Poly-3-hydroxybutyrate in Legionella pneumophila, an energy source for survival in low-nutrient environments.

Chloroform-soluble material was extracted from two strains of L. pneumophila serogroup 1 following growth in continuous culture. The purified material was identified as poly-3-hydroxybutyrate (PHB) by nuclear magnetic resonance spectroscopy and by gas chromatography-mass spectrometry.

Carbon load in aquatic ecosystems affects the diversity and biomass of water biofilm consortia and the persistence of the pathogen Campylobacter jejuni within them.

The influence of carbon load on autochthonous water microflora population distribution and diversity, and on the persistence of Campylobacter jejuni, was examined with a two-stage aquatic biofilm model. Serine was chosen since it is a carbon source utilised by C. jejuni and concentrations were chosen to reflect upper limits of amino acid load reported in surface water.

Analysis of coccal cell formation by Campylobacter jejuni using continuous culture techniques, and the importance of oxidative stress.

Campylobacter jejuni is an important human gastrointestinal pathogen. In hostile environments it may adapt its physiology to prolong survival, potentially including the adoption of a viable, non-culturable form and a change to coccal cell morphology. By independently controlling the individual parameters of continuous cultures of Camp.

Experimental campylobacter infection and diarrhoea in immunodeficient mice.

The responses of previously untested immunodeficient mouse strains to campylobacter infection are described. Three strains of adult immunodeficient mice (SCID-Beige, C.B-17-SCID-Beige and RAG-2) were inoculated intragastrically with Campylobacter jejuni NCTC 11168.

In vivo resistance to simian immunodeficiency virus superinfection depends on attenuated virus dose.

Infection of macaques with attenuated simian immunodeficiency virus (SIV) induces potent superinfection resistance that may be applicable to the development of an AIDS vaccine but little information exists concerning the conditions necessary for the induction of this vaccine effect. We report that only a high dose of attenuated SIVmac protected macaques against intravenous challenge with more virulent virus 15 weeks after primary infection. Three of four animals given 2000-20000 TCID50 of SIVmacC8, a molecular clone of SIVmac251(32H) with a 12 bp deletion in the nef gene, essentially resisted superinfection with uncloned SIVmac.

Molecular analysis of the malR gene of Clostridium butyricum NCIMB 7423, a member of the LacI-GalR family of repressor proteins.

Deletion of a region of DNA 5' to a previously characterised malQ gene of Clostridium butyricum resulted in increased production of the enzyme activity encoded by malQ, 4-alpha-glucanotransferase. Nucleotide sequence analysis revealed the presence of an open reading frame capable of encoding a protein of 335 amino acids. This protein was found to share 33% amino acid sequence identity with the Bacillus subtilis CcpA (catabolite control protein) repressor, 28% identity with the Streptomyces coelicolor MalR repressor, and 30%, 25%, and 21% amino acid identity with the Escherichia coli repressors GalR, LacI and MalI, respectively.

Airborne movement of anthrax spores from carcass sites in the Etosha National Park, Namibia.

Tests for airborne movement of anthrax spores downwind from three heavily contaminated carcass sites were carried out under a range of wind conditions. Anthrax spores were detected in just three of 43 cyclone or gelatin filter air samples taken at distances of 6, 12 and 18 m from the sites. In addition, nine positives resulted during sampling sessions in which the site was mechanically disturbed, with a further five positives being found in sessions subsequent to those in which the site had been disturbed.

Protective efficacy of a recombinant protective antigen against Bacillus anthracis challenge and assessment of immunological markers.

The efficacy of recombinant Bacillus anthracis Protective Antigen (rPA) produced in Bacillus subtilis and formulated in Alhydrogel or MPL-TDM-CWS (Ribi adjuvant) has been tested and compared to the licensed UK human vaccine in guinea pigs challenged by the aerosol route with the Ames strain of B. anthracis. rPA combined with the Ribi adjuvant was found to be the only formulation to provide 100% protection from challenge.

Comparison of the immunological and protective responses elicited by microencapsulated formulations of the F1 antigen from Yersinia pestis.

Purified native F1 antigen from Yersinia pestis was used to assess controlled-release vaccine delivery systems in poly(lactide-co-glycolide) (PLG) microparticles and liposomes. Antigen encapsulated in PLG microparticles induced high serum titres when injected i.p.

Methicillin-resistant Staphylococcus aureus versus the burn patient.

Methicillin-resistant Staphylococcus aureus (MRSA) has become a frequent cause of nosocomial infection, its increasing prevalence posing serious therapeutic and infection control problems within the hospital environment. MRSA is a major challenge to the burn patient, with potential to cause significant morbidity and mortality. Burn patients have been shown to become colonised and infected more readily than other patient groups.

Oral or parenteral administration of replication-deficient adenoviruses expressing the measles virus haemagglutinin and fusion proteins: protective immune responses in rodents.

The genes encoding the measles virus (MV) haemagglutinin (H) and fusion (F) proteins were placed under the control of the human cytomegalovirus immediate early promoter in a replication-deficient adenovirus vector. Immunofluorescence and radioimmune precipitation demonstrated the synthesis of each protein and biological activity was confirmed by the detection of haemadsorption and fusion activities in infected cells. Oral as well as parenteral administration of the H-expressing recombinant adenovirus elicited a significant protective response in mice challenged with MV.

