In silico prediction of Gallibacterium anatis pan-immunogens

The Gram-negative bacterium Gallibacterium anatis is a major cause of salpingitis and peritonitis in commercial egg-layers, leading to reduced egg production and increased mortality. Unfortunately, widespread multidrug resistance and antigenic diversity makes it difficult to control infections and novel prevention strategies are urgently needed. In this study, a pan-genomic reverse vaccinology (RV) approach was used to identify potential vaccine candidates. Firstly, the genomes of 10 selected Gallibacterium strains were analyzed and proteins selected on the following criteria; predicted surface-exposure or secretion, none or one transmembrane helix (TMH), and presence in six or more of the 10 genomes. In total, 42 proteins were selected. The genes encoding 27 of these proteins were successfully cloned in Escherichia coli and the proteins expressed and purified. To reduce the number of vaccine candidates for in vivo testing, each of the purified recombinant proteins was screened by ELISA for their ability to elicit a significant serological response with serum from chickens that had been infected with G. anatis. Additionally, an in silico prediction of the protective potential was carried out based on a protein property prediction method. Of the 27 proteins, two novel putative immunogens were identified; Gab_1309 and Gab_2312. Moreover, three previously characterized virulence factors; GtxA, FlfA and Gab_2156, were identified. Thus, by combining the pan-genomic RV approach with subsequent in vitro and in silico screening, we have narrowed down the pan-proteome of G. anatis to five vaccine candidates. Importantly, preliminary immunization trials indicated an in vivo protective potential of GtxA-N, FlfA and Gab_1309.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0080-0) contains supplementary material, which is available to authorized users.     

Evaluation of endoscopically obtained duodenal biopsy samples from cats and dogs in an adapter-modified Ussing chamber

This study was conducted to evaluate an adapter-modified Ussing chamber for assessment of transport physiology in endoscopically obtained duodenal biopsies from healthy cats and dogs, as well as dogs with chronic enteropathies. 17 duodenal biopsies from five cats and 51 duodenal biopsies from 13 dogs were obtained. Samples were transferred into an adapter-modified Ussing chamber and sequentially exposed to various absorbagogues and secretagogues. Overall, 78.6% of duodenal samples obtained from cats responded to at least one compound. In duodenal biopsies obtained from dogs, the rate of overall response ranged from 87.5% (healthy individuals; n = 8), to 63.6% (animals exhibiting clinical signs of gastrointestinal disease and histopathological unremarkable duodenum; n = 15), and 32.1% (animals exhibiting clinical signs of gastrointestinal diseases and moderate to severe histopathological lesions; n = 28). Detailed information regarding the magnitude and duration of the response are provided. The adapter-modified Ussing chamber enables investigation of the absorptive and secretory capacity of endoscopically obtained duodenal biopsies from cats and dogs and has the potential to become a valuable research tool. The response of samples was correlated with histopathological findings.     

Tenerife's freshwater macroinvertebrates: Status and threats (Canary Islands,Spain)

• 1 This paper lists 171 freshwater macroinvertebrate taxa from Tenerife (excluding chironomids) based on this study and other published information. Canarian endemism and a Mediterranean origin are dominant features in the data set. At least 14 species are Tenerife endemics. Endemism is especially high in the Diptera, Trichoptera and Coleoptera.
• 2 Extensive investigations were carried out at 57 sites in six different habitat categories and from at least two seasons. The most species-rich habitats were those with permanently running water, a biotope that is rare and diminishing in Tenerife.
• 3 The principal threat to freshwater environments is an increase in human exploitation of the water resources, which, if it is not moderated, will cause extinctions in the near future. Taxa that do not favour man-made canals are especially at risk—a risk that is increased by the small number of populations. These include the Canarian endemics Wormaldia tagananana, Tinodes canariensis (Trichoptera), Dixa tetrica (Diptera) and Baetis pseudorhodani (Ephemeroptera). Thirty-nine species were found at five or fewer sites.
• 4 Some species can survive in intermittent streams, although we found less than half of the taxa present in this habitat, indicating the extent of the diversity reductions that might be expected to follow the drying up of permanent streams.
• 5 Species living in the thin film of water in the madicolous habitats are similarly at risk. Three Canarian endemics were found exclusively in this habitat: Hydrotarsus pilosus, Limnebius punctatus (Coleoptera) and Satchelliella binunciolata (Diptera).
• 6 As a first step, protective measures are proposed for five of the most species-rich streams. If these were to be implemented, they would make a significant contribution towards maintaining biodiversity in the fresh waters of Tenerife.

     

Marennine,Promising Blue Pigments from a Widespread Haslea Diatom Species Complex

In diatoms, the main photosynthetic pigments are chlorophylls a and c, fucoxanthin, diadinoxanthin and diatoxanthin. The marine pennate diatom Haslea ostrearia has long been known for producing, in addition to these generic pigments, a water-soluble blue pigment, marennine. This pigment, responsible for the greening of oysters in western France, presents different biological activities: allelopathic, antioxidant, antibacterial, antiviral, and growth-inhibiting. A method to extract and purify marennine has been developed, but its chemical structure could hitherto not be resolved. For decades, H. ostrearia was the only organism known to produce marennine, and can be found worldwide. Our knowledge about H. ostrearia-like diatom biodiversity has recently been extended with the discovery of several new species of blue diatoms, the recently described H. karadagensis, H. silbo sp. inedit. and H. provincialis sp. inedit. These blue diatoms produce different marennine-like pigments, which belong to the same chemical family and present similar biological activities. Aside from being a potential source of natural blue pigments, H. ostrearia-like diatoms thus present a commercial potential for aquaculture, cosmetics, food and health industries.     

