Husbandry Protocols for Juvenile Loggerhead Sea Turtles (Caretta caretta) Based on Stress Response to Stocking Density and Dry-Dock Time.

When necessary, sea turtles are held captive for veterinarian care and research purposes. Protocols and basic guidelines have been described for husbandry of sea turtles with veterinarian needs but not considering physiological indicators of animal welfare. Because all sea turtle are imperiled species, monitoring their welfare is important. The aim of this study was to standardize husbandry protocols for loggerhead (Caretta caretta) juveniles held under seminatural conditions, based on circulating concentration of plasma corticosterone (Cort) and behavior. Two experiments were performed to analyze physiological and behavioral responses of the animals facing changes in stocking density and different dry-docking times. Cort analyses suggested that the number of animals per tank can be modified occasionally, without affecting their health and welfare. However, dry-docking time should be < 30 min, as indicated by the significant elevation of circulating Cort at ≥ 30 min, rising from 1.51- ng/ml to 5.28-ng/ml. Protocols tested did not affect behavioral responses, except for the breaths per move, which increased while Cort increased, despite differences exhibited by experimental animals in behavioral responses according to daily times (morning vs afternoon) and the sex of the animals.

Use of a RT-qPCR Method to Estimate Mycorrhization Intensity and Symbiosis Vitality in Grapevine Plants Inoculated with Rhizophagus irregularis.

Assessing the mycorrhization level in plant roots is essential to study the effect of arbuscular mycorrhizal fungi (AMF) on plant physiological responses. Common methods used to quantify the mycorrhization of roots are based on microscopic visualization of stained fungal structures within the cortical cells. While this method is readily accessible, it remains time-consuming and does not allow checking of the symbiosis vitality. The aim of this work is thus to develop an efficient method for assessing the intensity and vitality of mycorrhiza associated with grapevine through gene expression analyses by RT-qPCR. To this end, grapevine plants were inoculated with the AMF Rhizophagus irregularis (Ri). The relationship between mycorrhization level, assessed by microscopy, and expression of several fungus and grapevine genes involved in the symbiosis was investigated. In AMF-inoculated plants, transcript amounts of fungal constitutively-expressed genes Ri18S, RiTEF1α and RiαTub were significantly correlated to mycorrhization intensity, particularly Ri18S. Grapevine (VvPht1.1 and VvPht1.2) and AMF (GintPT, Ri14-3-3 and RiCRN1) genes, known to be specifically expressed during the mycorrhizal process, were significantly correlated to arbuscular level in the whole root system determined by microscopy. The best correlations were obtained with GintPT on the fungal side and VvPht1.2 on the plant side. Despite some minor discrepancies between microscopic and molecular techniques, the monitoring of Ri18S, GintPT and VvPht1.2 gene expression could be a rapid, robust and reliable method to evaluate the level of mycorrhization and to assess the vitality of AMF. It appears particularly useful to identify AMF-inoculated plants with very low colonization level, or with non-active fungal structures. Moreover, it can be implemented simultaneously with the expression analysis of other genes of interest, saving time compared to microscopic analyses.

Influence of Water Salinity on the Growth and Survivability of Asp Larvae Leuciscus aspius (Linnaeus, 1758) under Controlled Conditions.

The effect of water with a salinity 3, 5, 7, 9 and 11 ppt on the growth and survivability of asp Leuciscus aspius (L.) larvae was investigated. A control sample consisted of asp larvae reared up in freshwater (0 ppt). Larvae were fed for 21 days with nauplii of the brine shrimp Artemia salina. Water salinity was observed to have a considerable effect on the growth and survivability of asp larvae. In addition, saline water extended the life span of Artemia salina nauplii, which resulted in their prolonged availability to asp larvae. Asp larvae showed low tolerance to the salinity of water, reaching 9-11 ppt. Depending on the degree of salinity, the mean final weight of larvae varied from 122.6 to 139.4 mg, at body lengths from 23.8 to 25.6 mm, respectively. The best body length increments were recorded among asp larvae maintained in water with a salinity of 3 ppt. Depending on the level of water salinity, the final survivability of asp larvae ranged from 16.9 to 94.5%. The highest and increasing mortality was demonstrated among the larvae reared in water of the salinity equal to 11 ppt. It is not recommended to rear asp larvae in water with a salinity above 7 ppt due to the low survivability and large differences in the body size of the larvae that managed to survive.

