The survival of Pseudomonas aeruginosa during bromination in a model whirlpool spa

The susceptibility of Pseudomonas aeruginosa to bromination at 38°C in a controlled aquatic environment which simulated a whirlpool spa was evaluated with a laboratory and a naturally-occurring strain from a whirlpool spa. The laboratory strain of Ps. aeruginosa was more susceptible to disinfection with bromine than was the naturally-occurring strain.     

A guinea pig cage for easy cleaning.

A simple cage and rack system was designed for holding guinea pigs in a laboratory environment. The shoe box solid floor cage, made of plastic, has 2 holes for "outside of cage" watering and "outside of cage" feeding. All of the accessory equipments such as automatic watering devices and feeders were set on rack shelves to design the cage simply. This system has satisfactorily functioned for about one year in our laboratory and has substantially saved labor in the washing process.     

Genomic population structure associated with repeated escape of Salmonella enterica ATCC14028s from the laboratory into nature

Salmonella enterica serovar Typhimurium strain ATCC14028s is commercially available from multiple national type culture collections, and has been widely used since 1960 for quality control of growth media and experiments on fitness (“laboratory evolution”). ATCC14028s has been implicated in multiple cross-contaminations in the laboratory, and has also caused multiple laboratory infections and one known attempt at bioterrorism. According to hierarchical clustering of 3002 core gene sequences, ATCC14028s belongs to HierCC cluster HC20_373 in which most internal branch lengths are only one to three SNPs long. Many natural Typhimurium isolates from humans, domesticated animals and the environment also belong to HC20_373, and their core genomes are almost indistinguishable from those of laboratory strains. These natural isolates have infected humans in Ireland and Taiwan for decades, and are common in the British Isles as well as the Americas. The isolation history of some of the natural isolates confirms the conclusion that they do not represent recent contamination by the laboratory strain, and 10% carry plasmids or bacteriophages which have been acquired in nature by HGT from unrelated bacteria. We propose that ATCC14028s has repeatedly escaped from the laboratory environment into nature via laboratory accidents or infections, but the escaped micro-lineages have only a limited life span. As a result, there is a genetic gap separating HC20_373 from its closest natural relatives due to a divergence between them in the late 19^th century followed by repeated extinction events of escaped HC20_373.     

Spent media from cultures of environmental isolates of Escherichia coli can suppress the deficiency of biofilm formation under anoxic conditions of laboratory E. coli strains

The prevailing lifestyle of bacteria is sessile and they attach to surfaces in structures known as biofilms. In Escherichia coli, as in many other bacteria, biofilms are formed at the air-liquid interface, suggesting that oxygen has a critical role in the biofilm formation process. It has been reported that anaerobically growing E. coli laboratory strains are unable to form biofilms even after 96 h of incubation on Luria Bertani (LB) medium. After analyzing 22,000 transposon-induced and 26,000 chemically-induced mutants we failed to isolate an E. coli laboratory strain with the ability to form biofilm under anaerobic growth conditions. Notably, seven strains from a collection of E. coli isolated from different hosts and the environment had the ability to form biofilm in the absence of oxygen. Interestingly, spent medium from cultures of one strain, Souza298, can promote biofilm formation of E. coli laboratory strains growing under anaerobic conditions. Our results led us to propose that laboratory E. coli strains do not release (or synthesize) a molecule needed for biofilm formation under anoxic conditions but that they bear all the required machinery needed for this process.     

Biological properties of strains of tick-borne encephalitis virus isolated in the natural foci of the eastern part of the Russian plain

Strains of tick-borne encephalitis virus isolated in the natural foci of infection in the eastern part of the Russian Plain (the Kirov region) were examined for their biological properties. The strains examined were 69 strains isolated from ticks Ixodes persulcatus, 62 strains obtained from patients with the clinically manifest form of tick-borne encephalitis and 56 strains isolated from the blood of patients with the inapparent form of infection. Comparative studies on laboratory animals (albino mice, golden hamsters, suckling guinea pigs and other mammals) as well as comparative serologic studies provided evidence which suggested that all virus isolates from the Kirov region were antigenically identical with the strain "Sofin" isolated in the Far East and represent thus a single causative agent of the tick-borne encephalitis virus infection. This strain of virus is supposed to exist in two variants, in dependence on ecological conditions: one of these variants is the eastern variant (strain Sofin and strains from the Kirov region) and the other one is the western variant of tick-borne encephalitis virus.     