Prey‐type‐dependent foraging of young‐of‐the‐year fish in turbid and humic environments

Abstract – Fish, which are generally visual foragers, experiences reduced reaction distance in visually degraded environments, which has consequences for encounter rates with prey. Small prey is detected at shorter distances than larger prey, and piscivores are therefore predicted to be more strongly affected by visual degradation. In experiments, roach (Rutilus rutilus) were fed two plankton prey types and pike (Esox lucius) were fed Daphnia and larval roach, in clear water, algal turbid water and water coloured brown by dissolved organic matter (DOM). Planktivorous foraging in roach was not affected by visual degradation, while pike foraging on both Daphnia and larval roach was. Pike showed increased reaction distance to Daphnia in visually degraded water, while it was severely reduced with roach as prey even if the visual range was not reduced below pike reaction distances in clear water. Pike foraging on Daphnia was not affected, but when foraging on roach, the reduced search efficiency was counteracted by increased attack rates. However, there was no increase in movement and no difference between turbid and DOM treatments. Effects on piscivores will likely become more pronounced at later life stages as prey size and the reliance on long‐distance detection increases at the same time as changing climatic conditions may further deteriorate the visual conditions in future.     

Cold acclimation of winter and spring peas: carbon partitioning as affected by light intensity

Like most plants, pea (Pisum sativum L.) becomes tolerant to frost if it is first exposed to low non-freezing temperatures, a process known as cold acclimation. Cold acclimation is a complex process involving many physiological and metabolic changes. Two spring dry peas, two winter dry peas and one winter forage line were exposed to cold temperature in a controlled environment in two experiments, one using low light intensity and the other regular light intensity. Plants were harvested throughout the experiment and dry matter accumulation, water content, soluble and insoluble sugar concentrations were determined from shoot and root samples. Cold acclimation did not occur when temperatures were low if light intensity was low, even in winter peas. In contrast, with regular light intensity, the winter peas acquired more freezing tolerance than spring peas and a close relationship was found between the soluble sugar concentration of leaves just before the frost and the degree of freezing tolerance obtained by the different genotypes. Relationships between freezing tolerance and carbon partitioning between shoot and roots are discussed.     

Chemical communication in Ceratophysella sigillata (Collembola: Hypogastruridae): intraspecific reaction to alarm substances

When disturbed or damaged, the winter-active collembolan Ceratophysella sigillata releases a strong aromatic odour, which can be perceived even by humans. The efficiency of the substances released as an intraspecific alarm substance was investigated. C. sigillata showed distinct reactions to chemicals released by conspecifics; crushed conspecifics, cuticle washings (with pentane) and whole body extracts (with methyl acetate) were tested. Our results demonstrate that a very sensitive intraspecific avoidance reaction is present to the substances released by crushed C. sigillata as well as to washings and extracts: extracts from as little as half a springtail induced a significant avoidance reaction in conspecifics. Alarm substances were present in all instars of C. sigillata including eggs, and all instars of C. sigillata tested showed similar reactions to the extracts or crushed conspecifics.     

Composition rather than viscosity modifies the aroma compound retention of flavored stirred yogurt

The influence of thickening agents (modified starch/pectin mixture 0 and 7 g/L) and mechanical treatment (low, medium, and high) on the retention of esters (pentyl acetate and ethyl pentanoate), aldehydes (hexanal and (E)-2-hexenal), and a lactone (gamma-octalactone) in low-fat flavored stirred yogurts were investigated under equilibrium conditions. In the range studied, the thickening agent and mechanical treatment had little influence on aroma compound retention compared to the decreasing effect of increasing dairy protein concentration on aldehyde retention and the "salting out" effect of carbohydrates on esters. Moreover, experiments in dynamic mode (study of the release of hexanal when yogurts were heated) showed, in the conditions studied, that heat and mass transfer coefficients were not influenced by any of the studied factors (thickening agents and mechanical treatment). These results under static and dynamic conditions are not related to the significant decreasing effect of thickening agents on apple sensory scores associated with hexanal, observed in a previous sensory study. Thus, this sensory effect of thickening agents may be due to sensory interactions between perceptions rather than physicochemical interactions.     

Oxidative stability of bovine serum albumin- and sodium caseinate-stabilized emulsions depends on metal availability

Proteins often stabilize food emulsions and are also able to promote or delay lipid oxidation in complex systems. The purpose of this work was to investigate the relationship between metal ion availability and oxidative stability of oil-in-water emulsions stabilized by bovine serum albumin (BSA) or sodium caseinate (NaCas). Emulsions with similar and stable droplet size distributions were prepared with stripped sunflower oil (30 vol %) and protein solutions (20 g L(-)(1); pH = 6.5). In the absence of the water-soluble metal chelator EDTA, oxygen uptake, conjugated dienes, and volatile compounds developed faster in NaCas-stabilized emulsions than in those prepared with BSA. This effect is attributed to the chelating properties of NaCas and to electrostatic interactions that attract some metal ions at the interface where they could initiate lipid oxidation. When EDTA (100 muM) was present, oxidation was delayed to a greater extent in emulsions made with NaCas than in BSA stabilized emulsions. These conditions probably enabled NaCas to exert free-radical-scavenging activity.