Emerged macrophytes to the rescue: Perfluoroalkyl acid removal from wastewater and spiked solutions.

This study evaluated the potential for three emergent aquatic macrophytes to remove perfluoroalkyl acids (PFAAs) from contaminated waters in constructed wetland systems. Three plants (Iris pseudacorus L., Phragmites australis (Cav.) Trin. Ex Steud., and Typha latifolia L.) were exposed to an effluent from a tannery wastewater treatment plant (WWTP) that contained residual PFAAs, and to three spiked solutions with increasing concentrations of 11 perfluorocarboxylic acids (PFCAs) and three perfluorosulfonic acids (PFSAs) (500, 2500, and 5000 ng L-1, each). Thirty-six lightweight expanded clay aggregate- and vegetation-filled tanks (0.35 × 0.56 × 0.31 m) were exposed to the tested solutions at the Acque del Chiampo SpA WWTP in Arzignano (NE Italy). Throughout the experiment, PFAA concentrations and physicochemical water parameters were monitored via measures of the clay material, plastic tank inner surfaces, and below- and above-ground biomasses (after harvest). Vegetation growth was shown to be unaffected by increased PFAA levels in the spiked solutions. Alternatively, total biomass was significantly reduced when WWTP water was used, although we attribute this finding to the relatively high salinity that mainly restricted Typha and Iris development. The tested macrophytes were found to remove a significant PFAA mass from the contaminated waters (36% to ca. 80%, on average) when Phragmites was subjected to the highest PFAA concentrations. Such large accumulations were primarily associated with long C-chain PFAA stabilization in belowground biomass (26%, on average). Most PFAA translocations were observed in Typha, which accumulated mostly short perfluorinated C-chain PFBA, PFPeA, and PFHxA in the aboveground biomass (16%, on average). Despite some growth limitations, Iris was still the most efficient macrophyte for translocating PFBS under WWTP.

Lab-scale investigation of the ability of Polar Organic Chemical Integrative Sampler to catch short pesticide contamination peaks.

In this lab-scale study, the POCIS capacity to integrate short contamination peaks of variable intensity and duration was evaluated. POCIS were immersed for 14 days in tanks filled with tap water and spiked at different concentrations with 12 pesticides of various polarities (log Kow = 1.1-4.7) and classes (herbicides, fungicides, and insecticides). Concentrations were kept relatively constant at 1 µg L-1 and 5 µg L-1, respectively, in two "background" exposure tanks. Three contamination peaks of increasing intensity and decreasing duration were simulated (10 µg L-1 for 24 h, 40 µg L-1 for 6 h, and 60 µg L-1 for 1 h). This lab-scale study demonstrated that ten moderately polar compounds (2 < log Kow < 4) showed a linear uptake, as observed in previous studies, while a non-linear model fits the data of the two most polar pesticides (log Kow < 2). Depending on chemical polarity, some compounds exhibited a "burst effect" or "lag effect" during the first 3 days of exposure. After 14 days of exposure, contamination peaks appeared integrated for seven compounds, showing the ability of POCIS to catch very short pollution events and to provide acceptable time-weighted average concentration estimates under laboratory-controlled conditions.

Rearing of Bitterling (Rhodeus amarus) Larvae and Fry under Controlled Conditions for the Restitution of Endangered Populations.

