2003 Regional Ambient Fish Tissue Monitoring Program; Summary of Analyses

To supplement other environmental monitoring programs and to protect the health of people consuming fish from waters within this state, the state of Iowa conducts fish tissue monitoring. Since 1980, the Iowa Department of Natural Resources (IDNR), the United States Environmental Protection Agency Region VII (U.S. EPA), and the University of Iowa Hygienic Laboratory (UHL) have cooperatively conducted annual statewide collections and analyses of fish for toxic contaminants. Beginning in 1983, this monitoring effort became the Regional Ambient Fish Tissue Monitoring Program (the RAFT program). Currently, the RAFT program is the only statewide fish contaminant-monitoring program in Iowa. Historically, the data generated from the RAFT program have enabled IDNR to document temporal changes in contaminant levels and to identify Iowa lakes and rivers where high levels of contaminants in fish potentially threaten the health of Iowans consuming fish. The Iowa RAFT monitoring program incorporates three different but equally important types of monitoring sites: 1) status, 2) trend, and 3) follow-up.

2002 Regional Ambient Fish Tissue Monitoring Program Summary of Analyses

To supplement other environmental monitoring programs and to protect the health of people consuming fish from waters within this state, the state of Iowa conducts fish tissue monitoring. Since 1980, the Iowa Department of Natural Resources (IDNR), the United States Environmental Protection Agency Region VII (U.S. EPA), and the University of Iowa Hygienic Laboratory (UHL) have cooperatively conducted annual statewide collections and analyses of fish for toxic contaminants. Beginning in 1983, this monitoring effort became the Regional Ambient Fish Tissue Monitoring Program (the RAFT program). Currently, the RAFT program is the only statewide fish contaminant-monitoring program in Iowa. Historically, the data generated from the RAFT program have enabled IDNR to document temporal changes in contaminant levels and to identify Iowa lakes and rivers where high levels of contaminants in fish potentially threaten the health of fish consuming Iowans. The Iowa RAFT monitoring program incorporates three different but equally important types of monitoring sites: 1) status, 2) trend, and 3) follow-up.

2001 Regional Ambient Fish Tissue Monitoring Program; Summary of Analyses

To supplement other environmental monitoring programs and to protect the health of people consuming fish from waters within this state, the state of Iowa conducts fish tissue monitoring. Since 1980, the Iowa Department of Natural Resources (IDNR), the United States Environmental Protection Agency Region VII (U.S. EPA), and the University of Iowa Hygienic Laboratory (UHL) have cooperatively conducted annual statewide collections and analyses of fish for toxic contaminants. Beginning in 1983, this monitoring effort became the Regional Ambient Fish Tissue Monitoring Program (the RAFT program). Currently, the RAFT program is the only statewide fish contaminant-monitoring program in Iowa. Historically, the data generated from the RAFT program have enabled IDNR to document temporal changes in contaminant levels and to identify Iowa lakes and rivers where high levels of contaminants in fish potentially threaten the health of fish consuming Iowans.

Fish oil supplementation does not alter energy efficiency in healthy males

BACKGROUND AND AIMS: Fish oil (FO) supplementation prevents the development of obesity and insulin resistance, and upregulate the expression of UCP3 in skeletal muscle in rodents. This may represent indirect evidence that FO promotes fat oxidation and/or alter energy efficiency. The aim of this study was to evaluate whether such effects can be observed in humans. The metabolic effects of FO were assessed during exercise in order to obtain a direct measurement of energy efficiency. METHODS: Eight healthy male volunteers were studied with and without supplementation with 7.2 g/day FO (including 1.1 g/day eicosopentaenoic acid and 0.7 g/day decosahexaenoic acid) during 14 days. Their VO(2 max) was measured on cycle ergometer. Thereafter, energy metabolism (substrate oxidation, energy expenditure and energy efficiency) was assessed during a 30 min cycling exercise at 50% VO(2 max) performed 2 h 30 after a standardized, high carbohydrate breakfast. RESULTS: VO(2 max) was 38.6+/-2.2 after FO and 38.4+/-2.0 (mL x kg(-1) x min(-1)) in control conditions (NS). Basal plasma glucose, insulin and NEFA concentrations, and energy metabolism were similar with FO and in controls. During exercise, the increases in plasma NEFA concentrations, energy expenditure, glucose and lipid oxidation, and the decreases in glycaemia and insulinemia were not altered by FO intake. Energy efficiency was 22.4+/-0.6% after FO vs 21.8+/-0.7% in controls. In order to ascertain that the absence of effects of FO was not due to consumption of a carbohydrate meal immediately before exercise, 4 of the 8 subjects were re-studied in fasting conditions, FO also failed to alter energy efficiency in this subset of studies. CONCLUSION: FO supplementation did not significantly alter energy metabolism and energy efficiency during exercise in healthy humans.