Genomewide evolutionary rates in laboratory and wild yeast

As wild organisms adapt to the laboratory environment, they become less relevant as biological models. It has been suggested that a commonly used S. cerevisiae strain has rapidly accumulated mutations in the lab. We report a low-to-intermediate rate of protein evolution in this strain relative to wild isolates.     

The development of low temperature inactive (Lti) baker's yeast

The construction of a novel baker's yeast variety via traditional genetic techniques is described. The phenotype was designated "Lti" ("Low temperature inactive"). Lti mutations with the desired characteristics within a genetically well-defined haploid laboratory strain of Saccharomyces cerevisiae were isolated, and two different approaches were taken to obtain baker's yeast strains, which exhibit reduced fermenting activity at refrigeration temperatures. In a first approach, a chosen Lti strain carrying mutation lti9 was combined with other laboratory strains carrying defined MAL alleles. In a second approach, the same lti mutation was introduced in the genetic background of polyploid commercial baker's yeast strains that harbor important "industrial" properties. Lti strains arising from both approaches were characterized with specifically developed screening procedures. Strains of the "academic" Lti strain family displayed between 85% and 92% of the biomass yield of a commercial reference strain, whereas strains of the "industrial" Lti strain family showed a variation between 60% and 115%. Lti strains from both families varied strongly among each other in their activity in model doughs: at 8 degrees C they displayed activities between 5% and 30%, and at 30 degrees C between 40% and 113% of a commercial reference baker's yeast strain.     

Comparative analysis by polymerase chain reaction amplified minicircles of kinetoplast DNA of a stable strain of Trypanosoma cruzi from São Felipe, Bahia, its clones and subclones: possibility of predominance of a principal clone in this area

Molecular characterization of one stable strain of Trypanosoma cruzi, the 21 SF, representative of the pattern of strains isolated from the endemic area of S?o Felipe, State of Bahia, Brazil, maintained for 15 years in laboratory by serial passages in mice and classified as biodeme Type II and zymodeme 2 has been investigated. The kinetoplast DNA (kDNA) of parental strain, 5 clones and 14 subclones were analyzed. Schizodeme was established by comparative study of the fragments obtained from digestion of the 330-bp fragments amplified by polymerase chain reaction (PCR) from the variable regions of the minicircles, and digested by restriction endonucleases Rsa I and Hinf I. Our results show a high percentual of similarity between the restriction fragment length polymorphism (RFLP) for the parental strain and its clones and among these individual clones and their subclones at a level of 80 to 100%. This homology indicates a predominance of the same "principal clone" in the 21SF strain and confirms the homogeneity previously observed at biological and isozymic analysis. These results suggest the possibility that the T. cruzi strains with similar biological and isoenzymic patterns, circulating in this endemic area, are representative of one dominant clone. The presence of "principal clones" could be responsible for a predominant tropism of the parasites for specific organs and tissues and this could contribute to the pattern of clinico-pathological manifestations of Chagas's disease in one geographical area.     

The Genetic Basis of Malathion Resistance in Housefly (Musca domestica L.) Strains From Turkey

Organophosphate insecticide (parathion/diazinon) resistance in housefly (Musca domestica L.) is associated with the change in carboxylesterase activity. The product of MdE7 gene is probably playing a role in detoxification of xenobiotic esters. In our research, we have isolated, cloned and sequenced the MdE7 gene from five different Turkish housefly strains. High doses of malathion (600 g/fly) were applied in a laboratory environment for one year to Ceyhan1, Ceyhan2, Adana, and Ankara strains while no insecticide treatment was performed in the laboratory to Kirazli strain. Trp²⁵¹ Ser substitution was found in the product of MdE7 gene in all malathion-resistant and Kirazli stocks. In addition, we checked the malathion carboxylesterase (MCE), percent remaining activities in acetylcholinesterase (AChE), glutathion-S-transferase (GST), and general esterase activities in all five strains used in this study. In comparing with universal standard sensitive control WHO, a high level of MCE and GST activities were observed while lower level of general esterase activities was detected in the tested strains. In addition, a higher percent remaining activities in AChE than WHO susceptible strain were observed in all malathion-resistant strains.     