Among the several dozen European freshwater fish species, only European bitterling (Rhodeus amarus Bloch) and Rhodeus meridionalis belong to the group of ostrakophilous fish. The embryonic and larval development of the fish in this reproductive group until the time of the yolk sac resorption takes place in the gill cavity of river mussels (Anodonta sp. or Unio sp.). This paper presents the results of the European bitterling Rhodeus amarus being reared under controlled conditions. Bitterling larvae were caught together with river mussels in the natural environment and subsequently placed in a tank for behavioural observations. Bitterling larvae were seen swimming in the water within a week of placing the bivalves under controlled conditions. The bitterling larvae were 8.6 ± 0.11 mm long when they started to swim actively. The rearing was conducted in water at 20 and 26 ± 0.5 °C and lasted for 6.5 months (200 days) in both variants. Initially, the larvae were fed with live nauplii of Artemia salina and subsequently with fodder. The bitterlings in tanks with water at 26 ± 0.5 °C were 66.2 ± 3.0 mm long and weighed 3389 ± 548 mg. For comparison, bitterlings kept in water at 20 ± 0.5 °C were 64.48 ± 3.4 mm long and weighed 3242 ± 427 mg. No larval malformities or mortality were observed during the larvae and fry rearing. The bitterlings had well-developed secondary sexual characteristics and exhibited pre-spawning behaviour at the end of the rearing. This produced suitable bitterling stocking material to be used in the conservation of small or endangered populations.

Physiological and Immunological Status of Adult Honeybees (Apis mellifera) Fed Sugar Syrup Supplemented with Pentadecapeptide BPC 157.

Various factors contribute to a decline in diversity and number of bees. Here, an integrated approach in experimental BPC 157 therapy was implemented, combining laboratory-controlled and field study results. The aim of a study was to assess the effects of BPC 157 additional feeding of newly emerged worker honeybees on few biochemical and immunological parameters in hemolymph (glucose, trehalose, lipids, proteins, vitellogenin, glucose-oxidase (GOX)), and hypopharyngeal gland (HPG), in laboratory-controlled conditions. Additionally, to examine the physiological status of protein digestion, the enzymatic activity of leucine aminopeptidase (LAP) in the mid-guts of worker honeybees was analyzed. It was found that individual honeybees, in hoarding cages, following BPC 157 administration through carbohydrate food, showed positive physiological changes when compared to the control groups. Those results were complemented by strong and visible LAP activity, particularly noticeable in the apical parts of the epithelial cells in the mid-guts of young worker honeybees originated from treated hives, suggesting a link between alternative oral therapy with BPC 157 and honeybees' immunity.

Wheat Spike Blast Image Classification Using Deep Convolutional Neural Networks.

Wheat blast is a threat to global wheat production, and limited blast-resistant cultivars are available. The current estimations of wheat spike blast severity rely on human assessments, but this technique could have limitations. Reliable visual disease estimations paired with Red Green Blue (RGB) images of wheat spike blast can be used to train deep convolutional neural networks (CNN) for disease severity (DS) classification. Inter-rater agreement analysis was used to measure the reliability of who collected and classified data obtained under controlled conditions. We then trained CNN models to classify wheat spike blast severity. Inter-rater agreement analysis showed high accuracy and low bias before model training. Results showed that the CNN models trained provide a promising approach to classify images in the three wheat blast severity categories. However, the models trained on non-matured and matured spikes images showing the highest precision, recall, and F1 score when classifying the images. The high classification accuracy could serve as a basis to facilitate wheat spike blast phenotyping in the future.

Artificial Reproduction of Blue Bream (Ballerus ballerus L.) as a Conservative Method under Controlled Conditions.

The blue bream Ballerus ballerus (L.) is one of two species of the Ballerus genus occurring in Europe. The biotechnology for its reproduction under controlled conditions needs to be developed to conserve its local populations. Therefore, a range of experiments were performed to obtain valuable gametes from blue bream following hormonal stimulation with carp pituitary homogenate (CPH), Ovopel and Ovaprim. CPH and Ovopel were injected twice and Ovaprim-once, under a pectoral fin. The spawners were inspected 12 h after the last injection, and the gametes were collected. Eggs were collected from spawning females and fertilized with sperm from three males. The smallest number of ovulating blue bream (four females) was obtained from individuals stimulated with Ovaprim. There were six to nine ovulating females in the other groups, accounting for 60-90% of the females in the respective groups. The latency period was the shortest in the spawners stimulated with Ovopel. The fish in this group ovulated 14 h after the hormonal injection. Spawning was synchronized and all females spawned simultaneously. The highest average weight of eggs per fish (75.4 g) was obtained from the females stimulated with Ovopel, whereas the individuals stimulated with Ovaprim provided the smallest average amount of eggs (49.5 g). The study showed that blue bream spawners are not very sensitive to reproduction-related handling under controlled conditions.

