Genetic diversity and distribution patterns of diploid and polyploid hybrid water frog populations (Pelophylax esculentus complex) across Europe

Polyploidization is a rare yet sometimes successful way for animals to rapidly create geno‐ and phenotypes that may colonize new habitats and quickly adapt to environmental changes. In this study, we use water frogs of the Pelophylax esculentus complex, comprising two species (Pelophylax lessonae, genotype LL; Pelophylax ridibundus, RR) and various diploid (LR) and triploid (LLR, LRR) hybrid forms, summarized as P. esculentus, as a model for studying recent hybridization and polyploidization in the context of speciation. Specifically, we compared the geographic distribution and genetic diversity of diploid and triploid hybrids across Europe to understand their origin, maintenance and potential role in hybrid speciation. We found that different hybrid and parental genotypes are not evenly distributed across Europe. Rather, their genetic diversity is structured by latitude and longitude and the presence/absence of parental species but not of triploids. Highest genetic diversity was observed in central and eastern Europe, the lowest in the northwestern parts of Europe. This gradient can be explained by the decrease in genetic diversity during postglacial expansion from southeastern glacial refuge areas. Genealogical relationships calculated on the basis of microsatellite data clearly indicate that hybrids are of multiple origin and include a huge variety of parental genomes. Water frogs in mixed‐ploidy populations without any parental species (i.e. all‐hybrid populations) can be viewed as evolutionary units that may be on their way towards hybrid speciation. Maintenance of such all‐hybrid populations requires a continuous exchange of genomes between diploids and triploids, but scenarios for alternative evolutionary trajectories are discussed.     

Thermal and energetic consequences of nest location and breeding times in Water Pipits (Anthus spinoletta)

Summary The thermal environment has pronounced effects on the energy costs of thermoregulation and affects an animal's allocation of energy to self-maintenance and parental care. Consequently, the selection of reproductive periods, breeding habitats and nest-sites with a favourable microclimate can be advantageous, especially for birds breeding in harsh environments. In this study on Alpine Water Pipits (Anthus spinoletta), we evaluate the importance of spatial and temporal factors on thermoregulatory costs by combining laboratory measurements of metabolic rates under various temperatures with standard operative temperatures (T_es) recorded in the field in different microhabitats. Using these measurements we estimate the thermal and energetic consequences of nest locality and timing of reproduction. Our results show: (1) In the morning, T_es values were much higher on the east-north-east (ENE) slope of a valley than on the west-south-west (WSW) slope; in the afternoon this pattern was reversed. As a consequence, energy costs (E_hour) for thermoregulation on the ENE slope were up to 0.6 RMR (resting metabolic rate at night) lower than on the WSW slope during morning hours and about 0.8 RMR higher during afternoon hours. (2) During the incubation and nestling phases of first and second broods, total energy expenditure for thermoregulation in the daytime (E_daytime) was 0.2–0.3 RMR higher on the ENE slope than on the WSW slope. (3) Within slopes, E_daytime was lower during second broods than during first broods, with differences of 0.06–0.07 RMR during incubation and of 0.32 RMR during nestling care. These differences correspond to the flying costs of females incubating eggs (0.09 RMR) and rearing nestlings (0.25 RMR). We conclude that nest placement in relation to microclimate can improve the female's energy budget, both in terms of the total daily expenditure and its diurnal pattern. From thermal considerations alone, delaying breeding into mid-summer would be advantageous, but this advantage is probably outweighed by the reduced chances for second and replacement clutches and by the necessity to complete moult before migration.

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Thermische und energetische Konsequenzen von Brutplätzen und Brutzeiten bei Wasserpiepern (Anthus spinoletta)