Water loss and metabolic activity in bed bug eggs (Cimex lectularius)

Few studies have evaluated water loss and respiratory activity of insect eggs, particularly insects that are known to live within indoor environments. The present study quantifies water loss and respiratory activity for the eggs of a re‐emerging indoor pest of human environments Cimex lectularius (L.). Water loss is measured gravimetrically and calculated as a function of chorion permeability. For these studies, bed bug eggs are placed at 0% relative humidity and repeatedly weighed over 48 h. Temperature effects and bed bug strain differences on the standard metabolic rate (SMR) and respiratory quotient are measured using closed system respirometry. The SMR (; mL g^?1 h^?1) is measured for two field strain bed bugs and compared with a laboratory strain held at one temperature (25 °C). The standard metabolic rate is measured for Harlan (laboratory) strain bed bug eggs at six different temperatures (15, 20, 25, 30, 35 and 39 °C). Total water loss is not significantly different between all three strains. However, water loss across the chorion (chorion permeability) is significantly different between the Harlan laboratory strain and the two field collected strains. Standard metabolic rates for Harlan (laboratory) strain bed bug eggs increase with temperatures from 15 to 35 °C but decline at 39 °C. Overall, the Harlan bed bug eggs have the largest standard metabolic rates (0.18 ± 0.05 mL g^?1 h^?1) compared with the Epic Center strain eggs (0.14 ± 0.03 mL g^?1 h^?1) and Richmond strain eggs (0.16 ± 0.04 mL g^?1 h^?1), although this difference is not significant.     

Laboratory rat selection for the trait "the absence of audiogenic seizure proneness"

The hybrids between Krushinsky-Molodkina (KM) inbred strain, selected for high predisposition to audiogenic epilepsy (AE), and Wistar rats non-prone to audiogenic seizure were the initial population for selection. Rats were selected for the trait "the absence of audiogenic seizure proneness". The creation of such strain in which the significant proportion of animals develop no AE in response to sound and share partly the genetic background of the KM strain is very important for the correct use of RV strain as the laboratory model of seizure states. As alleles which determine the AE proneness are recessive the selection for the "opposite" trait proceeds necessarily slow.     

Divergent nutrition-related adaptations in two cockroach populations inhabiting different environments

Developmental, reproductive and size‐related variables were compared between two ecologically different strains of Periplaneta americana (L.). One strain was a laboratory‐reared culture, and the other a feral strain inhabiting an urban environment. The feral population was originally founded by escapees from the culture, but may also include immigrants from other urban populations. Both final‐instar nymphs and adults of the feral insects were heavier than their equivalents from the cultured population, this weight difference being due to a higher capacity for the storage of water and heavier fat‐free carcasses. Feral cockroaches also had heavier oothecae, which contained heavier offspring than those from cultured females. Feral animals had one or two more larval stadia and higher growth rates. Size‐related differences persisted in first and second filial generations reared under laboratory conditions on a nutritionally balanced diet, but were not apparent in first filial insects reared on a vegetable diet. Greater resistance to starvation was found in feral animals, and this was attributed primarily to their larger water stores. Feral animals were found to harbour a higher density of endosymbiotic bacteria in the fat body, which are known to enhance the efficiency of protein utilization. The data suggest that the characteristics of feral animals have been selected in the nutritionally harsh feral environment compared with the more benign culture conditions, with water availability playing a role.     

