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Many stenotherm marine species live in the cold and stable environment of the Antarctic Ocean which could be impacted by climate change in the upcoming years. To investigate this issue, gene expression analysis was carried out on Trematomus bernacchii specimens caught near Mario Zucchelli Station. Brain, gill and muscle tissues were sampled from naïve animals and from those kept in control (- 0.9°C) and experimental (+0.6°C) tanks for six hours, seven and twenty days, post acclimation. RNAseq data showed that the temperature increase mostly affected the brain, with a time dependent response. Immune response was up-regulated at mid-term; protein turnover and energy management were down-regulated and neuronal remodelling related genes were up-regulated at 20 days, where the highest number of differentially expressed genes was observed. Gill tissue showed a mild response after 20 days, mostly involving DNA replication. A great early-starting response to stabling was also observed across the entire experiment in brain and gills, with the latter being the most affected. The expression pattern clustering analysis showed that many synapse-related genes were down-regulated and energy related genes were up-regulated, while several binding processes were up-regulated at the latest time point in brain. The response in gills was milder and mostly involved cytoskeleton and glycolysis. No significant change was observed in muscle. These results identify the brain as the tissue most affected by both heat and stabling, evidencing its great sensitivity to the slightest environmental change, and highlight the importance of a careful experimental design when working with captive wild organisms.

Il presente studio si propone di approfondire la biologia ed il ciclo riproduttivo del gobione italiano (Gobio benacensis), specie endemica a corologia cisalpina inclusa nella categoria “Endangered” (EN) della lista rossa IUCN, con l’intento di fornire informazioni utili a redigere corretti piani gestionali volti alla conservazione della specie. Le indagini sono state condotte in un corso d’acqua del Bacino del Fiume Isonzo (Torrente Reca, Nordest Italia) ed hanno riguardato 86 esemplari catturati nell’aprile 2016 al fine di definirne la consistenza e la struttura della popolazione e 77 esemplari, catturati mensilmente e bimestralmente tra aprile 2016 e febbraio 2017, per la definizione del ciclo riproduttivo. Le curve di regressione del peso sulla lunghezza totale per i maschi (W=0,1253TL2,8770) e per le femmine (W=0,1555TL2,7554), confrontate mediante ANCOVA, non hanno denunciato differenze significative. I valori medi ± deviazione standard dell’indice gonadosomatico GSI per i maschi sono compresi tra 0,61±0,49 e 1,75±0,58, mentre per le femmine tra 1,02±0,88 e 14,15±2,80. L’indice epatosomatico HSI ha denunciato range compresi tra 0,05±0,02 e 2,68±1,26 e tra 0,25±0,19 e 1,75±0,58 rispettivamente per i maschi e per le femmine. Le analisi istologiche di ovari e testicoli, analizzati per la prima volta nella specie, hanno permesso di identificare gli stadi di maturazione, che unitamente all’analisi del GSI indicano che la specie depone le uova a più riprese durante il periodo riproduttivo compreso tra aprile e giugno. Nei maschi è stata, tuttavia, evidenziata una prolungata attività riproduttiva che si protrae fino al mese di agosto.

The aim of the present work is to study the biology and the reproductive cycle of the Italian gudgeon Gobio benacensis, which is included as “endangered” (EN) in the IUCN Redlist, due to the endangered status, information could be useful for the correct management of the species and for the scheduling of focused conservation plans. Analyses were carried out within a watercourse included in the Isonzo River Basin (Reca Stream, Northeast Italy) and regarded 86 specimen collected during April 2016 in order to assess population consistence and structure, and 77 specimens collected monthly and bimonthly between April 2016 and February 2017 to investigate the reproductive cycle. Regression between total length (TL) and body weight (W) obtained for males (W=0,1253TL2,8770) 54 2 and females (W=0,1555TL2,7554) were analyzed using the ANCOVA and did not show significant differences. Mean values of the gonadosomatic index (GSI) ranged from 0,61±0,49 and 1,75±0,58 for males and from 1,02±0,88 and 14,15±2,80 for females. Hepatosomatic index (HIS) showed ranges between 0,05±0,02 and 2,68±1,26 for males and between 0,25±0,19 e 1,75±0,58 for females. Analyses of ovaries and testicles, firstly performed for this species, allowed to identify maturation stages. The results show that G. benacensis females repeatedly spawn during the reproductive period between April and June and that males prolonged reproductive activity until August.

