Species and biotype identification ofSerratia strains associated with insects

Forty-eightSerratia strains associated with insects were, identified to species level and biotyped according to recent taxonomic schemes. Each strain was submitted to 36 biochemical tests, including 23 carbon source utilization tests. Twenty-eight strains were assigned to eight biotypes ofSerratia marcescens: A1a, A2a, and A6a (pigmented biotypes: 18 strains); and A3a, A3b, A4a, A5, and TCT (nonpigmented biotypes: 10 strains). However, biotypes A8a, A8b, and A8c, which are frequently involved in nosocomial infections, were not found in insects. Ninetten strains were identified asS. liquefaciens (S. proteamaculans) biotypes C1a (12 strains), C1c (4 strains), C1d (2 strains), and one atypicalS. liquefaciens strain. Only one strain was identified asS. marinorubra (a nonchitinolytic species). The recent emergence ofSerratia in human pathology calls for a reevaluation of the idea of usingSerratia to biologically control insects.     

Serratia ficaria sp. nov., a bacterial species associated with Smyrna figs and the fig waspBlastophaga psenes

Fourteen strains isolated from figs, caprifigs, and fig wasps collected in California and Tunisia, and from a small black ant in France, constitute a new DNA hybridization group that is 25–56% related toSerratia species, and 6–17% related to other species of Enterobacteriaceae. This homogeneous group (90% relatedness within the group) constitutes a new species that is namedSerratia ficaria sp. nov. (type strain, ICPB 4050, ATCC 33105). Strains of this species have a characteristic odor, similar to that ofS: odorifera andPseudomonas perolens. No strain ofS. ficaria has yet been recovered from clinical specimens.     

TheSerratia liquefaciens-S. proteamaculans-S. grimesii complex: DNA relatedness

Serratia liquefaciens andS. liquefaciens-like strains belonging to one of seven biotypes (C1ab, C1c, C1d, EB, RB, RQ, and Adc) were characterized by DNA relatedness (S1TCA method). These strains formed three distinct DNA relatedness groups: (i)S. liquefaciens sensu stricto (biotype C1ab); (ii)S. proteamaculans (biotypes C1c, EB, and RB); and (iii)S. grimesii (biotype C1d). Two biotypes were at the borderline of species level: Biotype RQ and biotype Adc were, respectively, related toS proteamaculans andS. grimesii. All of these strains were clearly distinct fromS. plymuthica and the otherSerratia species.     

Novel Endophytes of Rice form a Taxonomically Distinct Subgroup of Serratia marcescens

Six endophytic strains isolated from surface-sterilized rice roots and stems of different rice varieties grown in the Philippines were characterized. They were analyzed by physiological and biochemical tests, SDS-PAGE of whole-cell protein patterns, DNA-DNA hybridization and 16S rDNA sequencing. SDS-PAGE of whole-cell patterns showed that the six isolates fell into two subgroups which were similar but not identical in protein patterns to S. marcescens. The phylogenetic analysis of 16S rDNA sequences of two representative strains IRBG 500 and IRBG 501 indicated that they were closely related to S. marcescens(more than 99% identity). Physiological and biochemical tests corroborated that the isolates were highly related to each other and to S. marcescens. In cluster analysis, all six isolates were clustered together at 93% similarity level and grouped closely with Serratia marcescens at 86% similarity level. DNA-DNA hybridization studies revealed that the isolates shared high similarity levels with S. marcescens(≥86% DNA-DNA binding), indicating they belong to the same species. However, the isolates differed in several biochemical characteristics from the type strain. They produce urease and utilize urea and L(+) sorbose as a substrate, which is different from all known Serratia reference strains. These results suggest that the six endophytic isolates represent a novel, non-pigmented subgroup of S. marcescens.     