Assessment of captive rearing conditions on loggerhead hatchlings: Effect of handling frequency and stocking density.

Frequently, stranded sea turtles require rehabilitation under controlled conditions. Currently, few publications have described the conditions under which rehabilitation is to take place, particularly with respect to the hatchling life stage. To address this paucity of data, we conducted some experiments to assist rehabilitating facilities assess their handling of hatchlings. While in captivity, hatchlings are routinely handled, for example, for data collection and cleaning. Standardization of handling and housing protocols is necessary to define the most adequate rearing conditions to maintain hatchling welfare. Accordingly, the aim of this study was to assess plasma circulating corticosterone (Cort) concentration and growth, as a biomarker for the stress of hatchling loggerhead sea turtles (Caretta caretta) under controlled conditions. We performed two experiments to analyze handling frequency and stocking density. In both, Cort was measured and correlated with variations in animal weight and length. In handling experiments, Cort exhibited no significant increase when hatchlings were handled once a week, whereas Cort was significantly elevated when hatchlings were handled once every 2 weeks, suggesting that hatchlings have the ability to acclimate to frequent handling. However, hatchlings exhibited similar growth and mortality, regardless of handling regime. In stocking density experiments, hatchling isolation induced a significant elevation of Cort, in comparison with hatchlings placed with conspecifics at increasing densities. Growth increased in singly housed hatchlings, while mortality increased in tanks with three or more hatchlings. The results obtained suggest that Cort, growth, and mortality should be measured to assess hatchling welfare when kept under controlled conditions.

Rethinking temperature effects on leaf growth, gene expression and metabolism: Diel variation matters.

Plants have evolved to grow under prominently fluctuating environmental conditions. In experiments under controlled conditions, temperature is often set to artificial, binary regimes with constant values at day and at night. This study investigated how such a diel (24 hr) temperature regime affects leaf growth, carbohydrate metabolism and gene expression, compared to a temperature regime with a field-like gradual increase and decline throughout 24 hr. Soybean (Glycine max) was grown under two contrasting diel temperature treatments. Leaf growth was measured in high temporal resolution. Periodical measurements were performed of carbohydrate concentrations, carbon isotopes as well as the transcriptome by RNA sequencing. Leaf growth activity peaked at different times under the two treatments, which cannot be explained intuitively. Under field-like temperature conditions, leaf growth followed temperature and peaked in the afternoon, whereas in the binary temperature regime, growth increased at night and decreased during daytime. Differential gene expression data suggest that a synchronization of cell division activity seems to be evoked in the binary temperature regime. Overall, the results show that the coordination of a wide range of metabolic processes is markedly affected by the diel variation of temperature, which emphasizes the importance of realistic environmental settings in controlled condition experiments.

A Whole-Plant Screening Test to Select Freezing-Tolerant and Low-Dormant Genotypes.

Winter survival is a determinant factor for the persistence of perennials grown in northern climates. High winter survival cultivars, however, have lower yield due to their early transition into a dormant state in the fall. Here we describe a whole plant assay entirely performed indoor in growth chambers and walk-in freezers to identify low-dormant genotypes with superior freezing tolerance within populations of open pollinated species. Three successive freezing stresses are applied to a broad base of 3000 genotypes to progressively eliminate 97% of the population and to retain only the 3% best performing genotypes. This approach can be used to generate recurrently selected populations in different species.

Airborne signals and abiotic factors: the neglected side of the plant communication.

A relevant number of reports have examined the role of airborne signals in plant-plant communication, indicating that volatile organic compounds (VOCs) can prime neighboring plants against pathogen and/or herbivore attacks. Conversely, there is very limited information available on the possibility of the emission of VOCs by emitter plants under abiotic stress conditions, which may alert neighboring unstressed plants and prime these individuals (receivers) against the same stressors. The present opinion paper briefly reviews a few reports examining the effect of infochemicals produced by emitters on receiver plants subjected to abiotic stresses typical of global climate change. The ecological implications of these dynamics, as well as some concerns related to the potential roles of inter-plant communication in environmentally controlled experiments, have arisen. Some possible inter-plant communications applications (biomonitoring and biostimulation), mediated by airborne signals, and some directions for future studies on this topic, are also provided.