This chapter provides an overview of the main stress responses modulated by neuropeptides of the crustacean hyperglycemic hormone (CHH) family. The current scenario of upstream regulation of CHHs is presented here. Numerous sequence variants, splicing-isoforms, and posttranslational modifications of CHHs point to a wide functional diversification of these peptides.Other than the classical main CHH-mediated cascade resulting in energy mobilization, new aspects focused on CHH chirality are demanding deeper efforts on the characterization of their target and mode of action. Moreover, new CHH variants with immune-related functions have been recently described, broadening further the role of such a neuropeptides family.

Habituation consists of a decreased response to repeated stimulation and not involving fatigue or sensory adaptation1, and is common to many living entities, from single cells to the most complex organisms2-3. It has a considerable evolutionary value, since it allows an organism to disregard irrelevant repetitive stimuli. Here, we tested whether two different stimuli, with possible different social values, may have a different impact on habituation in individuals of crayfish Procambarus clarkii, a highly aggressive crustacean often fighting with conspecifics for the control of shelter and resources4. The stimuli consisted in a red upright Y-shaped figure, resembling a crayfish displaying its aggressive posture (high intensity stimulus condition), and a red downright Y-shaped figure, identical to the other but rotated of 180° and resembling the animals’ relaxed posture (low intensity stimulus condition). A habituation paradigm was used, in which two groups of 24 animals were presented with nine consecutive looming stimulations, thus creating the illusion of an approaching stimulus. Furthermore, a tenth looming presentation of a completely different stimulus (a circular blue moebius) was used to test for a spontaneous recovery of the response. As dependent measure, we recorded the duration of the alert reaction to the approaching shape (measured in video frames, recorded at 29 fps). Crayfish showed habituation in both conditions (p=0.002). The animals tested in the low intensity condition abruptly habituated to the repeating stimulus, and then showed a strong response recovery to the new stimulus (p=0.001). By contrast, crayfish in the high intensity condition maintained a consistently higher alertness during the administration of the repeating upright Y-shaped figure (M=11.61, SE=1.10) as compared to those in the low intensity condition (M=8.72, SE=1.21). Moreover, crayfish in the high intensity condition showed no spontaneous recovery to the approaching stimulus when it was changed in shape and colour (p=0.120). Our data confirm previous findings on looming in crayfish5-6 and indicate that they responded as if they faced real opponents7. Indeed, the more sustained pattern of response evoked by the upright stimulus seems to be due to a different social valence between the two schematic stimuli, with one stimulus being perceived as more threatening than the other. Our results show that P. clarkii is sensitive to the abstract visual representation of conspecifics.