Isolation and characterization of a Serratia marcescens with insecticidal activity from Polyphylla olivieri (Col.: Scarabaeidae)

Members of the genus Serratia are known for their abilities to infect insects. In this study, a red‐pigmented S. marcescens was isolated and characterized from the infected larvae of Polyphylla olivieri using bacterial cultivation, phylogenetic analysis as well as bioassays against larvae of the two insect pests, Plodia interpunctella and Ephestia kuehniella. Comparative 16S rRNA and groEL gene sequence BLAST analyses strongly suggested that the isolated strain should be placed in the genus Serratia, sharing high sequence similarities with several strain of S. marcescens associated with insects. Phylogenetic analysis placed the isolated bacterium with other S. marcescens bacteria in a clade with high bootstrapping values. To assess pathogenicity of the S. marcescens isolate, the bacterial cells were either injected into the haemolymph of the fifth‐instar larvae or added to the diets of insects. Survival curves of the control insects and those challenged with six different concentrations of S. marcescens showed that the S. marcescens isolate significantly reduced survival rates of the larvae. The LC₅₀s of the bacterium on P. interpunctella and E. kuehniella were 1992.26 and 1.09 × 10⁴ (CFU/μl) for injection bioassays at 6 h post‐injection, and 4.48 × 10⁴ and 1.96 × 10⁵ (CFU/10 μl) for feeding bioassays at 24 h post‐feeding, respectively. Injection of the bacterial culture supernatant into the larvae led to continuous bleeding from the site of injection, while injection of heat‐treated culture supernatant of the bacterium did not cause continuous bleeding. Together, our results showed the possibility of using this S. marcescens isolate in microbial control of the insect pests after addressing the safety concerns. Moreover, it might be considered as a source of useful bioactive molecules and genes with application in insect control and biotechnology via developing insect‐resistant plants.     

Isolation and identification of a novel endophytic bacterial strain with antifungal activity from wild blueberryVaccinium uliginosum

A strain of endophytic bacterium with a good anti-fungal ability was first isolated fromVaccinium uliginosum. It was identified asSerratia marcescens using the 16s rDNA sequence homology and through its physiological and biochemical characteristics. Through the alignment and cladistic analysis of homologous nucleotide sequences of knownSerratia marcescens, it was found to be a novel subspecies ofS. marcescens. An extensive suppressive spectrum of this endophytic strain ofS. marcescens against plant pathogenic fungi was observed. In particular, it inhibited the growth of the causal agents of blueberry leaf spot and of other ten plant fungal pathogens. The inhibition rate, among the different fungi, ranged from 22.7–70.5%. The study indicated the potential use of thisS. marcescens endophytic strain in the biological control of blueberry’s leaf spot as well as many other plant diseases. To our knowledge, this was the first time that an endophyticS. marcescens was isolated from a wild blueberry in China, and may also represent a novel introduction to the world.     

Coliforms and enterococci isolated from the intestinal tract of conventional mice

Coliforms and enterococci were isolated from the intestinal tract of infant (12-day-old) and adult (6-to 8-week-old) conventional mice. Eighty coliform isolates and eighty enterococcal strains were grouped according to their ability to ferment or hydrolyze various substrates. Sixty-one of the coliform isolates were identified asEscherichia coli. The remaining 19 strains were similar toE. coli, but did not produce-galactosidase. The enterococci belonged to two species:Streptococcus faecium andS. faecalis. Four biotypes ofS. faecium and two biotypes ofS. faecalis were detected. Xylosefermenting enterococci were isolated with a higher frequency from infant mice than from adults.     

Serratia marcescens SYBC08 catalase isolated from sludge containing hydrogen peroxide shows increased catalase production by regulation of carbon metabolism

