripening up to harvest, and specific and total phenols were measured by HPLC (High Pressure Liquid Chromatography). On the same olive samples (n = 450), spectral detections were performed using a portable NIR (Near Infrared)-AOTF (Acousto Optically Tunable Filter) device in diffuse reflectance mode (1100-2300 nm). Prediction models were developed for the main phenolic compounds (e.g., oleuropein, verbascoside, and 3,4-DHPEA-EDA) and total phenols using Partial Least Squares (PLS). Internal cross-validation (leave-one-out method) was applied for calibration and prediction models developed on the data sets relative to each single cultivar. Validation of the models obtained as the sum of the three sample sets (total phenols, n = 162; verbascoside, n = 162; oleuropein, n = 148; 3,4-DHPEA-EDA, n = 162) were performed by external sets of data. Obtained results in term of R(2) (in calibration, prediction and cross-validation) ranged between 0.930 and 0.998, 0.874-0.942, and 0.837-0.992, respectively. Standard errors in calibration (RMSEC), cross-validation (RMSECV), and prediction (RMSEP) were calculated obtaining minimum error in prediction of 0.68 and maximum of 6.33 mg/g. RPD ratios (SD/SECV) were also calculated as references of the model effectiveness. This work shows how NIR-AOTF can be considered a feasible tool for the on-field and nondestructive measurement of specific and total phenols in olives for oil production.