Cardiac reflexes in a warming world: thermal plasticity of barostatic control and autonomic tones in a temperate fish.

Thermal plasticity of cardiorespiratory function allows ectotherms like fish to cope with seasonal temperature changes and is critical for resilience to climate change. Yet, the chronic thermal effects on cardiovascular homeostatic reflexes in fish are little understood although this may have important implications for physiological performance and overall resilience to climate warming. We compared cardiac autonomic control and baroreflex regulation of heart rate in perch (Perca fluviatilis L.) from a reference area in the Baltic Sea at 18-19°C with conspecifics from the Biotest enclosure, a chronically heated ecosystem receiving warmed effluent water (24-25°C) from a nuclear power plant. Resting heart rate of Biotest fish displayed clear thermal compensation and was 58.3±2.3 beats min-1 compared with 52.4±2.6 beats min-1 in reference fish at their respective environmental temperatures (Q10=1.2). The thermally compensated heart rate of Biotest fish was a combined effect of elevated inhibitory cholinergic tone (105% in Biotest fish versus 70% in reference fish) and reduced intrinsic cardiac pacemaker rate. A barostatic response was evident in both groups, as pharmacologically induced increases and decreases in blood pressure resulted in atropine-sensitive bradycardia and tachycardia, respectively. Yet, the tachycardia in Biotest fish was significantly greater, presumably due to the larger scope for vagal release. Acclimation of Biotest fish to 18°C for 3 weeks abolished differences in intrinsic heart rate and autonomic tone, suggesting considerable short-term thermal plasticity of cardiovascular control in this species. The heightened hypotensive tachycardia in Biotest perch may represent an important mechanism of ectothermic vertebrates that safeguards tissue perfusion pressure when tissue oxygen demand is elevated by environmental warming.

Chronic environmental warming alters cardiovascular and haematological stress responses in European perch (Perca fluviatilis).

Environmental warming and acute stress increase cardiorespiratory activity in ectothermic animals like fish. While thermal acclimation can buffer the direct thermal effects on basal cardiorespiratory function during chronic warming, little is known about how acclimation affects stress-induced cardiorespiratory responses. We compared cardiovascular and haematological responses to chasing stress in cannulated wild European perch (Perca fluviatilis) from a reference area at natural temperature (16 °C) with perch from the 'Biotest enclosure'; an experimental system chronically warmed (22 °C) by effluents from a nuclear power plant. Routine blood pressure was similar, but Biotest perch had slightly higher resting heart rate (59.9 ± 2.8 vs 51.3 ± 2.9 beats min-1), although the Q 10 for heart rate was 1.3, indicating pronounced thermal compensation. Chasing stress caused hypertension and a delayed tachycardia in both groups, but the maximum heart rate increase was 2.5-fold greater in Biotest fish (43.3 ± 4.3 vs 16.9 ± 2.7 beats min-1). Moreover, the pulse pressure response after stress was greater in reference fish, possibly due to the less pronounced tachycardia or a greater ventricular pressure generating capacity and thermally mediated differences in aortic compliance. Baseline haematological status was also similar, but after chasing stress, the haematocrit was higher in Biotest fish due to exacerbated red blood cell swelling. This study highlights that while eurythermal fishes can greatly compensate routine cardiorespiratory functions through acclimation processes, stress-induced responses may still differ markedly. This knowledge is essential when utilising cardiorespiratory variables to quantify and compare stress responses across environmental temperatures, and to forecast energetic costs and physiological constraints in ectothermic animals under global warming.

Physiological constraints to climate warming in fish follow principles of plastic floors and concrete ceilings.

Understanding the resilience of aquatic ectothermic animals to climate warming has been hindered by the absence of experimental systems experiencing warming across relevant timescales (for example, decades). Here, we examine European perch (Perca fluviatilis, L.) from the Biotest enclosure, a unique coastal ecosystem that maintains natural thermal fluctuations but has been warmed by 5-10 °C by a nuclear power plant for over three decades. We show that Biotest perch grow faster and display thermally compensated resting cardiorespiratory functions compared with reference perch living at natural temperatures in adjacent waters. However, maximum cardiorespiratory capacities and heat tolerance limits exhibit limited or no thermal compensation when compared with acutely heated reference perch. We propose that while basal energy requirements and resting cardiorespiratory functions (floors) are thermally plastic, maximum capacities and upper critical heat limits (ceilings) are much less flexible and thus will limit the adaptive capacity of fishes in a warming climate.

