Culture and differentiation of rabbit intestinal organoids and organoid-derived cell monolayers.

Organoids emulate many aspects of their parental tissue and are therefore used to study pathogen-host interactions and other complex biological processes. Here, we report a robust protocol for the isolation, maintenance and differentiation of rabbit small intestinal organoids and organoid-derived cell monolayers. Our rabbit intestinal spheroid and monolayer cultures grew most efficiently in L-WRN-conditioned medium that contained Wnt, R-spondin and Noggin, and that had been supplemented with ROCK and TGF-β inhibitors.

Field Effectiveness of a Metofluthrin Fan-Based Emanator and Deet as Repellents Against Aedes vigilax in Southeast Queensland, Australia1.

A field study to compare a formulation containing 40% deet (N,N-diethyl-3-methyl benzamide) in ethanol (Bushman™) and a battery-powered fan emanator with a chemical strip containing 31.2% metofluthrin (OFF!® Clip-On™) was conducted at Redcliffe, Queensland, Australia, in February 2016. The 40% deet provided 100% protection against mosquitoes for 5 h until tests ceased, while the OFF! Clip-On device provided only 42.

Spatial Attention and Saccade Preparation Both Independently Contribute to the Discrimination of Oblique Orientations.

The extent to which the preparation of an eye movement and spatial attention both independently influence performance within the same task has long been debated. In a recent study that combined computational modelling with a dual-task, both saccade preparation and spatial cueing were revealed to separately contribute to the discrimination of targets oriented along the cardinal axis (horizontal and vertical). However, it remains to be seen whether and to what degree the same holds true when different perceptual stimuli are used.

The need for accuracy and validity in research on nightlife and drinking: A commentary on Devilly et al. and recommendations for future research.

Research on nightlife and drinking faces many unique challenges, and validity in research is an important concern. A recent publication by Devilly et al. entitled "SmartStart: Results of a large point of entry study into preloading alcohol and associated behaviours" contains definitions and assumptions about prior work that require more careful consideration.

Predicted global distribution of Burkholderia pseudomallei and burden of melioidosis.

Burkholderia pseudomallei, a highly pathogenic bacterium that causes melioidosis, is commonly found in soil in Southeast Asia and Northern Australia(1,2). Melioidosis can be difficult to diagnose due to its diverse clinical manifestations and the inadequacy of conventional bacterial identification methods(3). The bacterium is intrinsically resistant to a wide range of antimicrobials, and treatment with ineffective antimicrobials may result in case fatality rates (CFRs) exceeding 70%(4,5).

Predicted global distribution of and burden of melioidosis.

a highly pathogenic bacterium that causes melioidosis, is commonly found in soil in Southeast Asia and Northern Australia. Melioidosis can be difficult to diagnose due to its diverse clinical manifestations and the inadequacy of conventional bacterial identification methods. The bacterium is intrinsically resistant to a wide range of antimicrobials, and treatment with ineffective antimicrobials may result in case fatality rates (CFRs) exceeding 70%.

Griffith Edwards, the Addiction Research Unit and research on the criminal justice system.

Background: This paper reviews the early work of Griffith Edwards and his colleagues on alcohol in the criminal justice system and outlines the direction of research in this area in the Addiction Research Unit in the 1960s and 1970s. The paper outlines the link between that work and work undertaken in the more recent past in this area.

Methods: The key papers of the authors are reviewed and the impact of this work on policy and practice is discussed.

In situ hybridisation assay for localisation of rabbit calicivirus Australia-1 (RCV-A1) in European rabbit (Oryctolagus cuniculus) tissues.

Recently, a new lagovirus enzootic in Australian wild rabbits was identified and described as rabbit calicivirus Australia-1 (RCV-A1). Unlike the closely related Rabbit Haemorrhagic Disease Virus (RHDV), which causes fulminant hepatitis and rabbit death, RCV-A1 does not appear to induce any clinical disease. RCV-A1 has been postulated to act as an imperfect natural vaccine to RHDV thus reducing RHDV-induced rabbit mortality, which is detrimental for bio-control of rabbits in Australia.

