Estimation of foraging territories of Reticulitermes grassei through mark–release–recapture

Subterranean termites have highly cryptic life habits and their foraging activities are as a rule confined to below-ground level gallery systems. Mark–release–recapture (MRR) using fat-soluble histological dyes is a candidate method for the study of foraging dynamics and territories, but has not hitherto been standardized experimentally. A wide range of dye types and concentrations is reported in the literature. In this study, six potential dyes were evaluated at different concentrations for marking workers of Reticulitermes grassei (Clément) (Isoptera: Rhinotermitidae), under laboratory and simulated field conditions. Neutral red (1% wt/wt) was considered the most effective while showing acceptably low toxicity. In a subsequent field trial using dye impregnated into a wooden bait, the MMR procedure was used to map the foraging territory perimeter of a single colony. Assumptions inherent in the interpretation of MMR data are reviewed. To map the foraging territory perimeter by this method, two theoretical approaches are defined (a conservative and a non-conservative hypothesis). We show that the approach adopted may affect the estimate obtained by as much as 100%. Results are discussed in the context of the ecology and behaviour of subterranean termite colonies.     

Rabies virus glycoprotein expression in Drosophila S2 cells: Influence of re-selection on protein expression

The aim of this study was to achieve expression of recombinant rabies virus glycoprotein (rRVGP) in Drosophila S2 cells. For this, a cDNA coding for the selection hygromycin antibiotic and the cDNA encoding the RVGP protein under the control of the constitutive actin promoter (Ac) were cloned in an expression plasmid, which was transfected into S2 cells. S2 cell populations (S2AcRVGPHy) showed rRVGP expression in cell lysates, attaining concentrations up to 1.5 μg/10⁷ cells (705 μg/L). Of the transfected cells, 20% were shown to express the rRVGP. Cell subpopulations selected by limiting dilution expressed higher rRVGP yields and 90% of the cells were shown to express the rRVGP. Cell populations re-selected by addition of hygromycin were shown to express 10 times higher rRVGP yields. The data presented here show that Drosophila S2 cells can be efficiently transfected with an expression/selection plasmid for rRVGP expression, allowing its synthesis with a high degree of physical and biological integrity. The importance of subpopulation selection was indicated by the increasing rRVGP yields during these procedures.     

Interaction of chitosan with cell membrane models at the air-water interface

In this paper we employed phospholipid Langmuir monolayers as membrane models to probe interactions with chitosan. Using a combination of surface pressure--area and surface potential--area isotherms and rheological measurements with the pendent drop technique, we observed that chitosan interacts with phospholipid molecules at the air-water interface. We propose a model in which chitosan interacts with the phospholipids mainly through electrostatic interactions, but also including H-bonding and hydrophobic forces, depending on the phospholipid packing density. At large areas per molecule, chitosan in the subphase adsorbs onto the monolayer, expanding it. At small areas per molecule, chitosan is located in the subsurface. Indeed, a mixed chitosan-phospholipid monolayer can be transferred onto solid supports, even at high surface pressures. The effects of chitosan on the viscoelastic properties of phospholipid monolayers may be taken as evidence for the ability of chitosan to disrupt cell membranes.     

Mendelian randomization supports causality between maternal hyperglycemia and epigenetic regulation of leptin gene in newborns

Leptin is an adipokine that acts in the central nervous system and regulates energy balance. Animal models and human observational studies have suggested that leptin surge in the perinatal period has a critical role in programming long-term risk of obesity. In utero exposure to maternal hyperglycemia has been associated with increased risk of obesity later in life. Epigenetic mechanisms are suspected to be involved in fetal programming of long term metabolic diseases. We investigated whether DNA methylation levels near LEP locus mediate the relation between maternal glycemia and neonatal leptin levels using the 2-step epigenetic Mendelian randomization approach. We used data and samples from up to 485 mother-child dyads from Gen3G, a large prospective population-based cohort. First, we built a genetic risk score to capture maternal glycemia based on 10 known glycemic genetic variants (GRS₁₀) and showed it was an adequate instrumental variable (β = 0.046 mmol/L of maternal fasting glucose per additional risk allele; SE = 0.007; P = 7.8 × 10⁻¹¹; N = 467). A higher GRS₁₀ was associated with lower methylation levels at cg12083122 located near LEP (β = −0.072 unit per additional risk allele; SE = 0.04; P = 0.05; N = 166). Direction and effect size of association between the instrumental variable GRS₁₀ and methylation at cg12083122 were consistent with the negative association we observed using measured maternal glycemia. Lower DNA methylation levels at cg12083122 were associated with higher cord blood leptin levels (β = −0.17 log of cord blood leptin per unit; SE = 0.07; P = 0.01; N = 170). Our study supports that maternal glycemia is part of causal pathways influencing offspring leptin epigenetic regulation.     

Oceanicella actignis gen. nov., sp. nov., a halophilic slightly thermophilic member of the Alphaproteobacteria

Two isolates, designated PRQ-67^T and PRQ-68, with an optimum growth temperature of about 50 °C, growth range in medium containing between 1 and 9% NaCl and an optimum pH for growth between 7.5 and 8.0, were recovered from a shallow marine hot spring on a beach, Praia do Fogo, at Ribeira Quente, on the Island of São Miguel in the Azores. Comparisons of 16S rRNA gene sequences show these strains to be most closely related (93.1–94.7% similarity) to species of the genus Amaricoccus, within the family Rhodobacteraceae. Strains are non-pigmented and form non-motile pleomorphic cells that stain Gram-negative, are aerobic, oxidase and catalase positive. The major fatty acids are C_18:1ω7c and C_18:1ω7c 11-methyl. Ubiquinone 10 is the major respiratory quinone. Major polar lipids are phosphatidylcholine, phosphatidylglycerol and one aminolipid. Based on 16S rRNA gene sequence analysis, physiological and biochemical characteristics we describe a new species of a novel genus represented by strain PRQ-67^T (=DSM 22673^T = LMG 25334^T) for which we propose the name Oceanicella actignis.     

Survey of subterranean termites (Isoptera: Rhinotermitidae) in a managed silvicultural plantation in Portugal, using a line-intersection method (LIS)

Subterranean termites (Reticulitermes grassei) were surveyed over successive seasons in a managed eucalyptus plantation in southeastern Portugal for 26 months. Termite activity in seven diameter categories of lying dead wood was investigated by a modified line intersection method (LIS). Each item sampled was inspected and assessed for termite attack and for general (i.e. fungal) decay status using standard protocols. Line intersection is quantitative to the extent that it can link foraging and decay parameters to woody biovolume. It was found that termites selected items with larger diameter, the observed trend showing an exponential character with greater termite attack as diameter increased. Attack by termites was positively associated with prior decay by fungi. A clear positive relationship was shown between rainfall and total woody biovolume containing live termites, underlining the importance of moisture for termite activity. Subterranean termites appeared to be important wood decomposers in the woodland studied, with an average of 30% of lying dead wood branches showing signs of termite attack.