Bacterial SCP from methanol in Kuwait: Product revovery and composition

The rationale behind KISR's involvement in SCP is explained and emphasis towards the use of aerobic thermophilic/thermotolerant methanol-utilizing bacterial cultures is stressed. An attempt is made to correlate different pilot plant harvesting procedures for biomass recovery with product quality. The latter is expressed as crude and true protein content, moisture and ash content, mineral composition of the ash, and amino acids profiles. The most suitable harvesting procedures were found to be those using a mild heat-acid or acid-heat treatment. Both of these procedures gave high recovery efficiencies and suitable product nutrition.     

Physiological characteristics of four methylotrophic bacteria and their potential use in single-cell protein production

Summary Four methylotrophic bacteria, isolated at the Kuwait Institute for Scientific Research, were able to grow on methanol as the only carbon source at a maximum temperature of 44°C. An optimized medium composition was obtained through intensive chemostat studies varying both macro- and micro-element concentrations. Various batch and chemostat experiments were carried out at different pH, temperature, dilution rate and methanol concentrations. The results showed optimum pH around 6.8, at temperatures of 37 to 40°C, dilution rate 0.2–0.3 h^–1 and methanol was found to be inhibitory at concentrations above 20 g l^–1. The performance of all four bacteria under chemostat conditions was similar. Chemostat fermentation experiments using the optimized medium at 40°C, pH 6.8, dilution 0.2 h^–1 and 10 g methanol/l in the feed gave a biomass yield coefficient of 0.42–0.44 g/g methanol, 78–79% crude protein content, 58–62% total amino acid content and 10–11.5% nucleic acid content. In conclusion all four methylotrophic strains has good potential for use in the production of single-cell protein.

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Caractéristiques physiologiques de quatre bactéries méthylotrophes et leur emploi potentiel dans la production de protéine uni-celluliare

Résumé Quatre bactéries méthylotrophes, isolées à l'Institut pour la Recherche Scientifique du Kuweit, se sont révélées capables de croître sur méthanol comme seule source de carbone à une température maximum de 44°C. Des études intensives en chémostat de variation des concentrations tant en macro-qu'en micro-éléments, ont permis d'obtenir une composition optimum du milieu. Diverses expériences en milieu non renouvelé et en chémostat ont été effectuées à différents pH, températures, taux de dilution et concentration en méthanol. Les résultats ont montré un optimum de pH vers 6.8, de température entre 37 et 40°C, de taux de dilution de 0.2 à 0.3 h^–1. Le méthanol s'est révélé inhibiteur aux concentrations supérieures à 20 g l^–1. La performance des quatres bactéries dans les conditions du chémostat se sont révélées très semblables. Les expériences de fermentátion en chémostat, utilisant le milieu optimum à 40°C, pH 6.8, taux de dilution de 0.2 h^–1 et à 10 g de méthanol par litre dans l'alimentation ont produit un coefficient de rendement en biomasse de 0.42 à 0.44 g de cellules par g de méthanol contenant de 78 à 79% de protéines, de 58 à 62% d'acides aminés totaux et de 10 à 11.5% d'acides nucléiques. En conclusion, les quatre bactéries méthylotrophes présentent une bonne potentialité pour être utilisées dans la production de protéine uni-cellulaire.

     

Cooperative role of endogenous leucotrienes and platelet‐activating factor in ischaemia–reperfusion‐mediated tissue injury

Insufficient oxygen delivery to organs leads to tissue dysfunction and cell death. Reperfusion, although vital to organ survival, initiates an inflammatory response that may both aggravate local tissue injury and elicit remote organ damage. Polymorphonuclear neutrophil (PMN) trafficking to remote organs following ischaemia/reperfusion (I/R) is associated with the release of lipid mediators, including leucotriene (LT) B₄, cysteinyl‐LTs (CysLTs) and platelet‐activating factor (PAF). Yet, their potentially cooperative role in regulating I/R‐mediated inflammation has not been thoroughly assessed. The present study aimed to determine the cooperative role of lipid mediators in regulating PMN migration, tissue oedema and injury using selective receptor antagonists in selected models of I/R and dermal inflammation. Our results show that rabbits, pre‐treated orally with BIIL 284 and/or WEB 2086 and MK‐0571, were protected from remote tissue injury following I/R or dermal inflammation in an additive or synergistic manner when the animals were pre‐treated with two drugs concomitantly. The functional selectivity of the antagonists towards their respective agonists was assessed in vitro, showing that neither BIIL 284 nor WEB 2086 prevented the inflammatory response to IL‐8, C5a and zymosan‐activated plasma stimulation. However, these agonists elicited LTB₄ biosynthesis in isolated rabbit PMNs. Similarly, a cardioprotective effect of PAF and LTB₄ receptor antagonists was shown following myocardial I/R in mice. Taken together, these results underscore the intricate involvement of LTB₄ and PAF in each other's responses and provide further evidence that targeting both LTs and PAF receptors provides a much stronger anti‐inflammatory effect, regulating PMN migration and oedema formation.     