A high‐catalase‐producing strain, which was isolated from sludge containing hydrogen peroxide, was identified as Serratia marcescens SYBC08 by 16S rDNA sequence analysis. Serratia spp. was reported as non‐spore‐forming bacterium (except S. marcescens spp. sakuensis), but in our study electron microscopic observation revealed that the strain did produce spores. The content of the main fatty acid C_16:0 (14.8%) was significantly different from that of S. marcescens spp. sakuensis (33.2%) and S. marcescens spp. marcescens DSM 30121^T (34.8%), and the biochemical characteristics were not identical to those of S. marcescens spp. sakuensis. We speculate that the relatively high catalase activity and the spore structures may enable the strain to survive in a hydrogen peroxide environment. The most suitable carbon and nitrogen sources for the catalase production by S. marcescens SYBC08 were citric acid and corn steep liquor powder. A strategy of carbon metabolism regulation to enhance the catalase production was exploited. In the 7‐L fermenter, catalase production (20 353 U/mL) obtained in the presence of glucose and citric acid was 1.68‐ and 1.31‐fold higher than that obtained in the presence of glucose or citric acid, at equimolar carbon concentration. This production yield was much higher than that of many catalase‐producing strains, but only slightly lower than the production by Micrococcus luteus (34 601 U/mL). The results suggest that the new spore‐forming S. marcescens SYBC08 is a potential candidate for the production of catalase.     

Enterobacteria of emerging pathogenic significance from clinical cases in man and animals and detection of toads and wall lizards as their reservoirs

A total of 416 samples comprising faecal samples from diarrhoeic cases of man, calves, sheep and goats, and urine samples from patients with urinary tract infections, were examined for the presence of enterobacteria of emerging pathogenic significance.Citrobacter freundii from 20,C. intermedius biotype-a from four,Serratia marcescens (serotype 05:H13, bactericin type 16) from one andErwinia herbicola from two human stool samples were isolated. Only two urine samples yieldedC. freundii. Citrobacter freundii was isolated from 10 andC. intermedius biotype-a from two calves. From sheep and goats, two isolates ofC. freundii and three ofC. intermedius biotype-a were obtained. None of these samples yieldedEdwardsiella tarda orYersinia enterocolitica. The examination of 99 toads and 145 wall lizards revealed that toads were reservoirs forC. freundii, C. intermedius biotype-a andSalmonella brijbhumi, whereas wall lizards were reservoirs forC. freundii, C. intermedius biotype-a,E. herbicola, Enterobacter cloacae andSalmonella spp. These bacteria were present in the range of 2.0 × 10⁶ to 6.0 × 10¹¹ organisms per gram of intestinal contents. In addition, toads were carriers forEdwardsiella tarda (new serotypes 04167:H1 and 05159: nonmotile). None of the toads and wall lizards proved positive forC. intermedius biotype-b (C. koseri),S. marcescens andY. enterocolitica. C. freundii, C. intermedius biotype-a,E. herbicola andS. marcescens were resistant to penicillin and erythromycin whereasE. tarda isolates were also resistant to gentamycin, neomycin, colistin and sulfamethaxazole.     

Lack of genetic relatedness among animal and plant acholeplasmas by nucleic acid hybridization

The present study compared the genetic characteristics of three previously unclassified acholeplasmas of plant origin with those of the eight established species ofAcholeplasma. A radiolabeled DNA probe was derived from the lemon (L1) strain acholeplasma by using the nick translation method and hybridized to the excess unlabeled DNA isolated from the eight established species ofAcholeplasma and three new isolates from plants. Nucleic acid hydridization and serological tests confirmed that the L1 acholeplasma was distinct from all eight established species ofAcholeplasma but closely related to strains isolated from the flowers of grapefruit trees (GF1) and powder puff plants (PP2). The results suggest that these new acholeplasmas isolated exclusively from plants may represent a new species ofAcholeplasma.     

Hemolytic activity of Serratia marcescens

A cell-bound hemolytic activity was found in several strains of Serratia marcescens. One Serratia cell per ten erythrocytes was sufficient to cause complete lysis of human erythrocytes within 2 h in the liquid assay. The hemolytic activity resided in the membrane fraction and could be inactivated by incubating cells with proteases. The hemolytic activity was greatly enhanced in actively metabolizing Serratia cells and was partially controlled by the iron supply. Hemolysis was accompanied by degradation of erythrocyte membrane proteins (band 3 and 6, glycophorin) and was independent of the blood group. The exoprotease secreted by S. marcescens in large amounts was not involved in hemolysis. Comparison with various hemolytic strains of Escherichia coli showed that hemolysis of erythrocytes was more pronounced with S. marcescens than with E. coli. In contrast to hemolysis by E. coli, lysis of erythrocytes by S. marcescens was not enhanced by Ca²⁺ ions.Dedicated to Professor Dr. Gerhart Drews on the occasion of his 60th birthday     