A number of anthropogenic activities, including farming and urbanization, have a significant impact on the environment and can produce an irreversible damage at the population level. To assess the effects of environmental stressors as metal pollution on Apis mellifera ligustica populations, we analyzed the hypopharyngeal gland size and morphology of foraging bees along a urban-suburban gradient. Foraging bees are collected from beehives located in S. Giovanni (Trieste, “urban site”) and in Domio (Trieste, “suburban site”) over two full activity periods in early summer (July). Foraging bee heads from urban site (n = 4) and from suburban site (n = 2) were fixed and embedded in Epon 812-Araldite perform morphometrical analyses of hypopharyngeal glands, the minimum and maximum diameters of the acini was measure used ImageJ program. Diameters were expressed as mean±SE and differences were assessed by nonparametric statistics. Statistical analyses were performed using R version 3.0.1 software (R Development Core Team 2013). The statistical comparison showed a smaller diameter of the acini of hypopharyngeal gland of bees from suburban site (0.080 mm ± 0.003), compared to those of urban site (0.114 mm ± 0.004). Statistical analysis showed an highly significant difference in the size of the acini between the site of Domio (Wilcoxon rank sum test, p = 1.651x10-7) compared to the site of S. Giovanni. Metal contents were recorded for foraging bees (n = 20 from each site) and measured by inductively coupled plasma-mass spectrometry (ICP-MS). Recorded elements are As, Bi, Cd, Co, Cr, Cu, Ni, Pb, Sr, V, and Zn. They are accumulated in two different rank order Zn> Cu> Sr=Bi> Ni> Cr> Pb=Co> V>Cd >As in bees from urban site and Zn> Cu> Sr >Cr >Ni>B =Co=V>Pb>As>Cd in bees from suburban site. Significantly differences in concentration of Cd, Cr and Cu were recorded in bees from the two sites (Wilcoxon rank sum test p

The crustacean hyperglycaemic hormone (CHH), a pleiotropic neuropeptide, belongs to a family of structurally related peptides, having six cysteine residues in conserved positions forming three disulphide bridges, and regulating several physiological processes in crustaceans and insects. Structure-activity studies have shown that amidation of the C-terminus is important to confer biological activity to CHH. In this study we investigated the function of the d-Phe3 of the N-terminal motif of Pontastacus leptodactylus CHH by a mutational analysis. The d-Phe in position 3 was substituted by a d-Ala and the functionality of the mutated analogue (Glp-d-A-CHH) was tested by in vivo biological assays. The mutated analogue resulted far less active than its wild-type counterparts, either in d- (Glp-d-CHH) or l- (Glp-l-CHH) configuration. These results suggest that Phe3 is essential for the biological activity of P. leptodactylus CHH, demonstrating that also the N-terminus is involved in the binding with the receptor, and identifying in the Phe3 a hot spot for the peptide-receptor binding.

Main immune defences of the honey bees are the cellular responses represented by phagocytosis and melanisation. There are a number of factors that could impact on the honeybees immune system and, therefore, increase their susceptibility to disease and lower their survivorship such as: exposure to pesticides, in air, in pollen, in nectar and in water; fungicides from both field and in-hive treatments; varroacides; the pest Varroa destructor; antibiotics used in in-hive treatments; fungal pathogens such Nosema apis and the emerging Nosema ceranae (Botías et al., 2013); bacterial and viral infections. More important, it must be highlighted the interactions among different chemicals and their synergic effect with diseases in the immune suppression of individuals and on the colony. The present study use simple and reliable methods that can clearly describe the state of health of the bees (Gonzàles-Santoyo & Còrdoba-Aguilar, 2011; Pandley & Tiwari, 2012), such as total hemocyte counts (THCs) and the activities of the plasmatic phenoloxidase (PO) and its inactive form (proPO) to assess the immune competence of individuals. Specimens of Apis mellifera ligustica were collected from beehives located in S. Giovanni (Trieste, “control site”) and from hives placed in Domio (Trieste, “polluted site”) in summer and early autumn. Both in summer and in autumn the statistical comparison showed a greater number of circulating hemocytes in bees from the site of Domio (July: 1159333.3±123073.60 - mean ± SE, n = 15; October: 813333.3±50583.89 - mean ± SE, n = 15) compared to the numbers recorded in bees from S. Giovanni (July: 834166.7±55493.08 - mean ± SE, n = 12; October: 273333.3±33046.38 - mean ± SE, n = 12). The statistical analysis showed a trend towards significant difference in July (Wilcoxon rank sum test, p = 0.063) and an highly significant difference in October (Wilcoxon rank sum test, p < 0.00001). The honeybees from Domio presented an highly significant lower PO activity than those from S. Giovanni (ANCOVA: F3,206 = 38.73, p < 0.01, nS.Giovanni = 17, nDomio = 18). Also with regard to the enzymatic activity of the proPO we recorded a significantly higher one in the honeybees from S. Giovanni in comparison to those from Domio (ANCOVA: F3,206 = 23.66, p < 0.01, nS.Giovanni = 17, nDomio = 18). It should be noted that 52% of the bees collected from hives of Domio had 1 or 2 individuals of Varroa destructor on the tergites of the thorax. The higher activities of PO and proPO in the honeybees from S. Giovanni site is probably to be ascribed to the different quality of the environment in the 2 sites and thus it indicates a depression of non-specific immune competence and an increased susceptibility to Varroa destructor parasites in the honeybees from Domio.