"Australia is one of the darkest markets in the world": the global importance of Australian tobacco control.

"Australia is one of the darkest markets in the world...

Conservation of L and 3C proteinase activities across distantly related aphthoviruses.

The foot-and-mouth disease virus (FMDV) leader (L) proteinase is an important virulence determinant in FMDV infections. It possesses two distinct catalytic activities: (i) C-terminal processing at the L/VP4 junction; and (ii) induction of the cleavage of translation initiation factor eIF4G, an event that inhibits cap-dependent translation in infected cells. The only other member of the Aphthovirus genus, equine rhinitis A virus (ERAV), also encodes an L protein, but this shares only 32% amino acid identity with its FMDV counterpart.

Glycogen-accumulating organisms in laboratory-scale and full-scale wastewater treatment processes.

Laboratory-scale sequencing batch reactors (SBRs) as models for wastewater treatment processes were used to identify glycogen-accumulating organisms (GAOs), which are thought to be responsible for the deterioration of enhanced biological phosphorus removal (EBPR). The SBRs (called Q and T), operated under alternating anaerobic-aerobic conditions typical for EBPR, generated mixed microbial communities (sludges) demonstrating the GAO phenotype. Intracellular glycogen and poly-beta-hydroxyalkanoate (PHA) transformations typical of efficient EBPR occurred but polyphosphate was not bioaccumulated and the sludges contained 1.

On the evolution and molecular epidemiology of the potyvirus Papaya ringspot virus.

The potyvirus Papaya ringspot virus (PRSV) is found throughout the tropics and subtropics. Its P biotype is a devastating pathogen of papaya crops and its W biotype of cucurbits. PRSV-P is thought to arise by mutation from PRSV-W.

Cloning and expression of the phosphotriesterase gene hocA from Pseudomonas monteilii C11.

The cloning of a gene encoding the novel phosphotriesterase from Pseudomonas monteilii C11, which enabled it to use the organophosphate (OP) coroxon as its sole phosphorus source, is described. The gene, called hocA (hydrolysis of coroxon) consists of 501 bp and encodes a protein of 19 kDa. This protein had no sequence similarity to any proteins in the SWISS-PROT/GenBank databases.

Cloning, sequencing and expression of an alpha-amylase gene, amyA, from the thermophilic halophile Halothermothrix orenii and purification and biochemical characterization of the recombinant enzyme.

A recombinant clone expressing an amylase was identified from an Escherichia coli generated genomic library of the thermophilic, moderately halophilic, anaerobic bacterium Halothermothrix orenii by activity screening, and the gene encoding the enzyme was designated AmyA. The amyA gene was 1545 bp long, and encoded a 515 residue protein composed of a 25 amino acid putative signal peptide and a 490 amino acid mature protein. It possessed the five consensus regions characteristic of the alpha-amylase family and showed the greatest homology to the Bacillus megaterium group of alpha-amylases.

Filamentous Chloroflexi (green non-sulfur bacteria) are abundant in wastewater treatment processes with biological nutrient removal.

Most filamentous bacteria in biological nutrient removal (BNR) processes have not been identified beyond their morphotype and simple staining reactions. Furthermore, the majority of sludge filaments observed under the microscope do not hybridize to commonly used phylogenetic probes for well characterized bacterial phyla such as the Proteobacteria, Actinobacteria, Firmicutes and BACTEROIDETES: Specific 16S rRNA-targeted oligonucleotide probes were designed for the phylum Chloroflexi (green non-sulfur bacteria) and optimized for use in fluorescence in situ hybridization. Chloroflexi have been implicated in BNR systems by phylogenetic identification of filamentous bacteria isolated by micromanipulation from sludge and culture-independent molecular phylogenetic surveys.

Functional analysis of microbial communities in aerobic-anaerobic sequencing batch reactors fed with different phosphorus/carbon (P/C) ratios.