Parasitic fauna in hybrid tambacu from fish farms

The objective of this work was to evaluate the parasitic fauna of hybrid tambacu (Colossoma macropomum x Piaractus mesopotamicus) from fish farms and the host-parasite relationship. A hundred and fourteen fish were collected from four fish farms in Macapá, in the state of Amapá, Brazil, 80.7% of which were infected by: Ichthyophthirius multifiliis (Ciliophora); Piscinoodinium pillulare (Dinoflagellida); Anacanthorus spatulatus, Notozothecium janauachensis, and Mymarothecium viatorum (Monogenoidea); Neoechinorhynchus buttnerae (Acanthocephala); Cucullanus colossomi (Nematoda); Perulernaea gamitanae (Lernaeidae); and Proteocephalidae larvae (Cestoda). A total of 8,136,252 parasites were collected from the examined fish. This is the first record of N. buttnerae, C. colossomi, N. janauachensis, M. viatorum, and Proteocephalidae for hybrid tambacu in Brazil. Ichthyophthirius multifiliis was the most prevalent parasite, whereas endohelminths were the less. A positive correlation was observed between number of I. multifiliis and total length and weight of fish, as well as between number of P. gamitanae and total length. The infection by I. multifiliis had association with the parasitism by Monogenoidea. Low water quality contributes to high parasitism of hybrid tambacu by ectoparasites, which, however, does not influence the relative condition factor of fish.

Study of fish swimming activity using Acoustical Doppler Velocimetry (ADV) techniques

Peer reviewed

Fish oil replacement by soybean oil in the diet of fat snook juveniles

The objective of this work was to evaluate fish oil replacement by soybean oil in diets, as for the effects on the performance and body composition of juveniles of fat snook (Centropomus parallelus). The experiment was carried out in a randomized block design, with three treatments (lipid sources) and six replicates, in a 60-day period. Fat snook juveniles (24.17±0.28g) were distributed in 18 experimental tanks of 200 L each, equipped with aeration and heating systems, under continuous water renovation (800% per day). Three isoproteic (44% CP) and isoenergetic (4,635 kcal CE kg-1) diets were formulated to comprise three replacement rates (0, 50, and 100%) of fish oil by soybean oil. Biometric analyses were done to evaluate fish performance, and two entire specimens from each replicate were used for body composition analyses. The zootechnical indices of weight gain (38.68±5.41 g), feed conversion (1.38±0.10), and specific growth at 1.70±0.18% weight gain per day were considered satisfactory. Lipid source substitution does not affect the performance and body composition of fat snook juveniles, which suggests that soybean oil can replace fish oil in diet formulation.

Screening for ALK in non-small cell lung carcinomas: 5A4 and D5F3 antibodies perform equally well, but combined use with FISH is recommended.

