Development of a peptide nucleic acid array platform for the detection of genetically modified organisms in food

Two previously developed platforms, a multiplex polymerase chain reaction (PCR) and a peptide nucleic acid (PNA) array, the former allowing for the simultaneous detection of five transgenes and two endogenous controls in food and feed matrices and the latter for the assessment of the identity of amplified PCR products, were combined in order to develop a PNA array device for the screening of genetically modified organisms (GMOs) in food. PNA probes were opportunely designed, synthesized, and deposited on commercial slides. The length of the probes as well as the distance of the probes from the surface were evaluated and found to be critical points. The most suitable probes were found to be 15-mer PNAs linked to the slide surface by means of two 2-(2-aminoethoxy)ethoxyacetic acids as spacers. The device was tested on a model system constituted by flour samples containing a mixture of standards at known concentrations of transgenic material, in particular Roundup Ready soybean and Bt11, Bt176, Mon810, and GA21 maize: The DNA was amplified using the specific multiplex PCR method and tested on the PNA array. The method proposed was found to be able to correctly identify every GMO present in the tested samples.     

Structural basis for antioxidant activity of trans-resveratrol: ab initio calculations and crystal and molecular structure

From the experimental crystal structure and ab initio calculations on resveratrol and its derivatives, structural features of mechanistic importance are described. The molecular structure reveals the relative coplanarity of the trans-stilbene skeleton, and the molecular packing in the solid state shows an extensive hydrogen bond network that elucidates the flip-flop motion of the three hydroxyl groups that alternately form and break H bonds with each of the neighboring phenolic oxygens. The dynamic behavior provoked by the alternation of hydrogen bond formation and breaking can result in the ready mobility of up to three hydrogen atoms per resveratrol molecule that can be transferred to reactive oxidants that are rich in electron density. In addition, theoretical studies confirm the planarity of resveratrol as well as for half of the molecule of a condensation dimeric derivative of resveratrol, trans-sigma-viniferin. Furthermore, these studies show the p-4'-OH group to be more acidic compared to the other two m-OH groups. These features correlate with the biological activity of resveratrol as an antioxidant and support earlier studies showing H-atom transfer to be the dominant mechanism by which phenolic antioxidants intercept free radicals.     

Electrical resistivity tomography as a non‐destructive method for mapping root biomass in an orchard

Multi‐electrode soil electrical resistivity (ρ) tomography was used for the non‐invasive study of tree roots in situ and their spatial distribution in an agricultural soil. The quantitative relations of ρ and root biometry and the contribution of different root size classes were investigated with two‐ and three‐dimensional 48‐electrode tomograms in an orchard in southern Italy on a Typic haploxeralf fine, mixed termic soil. Root biomass density (RD) and root length density (RLD) were measured destructively on coarse (>2 mm diameter) and fine roots, and soil paste electrical conductivity, water content, stone content, texture, organic matter and pH were measured on soil samples taken up to 0.48‐m deep. Areas of large ρ values (up to 460 ohm m) were found close to tree trunks and variability in ρ was related to RD (0–0.137 Mg m^?3) only; the resistive response was from coarse roots. The effect of other soil variables on ρ was overshadowed by the presence of roots and therefore no significant multivariate relationship was found. A highly significant ρ‐RD gamma GLM model used to fit positively skewed data provides a useful framework for regression analysis when ρ is dominated by roots. Soil electrical resistivity is promising as a proxy for RD in orchards, but not for RLD, and the effect of tree roots on ρ needs to be taken into account in electrical surveys of soils.     