Isolation over 35 years in a heated biotest basin causes selection on MHC class IIß genes in the European perch (Perca fluviatilis L.).

Genes that play key roles in host immunity such as the major histocompatibility complex (MHC) in vertebrates are expected to be major targets of selection. It is well known that environmental conditions can have an effect on host-parasite interactions and may thus influence the selection on MHC. We analyzed MHC class IIß variability over 35 years in a population of perch (Perca fluviatilis) from the Baltic Sea that was split into two populations separated from each other. One population was subjected to heating from cooling water of a nuclear power plant and was isolated from the surrounding environment in an artificial lake, while the other population was not subjected to any change in water temperature (control). The isolated population experienced a change of the allelic composition and a decrease in allelic richness of MHC genes compared to the control population. The two most common MHC alleles showed cyclic patterns indicating ongoing parasite-host coevolution in both populations, but the alleles that showed a cyclic behavior differed between the two populations. No such patterns were observed at alleles from nine microsatellite loci, and no genetic differentiation was found between populations. We found no indications for a genetic bottleneck in the isolated population during the 35 years. Additionally, differences in parasitism of the current perch populations suggest that a change of the parasite communities has occurred over the isolation period, although the evidence in form of in-depth knowledge of the change of the parasite community over time is lacking. Our results are consistent with the hypothesis of a selective sweep imposed by a change in the parasite community.

By-catch of grey seals (Halichoerus grypus) in Baltic fisheries--a Bayesian analysis of interview survey.

Baltic seals are recovering after a population decline. The increasing seal stocks cause notable damage to fisheries in the Baltic Sea, with an unknown number of seals drowning in fishing gear every year. Thus, sustainable seal management requires updated knowledge of the by-catch of seals--the number of specimens that die in fishing gear. We analyse the by-catch of grey seals (Halichoerus grypus) in Finland, Sweden, and Estonia in 2012. We collect data with interviews (35 in Finland, 54 in Sweden, and 72 in Estonia) and analyse them with a hierarchical Bayesian model. The model accounts for variability in seal abundance, seal mortality and fishing effort in different sub-areas of the Baltic Sea and allows us to predict the by-catch in areas where interview data was not available. We provide a detailed description of the survey design and interview methods, and discuss different factors affecting fishermen's motivation to report by-catch and how this may affect the results. Our analysis shows that the total yearly by-catch by trap and gill nets in Finland, Sweden and Estonia is, with 90% probability, more than 1240 but less than 2860; and the posterior median and mean of the total by-catch are 1550 and 1880 seals, respectively. Trap nets make about 88% of the total by-catch. However, results also indicate that in one of the sub-areas of this study, fishermen may have underreported their by-catch. Taking the possible underreporting into account the posterior mean of the total by-catch is between 2180 and 2380. The by-catch in our study area is likely to represent at least 90% of the total yearly grey seal by-catch in the Baltic Sea.

Genetic structure and dynamics of a small introduced population: the pikeperch, Sander lucioperca, in the Rhône delta.

Genetic data on introduced populations may help us to understand how these species succeed in colonising new territories. The pikeperch is a predatory fish widely introduced in Europe and has at times been considered as an invasive species. However, little is known about the genetics of both native and introduced populations. In the present study, we surveyed an introduced pikeperch population from the Rhône River delta, a habitat that has been highly modified for agricultural purposes. Using six microsatellites, we genotyped 93 individuals distributed among four hydraulically connected water bodies: the Rhône River, an irrigation canal, a drainage canal and a brackish lagoon. Population isolation were revealed by significant genetic distances and bottleneck highlighted by population monitoring. However, values of allelic richness and unbiased expected heterozygosity observed in these populations were similar, or even higher, compare to 18 native populations from the Baltic Sea drainage. It may be explained by multiple introductions in the Rhône drainage but also by demographic strategy that would have facilitated population persistence in this fragmented habitat. Similarly, heterozygote deficits (revealed by F(IS) values) have been detected, but were also found in native populations suggesting that mating among relatives could also result from a mating behavior of the species, maybe reinforce here by the reduced carrying capacity of the artificial canals and their respective isolation. Despite harsh environmental conditions and suspected inbreeding, the pikeperch has successfully maintained viable populations in the Rhône delta. Our study suggests that one of the factors in this invasive success, apart from its ecology, could be the maintenance of a good level of genetic diversity in introduced pikeperch populations. This genetic diversity probably stem from both its popularity as game fish and food resource which led to numerous stocking and an increasing propagule pressure and the reproductive strategy of the species.

Condition-based use of antimicrobials in cats in Finland: results from two surveys.