Investigating the root plasticity response of Centaurea jacea to soil water availability changes from isotopic analysis.

Root water uptake is a key ecohydrological process for which a physically based understanding has been developed in the past decades. However, due to methodological constraints, knowledge gaps remain about the plastic response of whole plant root systems to a rapidly changing environment. We designed a laboratory system for nondestructive monitoring of stable isotopic composition in plant transpiration of a herbaceous species (Centaurea jacea) and of soil water across depths, taking advantage of newly developed in situ methods. Daily root water uptake profiles were obtained using a statistical Bayesian multisource mixing model. Fast shifts in the isotopic composition of both soil and transpiration water could be observed with the setup and translated into dynamic and pronounced shifts of the root water uptake profile, even in well watered conditions. The incorporation of plant physiological and soil physical information into statistical modelling improved the model output. A simple exercise of water balance closure underlined the nonunique relationship between root water uptake profile on the one hand, and water content and root distribution profiles on the other, illustrating the continuous adaption of the plant water uptake as a function of its root hydraulic architecture and soil water availability during the experiment.

Desove y desarrollo intracapsular del caracol marino Hexaplexnigritus (Neogastropoda: Muricidae) en laboratorio / Spawning and intracapsular development of the marine snail Hexaplex nigritus (Neograstropoda: Muricidae) in the laboratory

Introducción: El caracol marino Hexaplex nigritus es un murícido fuertemente explotado en el Golfo de California para consumo y artesanías. Cuando se reproducen, los adultos se agregan en forma de arrecifes artificiales facilitando su identificación y extracción, lo cual, ha mermado su población en dicha zona. Objetivo: Investigar el desove y desarrollo intracapsular y larvario de esta especie como herramientas para su producción en cautiverio con fines de repoblamiento. Métodos: Se recolectaron 18 progenitores que fueron mantenidos en un sistema cerrado con parámetros y alimentación controladas durante abril 2017 hasta septiembre 2018. Se registró: crecimiento de progenitores, desove de masas ovígeras, cantidad de cápsulas y número de embriones y larvas dentro de las cápsulas. La temperatura de incubación fue de 28.2 ± 1.5 °C. Las principales estructuras morfológicas de embriones y larvas fueron documentadas conforme su desarrollo. Resultados: El desarrollo embrionario intracapsular y de la larva velígera hasta su asentamiento tuvieron una duración de 14-15 y 31-32 días, respectivamente. Cada hembra de 80-88 mm de longitud produjo alrededor de 4 masas ovígeras, 627 cápsulas y 663 993 larvas velígeras extracapsulares por año. Conclusiones: La fácil adaptación y el potencial reproductivo mostrado por H. nigritus en laboratorio se presentan como estrategias promisorias para su repoblación y conservación.

Introduction: The marine snail Hexaplex nigritus is a heavily exploited muricid in the Gulf of California for consumption and handcrafts. When they reproduce, adults aggregated in the form of artificial reefs facilitating their identification and extraction, situation that has reduced their population in that area. Objective: In order to investigate the spawning and intracapsular and larval development of this species as tools for its production in captivity for repopulation purposes. Methods: Eighteen brooders were collected and kept in a closed system with controlled parameters and feeding from April 2017 to September 2018. Growth of parents, spawning of ovigerous masses, number of capsules, and number of embryos and larvae within the capsules, were registered. The main morphological structures of embryos and larvae were documented according to their development. Results: Intracapsular embryonic development and veliger larva until its establishment lasted 14-15 and 31-32 days, respectively. Each 80-88 mm length female produced around 4 ovigerous masses, 627 capsules and 663 993 extracapsular veliger larvae per year. Conclusions: Easy adaptation and reproductive potential shown by H. nigritus in the laboratory are presented as promising strategies for its repopulation and conservation.