Towards a standardised surveillance for Trichinella in the European Union

Each year, more than 167 million pigs in the European Union (EU) are tested for Trichinella spp. under the current meat hygiene regulations. This imposes large economic costs on countries, yet the vast majority of these pigs test negative and the public health risk in many countries is therefore considered very low. This work reviewed the current Trichinella status across the EU as well as the national level of monitoring and reporting. It also reviewed which animal species were affected by Trichinella and in which species it should be surveyed. This information was used to design a cost-effective surveillance programme that enables a standardised monitoring approach within the EU. The proposed surveillance programme relies on identifying sub-populations of animals with a distinct risk. Low-risk pigs are finisher pigs that originate from so-called controlled housing. All other pigs are considered high-risk pigs. Controlled housing is identified by the application of a specific list of management and husbandry practices. We suggest that member states (MS) be categorised into three classes based on the confidence that Trichinella can be considered absent, in the specified sub-population of pigs above a specified design prevalence which we set to 1 per million pigs. A simple and transparent method is proposed to estimate this confidence, based on the sensitivity of the surveillance system, taking into account the sensitivity of testing and the design prevalence. The probability of detecting a positive case, if present, must be high (>95 or >99%) to ensure that there is a low or negligible risk of transmission to humans through the food chain. In MS where the probability of a positive pig is demonstrated to be negligible, testing of fattening pigs from a sub-population consisting of pigs from controlled housing can be considered unnecessary. Furthermore, reduced testing of finishers from the sub-population consisting of pigs from non-controlled housing might even be considered, if conducted in conjunction with a proportionate sampling scheme and a risk-based wildlife surveillance programme where applicable. The proposed surveillance programme specifies the required number of samples to be taken and found negative, in a MS. A MS with no data or positive findings will initially be allocated to class 1, in which all pigs should be tested. When a MS is able to demonstrate a 95% or 99% confidence that Trichinella is absent, the MS will be allocated to class 2 or 3, in which the testing requirement is lower than in class 1.     

Impacts and adaptation of European crop production systems to climate change

The studies on anthropogenic climate change performed in the last decade over Europe show consistent projections of increases in temperature and different patterns of precipitation with widespread increases in northern Europe and decreases over parts of southern and eastern Europe. In many countries and in recent years there is a tendency towards cereal grain yield stagnation and increased yield variability. Some of these trends may have been influenced by the recent climatic changes over Europe.A set of qualitative and quantitative questionnaires on perceived risks and foreseen impacts of climate and climate change on agriculture in Europe was distributed to agro-climatic and agronomy experts in 26 countries. Europe was divided into 13 Environmental Zones (EZ). In total, we had 50 individual responses for specific EZ. The questionnaires provided both country and EZ specific information on the: (1) main vulnerabilities of crops and cropping systems under present climate; (2) estimates of climate change impacts on the production of nine selected crops; (3) possible adaptation options as well as (4) adaptation observed so far. In addition we focused on the overall awareness and presence of warning and decision support systems with relevance for adaptation to climate change.The results show that farmers across Europe are currently adapting to climate change, in particular in terms of changing timing of cultivation and selecting other crop species and cultivars. The responses in the questionnaires show a surprisingly high proportion of negative expectations concerning the impacts of climate change on crops and crop production throughout Europe, even in the cool temperate northern European countries.The expected impacts, both positive and negative, are just as large in northern Europe as in the Mediterranean countries, and this is largely linked with the possibilities for effective adaptation to maintain current yields. The most negative effects were found for the continental climate in the Pannonian zone, which includes Hungary, Serbia, Bulgaria and Romania. This region will suffer from increased incidents of heat waves and droughts without possibilities for effectively shifting crop cultivation to other parts of the years. A wide range of adaptation options exists in most European regions to mitigate many of the negative impacts of climate change on crop production in Europe. However, considering all effects of climate change and possibilties for adaptation, impacts are still mostly negative in wide regions across Europe.     

An epidemiological study of canine filarioses in north-west Italy: What has changed in 25 years?

A field survey aimed at updating knowledge of the distribution and prevalence of filarial infections in the Piedmont region was undertaken. Blood samples were collected from 2628 dogs (10–15 dogs/100 km²) which had remained in the local area, were kept in the open overnight, were more than 2 years old and had not previously been treated with microfilaricidal drugs. Infection was diagnosed by a modified Knott test. Comparison of the results with those from a similar survey carried out 25 years ago showed a 3-fold increase in the size of the endemic area, with the colonization of new habitats, and a 4-fold increase in the prevalence of Dirofilaria immitis infection.     