Two surveys were carried out to investigate how antimicrobials are used in cats in Finland. Information was retrieved from 419 prescriptions and 311 questionnaire sheets concerning antimicrobial treatment for feline patients. Infected wounds, skin disorders and urinary tract infections were the most common reasons for antimicrobial treatment in cats. Compliance with prudent use guidelines was good. beta-Lactams such as amoxycillin and amoxycillin with clavulanic acid were frequently used. Fluoroquinolones were used judiciously. The median length of treatment was 10 days for most conditions. Diagnosis was mainly based on clinical signs, and bacteriological culture and sensitivity testing was rarely performed. This study provides basic descriptive information on how antimicrobials are used in cats and that could help when revising guidelines for the condition-based use of antimicrobials in animals.

Genetic differentiation among European whitefish ecotypes based on microsatellite data.

The amount of genetic differentiation at DNA microsatellite loci in European whitefish (Coregonus lavaretus) was assessed among ecotypes, populations and run-timing types. The magnitude of genetic changes potentially caused by hatchery broodstock rearing were also compared with those observed in corresponding natural populations. A total of 35 populations were studied, including 33 Coregonus lavaretus populations and two samples of Coregonus peled. Five of the six whitefish ecotypes in Finland were represented within C. lavaretus populations. Genetic diversity among C. lavaretus populations proved to be high compared to two C. peled populations. The genetic D(A) distance between these two species was as high as 0.86. The genetic differentiation among ecotypes was generally low and thus gives support for the hypothesis of one native European whitefish species in Scandinavia. Among the ecotypes the northern, large sparsely-rakered, bottom-dwelling whitefish was most unique. Thus, observed genetic differences in quantitative traits have either developed independently of phylogenetic lineages, or have mixed and later changed according to environments and selection pressures. Overall genetic distances between the anadromous whitefish populations along the Finnish coast, especially in the Bothnian Bay area, were small. Populations of this area have been heavily influenced by human activities, and they also have the highest probability of mixing by natural means. In two cases, the Rivers Iijoki and Tornionjoki, statistically significant genetic differences could be observed between summer- and autumn-run spawning-time types. Wild populations had slightly higher allelic diversity than hatchery-reared populations of corresponding rivers. Although some reduction in genetic diversity during hatchery rearing is possible, it is an important aid in maintaining endangered populations.

Pim-1 kinase phosphorylates RUNX family transcription factors and enhances their activity.

BACKGROUND: The pim family genes encode oncogenic serine/threonine kinases which in hematopoietic cells have been implicated in cytokine-dependent signaling as well as in lymphomagenesis, especially in cooperation with other oncogenes such as myc, bcl-2 or Runx family genes. The Runx genes encode alpha-subunits of heterodimeric transcription factors which regulate cell proliferation and differentiation in various tissues during development and which can become leukemogenic upon aberrant expression. RESULTS: Here we have identified novel protein-protein interactions between the Pim-1 kinase and the RUNX family transcription factors. Using the yeast two-hybrid system, we were able to show that the C-terminal part of human RUNX3 associates with Pim-1. This result was confirmed in cell culture, where full-length murine Runx1 and Runx3 both coprecipitated and colocalized with Pim-1. Furthermore, catalytically active Pim-1 kinase was able to phosphorylate Runx1 and Runx3 proteins and enhance the transactivation activity of Runx1 in a dose-dependent fashion. CONCLUSION: Altogether, our results suggest that mammalian RUNX family transcription factors are novel binding partners and substrates for the Pim-1 kinase, which may be able to regulate their activities during normal hematopoiesis as well as in leukemogenesis.

Expression of human pim family genes is selectively up-regulated by cytokines promoting T helper type 1, but not T helper type 2, cell differentiation.

Cytokines are the most important inducers of T helper (Th) cell differentiation. Interleukin-12 (IL-12) and interferon-alpha (IFN-alpha) are responsible for human Th1-cell differentiation, while IL-4 is the critical cytokine promoting Th2-cell development. These two subsets of cells co-ordinate immunological responses to pathogens as well as autoimmune or allergic reactions. The pim family of proto-oncogenes encodes serine/threonine-specific kinases involved in cytokine-mediated signalling pathways in haematopoietic cells. Here we demonstrate that expression of pim-1 and pim-2 mRNAs is selectively up- or down-regulated in human cord-blood-derived CD4+ cells freshly induced to polarize towards Th1 or Th2 cells, respectively, whereas their expression is inhibited in both cell types by the immunosuppressive transforming growth factor beta (TGF-beta). Moreover, the Th1-specific cytokines IL-12 and IFN-alpha, but not the Th2-specific cytokine IL-4, transiently up-regulate pim-1 and pim-2 mRNA expression in human peripheral blood T cells and natural killer cells. In addition, the Pim-1 protein levels are strongly up-regulated by Th1-specific cytokines in all of these cell types. Taken together, our results suggest that pim genes and their protein products are involved in the early differentiation process of T helper cells.