The main immune defences of the honeybees are the cellular responses represented by phagocytosis and melanisation. There are a number of factors that could impact on the honeybees’ immune system and, therefore, increase their susceptibility to disease and lower their survivorship, such as: exposure to pesticides in the air, pollen, nectar and water; fungicides from both field and in-hive treatments; varroacides; the pest Varroa destructor; antibiotics used in in-hive treatments; fungal pathogens such Nosema apis and the emerging Nosema ceranae; bacterial and viral infections. More important, we highlight the interactions among different chemicals and their synergistic effects with diseases in the immune suppression of individuals and on the colony. The present study uses methodologies that can describe the health status of the honeybees, such as total hemocyte counts (THCs), the activities of the plasmatic phenoloxidase (PO) and its inactive form (proPO), to assess the immune competence of individuals. Specimens of Apis mellifera ligustica were collected in summer and early autumn from beehives located in S. Giovanni (Trieste, “control site”) and from hives placed in Domio (Trieste, “polluted site”). In both seasons the statistical comparison showed a greater number of circulating hemocytes in honeybees from Domio compared to the numbers recorded in those from S. Giovanni. The Domio’s honeybees showed a lower PO and proPO activities than those from S. Giovanni. It should be noted that 52% of the bees collected from the hives in Domio had one or two individuals of Varroa destructor on the tergites of the thorax. The higher PO and proPO activities recorded in the bees from S. Giovanni site are probably due to the different quality of the environment in the two sampling sites indicating a depression of non-specific immune competence and an increased susceptibility to Varroa destructor parasites in the honeybees from the polluted site of Domio.

Carabus Linnaeus, 1758 species are polyphagous and prey on every kind of food (earthworms, snails, caterpillars and fruit). Some macromorphological differentiations within the genus are often interpretated as related to specialised modes of nutrition. This study examined, in laboratory rearing conditions, larval development and survival under different diets (meat, earthworm, snail and fruit) for Carabus lefebvrei Dejean, 1826. The survival of animals varied significantly with diet. Our results on the survival and duration of development during laboratory rearing revealed that the pre-imaginal development of C. lefebvrei was influenced differently by various diets. The pure-snail diet was essential for fast larval development and adult survival, while the meat and mixed diets were unsuitable for proper development and survival.

La review descrive l'identificazione molecolare dei neuropeptidi dei crostacei essenziali per la crescita e la riproduzione

Five polymerase chain reaction (PCR) products which could not be reliably typed by allele-specific oligonucleotide (ASO) probing at the human leukocyte antigen (HLA) DQA1 locus were analyzed by polyacrylamide gel electrophoresis and direct sequencing. The first method revealed the preferential amplification of only one of the two alleles in two cases. Direct sequencing of PCR products allowed unambiguous genetic typing but a high number of artifacts was observed. Several of these artifacts occurred in the sequences recognized by the ASOs. This finding provides an explanation for the mistyping in the ASO probing procedure because Taq polymerase errors both created new genetic specificities and eliminated site-specific polymorphisms. Reversed-phase HPLC-MS of the five forensic templates showed a high degree of DNA damage. These data together indicate that the risk of mistyping when using the ASO probing procedure cannot be neglected in the forensic analysis of damaged DNA samples.

HUM. GENET.