Evidence for quantitative trait locus for conformation traits on chromosome 19 in beef cattle

A total of 110 F₁ crossbred individuals of Piemontese × Chianina cattle and 75 F₂ intercross were genotyped for the DNA marker IDVGA-46, composed of an (AC) repeat, that showed a polymorphism of three alleles: 205, 207 and 229 base pairs. Association of marker polymorphism to beef conformation measures was tested with a linear mixed model which included the fixed effect of the marker genotype separated according to the origin of the allele: whether inherited from Piemontese or from Chianina and random additive genetic effect of the animals. Carriers of allele 205, when inherited from Chainina, were larger than carriers of 207 and showed a globally better body structure. No significant association was found in the animals that had inherited the alleles from Piemontese. It would be interesting to verify, in the chromosome portions flanking the mentioned marker (chromosome 19; q16 band), the existence of eventual coding sequences influencing growth and conformation.     

Performance of crosses among Hereford, Angus, and Simmental cattle with different levels of Simmental breeding: VI. Maternal heterosis of 3- to 8-year-old dams and the dominance model.

Data from Hereford, 25% Simmental-75% Hereford, 50% Simmental-50% Hereford, and 75% Simmental-25% Hereford dams were used to estimate maternal heterosis and level of agreement with the dominance model. Cows were located at the Northern Agricultural Research Center near Havre, MT and were managed consistent with practices for western range environments. Sample halves of dam breed groups were bred to Charolais and Tarentaise sires to produce calves at 3 to 8 yr of age. There were 766 exposures to breeding that resulted in 581 calves. Breed group means for most traits supported the dominance model. Maternal heterosis was estimated by regression techniques for 22 cow and calf traits. Maternal heterosis was not significant for day of conception, number of services, gestation length, or calving difficulty. Estimates of maternal heterosis for calf growth traits ranged from .7% for weaning height to 5.2% for weaning weight and 7.5% for weaning condition score. Calf weight per unit of cow weight at weaning showed significant maternal heterosis (7.1%). Higher levels of maternal heterosis were exhibited for milk production (8.2 to 11.1%) and the negative, but nonsignificant, estimate of maternal heterosis for early minus late milk production suggested more persistent lactation for crossbred cows. Maternal heterosis was 11.5% for proportion of dams that calved and 10.4% for proportion of dams that weaned calves. Calf weaning weight per cow exposed to breeding, a characteristic combining calf growth and dam reproduction, exhibited 17.9% maternal heterosis.     

Effect of infectious bovine rhinotracheitis virus immunization on viral shedding in challenge-exposed calves treated with dexamethasone

Calves not vaccinated with infectious bovine rhinotracheitis virus (IBRV) became latently infected when challenge exposed and treated with dexamethasone (DM). Calves that shed IBRV after DM treatment were considered to be latently infected. Vaccination with a temperature-sensitive intranasal vaccine or with formalinized IBRV in Freund's complete adjuvant (IBRV-FCA) protected some, but not all, calves against latent infection--indicating a role for the immune response in preventing latent infection. That all latently infected calves were not detected after DM treatment was indicated by the fact that after a 2nd DM treatment of 3 calves treated 6 months previously and not found to shed virus, 1 of the calves was latently infected. Latently infected calves were inoculated with successive doses of IBRV-FCA and treated with DM. Nonvaccinated calves shed virus, whereas vaccinated calves similarly treated did not shed virus. Because both groups had a comparable cell-mediated immune response, as determined by blastogenic response to IBRV, but the vaccinated group had significantly higher virus-neutralizing antibody titers, a role for humoral antibody in preventing viral shedding was indicated.     

Leishmania infantum and Neospora caninum simultaneous skin infection in a young dog in Italy.

Leishmania infantum, the agent of canine leishmaniasis in Mediterranean countries, and Neospora caninum, a recently recognized protozoal pathogen in dogs, were diagnosed in a 9-month-old Argentine Dogo dog. Both skin lesions and neurological signs were present. Histopathology of cutaneous lesions revealed a suppurative, diffuse dermatitis with numerous intracellular protozoa. Serology was positive for both L. infantum (1:640) and N. caninum (1:800). Double-label immunohistochemical staining of skin samples with hyperimmune serum from L. infantum-infected dogs was positive for protozoa within macrophages, while the polyclonal antibody specific for N. caninum showed positive reactions for protozoa in endothelial cells and fibroblasts. Transmission electron microscopy (TEM) confirmed the infection with both protozoa. This is, to the authors' knowledge, the first case of simultaneous infection with L. infantum and N. caninum in a dog. It is possible that the immunosuppressive effects of Leishmania infection or long-term steroid therapy may have been a contributing factor to the development of N. caninum in this dog.