Pim-1 kinase enhances NFATc activity and neuroendocrine functions in PC12 cells.

The activity of NFATc family transcription factors is tightly regulated in T cells via signaling pathways initiated by stimulation of the T cell receptor or its downstream effectors such as the Pim-1 serine/threonine kinase. Here, we demonstrate that NFATc-dependent transcription is inducible also in NGF-differentiated rat PC12 pheochromocytoma cells treated with phorbol esthers, calcium ionophores and/or forskolin and that the Pim-1 kinase can further potentiate the effects of these agents. PC12 cells share many characteristics with sympathetic neurons and can be induced to produce and release catecholamines, such as dopamine and noradrenaline, and inflammatory cytokines, such as interleukin 6. Interestingly, Pim-1 can synergize with forskolin-induced signaling pathways to stimulate also neuroendocrine functions of PC12 cells.

Pim-1 kinase promotes inactivation of the pro-apoptotic Bad protein by phosphorylating it on the Ser112 gatekeeper site.

Constitutive expression of the Pim-1 kinase prolongs survival of cytokine-deprived FDCP1 cells, partly via maintenance of Bcl-2 expression. Here, we show that Pim-1 colocalizes and physically interacts with the pro-apoptotic Bad protein and phosphorylates it in vitro on serine 112, which is a gatekeeper site for its inactivation. Furthermore, wild-type Pim-1, but not a kinase-deficient mutant, enhances phosphorylation of this site in FDCP1 cells and protects cells from the pro-apoptotic effects of Bad. Our results suggest that phosphorylation of Bad by Pim-1 is one of several mechanisms via which the Pim-1 kinase can enhance Bcl-2 activity and promote cell survival.

Pim-1 kinase inhibits STAT5-dependent transcription via its interactions with SOCS1 and SOCS3.

Signal transducer and activator of transcription 5 (STAT5) plays a critical role in cytokine-induced survival of hematopoietic cells. One of the STAT5 target genes is pim-1, which encodes an oncogenic serine/threonine kinase. Here we demonstrate that Pim-1 inhibits STAT5-dependent transcription in cells responsive to interleukin-3, prolactin, or erythropoietin. Ectopic expression of Pim-1 in cytokine-dependent FDCP1 myeloid cells results in reduced tyrosine phosphorylation and DNA binding of STAT5, indicating that Pim-1 interferes already with the initial steps of STAT5 activation. However, the Pim-1 kinase does not directly phosphorylate or bind to STAT5. By contrast, Pim-1 interacts with suppressor of cytokine signaling 1 (SOCS1) and SOCS3 and potentiates their inhibitory effects on STAT5, most likely via phosphorylation-mediated stabilization of the SOCS proteins. Thus, both Pim and SOCS family proteins may be components of a negative feedback mechanism that allows STAT5 to attenuate its own activity.

Prey depletion by the foraging of the Eurasian treecreeper, Certhia familiaris, on tree-trunk arthropods.

We examined to what extent breeding insectivorous treecreepers, Certhia familiaris, affect the abundance and the mean size of their prey population on the surface of tree trunks. In order to determine foraging pressure on tree trunks, we observed the parent birds' foraging behaviour in marked squares (25×25 m) at a short (10 m) and long distance (90 m) from the nest, when the nestlings were near fledging. Immediately after fledging, we measured the remaining food abundance on a sample of tree trunks near to and far from the nest. All arthropods longer than 1 mm were collected and identified to family level and their length was measured. Treecreepers foraged for significantly less time on trunks far from the nest than on trunks close to the nest (5 s vs 186 s). Consequently, treecreepers were found to deplete food abundance on trunks close to the nest. During the nesting period, parent birds removed almost twice as many spiders and other arthropods (excluding Formica ants) from the tree trunks close to the nest than from those further away. The size distribution of arthropods was the same between frequently and seldom-used tree trunks, suggesting that arthropod consumption by treecreepers was not size selective. Our direct measurements on the abundance of arthropods provide rare evidence for the ability of predators to deplete their food resources to a large extent.