Onthophagus landolti and Canthon indigaceus chevrolati (Coleoptera: Scarabaeinae) are attracted to the feces of ivermectin-treated cattle in the Mexican tropics / Onthophagus landolti y Canthon indigaceus chevrolati (Coleoptera: Scarabaeinae) son atraídos por las heces de ganado tratado con ivermectina en el trópico mexicano

Abstract Ivermectin (IVM) is widely used for parasite control in livestock in the tropics. Residual IVM in feces conserves its insecticide activity for weeks and can harm dung beetle (DB) species. Attraction to the feces of IVM-treated cattle was tested using the DB species Onthophagus landolti (Harold) and Canthon indigaceus chevrolati (Harold) as models. Experiments were done under controlled laboratory conditions, semi-controlled field conditions and uncontrolled field conditions. Olfactometers were used in the controlled and semi-controlled trials. The control treatment was baited IVM-free feces, and the experimental treatments were the feces of cattle treated with 1 % IVM (subcutaneous administration; single, 0.2 mg/kg bw dosage) and collected at 5, 14, 21 and 28 days post-treatment. The uncontrolled field trial involved pitfall traps baited with IVM-free feces or feces from IVM-treated cattle collected five days post-treatment. Under controlled and semi-controlled conditions, the feces of IVM-treated cattle (at 5, 14, 21 or 28 days post-treatment) attracted more O. landolti and C. i. chevrolati individuals than IVM-free feces (P < 0.05). The same response occurred under uncontrolled conditions. This clear attraction for IVM-containing cattle feces by the studied DB species highlights that incorrect IVM use may pose a risk to DB communities in cattle production systems.(AU)

Resumen La ivermectina (IVM) es ampliamente utilizada para el control de parásitos en el ganado en los trópicos. La IVM residual en las heces conserva su actividad insecticida durante semanas y puede dañar diversas especies de escarabajos estercoleros. La atracción a las heces del ganado tratado con IVM se probó usando a las especies Onthophagus landolti (Harold) y Canthon indigaceus chevrolati (Harold) como modelos de estudio. Los experimentos se realizaron bajo condiciones de laboratorio controladas, condiciones de campo semicontroladas y condiciones de campo no controladas. Se utilizaron olfatómetros en los ensayos controlados y semicontrolados. El tratamiento de control consistió en heces exentas de IVM, y los tratamientos experimentales fueron heces de ganado tratado con IVM al 1 % (administración subcutánea, dosis única, 0.2 mg / kg pv) las cuales se recogieron a los 5, 14, 21 y 28 días después del tratamiento al ganado. El ensayo de campo no controlado incluyó trampas de caída libre o pitfall cebadas con heces libres de IVM y con heces de ganado tratado con IVM recogido cinco días después del tratamiento. En condiciones controladas y semicontroladas, las heces del ganado tratado con IVM (a los 5, 14, 21 y 28 días después del tratamiento) atrajeron más individuos O. landolti yC. i. chevrolati que las heces sin IVM (P < 0.05). La misma respuesta ocurrió bajo condiciones no controladas. Esta clara atracción a las heces de ganado que contienen IVM por las especies estudiadas pone de relieve que el uso incorrecto de IVM puede plantear un riesgo para las comunidades de escarabajos estercoleros en los sistemas de producción ganadera.(AU)

Evaluation of nanoparticle emissions from a laser printer in an experimental chamber and estimation of the human particle dose.

The aim of this study was to evaluate the nanoparticle emissions from a laser printer in a chamber in conjunction with emissions from printers in a print room (PR) and to characterize the processes that lead to increased nanoparticle concentrations, as well as to estimate the human particle dose of the printers' users. Measurements were conducted in a small stainless steel environmental chamber under controlled conditions, where the evolution of particle size distributions (PSDs) with time and printed pages was studied in detail. Printer was generating nanoparticles (vast majority ˂ 50 nm with mode on ~ 15 nm) primarily during cold startup. Previously, 1-week sampling was also done in a PR at the Technical University of Crete, where the tested laser printer is installed along with three other printers. Similarly, as it was observed in the chamber study, printers' startup on any given day was characterized by a sharp increase in particle number (PN) concentrations. Average measured PN concentrations during printing hours in PR (5.4 × 103 #/cm3) is similar to the one observed in chamber measurements (6.7 × 103 #/cm3). The ExDoM2 dosimetry model was further applied to calculate the deposition of particles in the human respiratory tract. More precisely, the increase in particle dose for an adult Caucasian male was 14.6- and 24.1-fold at printers' startup, and 1.2- and 5.2-fold during printing in the PR and experimental chamber, respectively, compared to the exposure dose at background concentrations (BCs).