OBJECTIVES: Immunohistochemistry (IHC) has become a promising method for pre-screening ALK-rearrangements in non-small cell lung carcinomas (NSCLC). Various ALK antibodies, detection systems and automated immunostainers are available. We therefore aimed to compare the performance of the monoclonal 5A4 (Novocastra, Leica) and D5F3 (Cell Signaling, Ventana) antibodies using two different immunostainers. Additionally we analyzed the accuracy of prospective ALK IHC-testing in routine diagnostics. MATERIALS AND METHODS: Seventy-two NSCLC with available ALK FISH results and enriched for FISH-positive carcinomas were retrospectively analyzed. IHC was performed on BenchMarkXT (Ventana) using 5A4 and D5F3, respectively, and additionally with 5A4 on Bond-MAX (Leica). Data from our routine diagnostics on prospective ALK-testing with parallel IHC, using 5A4, and FISH were available from 303 NSCLC. RESULTS: All three IHC protocols showed congruent results. Only 1/25 FISH-positive NSCLC (4%) was false negative by IHC. For all three IHC protocols the sensitivity, specificity, positive (PPV) and negative predictive values (NPV) compared to FISH were 96%, 100%, 100% and 97.8%, respectively. In the prospective cohort 3/32 FISH-positive (9.4%) and 2/271 FISH-negative (0.7%) NSCLC were false negative and false positive by IHC, respectively. In routine diagnostics the sensitivity, specificity, PPV and NPV of IHC compared to FISH were 90.6%, 99.3%, 93.5% and 98.9%, respectively. CONCLUSIONS: 5A4 and D5F3 are equally well suited for detecting ALK-rearranged NSCLC. BenchMark and BOND-MAX immunostainers can be used for IHC with 5A4. True discrepancies between IHC and FISH results do exist and need to be addressed when implementing IHC in an ALK-testing algorithm.