Fluorescence in situ hybridization (FISH) was used to analyse the community composition of a sequencing batch reactor (SBR) operating with aerobic-anaerobic cycling and fed acetate as its sole carbon source. Phosphorus was removed from the SBR microbiologically. Marked shifts in the community structure occurred as the phosphorus/carbon (P/C) ratio in the feed was changed.

Prospecting for novel lipase genes using PCR.

A PCR method suitable for the isolation of lipase genes directly from environmental DNA is described. The problems associated with the low levels of similarity between lipase genes were overcome by extensive analysis of conserved regions and careful primer design. Using this method, a lipase gene (oli-lipase) was isolated directly from environmental DNA.

Cloning and characterization of a novel haemolysin in Vibrio cholerae O1 that does not directly contribute to the virulence of the organism.

A previously undescribed haemolysin, distinct from the major Vibrio cholerae O1 El Tor haemolysin, HlyA, was cloned from the O1 classical biotype strain Z17561. This novel haemolysin showed 71.5% overall similarity to the delta-thermostable direct haemolysin of Vibrio parahaemolyticus, and so it has been termed V.

Molecular analysis of a haemagglutinin of Haemophilus paragallinarum.

The gene encoding a haemagglutinin of H. paragallinarum, hagA, has been identified and the full-length nucleotide sequence determined. A approximately 39 kDa protein, recognized by an anti-haemagglutinin monoclonal antibody, mAb4D, was purified from H.

The genes encoding virulence-associated proteins and the capsule of Streptococcus pneumoniae are upregulated and differentially expressed in vivo.

The polysaccharide capsule of Streptococcus pneumoniae and several well-characterized virulence proteins are known to contribute to the pathogenesis of pneumococcal disease. However, there is a paucity of data on the expression of their respective genes in vivo. In this study, the relative abundance of the mRNA transcripts of the genes encoding pneumolysin (ply), pneumococcal surface protein A (pspA), pneumococcal surface antigen A (psaA) and choline-binding protein A (cbpA), and of the first gene of the capsular polysaccharide biosynthesis locus (cps2A), was measured in virulent type 2 pneumococci harvested from the blood of BALB/c mice at 12 h and 24 h following intraperitoneal infection.

Identification of structural determinants of the first transmembrane domain of the small envelope protein of duck hepatitis B virus essential for particle morphogenesis.

The envelope of duck hepatitis B virus (DHBV) consists of the small (S) and large (L) envelope proteins, which share a common C-terminal multispanning transmembrane region but differ by the long N-terminal pre-S domain of L, which is essential for interactions with both the receptor and nucleocapsid. To achieve these dual functions, L acquires mixed topologies through S-dependent post-translational translocation of its pre-S domain. This study has examined the role of S in this unusual mechanism of translocation by analysis of the alpha-helical transmembrane domains and their potential to engage in lateral interactions for envelope assembly.

A time to kill: viral manipulation of the cell death program.

Many viruses have as part of their arsenal the ability to modulate the apoptotic pathways of the host. It is counter-intuitive that such simple organisms would be efficient at regulating this the most crucial pathway within the host, given the relative complexity of the host cells. Yet, viruses have the potential to initiate or stay the onset of programmed cell death through the manipulation of a variety of key apoptotic proteins.

Reassessment of major products of N2 fixation by bacteroids from soybean root nodules.

NH3/ was the principal product from soybean bacteroids, prepared by various procedures, when assayed in solution in a flow chamber under N2 fixation conditions. In addition, small quantities of alanine were produced (reaching 20% of NH3/ under some conditions). Some 15N was assimilated by bacteroids purified from soybean root nodules on Percoll density gradients and shaken with 15N2 and 0.

The genetic structure of Escherichia coli populations in primary and secondary habitats.

Escherichia coli were recovered from the members of two two-person households and their associated septic tanks. The E. coli were isolated using selective and non-selective isolation techniques, characterized using the method of multi-locus enzyme electrophoresis and screened for the presence of virulence factors associated with extra-intestinal disease by using PCR.