Atrial fibrillation pacing decreases intravascular shear stress in a New Zealand white rabbit model: implications in endothelial function

Atrial fibrillation (AF) is characterized by multiple rapid and irregular atrial depolarization, leading to rapid ventricular responses exceeding 100 beats per minute (bpm). We hypothesized that rapid and irregular pacing reduced intravascular shear stress (ISS) with implication to modulating endothelial responses. To simulate AF, we paced the left atrial appendage of New Zealand White rabbits (n = 4) at rapid and irregular intervals. Surface electrical cardiograms were recorded for atrial and ventricular rhythm, and intravascular convective heat transfer was measured by microthermal sensors, from which ISS was inferred. Rapid and irregular pacing decreased arterial systolic and diastolic pressures (baseline, 99/75 mmHg; rapid regular pacing, 92/73; rapid irregular pacing, 90/68; p < 0.001, n = 4), temporal gradients ( ${\partial\tau/\partial t}$ from 1,275 ± 80 to 1,056 ± 180 dyne/cm² s), and reduced ISS (from baseline at 32.0 ± 2.4 to 22.7 ± 3.5 dyne/cm²). Computational fluid dynamics code demonstrated that experimentally inferred ISS provided a close approximation to the computed wall shear stress at a given catheter to vessel diameter ratio, shear stress range, and catheter position. In an in vitro flow system in which time-averaged shear stress was maintained at ${{\tau_{\rm avg}} = 23 \pm 4\, {\rm dyn}\, {\rm cm}^{-2} {\rm s}^{-1}}$ , we further demonstrated that rapid pulse rates at 150 bpm down-regulated endothelial nitric oxide, promoted superoxide (O ₂ ^.? ) production, and increased monocyte binding to endothelial cells. These findings suggest that rapid pacing reduces ISS and ${\partial\tau/\partial t}$ , and rapid pulse rates modulate endothelial responses.     

Insecticide resistance in dengue vectors from hotspots in Selangor,Malaysia

BackgroundIn Malaysia, dengue remains a top priority disease and usage of insecticides is the main method for dengue vector control. Limited baseline insecticide resistance data in dengue hotspots has prompted us to conduct this study. The present study reports the use of a map on the insecticide susceptibility status of Aedes aegypti and Aedes albopictus to provide a quick visualization and overview of the distribution of insecticide resistance.Method and resultsThe insecticide resistance status of Aedes populations collected from 24 dengue hotspot areas from the period of December 2018 until June 2019 was proactively monitored using the World Health Organization standard protocol for adult and larval susceptibility testing was conducted, together with elucidation of the mechanisms involved in observed resistance. For resistance monitoring, susceptibility to three adulticides (permethrin, deltamethrin, and malathion) was tested, as well as susceptibility to the larvicide, temephos. Data showed significant resistance to both deltamethrin and permethrin (pyrethroid insecticides), and to malathion (organophosphate insecticide) in all sampled Aedes aegypti populations, while variable resistance patterns were found in the sampled Aedes albopictus populations. Temephos resistance was observed when larvae were tested using the diagnostic dosage of 0.012mg/L but not at the operational dosage of 1mg/L for both species.ConclusionThe present study highlights evidence of a potential threat to the effectiveness of insecticides currently used in dengue vector control, and the urgent requirement for insecticide resistance management to be integrated into the National Dengue Control Program.     

A mutant <Emphasis Type="SmallCaps">l</Emphasis>-asparaginase II signal peptide improves the secretion of recombinant cyclodextrin glucanotransferase and the viability of <Emphasis Type="Italic">Escherichia coli</Emphasis>

l-Asparaginase II signal peptide was used for the secretion of recombinant cyclodextrin glucanotransferase (CGTase) into the periplasmic space of E. coli. Despite its predominant localisation in the periplasm, CGTase activity was also detected in the extracellular medium, followed by cell lysis. Five mutant signal peptides were constructed to improve the periplasmic levels of CGTase. N1R3 is a mutated signal peptide with the number of positively charged amino acid residues in the n-region increased to a net charge of +5. This mutant peptide produced a 1.7-fold enhancement of CGTase activity in the periplasm and significantly decreased cell lysis to 7.8% of the wild-type level. The formation of intracellular inclusion bodies was also reduced when this mutated signal peptide was used as judged by SDS–PAGE. Therefore, these results provide evidence of a cost-effective means of expression of recombinant proteins in E. coli.     