Regulation of bistable ICEclc transfer in pseudomonas knackmussii B13

L'élément génétique intégratif et conjugatif auto-transférable de 103 kb qui se trouve dans le génome de Pseudomonas knackmussii B13 (ICEc/c) confère la capacité de dégrader le 3-chlorobenzoate et le 2-aminophénol. L'élément ICE c/c peut être transféré par conjugaison de la souche B13 à diverses bêta- et gamma- protéobactéries. Seule une sous-population de 3 à 5% des cellules transfère l'élément, les cellules dites "compétentes pour le transfert". L'acquisition de la compétence pour le transfert est vraisemblablement la conséquence d'une régulation bistable, conduisant une partie des cellules au transfert de l'élément ICE c/c tandis que, dans les autres, l'élément reste quiescent et ne se transfère pas. À ce jour, les mécanismes et les acteurs moléculaires qui régulent l'activation bistable de l'élément sont restés inconnus. Mon travail de doctorat visait à identifier les éléments bistables du régulon de la compétence pour le transfert et d'analyser les fondements moléculaires de la bistabilité de l'élément ICE c/c chez P. knackmussii. Le premier chapitre introduit le thème du transfert génétique horizontal avec un accent particulier sur les éléments intégratifs et conjugatifs (ICE) et ICEcIc. L'état actuel des connaissances sur l'organisation génétique, la régulation, l'intégration et le transfert de différents modèles de ICEs est exposé en détail. En outre, je m'étends sur les phénomènes d'hétérogénéité et de bistabilité phénotyplques, qu'on peut distinguer dans une population isogénique dans des conditions de culture homogènes, et qui sont susceptibles de jouer un rôle dans le transfert de l'élément ICE c/c, dans la mesure où il ne s'active et n'est transférable que dans une très petite sous-population de cellules. Dans le chapitre 2, je présente une analyse globale des régions promotrices minimales des gènes appartenant au régulon de la compétence pour le transfert de l'élément ICE c/c. Nous avons étudié les caractéristiques d'expression des promoteurs et, s'ils s'avéraient bistables, leur activation dans le temps par comparaison avec le mutant lntB13. Pour ce faire, nous avons utilisé des fusions de promoteurs avec des gènes rapporteurs et testé l'expression bistable chez P. knackmussii par microscopie à épifluorescence. Pour six promoteurs présentant une expression bistable, nous avons employé de la microscopie temporelle pour déterminer la chronologie de leur expression par rapport à Pint et PinR. Parmi eux, nous avons identifié deux gènes exprimés précocement et trois gènes exprimés tardivement dans le processus d'acquisition de la compétence de transfert. Dans le chapitre 3, j'expose une analyse d'expression génétique pour l'un des groupes de gènes dont la transcription est la plus élevée dans la région conservée de ICE c/c, les gènes orf81655-orf68241 contenus dans une région de 14 kb. Nous montrons d'abord que cet opéron fait partie du même régulon bistable que intB13 et inrR et analysons les caractéristiques génétiques qui conduisent à une transcription élevée. Nous étudions les fonctions biologiques de ce groupe de gènes par des délétlons ciblées et montrons que certaines d'entre elles empêchent le transfert de l'élément. Nous approfondissons la caractérlsatlon de I'orf8l655 en construisant une fusion transcrlptionnelle avec le gène codant pour la protéine fluorescente verte (egfp) (en utilisant le système minl-Tn5). L'expression de Vorf81655 dans des cellules individuelles est comparée au signal mesuré par hybridation in situ en fluorescence (FISH) sur le ARN messager du gène. En utilisant FISH, des délétlons du promoteur et de l'analyse directe de transcription, nous avons localisé la région promotrice du groupe de gènes. En outre, nous avons utilisé des mutations dirigées pour comprendre la bistabilité de cette région promotrice, caractérisée par une transcription très élevée et une traduction lente de l'ARN messager.  Dans le chapitre 4, nous nous efforçons de comprendre comment la bistabilité est générée au sein du régulon te de l'élément ICE c/c. Pour ce faire, nous avons tenté de reconstituer une expression bistable, dans un hôte qui ne présente pas de bistabilité naturellement, à partir d'éléments génétiques individuels. L'hôte choisi est Pseudomonas putida dans lequel nous avons introduit une copie unique de Pint, PinR ou PaipA fusionnés à la egfp, construits qui permettent d'observer l'apparition de bistabilité. Nous avons ensuite construit différents assemblages de composants génétiques de l'élément ICE c/c, en nous concentrant sur la région parA-inrR. En effet, nous avons pu démontrer qu'une expression bistable apparaît dans P. putida grâce à ces éléments en l'absence de l'élément ICE c/c complet. À noter que la plupart des construits génétiques activent PaipA ou P|,,R, mais qu'un seul recrée la bistabilité de Pint, ce qui suggère que la région parA-inrR permet à la fois d'engendrer la bistabilité et d'opérer la transition entre les promoteurs précoces et les promoteurs tardifs du régulon de la bistabilité. Dans le chapitre 5, nous concluons sur une discussion de la pertinence de nos résultats et sur de futures perspectives de recherche. -- The 103-kb self-transmissible integrative and conjugative element (ICE) of Pseudomonas knackmussii B13 (ICEc/c) confers the capacity to degrade 3- chlorobenzoate and 2-aminophenol. ICEc/c can be conjugated from strain B13 to a variety of Beta- and Gammaproteobacteria. Interestingly, ICE c/c transfer is observed in a subpopulatlon of cells (3-5%) only, the so-called 'transfer competent' cells. The formation of transfer competence (tc) is thought to be the consequence of a 'bistable' decision, which forces those cells to follow the developmental path which leads to ICEc/c transfer, whereas in others ICE c/c remains silent and does not transfer. So far, the mechanisms and molecular partners generating this bistable transfer activation in cells of P. knackmussii B13 remain mostly unidentified. This thesis aimed at understanding the extent of the tc bistability regulon and to dissect the molecular basis of bistabillty formation of ICEc/c in P. knackmussii. The first chapter is a general Introduction on horizontal gene transfer (HGT) with particular emphasis on ICEs and ICE c/c. The emphasis is made on the current knowledge about the HGT gene organization, regulation and specific integration and transfer aspects of the different ICEs models. Furthermore, I focus on the phenomena of phenotypic heterogeneity and bistability (the property of two distinguishable phenotypes existing within an isogenic population under homogeneous conditions), which may play a particular role in ICEc/c behaviour, since ICE activation and transfer only occurs in a very small subpopulation of cells. In Chapter Two, I focus on a global analysis of the different core promoters that might belong to the ICEc/c tc pathway regulon. We studied both expression patterns of ICEc/c promoters and, once being identified as "bistable", their temporal activation compared to that of intB13. In order to do this, we used promoter reporter fusions and tested blstability expression in P. knackmussii using epifluorescence microscopy. For the 6 promoters that showed bistable expression, we used time-lapse microscopy to study the timing of promoter expression in comparison to that of P,,,t or PlnR. We could establish two "early" and 3 "late" phase promoters in the process of transfer competence. In Chapter Three, I focused my attention on analysis of gene expression of one of the most highly transcribed gene clusters in the conserved core region of ICEc/c, a 14-kb gene cluster formed by the genes orf81655-orf68241. First we showed that this operon is part of the same bistability 'regulon' as intB13 and inrR, and analysed the genetic features that lead to high transcription. We studied the potential biological function of this cluster for ICE c/c by making specific gene deletions, showing that some interrupt ICEc/c transfer. We further analysed the orfdl655 promoter by constructing transcriptional egfp fusion reporter strains using the miniTn5 delivery system. Expression of the orf81655 promoter in single cells was compared to signals measured by Fluorescence In Situ Hybridization (FISH) on orfSl655 mRNA. We localized the promoter region of the gene cluster using FISH, promoter deletions, and by direct transcript analysis. We further used site-directed mutagenesis to understand the bistability character of the promoter region and the extremely high transcription but low translation from this mRNA. In Chapter Four, we set out to understand how bistability is generated in the tc pathway of ICEc/c. For this we tried rebuilding bistable expression from ICEc/c individual gene components in a host, which normally does not display bistability. As host we used P. putida without ICEc/c but with a single copy Pint-, PlnR- or PalpA- egfp fusion that enabled us to verify bistability formation. Subsequently, we built different assemblages of ICEc/c gene components, focusing on the parA-inrR region. Indeed, we found that bistable expression can be build from those components in P. putida without ICEc/c. Interestingly, most genetic constructs activated PaipA or PlnR, but only one resulted in bistable activation of PinT. This suggests that the parA-inrR region acts as a bistability "generator", but also as a bistability "relay" from early to late promoters in the tc pathway hierarchy. In the final fifth chapter, we conclude with a discussion of the relevance of the present thesis and the resulting perspectives for future studies.

