Ringed seal (Pusa hispida) seasonal movements,diving, and haul‐out behavior in the Beaufort,Chukchi, and Bering Seas (2011–2017)

Continued Arctic warming and sea‐ice loss will have important implications for the conservation of ringed seals, a highly ice‐dependent species. A better understanding of their spatial ecology will help characterize emerging ecological trends and inform management decisions. We deployed satellite transmitters on ringed seals in the summers of 2011, 2014, and 2016 near Utqia?vik (formerly Barrow), Alaska, to monitor their movements, diving, and haul‐out behavior. We present analyses of tracking and dive data provided by 17 seals that were tracked until at least January of the following year. Seals mostly ranged north of Utqia?vik in the Beaufort and Chukchi Seas during summer before moving into the southern Chukchi and Bering Seas during winter. In all seasons, ringed seals occupied a diversity of habitats and spatial distributions, from near shore and localized, to far offshore and wide‐ranging in drifting sea ice. Continental shelf waters were occupied for >96% of tracking days, during which repetitive diving (suggestive of foraging) primarily to the seafloor was the most frequent activity. From mid‐summer to early fall, 12 seals made ~1‐week forays off‐shelf to the deep Arctic Basin, most reaching the retreating pack‐ice, where they spent most of their time hauled out. Diel activity patterns suggested greater allocation of foraging efforts to midday hours. Haul‐out patterns were complementary, occurring mostly at night until April‐May when midday hours were preferred. Ringed seals captured in 2011—concurrent with an unusual mortality event that affected all ice‐seal species—differed morphologically and behaviorally from seals captured in other years. Speculations about the physiology of molting and its role in energetics, habitat use, and behavior are discussed; along with possible evidence of purported ringed seal ecotypes.     

Recognizing the enemy: do predator cues influence hatching in Neotropical annual killifish?

Annual fish species have evolved complex adaptations to survive in temporary wetlands. The main adaptation of these fish is the ability to produce embryos that survive dry periods. Embryo development of this fish can show variation at multiple levels influenced by many environmental factors, such as photoperiod and temperature. Predator cues are another factor that can influence the embryonic stage. One way in which annual fish could adapt to predators is by using risk-spreading strategies (through bet-hedging). Nonetheless, this strategy depends on the coevolutionary history between predators and preys and on the degree of environmental unpredictability, resulting in different responses across different species. This study investigated the influence of predator cues on the embryonic development and hatching of two Austrolebias species that inhabit ponds that present differences in hydroperiod and the risk of predator presence. The results confirmed a differentiated response between the two annual fish species tested, corroborating the modulation of hatching against the risk of predation by native predatory fish. The authors further showed that development times varied between the two annual fish species, regardless of the presence of predators. They highlight that the variation in embryonic development is strongly affected by different levels of hydroperiod unpredictability faced by the two species. To unravel finer-scale local adaptations in the annual fish embryo development, future studies should focus on a region with greater spatial gradient.     

Reducing Biotic and Abiotic Land‐Use Legacies to Restore Invaded,Abandoned Citrus Groves

Land‐use legacies associated with agriculture, such as increased soil fertility and elevated soil pH, promote invasions by non‐native plant species on former agricultural lands. Restoring natural soil conditions (i.e. low fertility and low pH) may be an effective, long‐term method to control and reduce the abundance of non‐native and ruderal species that invade abandoned agricultural lands. In this study, we examined how soil manipulation treatments of lowering soil fertility with carbon additions and lowering soil pH by applying sulfur affect non‐native and ruderal native plant species abundance in two former citrus groves in central Florida. Non‐native plant biomass was removed by one of two methods (tilling or topsoil removal), and was combined with a soil amendment of sulfur, carbon, sulfur + carbon, or none. The biomass removal treatments significantly decreased non‐native abundance, with topsoil removal as the most effective. Carbon additions did not affect soil fertility or vegetation. Sulfur and sulfur + carbon additions significantly decreased soil pH in both groves for at least 1 year post‐treatment; however, we did not see a significant vegetation response. Overall, our results suggest that removing vegetation by tilling and topsoil removal is an effective method for reducing non‐target species cover. Although we did not see a response of vegetation to our treatments, we were able to restore the initial soil characteristics, which can be a first step toward complete restoration.     

Electrophoresis in Gels Suspended in Hydrophobic Solvents

Protein components in mixtures may be separated by zone electrophoresis in columns of gel suspended in hydrophobic solvents. The method is useful for the location of specific antibodies in IgG elec-trophoretic spectra.     

Entomological surveillance,spatial distribution,and diversity of Culicidae (Diptera) immatures in a rural area of the Atlantic Forest biome,State of São Paulo,Brazil

Because of the high adaptive capacity of mosquitoes, studies that focus on transitional environments become very important, such as those in rural areas, which are considered as bridges between wild diseases and human populations of urban areas. In this study, a survey of the existing species of mosquitoes was performed in an Atlantic Forest area of the city of Santa Bárbara d'Oeste, São Paulo state, Brazil, using traps for immatures and analyzing the frequency and distribution of these insects over the sampling months. Five mosquito species were found: Aedes albopictus (the most frequent species), Aedes aegypti, Aedes fluviatilis, Culex quinquefasciatus, and Toxorhynchites theobaldi. The 4,524 eggs collected in ovitraps showed the presence of the tribe Aedini. Aedes aegypti and Ae. albopictus were identified after larval hatching in the laboratory, with different spatial distributions: the first of which coincides with the area of greatest diversity calculated using the Simpson index, while the second does not. The association of ecological analysis of spatial diversity with simple methods of data collection enables the identification of possible epidemiological risk situations and is a strategy that may be implemented to monitor ecological processes resulting from the interaction among different species of mosquitoes.     