Diversity and biogeochemical structuring of bacterial communities across the Porangahau ridge accretionary prism, New Zealand

Sediments from the Porangahau ridge, located off the northeastern coast of New Zealand, were studied to describe bacterial community structure in conjunction with differing biogeochemical regimes across the ridge. Low diversity was observed in sediments from an eroded basin seaward of the ridge and the community was dominated by uncultured members of the Burkholderiales. Chloroflexi/GNS and Deltaproteobacteria were abundant in sediments from a methane seep located landward of the ridge. Gas-charged and organic-rich sediments further landward had the highest overall diversity. Surface sediments, with the exception of those from the basin, were dominated by Rhodobacterales sequences associated with organic matter deposition. Taxa related to the Desulfosarcina/Desulfococcus and the JS1 candidates were highly abundant at the sulfate-methane transition zone (SMTZ) at three sites. To determine how community structure was influenced by terrestrial, pelagic and in situ substrates, sequence data were statistically analyzed against geochemical data (e.g. sulfate, chloride, nitrogen, phosphorous, methane, bulk inorganic and organic carbon pools) using the Biota-Environmental matching procedure. Landward of the ridge, sulfate was among the most significant structuring factors. Seaward of the ridge, silica and ammonium were important structuring factors. Regardless of the transect location, methane was the principal structuring factor on SMTZ communities.     

Geomicrobial characterization of gas hydrate-bearing sediments along the mid-Chilean margin

Bacterial diversity in eight sediment cores from the mid-Chilean margin was studied using length heterogeneity (LH)-PCR, and described in relation to in situ geochemical conditions. DNA from the sulfate-methane transition (SMT) of three cores [one containing methane gas; two proximal to a gas hydrate mound (GHM)] was cloned and sequenced. Clones related to uncultured relatives of Desulfosarcina variabilis were found in all clone libraries and dominated one. Desulfosarcina variabilis related clones were similar to phylotypes observed at the SMT in association with anaerobic methane oxidation in the Eel River basin, Cascadia margin and the Gulf of Mexico. The LH-PCR amplicon associated with D. variabilis clones matched the amplicon that dominated most SMT samples, indicating environmental selection for D. variabilis relatives. Clones related to the Verrucomicrobia dominated the library for the methane gas-containing core. Uncultured Treponema relatives dominated the library for the core obtained on the edge of a GHM. Statistical analysis using geochemical data to describe variance in LH-PCR data revealed that stable carbon isotope ratios of dissolved inorganic carbon are the principal structuring factor on SMT communities. These data suggest that D. variabilis relatives are involved in anaerobic oxidation of methane at the SMT in Chilean margin sediments.     

CNIH4 Interacts with Newly Synthesized GPCR and Controls Their Export from the Endoplasmic Reticulum

The molecular mechanisms regulating G protein‐coupled receptors (GPCRs) trafficking from their site of synthesis in the endoplasmic reticulum (ER) to their site of function (the cell surface) remain poorly characterized. Using a bioluminescence resonance energy transfer‐based proteomic screen, we identified a novel GPCR‐interacting protein; the human cornichon homologue 4 (CNIH4). This previously uncharacterized protein is localized in the early secretory pathway where it interacts with members of the 3 family of GPCRs. Both overexpression and knockdown expression of CNIH4 caused the intracellular retention of GPCRs, indicating that this ER‐resident protein plays an important role in GPCR export. Overexpression of CNIH4 at low levels rescued the maturation and cell surface expression of an intracellularly retained mutant form of the β2‐adrenergic receptor, further demonstrating a positive role of CNIH4 in GPCR trafficking. Taken with the co‐immunoprecipitation of CNIH4 with Sec23 and Sec24, components of the COPII coat complex responsible for ER export, these data suggest that CNIH4 acts as a cargo‐sorting receptor, recruiting GPCRs into COPII vesicles .      

Deep-sea shipwrecks represent island-like ecosystems for marine microbiomes

Biogeography of macro- and micro-organisms in the deep sea is, in part, shaped by naturally occurring heterogeneous habitat features of geological and biological origin such as seeps, vents, seamounts, whale and wood-falls. Artificial features including shipwrecks and energy infrastructure shape the biogeographic patterns of macro-organisms; how they influence microorganisms is unclear. Shipwrecks may function as islands of biodiversity for microbiomes, creating a patchwork of habitats with influence radiating out into the seabed. Here we show microbiome richness and diversity increase as a function of proximity to the historic deep-sea shipwreck Anona in the Gulf of Mexico. Diversity and richness extinction plots provide evidence of an island effect on microbiomes. A halo of core taxa on the seabed was observed up to 200 m away from the wreck indicative of the transition zone from shipwreck habitat to the surrounding environment. Transition zones around natural habitat features are often small in area compared to what was observed at Anona indicating shipwrecks may exert a large sphere of influence on seabed microbiomes. Historic shipwrecks are abundant, isolated habitats with global distribution, providing a means to explore contemporary processes shaping biogeography on the seafloor. This work is a case study for how built environments impact microbial biodiversity and provides new information on how arrival of material to the seafloor shapes benthic microbiomes.Subject terms: Biogeography, Microbial biooceanography