Predicting mature cork biomass with t years of growth from one measurement taken at any other age

Cork is a natural product that is extracted from the outer bark of the cork oak tree. According to Portuguese legislation, the interval between two consecutive cork extractions on the same tree must be equal or greater than 9 years. Although the majority of the cork oak stands are debarked at the end of this period, this rotation may not be the optimum in many cases. The existing models for cork weight prediction can only be used for trees debarked at a 9 years or, in one model, at a 10-years rotation period, since the data used for its development was limited to these growth periods. The development of a method that allows for the prediction of the mature cork biomass with t years of growth, based in one measurement taken at any other age, was the main objective of this work. The method is based on the knowledge that the density of the cork tissue is constant between the inner and outer cork rings, being significantly different from density of the cork back. It can be implemented using two different equations that were developed during this work: a model to estimate cork biomass with 9 years of age and a model to estimate the cork back weight proportion at 9 years of age. For the first model, four different alternative models were developed, considering different levels of information collected during forest inventory. The model to estimate the cork back weight proportion leads to the biomass of cork tissue. Cork biomass at t years is obtained by decreasing or increasing the biomass of cork tissue according to the difference in cork thickness between t and 9 years of growth. The proposed method was evaluated by comparing the observed and estimated values of cork biomass from an independent data set with corks with 9, 10 and 11 years of age. The results showed similar precision for corks with 9, 10 or 11 years of age. As expected the precision of the predictions increases when the model to estimate cork biomass with 9 years of age uses more information. The presented method should be an important tool for cork oak stand management, for the prediction of the evolution of carbon stocks in cork oak stands, and will allow analyzing the impact in cork biomass production of decreasing or extending the interval between two consecutive cork extractions.     

Antixenosis resistance to leaf miner Leucoptera coffeella in Coffea species

The coffee leaf miner, Leucoptera coffeella, is the most important pest of Coffea arabica plantations in Brazil, and all cultivars are susceptible to the insect. However, distinct reactions have been observed in other species, such as C. congensis and C. canephora. In this work, the occurrence of antixenotic response was surveyed in individuals of Coffea species, selected for resistance to the leaf miner at both field and laboratory conditions. Tests performed on young plants from C. arabica cultivars Icatu Precoce IAC 3282, Mundo Novo IAC 515-20, Tupi IAC 1669-33 and Catuaí Amarelo IAC 62 demonstrated that these were preferred for oviposition when pooled with seedlings from C. canephora cv. Apoat? IAC 2258. Apoat? was also preferred over C. racemosa for oviposition. Similar results were observed in tests using detached leaves, and higher amount of eggs was counted in the cv Obat? IAC 1669-20 and Catuaí Vermelho IAC 81, and lower oviposition frequency was observed in C. canephora cv. Guarini IAC 1598. C. congensis showed an oviposition frequency between C. arabica and C. canephora. Also, the results suggest that the frequency of oviposition is co-related with the plant resistance level, during larvae infection.     

Almond allergens: molecular characterization, detection, and clinical relevance

Almond ( Prunus dulcis ) has been widely used in all sorts of food products (bakery, pastry, snacks), mostly due to its pleasant flavor and health benefits. However, it is also classified as a potential allergenic seed known to be responsible for triggering several mild to life-threatening immune reactions in sensitized and allergic individuals. Presently, eight groups of allergenic proteins have been identified and characterized in almond, namely, PR-10 (Pru du 1), TLP (Pru du 2), prolamins (Pru du 2S albumin, Pru du 3), profilins (Pru du 4), 60sRP (Pru du 5), and cupin (Pru du 6, Pru du γ-conglutin), although only a few of them have been tested for reactivity with almond-allergic sera. To protect sensitized individuals, labeling regulations have been implemented for foods containing potential allergenic ingredients, impelling the development of adequate analytical methods. This work aims to present an updated and critical overview of the molecular characterization and clinical relevance of almond allergens, as well as review the main methodologies used to detect and quantitate food allergens with special emphasis on almond.     

Ecotoxicological Assessment of Contaminated River Sites as a Proxy for the Water Framework Directive: an Acid Mine Drainage Case Study

Metal contamination of freshwater bodies resulting from mining activities or deactivated mines is a common problem worldwide such as in Portugal. Bra?al (galena ore) and Palhal (pyrrhotite, chalcopyrite, galena, sphalerite, and pyrite ore), located in a riverside position, are both examples of deactivated mining areas lacking implemented recovery plans since their shutdown in the early mid-1900s. In both mining areas, effluents still flow into two rivers. The purpose of this work was to evaluate the potential hazard posed by the mining effluents to freshwater communities. Therefore, short- and long-term ecotoxicological tests were performed on elutriates from river sediments collected at each site using standard test organisms that cover different functional levels (Vibrio fischeri, Pseudokirchneriella subcapitata, Lemna minor, and Daphnia sp.). The results show that elutriates from the sediments of Palhal were very toxic to all tested species, while in contrast, elutriates from Bra?al showed generally no toxicity for the tested species. Our study highlights the usefulness of using an ecotoxicological approach to help in the prioritization/scoring of the most critical areas impacted by deactivated mines. This ecotoxicological test battery can provide important information about the ecological status of each concerning site before investing in the application of time-consuming and costly methods defined by the Water Framework Directive or can stand as a meaningful complementary analysis.     