Zusammenfassung Die Umgebungstemperatur beeinflusst den Energieaufwand für die Thermoregulation und bestimmt somit, wie viel Energie ein Tier für die Selbsterhaltung benötigt und wie viel es für Brutpflege aufwenden kann. Daher ist es vorteilhaft, für die Fortpflanzung Zeiten und Orte mit besonders günstigen mikroklimatischen Bedingungen zu wählen. Das gilt insbesondere für Vögel, die in unwirtlichen Biotopen wie z. B. alpinen Regionen brüten. Wir untersuchten den Einfluss von Brutzeit und -ort auf die energetischen Kosten der Thermoregulation am Beispiel einer Population des Wasserpiepers (Anthus spinoletta) im Dischmatal bei Davos (Schweiz). Dazu verknüpften wir drei verschiedene Datensätze: a) Ruhestoffwechsel (RMR), gemessen an gefangenen Vögeln unter verschiedenen Temperaturen, b) Umgebungs- und Nesttemperaturen im Freiland während der Brutzeit und c) standardisierte operative Temperaturen (T_es), ermittelt mit Kupfervögeln in verschiedenen Mikrohabitaten.Die Ergebnisse zeigen: (1) In den Morgenstunden sind T_es-Werte am Ost-Nord-Ost-Hang (ONO) deutlich höher als am West-Süd-West-Hang (WSW); am Nachmittag sind die Verhältnisse umgekehrt. Als Folge davon sind die Energiekosten (E_hour) der Thermoregulation am ONO-Hang in den Morgenstunden bis zu 0.6 RMR niedriger und nachmittags ca. 0.8 RMR höher als am WSW-Hang. (2) In der Bebrütungs- und Nestlingsphase von Erst-und Zweitbruten war der gesamte Energieaufwand für Thermoregulation während der Tagesstunden (E_daytime) am ONO-Hang 0.2–0.3 RMR höher als am WSW-Hang. (3) Innerhalb jedes Hanges war E_daytime bei Zweitbruten niedriger als bei Erstbruten: 0.06–0.07 RMR während der Bebrütung der Eier und 0.32 RMR während der Nestlingsphase. Diese Größenordnungen entsprechen den Flugkosten von Weibchen während der Bebrütungszeit (0.09 RMR) bzw. der Jungenaufzucht (0.25 RMR).Unsere Ergebnisse belegen, dass die Wahl von klimatisch günstigen Habitaten die Energiebilanz von Brutvögeln verbessern kann, sowohl in Hinblick auf den täglichen Gesamtaufwand als auch bezüglich der Verteilung des Energieverbrauchs über den Tag. Aus thermoregulatorischen Gründen wäre auch eine zeitliche Verlagerung des Brutgeschäfts in die Sommermonate (Juli, August) vorteilhaft. Das würde jedoch die Möglichkeiten für Ersatz-und Zweitbruten verringern und die Zeit für die Mauser vor dem Herbstzug verkürzen. Da in unserem Untersuchungsgebiet diese Faktoren für eine erfolgreiche Fortpflanzung eine größere Bedeutung haben als der Energieaufwand für die Thermoregulation, ist der frühe — energetisch 6–32 % teurere — Brutbeginn der Vögel sinnvoll.

     

Selection through predation, snowfall and microclimate on nest-site preferences in the Water Pipit Anthus spinoletta

Nest-site characteristics can have a strong impact on reproductive success in birds. Nest sites should simultaneously protect from predators, offer shelter and provide a favourable microclimate. We studied the relationship between three agents of natural selection (predators [i.e. Adders and birds/mammals], snowfall and microclimate), nest-site characteristics and reproductive success to determine whether these influenced preference for specific nest-site characteristics in the Water Pipit Anthus spinoletta . Pooled over all nests, the relative importance as agents of natural selection decreased from mammalian/avian predation (15% of all nests) through Adder predation (12%) to snowfall (7%), but there were clear differences in space and time. Predation by Adders selected for nest sites surrounded by few medium-sized shrubs. Selection by mammalian and avian predators favoured no specific nest-site characteristics. Protection from snowfall was best in nests surrounded by relatively few medium-sized shrubs. Microclimate had a strong influence on nestling survival and duration of nestling period. In nests on ENE-facing slopes, where maximum temperatures were reached in the morning, nestling survival was higher than on WSW-facing slopes, where temperature maxima occurred in the afternoon. Our results indicate that weak, but significant, directional selection is acting on preference for certain nest-site characteristics through effects on survival and development of nestlings. As predation and snowfall are unpredictable, the evolution of an optimal nest placement strategy is unlikely on a small scale. On a larger scale, however, choice of one breeding area over another may be favoured because of predictable differences between locations in terms of survival and nestling development.     

Endothelial NO-synthase intron 4 polymorphism is associated with disturbed in vivo nitric oxide production in individuals prone to type 2 diabetes.