From isolates to a synthetic laboratory population: maintenance of variability in the nematode Haemonchus contortus

Haemonchus contortus isolates were collected from goats of five locations with different climatic characteristics in Guadeloupe archipelago. They were investigated for morphology, morphometrics and allozyme diversity after passage in immunosuppressed lambs using long acting corticoids. The basic aim of the work was to construct a synthetic strain in laboratory conditions which was representative of the isolates. The isolates were only slightly different although climatic conditions were very different. The resemblance of isolates might be due to the practice of goat exchanges between farms or to their insular origin. However the isolate from a smaller island (Les Saintes) was different (mostly on morphometrics) from all the others originating from Guadeloupe main Island. The first assemblage in laboratory resulting from the installation from a mixture of the five isolates was not very representative, whereas the next generation (synthetic strain) resembled all the isolates as shown from allozyme study. Female fecundity and length in established synthetic strain were lower than that recorded in isolates, indicating a decrease in fitness, possibly due to the stability of experimental environment. The representativity of the synthetic strain was good but the strain could still evaluate on further passages and should be evaluated on a large number of generations maintained in laboratory.     

Antagonistic effect of enterocin CCM 4231 from Enterococcus faecium on "bryndza", a traditional Slovak dairy product from sheep milk

"Bryndza" is a traditional Slovak dairy product (type of soft cheese) made from sheep cheese which was ripened for 14 days. Because its manufacture, transporting and/or storing represent conditions which facilitate contamination, the effect of enterocin CCM4231 in "bryndza" was investigated with the aim to reduce the contaminant agents. "Bryndza" was divided into equal portions (50 g). The experimental sample (ES) as well as the control sample one (C1) were inoculated with Listeria innocua Li1 strain. The other control samples C2 and C3 were without Li1 strain. C3 control was selected as a reference control. ES and C2 portions were treated with purified enterocin CCM4231 in a concentration of 6400 AU/ml. Before the experimental inoculation, "bryndza" was checked for the presence of contaminant agents. The experiment lasted 1 week and the samples were stored in the refrigerator at 4 degrees C. Sampling was performed on day 1, on day 4 and on day 7. The control samples C2 and C3 were checked only on day 1 and then after 1 week. The following contaminant agents were detected in "bryndza" before its experimental inoculation with L. innocua Li1 strain: Escherichia coli in the amount 10(3) cfu/ml/g, Staphylococcus aureus (10(2) cfu/ml/g) and enterococci (10(4) cfu/ml/g). In the control sample C2, the number of E. coli was reduced to 10(2) cfu/ml/g. Enterococci and staphylococci were totally eliminated there. Concerning C3 control, natural decrease of bacteria was found and/or their unchanged counts. The value of pH (5) was stable during the whole experiment. In the experimental sample inoculated with Li1 strain, its counts were decreased immediately after enterocin CCM4231 addition approximately by one order of magnitude. This inhibitory effect was also detectable on day 4 by the difference of one order of magnitude between ES and C1. On day 7, 10(3) cfu/ml/g of Li1 strain were detected in both samples (ES, C1). The difference by one order of magnitude indicated, an inhibitory effect of enterocin CCM4231 in "bryndza". However, bacteriocin activity was not determined by laboratory analyses.     

The activity of streptomyces phage-virus in fresh water

Streptomyces (Actinomycetales) host strains and their virulent phages were obtained from a restricted locality, both fresh waters and soils being sampled. There was some evidence that one category of host strains was confined to fresh water and was susceptible to phage there. There were several instances where a particular host strain elicited phages both from fresh water and from soil and the phages from these two environments subsequently exhibited differing polyvalency ranges in the laboratory. The possible significance of this finding is that it hints at the existence of phage ecotypes, with properties and polyvalency ranges adapted to the immediate environment.     

Isolation and Characterization of a Fucoidan-Degrading Marine Bacterial Strain and Its Fucoidanase

A marine bacterial strain that degraded fucoidan from Kjellmaniella crassifolia (class Phaeophyceae, order Laminariales, family Laminariaceae) was isolated in our laboratory. The strain was gram-negative, ubiquinone 8 was the predominant respiratory quinone, and the GC-content of its genomic DNA was 36%. The cells of the strain were rod-shaped (2.0 m long × 1.0 m wide), and each cell was motile by means of one polar flagellum. Phylogenetic analysis of its 16S ribosomal DNA sequence indicated that it was a member of the family Alteromonadaceae. It produced a type of extracellular fucoidanase, an endosulfated fucan-digesting enzyme. The enzyme was purified with 3500-fold purity at 12.0% yield. Optimum conditions for the enzyme reaction were approximately pH 6.5 to 8.0 and temperature 30° to 35°C. The enzyme was activated by calcium ions, and maximum activity was observed in the presence of greater than 30 mM calcium ion.     