Forest fuel management as a conservation tool for early successional species under agricultural abandonment: The case of Mediterranean butterflies

In cultural landscapes there are often negative biodiversity consequences of agricultural abandonment and subsequent scrub and forest encroachment, due to homogenization and the loss of early-successional habitats. The common forestry practice of removing understory vegetation to prevent fire hazard (fuel management) probably has the side-effect of ameliorating these consequences, but it is uncertain whether it effectively restores habitats for early-successional species. Here we examine the influence of time since fuel management and management frequency on butterfly assemblages, using a chronosequence of cork oak (Quercus suber) stands spanning about 70 years. Overall species richness increased immediately after management and abundances peaked about 2–3 years later, while both declined thereafter for about 10–20 years to pre-disturbance levels. Richness and abundances were also much higher in recurrently managed stands. Most life history groups showed successional trends similar to the overall species richness and abundances, though consistent positive effects of fuel management were only observed for species with univoltine life cycle, herbaceous layer feeding, larval overwintering, and intermediate body size. Individual species were largely associated with recent and recurrent management, though a few specialists occurred most often in undisturbed stands. These findings suggest that fuel management at <10 years intervals is strongly positive for butterfly assemblages in landscapes under land abandonment. However, to maintain the overall forest biodiversity it is critical that patches of undisturbed habitat are also retained at the landscape scale.     

A modified agglutination test for the diagnosis of Besnoitia besnoiti infection

Bovine besnoitiosis is caused by Besnoitia besnoiti, an obligate intracellular apicomplexan parasite. Affected animals present cutaneous and systemic manifestations and the disease may lead to considerable economic losses. Although generally associated to tropical and subtropical areas, bovine besnoitiosis is now considered an emergent disease in Europe, due to the increasing number of new cases and apparent geographical expansion. In this study we evaluated the performance of a modified agglutination test (B-MAT) in the serodiagnosis of bovine besnoitiosis in comparison to the indirect immunofluorescent-antibody test (IFAT). To establish optimal protocol conditions we used bovine sera with a known infection status for B. besnoiti infection. Positive animals (n=36) presented B. besnoiti dermal cysts and anti-B. besnoiti specific antibodies, as determined by the indirect immunofluorescence test (IFAT). Negative animals (n=103) were from non-endemic areas in Portugal and negative by the IFAT. From here, we evaluated the sensitivity and specificity of the B-MAT relative to the IFAT with a panel of sera from herds with history of bovine besnoitiosis in Portugal, Spain and France (n=402), using three serum dilutions (1:80, 1:160, 1:320). Considering the positive cut-off at 1:160 serum dilution, the B-MAT showed an almost perfect test agreement with the IFAT; (κ=0.968; 95% CI: 0.941-0.996) with a relative sensitivity of 97.2% (95% CI: 94.1-100%) and a relative specificity of 99.3% (95% CI: 98.4-100%). As a simple and inexpensive technique the B-MAT represents a valuable tool for the diagnosis and study of the epidemiology of bovine besnoitiosis.     

Microtomographic characterization of calcifications in canine mammary tumours

The present work describes the microtomographic characterization of macro‐ and microcalcifications present in excised canine mammary glands. In human breast cancer, microcalcifications are highly relevant for diagnosis and prognosis, often being the sole element determining biopsy. Canine mammary tumours are considered a model for human breast cancer, but the morphological features of calcifications had still to be studied in this species. The objective of this research is to contribute to the characterization of the mineralization features of the canine mammary gland. In the present study, the excised mammary glands of 33 bitches underwent fluoroscopic examination. In 30 of the samples, the presence of calcification was suspected, and multiple biopsies were taken of these areas. Biopsy fragments underwent microtomographic scanning. Microcalcifications were found in non‐neoplastic glandular tissue, benign and malign lesions, as it is known to happen in humans. Qualitative evaluation regarding morphology of the imaged calcifications showed similarities to breast cancer findings, based on the BI‐RADS 2013 classification, such as pleomorphism and shape. No differences in the quantitative morphological parameters of volume, surface, surface/volume, SMI and structure thickness were found when macrocalcifications were considered. However, although significant differences existed in these parameters between microcalcifications from malignant canine mammary tumours and the two other groups, none were found between non‐neoplastic and benign tumours. Findings further support the use of this spontaneous animal model for the study of human breast cancer, considering how clinically relevant microcalcifications are in humans.     

Detection of the root lesion nematode Pratylenchus penetrans in potato tubers

The root lesion nematode Pratylenchus penetrans parasitizes a wide range of economically important crops, including potato (Solanum tuberosum). Damage by P. penetrans impacts not only the potato yield but can also reduce the tuber quality. Detailed information on tuber infection by P. penetrans is scarce for most cultivars and molecular detection of nematodes from infected tubers is needed. The objective of this study was to assess tuber symptomatology due to P. penetrans infection in 10 potato cultivars and to provide an accurate molecular methodology for nematode detection using tuber peels. Sprouts of certified potato seed from cultivars Agata, Agria, Camel, Désirée, Dirosso, Kennebec, Laura, Picasso, Royata, and Stemster were planted in 2 L pots, and soil was inoculated with 4 P. penetrans/g of soil. Sixty days after inoculation, tubers were harvested, inspected for lesions, and the number of nematodes/g of potato peel assessed. Observations of tubers with symptoms showed the presence of P. penetrans in superficial layers of peels around the lenticels and injured necrotic tissue. Different nematode stages were detected in tubers of all inoculated cultivars, varying from 4 to 46 nematodes/g of potato peel. Species-specific primers showed suitable sensitivity and reproducibility for the detection of P. penetrans in tuber potato peel samples. The molecular detection of P. penetrans directly from tuber peels can facilitate routine nematode inspections of potato seed tubers or cull potatoes for nematode detection, and prevent further dissemination of this species.