Macroinvertebrate assemblages on reed beds, with special attention to Chironomidae (Diptera), in Mediterranean shallow lakes.

Macroinvertebrates associated to reed-beds (Phragmites australis) in six shallow natural water bodies along the 220 km of coast of the Comunidad Valenciana (Spain) were studied. These sites were selected to reflect different trophic states, but also, and due to the natural variability of mediterranean wetlands, they greatly differ in salinity and hydroperiod. To unify the sampling, reed bed was chosen to provide data from a habitat common to all wetlands, including the most eutrophic ones where submerged macrophytes have disappeared due to water turbidity. Individual submerged stems of Phragmites australis were sampled along with the surrounding water. The animal density found refers to the available stem surface area for colonization. Forty-one taxa were recorded in total, finding Chironomidae to be the most important group, quantitatively and qualitatively. In freshwater sites it was observed an increase in macroinvertebrate"s density at higher trophic states. Nevertheless each studied region had a different fauna. The PCA analysis with macroinvertebrate groups distinguished three types of environment: freshwaters (characterized by swimming insect larvae, collectors and predators, oligochaetes and Orthocladiinae), saline waters (characterized by crustaceans and Chironominae) and the spring pool, which shares both taxa. Chironomids were paid special attention for being the most abundant. A DCA analysis based on the relative abundance of Chironomids reveals salinity as the main characteristic responsible for its distribution, but trophic state and hydrological regime were also shown to be important factors.

Nutrient enrichment and water depth modify consumer control in rocky shore macroalgal communities