Phylogenetic relationships of Italian Bellevalia species (Asparagaceae), inferred from morphology,karyology and molecular systematics

The seven Bellevalia species and subspecies known from Italy, representing about 10% of the genus and three out of six sections, were studied. An integrated morphological, karyological and molecular approach was used to infer phylogenetic and systematic relationships among them. B. romana (the generitype) is the most distinctive species on karyotype asymmetry grounds. B. boissieri and B.dubia, usually considered as subspecies of one species (the latter endemic to Sicily), deserve specific status based on biparental nrDNA markers (internal transcribed spacer, ITS), since they do not form a single clade. The allotetraploid endemic B. pelagica, morphologically similar to B. romana, is sister to the latter under parsimony, both in morphological and ITS trees; it is also related with B. dubia, based on karyotype asymmetry and a uniparental cpDNA marker (trnL^(UAA)–trnF^(GAA) IGS (intergenic spacer)). A second allotetraploid endemic, B. webbiana, is closely related, on morphological, karyological and molecular grounds, with B. boissieri and B. ciliata, and also with B. trifoliata, three species that might all involved in its origin. B. sect. Conicae Feinbr. and sect. Nutantes Feinbr. are here typified, the former (type: B. ciliata) is most likely a synonym of the latter (type: B. trifoliata).     

LC-MS characterization and purity assessment of a prototype bispecific antibody

Bispecific IgG asymmetric (heterodimeric) antibodies offer enhanced therapeutic efficacy, but present unique challenges for drug development. These challenges are related to the proper assembly of heavy and light chains. Impurities such as symmetric (homodimeric) antibodies can arise with improper assembly. A new method to assess heterodimer purity of such bispecific antibody products is needed because traditional separation-based purity assays are unable to separate or quantify homodimer impurities. This paper presents a liquid chromatography-mass spectrometry (LC-MS)-based method for evaluating heterodimeric purity of a prototype asymmetric antibody containing two different heavy chains and two identical light chains. The heterodimer and independently expressed homodimeric standards were characterized by two complementary LC-MS techniques: Intact protein mass measurement of deglycosylated antibody and peptide map analyses. Intact protein mass analysis was used to check molecular integrity and composition. LC-MS^E peptide mapping of Lys-C digests was used to verify protein sequences and characterize post-translational modifications, including C-terminal truncation species. Guided by the characterization results, a heterodimer purity assay was demonstrated by intact protein mass analysis of pure deglycosylated heterodimer spiked with each deglycosylated homodimeric standard. The assay was capable of detecting low levels (2%) of spiked homodimers in conjunction with co-eluting half antibodies and multiple mass species present in the homodimer standards and providing relative purity differences between samples. Detection of minor homodimer and half-antibody C-terminal truncation species at levels as low as 0.6% demonstrates the sensitivity of the method. This method is suitable for purity assessment of heterodimer samples during process and purification development of bispecific antibodies, e.g., clone selection.     

Pattern of Social Interactions after Group Integration: A Possibility to Keep Stallions in Group

Horses are often kept in individual stables, rather than in outdoor groups, despite such housing system fulfilling many of their welfare needs, such as the access to social partners. Keeping domestic stallions in outdoor groups would mimic bachelor bands that are found in the wild. Unfortunately, the high level of aggression that unfamiliar stallions display when they first encounter each other discourages owners from keeping them in groups. However, this level of aggression is likely to be particularly important only during group integration, when the dominance hierarchy is being established, whereas relatively low aggression rates have been observed among stable feral bachelor bands. We investigated the possibility of housing breeding stallions owned by the Swiss National Stud in groups on a large pasture (5 stallions in 2009 and 8 stallions in 2010). We studied the pattern of agonistic, ritual and affiliative interactions after group integration (17–23 days), and the factors influencing these interactions (time after group integration, dominance rank, age or experience of group housing). We found that stallions displayed generally more ritual than agonistic and than affiliative interactions. The frequency of agonistic and ritual interactions decreased quickly within the first three to four days. The frequency of affiliative interactions increased slowly with time before decreasing after 9–14 days. A stable hierarchy could be measured after 2–3 months. The highest-ranking males had less ritual interactions than the lowest-ranking. Males had also less agonistic, ritual and affiliative interactions if they had already been housed in a group the previous year. Therefore, we found that breeding stallions could be housed together on a large pasture, because the frequency of agonistic interactions decreased quickly and remained at a minimal level from the fourth day following group integration. This housing system could potentially increase horse welfare and reduce labour associated with horse management.     

The Embedding Problem for Markov Models of Nucleotide Substitution

Continuous-time Markov processes are often used to model the complex natural phenomenon of sequence evolution. To make the process of sequence evolution tractable, simplifying assumptions are often made about the sequence properties and the underlying process. The validity of one such assumption, time-homogeneity, has never been explored. Violations of this assumption can be found by identifying non-embeddability. A process is non-embeddable if it can not be embedded in a continuous time-homogeneous Markov process. In this study, non-embeddability was demonstrated to exist when modelling sequence evolution with Markov models. Evidence of non-embeddability was found primarily at the third codon position, possibly resulting from changes in mutation rate over time. Outgroup edges and those with a deeper time depth were found to have an increased probability of the underlying process being non-embeddable. Overall, low levels of non-embeddability were detected when examining individual edges of triads across a diverse set of alignments. Subsequent phylogenetic reconstruction analyses demonstrated that non-embeddability could impact on the correct prediction of phylogenies, but at extremely low levels. Despite the existence of non-embeddability, there is minimal evidence of violations of the local time homogeneity assumption and consequently the impact is likely to be minor.