Extended survival and persistence of Campylobacter spp. in water and aquatic biofilms and their detection by immunofluorescent-antibody and -rRNA staining.

In water microcosm experiments, the survival times of Campylobacter isolates differed by up to twofold, as determined by culturing; this difference increased to fourfold when particular combinations of temperature and oxygenation were used. The mean survival times were much longer at 4 and 10 degrees C (202 and 176 h, respectively) than at 22 and 37 degrees C (43 and 22 h, respectively). The influence of anaerobiosis on survival time was less dramatic and differed considerably between isolates.

Molecular analysis of a Clostridium butyricum NCIMB 7423 gene encoding 4-alpha-glucanotransferase and characterization of the recombinant enzyme produced in Escherichia coli.

An Escherichia coli clone was detected in a Clostridium butyricum NCIMB 7423 plasmid library capable of degrading soluble amylose. Deletion subcloning of its recombinant plasmid indicated that the gene(s) responsible for amylose degradation was localized on a 1.8 kb NspHI-Scal fragment.

Characterization of novel long-chain 1,2-diols in Thermus species and demonstration that Thermus strains contain both glycerol-linked and diol-linked glycolipids.

In this study, we purified and characterized tetra- and triglycosyl glycolipids (GL-1 and GL-2, respectively) from two different colonial forms of Thermus scotoductus X-1, from T. filiformis Tok4 A2, and from T. oshimai SPS-11.

Characterisation of the meningococcal transferrin binding protein complex by photon correlation spectroscopy.

Photon correlation spectroscopy demonstrated for the first time that co-purified meningococcal TbpA+B form a complex in solution. This structure bound hTf and the resultant species underwent partial dissociation after exposure to additional hTf or following prolonged incubation. Purified TbpA and TbpB had similar apparent sizes but showed distinctive size profiles suggesting that TbpA forms a largely homogeneous population while TbpB may produce more variable particle sizes under these conditions.

Experimentally assessed public health risks associated with pigs from farms experiencing anthrax.

Following an outbreak of anthrax in an intensive pig rearing unit in north Wales in 1989 a study was initiated by the Ministry of Agriculture, Fisheries and Food to assess public health risks during such an outbreak. Of 50 pigs infected by the addition of Bacillus anthracis spores to their feed, two died of anthrax six and eight days later. The remainder were observed for 21 days and exhibited only mild and transient clinical signs of disease.

Macaques infected with attenuated simian immunodeficiency virus resist superinfection with virulence-revertant virus.

Macaques infected with attenuated simian immunodeficiency virus (SIVmac) can resist superinfection challenge with virulent virus, showing the potential of live attenuated virus as an AIDS vaccine. Superinfection resistance does not, however, prevent the generation of virulent virus in vivo, suggesting that such virus may circumvent the resistance effect. Here, we show that three macaques already infected with the attenuated molecular clone SIVmacC8 were resistant to superinfection with virulent virus that arose in vivo following repair of a 12 bp attenuating lesion in the nef/3' LTR.

Anti-major histocompatibility complex antibody responses to simian B cells do not protect macaques against SIVmac infection.

Macaques have been protected against infection with human cell-grown SIVmac by immunization with antigens encoded by the human major histocompatibility complex (MHC). Here, we investigated the efficacy of alloimmunization with simian B cells expressing high levels of MHC class I and class II molecules to confer protection against systemic challenge with simian-grown SIVmac. Eight rhesus macaques were vaccinated with glutaraldehyde-fixed and beta-propiolactone-inactivated herpesvirus papio-transformed B cells.

Poly(DL-lactide-co-glycolide)-encapsulated plasmid DNA elicits systemic and mucosal antibody responses to encoded protein after oral administration.

We have developed a method for the encapsulation of plasmid DNA in poly(DL-lactide-co-glycolide) microparticles. Encapsulated DNA, expressing the insect protein luciferase under the transcriptional control of the human cytomegalovirus immediate early promoter, was administered to mice by intraperitoneal injection or oral gavage. Intraperitoneal injection of encapsulated DNA elicited good serum IgG and IgM responses, and a modest IgA response.

Barriers to hepatitis C transmission within breathing systems: efficacy of a pleated hydrophobic filter.

It has been suggested that breathing circuits contaminated with body fluids may provide a route of nosocomial patient-to-patient transmission of the hepatitis C virus. Thus, a number of authorities have recommended the use of breathing circuit filters to minimize such risks. The present study sought to simulate a humidified breathing circuit and evaluate two different designs of breathing circuit filters to determine their efficacy in preventing passage of the hepatitis C virus.

Degradation of 3-chlorobenzoate by thermophilic micro-organisms.

Thermophilic (75 degrees C), anaerobic biodegradation of chlorobenzoates was investigated using different inocula from geothermal and non-geothermal environments. Microbial dehalogenation of 3-chlorobenzoate (0.5 mmol l-1 was achieved by two mixed cultures growing anaerobically at 75 degrees C.

Changes with growth rate in the membrane lipid composition of and amino acid utilization by continuous cultures of Campylobacter jejuni.

Methods and media (defined and complex) are described which permit studies designed to determine the influence of single environmental factors on the survival and virulence of Campylobacter jejuni. The effect of growth rate on selected physiological traits (amino acid utilization, membrane lipid composition, motility, cell morphology) was studied in continuous culture. In both media, growth was at the expense of amino acid (serine, aspartate, glutamate and proline) catabolism.