Insulin resistance, as well as vascular disease, both share a relevant genetic background taking the influence of a positive family history of these disorders. On the other hand, insulin resistance is associated with a proatherosclerotic disturbance in nitric oxide dependent vasodilation, probably contributing to the link between these two disorders. We examined the association between nitric oxide dependent vasodilation (measured with high resolution ultrasound at 13 MHz) and three relevant NO-synthase (eNOS)-polymorphisms in 200 insulin resistant subjects participating in the Tuebinger Lifestyle Intervention Program (TULIP). This study revealed that carriers of the eNOS intron 4 polymorphism (aa 2.16%; ab 24.2%; bb 73.2%) show significantly worse endothelial, and thereby eNOS dependent vasodilation (p=0.03, multivariate ANOVA), as compared to wildtype carriers. The 5' UTR T-786C and the G894 T polymorphism did not show any influence on eNOS-activity. In subjects at increased risk to develop type 2 diabetes, the eNOS intron 4 polymorphism is independently associated with endothelial function as indicated by disturbed endothelial NO production. Due to the high prevalence and the relatively strong effect, this polymorphism might help to identify subjects at increased risk for atherosclerosis associated with overweight and insulin resistance.     

Sire breed comparisons for meat and eating quality traits in Australian pig populations

Sire breed comparisons for carcase (n = 1169), meat and eating quality (n = 686) traits were obtained using data recorded on progeny of known pedigree sired by Duroc (DU), Large White (LW), Landrace (LR) and Duroc Synthetic (DS) boars from crossbred (LW/LR) sows. Animals were reared in eco-shelters in large single-sex contemporary groups and slaughtered on an age constant basis at 22 weeks of age. Compared to progeny from other sire groups, animals sired by purebred Duroc boars tended to have improved eating quality traits (higher intramuscular fat and lower shear force) at the expense of poorer carcase characteristics (higher subcutaneous and belly fat). Animals sired by DS boars tended to be heavier and leaner than those sired by LW, LR or DU boars; intramuscular fat but not belly fat was correspondingly lower, while tenderness was generally consistent with that of the DU progeny. Significant variability of sire progeny groups within sire breed suggests that sire breed selection, potentially used for improving traits such as meat quality in commercial progeny, will be less accurate in the absence of sire-specific information, which is typically poorly recorded in this class of traits.     

Genetic aspects of feed efficiency and reduction of environmental footprint in broilers: a review

Currently, optimization of feed efficiency is one of the main challenges in improvement programs of livestock and poultry genetics. The objective of this review is to present the genetic aspects of feed efficiency related traits in meat-type chicken and possible ways to reduce the environmental impact of poultry meat production with effective breeding. Basic measures of feed efficiency are defined and the genetic background of these traits, including a review of heritabilities is described. Moreover, a number of genomic regions and candidate genes determining feed efficiency traits of broilers that were detected over the past decades are described. Classical and genomic selection strategies for feed efficiency in the context of its relationships with other performance traits are discussed as well. Finally, future strategies to improve feed digestibility are described as it is expected that they will decrease wastes and greenhouse gas emission. Further genetic improvement of feed efficiency, should be examined jointly with appropriate feeding strategies in broilers.     

Liquid chromatography/electrospray ionisation-mass spectrometry method for the quantification of sphingosine and sphinganine in cell cultures exposed to fumonisins

Fumonisins, mycotoxins produced by Fusarium verticillioides, are potent inhibitors of the de novo sphingolipid biosynthesis via inhibition of the key enzyme ceramide synthase. The cellular response to a fumonisin exposure is obvious as an alteration of the ratio of the sphingoid bases sphingosine (SO) and sphinganine (SA). We developed a new column liquid chromatography/electrospray ionisation-mass spectrometry (LC-ESI-MS) method for the rapid, simultaneous and quantitative determination of these bases in cell cultures of immortalised human kidney epithelial cells (IHKE cells). For sample preparation, cell lysates were only diluted, centrifuged and directly used for LC-MS measurements. Quantification was carried out using phytosphingosine (PSO) as an internal standard. Detecting the protonated molecule [M+H](+) signals of SO (m/z 300) and SA (m/z 302) in the selected ion monitoring (SIM) mode, detection limits of 10 pg for SO (signal-to-noise ratio S/N=3:1) and 25 pg for SA (S/N=3:1) were established. The average recovery for SO and SA was higher than 90% for control IHKE-cells, respectively. The developed LC-ESI-MS method allows the sensitive, selective and rapid monitoring of sphingosine and sphinganine in cell matrices with a drastically reduced time for sample preparation.