Transcriptomic comparison between two Vitis vinifera L. varieties (Trincadeira and Touriga Nacional) in abiotic stress conditions.

BACKGROUND: Predicted climate changes announce an increase of extreme environmental conditions including drought and excessive heat and light in classical viticultural regions. Thus, understanding how grapevine responds to these conditions and how different genotypes can adapt, is crucial for informed decisions on accurate viticultural actions. Global transcriptome analyses are useful for this purpose as the response to these abiotic stresses involves the interplay of complex and diverse cascades of physiological, cellular and molecular events. The main goal of the present work was to evaluate the response to diverse imposed abiotic stresses at the transcriptome level and to compare the response of two grapevine varieties with contrasting physiological trends, Trincadeira (TR) and Touriga Nacional (TN). RESULTS: Leaf transcriptomic response upon heat, high light and drought treatments in growth room controlled conditions, as well as full irrigation and non-irrigation treatments in the field, was compared in TR and TN using GrapeGene GeneChips®. Breakdown of metabolism in response to all treatments was evidenced by the functional annotation of down-regulated genes. However, circa 30 % of the detected stress-responsive genes are still annotated as «Unknown¼ function. Selected differentially expressed genes from the GrapeGene GeneChip® were analysed by RT-qPCR in leaves of growth room plants under the combination of individual stresses and of field plants, in both varieties. The transcriptomic results correlated better with those obtained after each individual stress than with the results of plants from field conditions. CONCLUSIONS: From the transcriptomic comparison between the two Portuguese grapevine varieties Trincadeira and Touriga Nacional under abiotic stress main conclusions can be drawn: 1. A different level of tolerance to stress is evidenced by a lower transcriptome reprogramming in TN than in TR. Interestingly, this lack of response in TN associates with its higher adaptation to extreme conditions including environmental conditions in a changing climate; 2. A complex interplay between stress transcriptional cascades is evidenced by antagonistic and, in lower frequency, synergistic effects on gene expression when several stresses are imposed together; 3. The grapevine responses to stress under controlled conditions are not fully extrapolated to the complex vineyard scenario and should be cautiously considered for agronomic management decision purposes.

The Effect of Temperature and Laboratory Rearing Conditions on the Development of Dermestes maculatus (Coleoptera: Dermestidae).

Experiments were conducted to study the life cycle of Dermestes maculatus and to establish the total developmental time and the developmental time of immature stages, in relation with six different temperatures. We also analyzed the variations in size, morphology, and other indicators of temporal variation during life cycle of D. maculatus, in relation with temperature. One hundred larvae were selected per experiment, reared individually. The remaining larvae were reared to evaluate and establish temporal variations among the instars (length, cephalic width, and dry weight). In all trials, survivorship was greater than 50% and seven larval instars were found. Data of the average developmental time of immature stages and of the total cycle, at different temperatures, are provided. This is of relevance when estimating particularly, a minimum PMI. No relation between morphometric parameters and temperature was found, suggesting that other random factors may have been involved. Thus, this indicates that the method of isomegalen diagrams could not be used for calculating PMI.

Marks caused by the scavenging activity of Necrobia rufipes (Coleoptera: Cleridae) under laboratory conditions.

Insects are an important group involved in carrion consumption and are thus of forensic interest. In the laboratory we studied the taphonomic marks that Necrobia rufipes (Cleridae) can produce. Pig trotters were exposed to adult beetles at 21 ± 3 °C and 12:12 h day/night cycle. We made observations and took photographs every 4-5 days for 12 months. Marks were noted after a month. We found scratches, pits, holes, and tunnels in several kinds of tissue such as integumental, connective and muscular. This work contributes preliminary data of significant application in biology, ecology, anthropology and forensics. Until now, no study has provided taphonomic information with N. rufipes.