Nisin treatment for inactivation of Salmonella species and other gram-negative bacteria.

Nisin, produced by Lactococcus lactis subsp. lactis, has a broad spectrum of activity against gram-positive bacteria and is generally recognized as safe in the United States for use in selected pasteurized cheese spreads to control the outgrowth and toxin production of Clostridium botulinum. This study evaluated the inhibitory activity of nisin in combination with a chelating agent, disodium EDTA, against several Salmonella species and other selected gram-negative bacteria. After a 1-h exposure to 50 micrograms of nisin per ml and 20 mM disodium EDTA at 37 degrees C, a 3.2- to 6.9-log-cycle reduction in population was observed with the species tested. Treatment with disodium EDTA or nisin alone produced no significant inhibition (less than 1-log-cycle reduction) of the Salmonella and other gram-negative species tested. These results demonstrated that nisin is bactericidal to Salmonella species and that the observed inactivation can be demonstrated in other gram-negative bacteria. Applications involving the simultaneous treatment with nisin and chelating agents that alter the outer membrane may be of value in controlling food-borne salmonellae and other gram-negative bacteria.     

Oxygen and temperature effects on acetaldehyde-induced catabolite inactivation in Pichia pastoris

Summary Exposure of methylotrophic yeasts to other carbon sources after growth on methanol results in catabolite inactivation. As a result, peroxisomes are rapidly degraded effectively disabling the metabolic pathway initiated by alcohol oxidase in favour of a more energetically favourable route. A model equation has been developed to describe the effect of temperature, dissolved oxygen concentration and acetaldehyde (catabolite) concentration on catabolite inactivation in Pichia pastoris. When pre-exposed to 4 g/l acetaldehyde at 30°C, the rate of conversion of ethanol to acetaldehyde decreased by 75%. Inactivation was reduced to 45% at 30°C by reducing the dissolved oxygen concentration. At high dissolved oxygen concentration, enzyme function was only inactivated by 20% if the temperature during the period of exposure to acetaldehyde was reduced to 5°C. The influence of acetaldehyde can be eliminated completely by operating at 5°C and low dissolved oxygen concentrations. Application of these findings to process design has enabled us to conduct preliminary reactions in laboratory-scale reactors that have yielded acetaldehyde concentrations greater than 3 M (130 g/l) in 4 h. Offsprint requests to: S. J. B. Duff     

Changes in biofilms induced by flow regulation could explain extinctions of aquatic snails in the lower River Murray,Australia

Notopala sublineata, Notopala hanleyi(Viviparidae) andThiara balonnensis (Thiaridae) are prosobranchgastropodsthat were once abundant in the lower River Murray. These andothersnail taxa have declined markedly over the last 50 years,coinciding with increased flow regulation by dams and weirs.Inthis paper we speculate that the decline may be linked tochangesin the nature of food resources. Before regulation, most ofthebiofilm biomass in the lower Murray probably was microbial, asfluctuating water levels and high turbidity would havemaintainedthese communities in a state of early succession. Bystabilisingseasonal water levels, we suggest that regulation has promotedthegrowth of filamentous algae, perhaps at the expense ofbacteria.Evidence from gut and faecal pellet analysis, and fromanalysis ofcarbon stable-isotopes, suggests that the gastropod taxa aredetritivores, feeding mainly on amorphous organic detritus.Algaehave a relatively high C:N ratio (low nutritional value) andmay bean inadequate food to maintain female growth and reproduction,especially in viviparous snails.     

Improvement of the total turnover number and space-time yield for chloroperoxidase catalyzed oxidation

Chloroperoxidase from Caldariomyces fumago was applied for the oxidation of indole to oxindole using hydrogen peroxide as the oxidant in aqueous t-butyl alcohol medium. Different ways of adding the oxidant, various reactor types, and the use of a hydrogen peroxide-stat were compared, resulting in a 20-fold increase of the total turnover number (ttn) and space-time yield (sty). The highest ttn of >860,000 was obtained in a fed-batch reactor, whereas the highest sty of 120 g/(L . d) was reached in a continuously operated enzyme membrane reactor. The results were compared to other enzyme systems already established for the synthesis of amino acids and carbohydrates. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 283-288, 1997.     