The objective of my thesis is to assess mechanisms of ecological community control in macroalgal communities in the Baltic Sea. In the top-down model, predatory fish feed on invertebrate mesograzers, releasing algae partly from grazing pressure. Such a reciprocal relationship is called trophic cascade. In the bottom-up model, nutrients increase biomass in the food chain. The nutrients are first assimilated by algae and, via food chain, increase also abundance of grazers and predators. Previous studies on oceanic shores have described these two regulative mechanisms in the grazer - alga link, but how they interact in the trophic cascades from fish to algae is still inadequately known. Because the top-down and bottom-up mechanisms are predicted to depend on environmental disturbances, such as wave stress and light, I have studied these models at two distinct water depths. There are five factorial field experiments behind the thesis, which were all conducted in the Finnish Archipelago Sea. In all the experiments, I studied macroalgal colonization - either density, filament length or biomass - on submerged colonization substrates. By excluding predatory fish and mesograzers from the algal communities, the studies compared the strength of the top-down control to natural algal communities. A part of the experimental units were, in addition, exposed to enriched nitrogen and phosphorus concentrations, which enabled testing of bottom-up control. These two models of community control were further investigated in shallow (<1 m) and deep (ca. 3 m) water. Moreover, the control mechanisms were also expected to depend on grazer species. Therefore different grazer species were enclosed into experimental units and their impacts on macroalgal communities were followed specifically. The community control in the Baltic rocky shores was found to follow theoretical predictions, which have not been confirmed by field studies before. Predatory fish limited grazing impact, which was seen as denser algal communities and longer algal filaments. Nutrient enrichment increased density and filament length of annual algae and, thus, changed the species composition of the algal community. The perennial alga Fucus vesiculosusA and the red alga Ceramium tenuicorne suffered from the increased nutrient availabilities. The enriched nutrient conditions led to denser grazer fauna, thereby causing strong top-down control over both the annual and perennial macroalgae. The strength of the top-down control seemed to depend on the density and diversity of grazers and predators as well as on the species composition of macroalgal assemblages. The nutrient enrichment led to, however, weaker limiting impact of predatory fish on grazer fauna, because fish stocks did not respond as quickly to enhanced resources in the environment as the invertebrate fauna. According to environmental stress model, environmental disturbances weaken the top-down control. For example, on a wave-exposed shore, wave stress causes more stress to animals close to the surface than deeper on the shore. Mesograzers were efficient consumers at both the depths, while predation by fish was weaker in shallow water. Thus, the results supported the environmental stress model, which predicts that environmental disturbance affects stronger the higher a species is in the food chain. This thesis assessed the mechanisms of community control in three-level food chains and did not take into account higher predators. Such predators in the Baltic Sea are, for example, cormorant, seals, white-tailed sea eagle, cod and salmon. All these predatory species were recently or are currently under intensive fishing, hunting and persecution, and their stocks have only recently increased in the region. Therefore, it is possible that future densities of top predators may yet alter the strengths of the controlling mechanisms in the Baltic littoral zone.

Is response to fire influenced by dietary specialization and mobility? A comparative study with multiple animal assemblages

Fire is a major agent involved in landscape transformation and an indirect cause of changes in species composition. Responses to fire may vary greatly depending on life histories and functional traits of species. We have examined the taxonomic and functional responses to fire of eight taxonomic animal groups displaying a gradient of dietary and mobility patterns: Gastropoda, Heteroptera, Formicidae, Coleoptera, Araneae, Orthoptera, Reptilia and Aves. The fieldwork was conducted in a Mediterranean protected area on 3 sites (one unburnt and two burnt with different postfire management practices) with five replicates per site. We collected information from 4606 specimens from 274 animal species. Similarity in species composition and abundance between areas was measured by the Bray-Curtis index and ANOSIM, and comparisons between animal and plant responses by Mantel tests. We analyze whether groups with the highest percentage of omnivorous species, these species being more generalist in their dietary habits, show weak responses to fire (i.e. more similarity between burnt and unburnt areas), and independent responses to changes in vegetation. We also explore how mobility, i.e. dispersal ability, influences responses to fire. Our results demonstrate that differences in species composition and abundance between burnt and unburnt areas differed among groups. We found a tendency towards presenting lower differences between areas for groups with higher percentages of omnivorous species. Moreover, taxa with a higher percentage of omnivorous species had significantly more independent responses of changes in vegetation. High- (e.g. Aves) and low-mobility (e.g. Gastropoda) groups had the strongest responses to fire (higher R scores of the ANOSIM); however, we failed to find a significant general pattern with all the groups according to their mobility. Our results partially support the idea that functional traits underlie the response of organisms to environmental changes caused by fire.