The basis of decreased recombination in certain outcrosses of Neurospora crassa.

Crossing over in a multiply marked segment of linkage group I was conspicuously reduced in outcrosses between a marked laboratory strain and each of six unrelated wild-collected strains, compared with crosses between inbred laboratory strains. The marked chromosome segment was transferred intact from the inbred strain to one of the wild-collected strains by seven recurrent backcrosses, and conversely, the corresponding segment of the wild strain was transferred to the inbred background by backcrossing to the multiply marked laboratory strain. Recombination was then monitored in crosses from parents having the marked and unmarked chromosome segments from the same or from unrelated sources. Meiotic crossing-over in the marked segment remained low in crosses between parents that were dissimilar with respect to genetic background, but crossing over was restored to a high level when the genetic background of both parents was that of the inbred laboratory strain, regardless of the source of the marked segments. Reduced recombination in outcrosses was therefore not due to heterologies in the marked segment but must be attributed to modifiers that are unlinked or distant from the monitored region.     

The use of mitochondrial mutants in the isolation of hybrids involving industrial yeast strains

Summary Methods for the isolation of hybrids in which one or both of the parental strains are industrial yeasts, using mitochondrial mutations as markers for the selection and isolation of the hybrids, are described. The systems used included crosses of industrial strains with auxotrophic laboratory strains which also carried a mitochondrial antibiotic resistance mutation, crosses using an auxotrophic laboratory strain and a petite mutant of an industrial strain carrying a rescuable antibiotic resistance mutation, and crosses using a petite mutant of an industrial strain, carrying a rescuable mitochondrial mutation for antibiotic resistance and a respiratory-competent industrial strain which carried some other marker.     

Heritability of flight distance for Cydia pomonella

Cydia pomonella L. (Lepidoptera: Tortricidae) is considered to be rather sedentary, but some individuals undertake flights of several kilometres in the field. This paper investigates the genetic influence on this variability. The flight capacity was measured in the laboratory by a flight mill and its heritability was estimated for two different strains. The laboratory strain was kept for more than 45 generations and the field strain from Embrach (northern Switzerland) was recently collected in the field.The multiple-trait-restricted-maximum-likelihood method was used for the estimation of genetic variances and covariances. A mixed full-sib/half-sib design was applied for the field strain and a full-sib design for the laboratory strain. The heritability of total distance was 0.57 for the field strain and 0.37 for the laboratory strain (both sexes). In addition, a heritability of 0.38 for total distance was estimated by parent-offspring regression for the laboratory strain. All three values were significantly different from zero P<0.05 and show that there is a significant additive genetic influence on flight capacity.The genetic correlations between total distance and other flight traits (total duration, flight velocity, longest flight) were between 0.84 and 1.00 for both strains and suggest that these traits actually belong to a single one. High genetic correlations were also found between total distance and the morphological traits body weight and wing length for the field strain, whereas a negative correlation was found between total flight distance and body weight for the laboratory strain. This difference between the two strains was interpreted as a possible trade-off between flight capacity and fecundity.     

The use of theVibrio harveyi luminescence mutagenicity assay as a rapid test for preliminary assessment of mutagenic pollution of marine sediments

Mutagenic pollution of the natural environment is currently one of the most serious environmental problems. It includes the pollution of marine sediments. Therefore, rapid detection of the presence of mutagens is an important issue. Recently, we have developed a novel microbiological assay for rapid assessment of mutagenicity of samples from the natural environment. This assay is based on bioluminescence of a mutant Vibrio harveyi strain, and was shown to be useful in testing samples of marine water and plant tissues. Here we demonstrate the usefulness of this assay in preliminary assessment of mutagenic pollution of marine sediments. Mutagenicity of environmental samples taken from the Baltic Sea, is documented and compared here with a commercially available standard sediment sample (IAEA 383), which contains known amounts of mutagenic compounds. The whole procedure, from obtaining a sample in the laboratory to getting final results, is very short (less than 4 h).