A comparison of solution and membrane-bound DNA × DNA hybridization,as used to infer phylogeny

A new method of membrane-bound DNA × DNA hybridization was devised to accommodate the study of small quantities of DNA obtained from museum specimens for phylogeny reconstruction. Membranebound, single-stranded target genomic DNAs were competitively hybridized with a total genomic DNA probe to form hybrid duplexes required for the DNA dissociation experiments. We compared the thermal elution profiles derived from dissociating duplexes made with probes of whole genomic, single-copy, and repetitive DNA, as well as solution DNA × DNA hybridization using sc tracer. Quantitatively, pairwise indices of genetic distance derived from duplexes made with genomic probes depended entirely on hybridization of repetitive sequences, but a parallel set of experiments using repetitive and sc probes produced qualitatively similar results. The indices of genetic distance generated by the membrane-bound hybrids form an internally consistent, resolved tree which is in agreement with the solution DNA × DNA hybridization trials and traditional views of the phylogeny of the taxa under study.Correspondence to: P. Houde     

ultraviolet radiation from F and K stars and implications for planetary habitability

Now that extrasolar planets have been found, it is timely to ask whether some of them might be suitable for life. Climatic constraints on planetary habitability indicate that a reasonably wide habitable zone exists around main sequence stars with spectral types in the early-F to mid-K range. However, it has not been demonstrated that planets orbiting such stars would be habitable when biologically-damaging energetic radiation is also considered. The large amounts of UV radiation emitted by early-type stars have been suggested to pose a problem for evolving life in their vicinity. But one might also argue that the real problem lies with late-type stars, which emit proportionally less radiation at the short wavelengths ( < 200 nm) required to split O2 and initiate ozone formation. We show here that neither of these concerns is necessarily fatal to the evolution of advanced life: Earth-like planets orbiting F and K stars may well receive less harmful UV radiation at their surfaces than does the Earth itself.     

Antagonistic natural selection revealed by molecular sex identification of nestling collared flycatchers

Natural selection may act in different directions during different life-history stages, or in different directions on different classes of individuals. Antagonistic selection of this kind may be an important mechanism by which additive genetic variation for quantitative traits is maintained, and can prevent populations or species reaching local adaptive peaks. This paper reports the results of a study of viability selection on morphological traits of nestling collared flycatchers Ficedula albicollis . Analyses performed without knowledge of the sex of nestlings suggested that no selection was occurring on these traits. However, using molecular sex identification with the avian CHD gene, it is shown that selection acts in different directions on male and female body size from fledging to breeding, apparently favouring relatively small males and large females. The results suggest that differential selection on male and female nestlings may contribute to purely phenotypic sexual size dimorphism in this species. These findings highlight the potential of newly developed molecular sexing techniques to reveal the consequences of an individual's gender for many aspects of its life history in taxa where gender cannot be determined on the basis of external appearance.     

Early evolution of chili peppers (Capsicum)

An hypothesis is advanced based upon geographical information and data from horizontal starch gel electrophoresis regarding the place and mode of evolution of the chili peppers (Capsicum). The hypothesis suggests a major portion of the genusCapsicum arose in a nuclear area in south central Bolivia with subsequent migration into the Andes and lowland Amazonia accompanied by radiation and speciation.     

Role of Intestinal Microflora in the Metabolism of Guanidinosuccinic Acid

Among a variety of bacteria isolated from the gastrointestinal tracts of rats and humans, only streptococci of group N are capable of degrading guanidinosuccinic acid added to their culture medium. The urinary excretion of guanidinosuccinic acid by germfree rats is greater than that of conventional rats. The excretion of this compound by gnotobiotic rats correlates with the capacity of their intestinal microflora to degrade guanidinosuccinic acid in culture. Thus, guanidinosuccinic acid excretion is low in rats infected exclusively with Streptococcus faecalis, and the excretion is not altered when germfree rats are infected with an organism unable to degrade guanidinosuccinic acid (Lactobacillus). These findings suggest that the intestinal microflora, particularly Streptococcus, play a role in the metabolism of guanidinosuccinic acid by the host.     

Transcription of the Adenovirus Genome by an α-Amanitine-Sensitive Ribonucleic Acid Polymerase in HeLa Cells

The properties of the ribonucleic acid (RNA) polymerase activity which transcribes the major portion of the adenovirus genome were studied. Nuclei were prepared from infected cells and incubated in vitro. Virus-specific RNA was determined by hybridization to adenovirus deoxyribonucleic acid (DNA). Adenovirus DNA is transcribed principally by an activity which resembles closely polymerase II of the host cell. This activity is inhibited by alpha-amanitine and stimulated by (NH(4))(2)SO(4). Its product is high-molecular-weight heterogeneous RNA. The polymerase activity measured early in infection (3 to 5 hr) resembles that found late in infection (16 to 18 hr).