Tectonic-palaeoenvironmental forcing of clay-mineral assemblages in nonmarine settings: the Oligocene-Miocene As Pontes Basin (Spain)

Two small, alluvial-lacustrine subbasins developed during the early restraining overstep stages of the Oligocene-Miocene As Pontes strike-slip Basin (NW Spain). Later, the basin evolved into a restraining bend stage and an alluvial-swamp-dominated depositional framework developed. The palaeobiological record demonstrates that the Oligocene-Miocene palaeoclimate in NW Spain was subtropical, warm and humid to subhumid.

AMP-activated protein kinase plays an important evolutionary conserved role in the regulation of glucose metabolism in fish skeletal muscle cells

AMPK, a master metabolic switch, mediates the observed increase of glucose uptake in locomotory muscle of mammals during exercise. AMPK is activated by changes in the intracellular AMP:ATP ratio when ATP consumption is stimulated by contractile activity but also by AICAR and metformin, compounds that increase glucose transport in mammalian muscle cells. However, the possible role of AMPK in the regulation of glucose metabolism in skeletal muscle has not been investigated in other vertebrates, including fish. In this study, we investigated the effects of AMPK activators on glucose uptake, AMPK activity, cell surface levels of trout GLUT4 and expression of GLUT1 and GLUT4 as well as the expression of enzymes regulating glucose disposal and PGC1α in trout myotubes derived from a primary muscle cell culture. We show that AICAR and metformin significantly stimulated glucose uptake (1.6 and 1.3 fold, respectively) and that Compound C completely abrogated the stimulatory effects of the AMPK activators on glucose uptake. The combination of insulin and AMPK activators did not result in additive nor synergistic effects on glucose uptake. Moreover, exposure of trout myotubes to AICAR and metformin resulted in an increase in AMPK activity (3.8 and 3 fold, respectively). We also provide evidence suggesting that stimulation of glucose uptake by AMPK activators in trout myotubes may take place, at least in part, by increasing the cell surface and mRNA levels of trout GLUT4. Finally, AICAR increased the mRNA levels of genes involved in glucose disposal (hexokinase, 6-phosphofructokinase, pyruvate kinase and citrate synthase) and mitochondrial biogenesis (PGC-1α) and did not affect glycogen content or glycogen synthase mRNA levels in trout myotubes. Therefore, we provide evidence, for the first time in non-mammalian vertebrates, suggesting a potentially important role of AMPK in stimulating glucose uptake and utilization in the skeletal muscle of fish.

Bat assemblages in the Nietoperek bat reserve (Western Poland) and their conservation strategies

Międzyrzecz Fortified Front, were Natura 2000 site PLH080003 Nietoperek is situated, was built by the Germans in the 1930s and during the World War II. It is composed of above ground bunkers connected by underground tunnels of ca. 32 km total length. Nietoperek is the eighth largest bats hibernation site in EU. Monthly censuses were carried out from October to April during three consecutive winter seasons (2011/12 – 2013/14) in area covering ca. 30% of the undergrounds. The aims of the study were: (1) to describe changes in numbers of each species in the course of hibernation season, (2) to suggest deadlines for counting particular bat species to obtain maximal numbers and (3) to describe negative impact of tourism on hibernating bats. The results will be useful for restriction of winter tourism in Nietoperek. The total number of bats observed during the study was 37869 individuals of 9 species. Because of difficulties in distinguishing without handling M. mystacinus and M. brandtii were treated as one group. M. myotis constituted from 53% (first season) to 64% (last season) of all hibernating bats. The maximal numbers of individuals were observed in November (first two seasons) and in December (third season). M. daubentonii constituted from 27% (first season) to 21% (last season) and M. nattereri from 10% (first season) to 11% (second season) of all bats. During the three seasons the maximal numbers of M. daubentonii and M. nattereri were observed in November and December respectively. B. barbastellus and P. auritus constituted from 4% (first season) to 2% (last season) of the multi species colony. The maximal numbers of B. barbastellus were observed in January and P. auritus in January (first and second seasons) and in December (third season). Results indicated that the best period for counting maximal numbers of M. myotis and M. daubentonii is November, for M. nattereri is December and for B. barbastellus and P. auritus is January. The study undertaken in the part visited by tourists in winter (total length of 900 m) proved negative effect caused by human disturbance with 23% decline of total bat numbers.