Hemolysin as a marker for Serratia

All Serratia marcescens strains (total of 33) of different sources were hemolytic including clinical strains previously classified as being nonhemolytic. DNA fragments of the two hemolysin genes hybridized with the chromosomal DNA of S. marcescens, S. liquefaciens, S. kiliensis, S. grimesii, S. proteamaculans, S. plymutica, S. rubridaea which were also hemolytic. The restriction pattern of the hemolysin locus differed in each strain. S. ficaria and S. marinorubra expressed a different hemolysin which was much smaller than the S. marcescens hemolysin since it diffused through dialysis membranes. The DNA of the latter strains did not hybridize with the S. marcescens hemolysin DNA probes. Some S. marcescens strains, S. kiliensis and S. liquefaciens also expressed in addition the small hemolysin. No hybridization was found with DNA of Escherichia coli, Salmonella typhimurium, Proteus mirabilis, Proteus vulgaris, Citrobacter freundii, Enterobacter cloacae, Klebsiella arerogenes, Klebsiella pneumoniae, Shigella dysenteriae, Yersinia enterocolitica, Yersinia pseudotuberculosus, Listeria sp., Aeromonas sp., Legionella sp. and a Meningococcus sp., indicating that the hemolysin DNA probes are specific for Serratia, or that the hemolysin genes occur rarely in genera other than Serratia.     

Biochemical,serological, and phage typing characteristics of 459Yersinia strains isolated from a terrestrial ecosystem

The origins of human contamination withYersinia enterocolitica are still unknown. We have investigated the major components of a terrestrial ecosystem (soil, earthworms, field voles, shrews, crops, hares, rabbits, and birds) for the presence ofYersinia. Four hundred fifty-nine strains ofYersinia were isolated. We report the first isolations of typicalY. enterocolitica belonging to classical or new biotypes and ofY. enterocolitica-like organisms (sucrose negative; rhamnose positive; melibiose and rhamnose positive) from soil samples, earthworms, crops, and birds. Sucrose-negativeY. enterocolitica strains and biotypes 1, 2, and 3, usually associated with human nonmesenteric syndromes, are predominant in soil, which can be considered as a reservoir for these biotypes.Y. enterocolitica serogroups O∶3 and O∶9, strains of which are responsible in Europe for human mesenteric syndromes, were not found in this study. The epidemiology ofY. enterocolitica infections is discussed.     

Selective blockage of Serratia marcescens ShlA by nickel inhibits the pore‐forming toxin‐mediated phenotypes in eukaryotic cells

Serratia marcescens is an opportunistic pathogen with increasing incidence in clinical settings. This is mainly attributed to the timely expression of a wide diversity of virulence factors and intrinsic and acquired resistance to antibiotics, including β‐lactams, aminoglycosides, quinolones, and polypeptides. For these reasons, S. marcescens has been recently categorised by the World Health Organization as one priority to strengthen efforts directed to develop new antibacterial agents. Therefore, it becomes critical to understand the underlying mechanisms that allow Serratia to succeed within the host. S. marcescens ShlA pore‐forming toxin mediates phenotypes that alter homeostatic and signal transduction pathways of host cells. It has been previously demonstrated that ShlA provokes cytotoxicity, haemolysis and autophagy and also directs Serratia egress and dissemination from invaded nonphagocytic cells. However, molecular details of ShlA mechanism of action are still not fully elucidated. In this work, we demonstrate that Ni²⁺ selectively and reversibly blocks ShlA action, turning wild‐type S. marcescens into a shlA mutant strain phenocopy. Combined use of Ni²⁺ and calcium chelators allow to discern ShlA‐triggered phenotypes that require intracellular calcium mobilisation and reveal ShlA function as a calcium channel, providing new insights into ShlA mode of action on target cells.     

Size-dependent foraging efficiency,cannibalism and zooplankton community structure

Only recently has the importance of positive interactions among plant species in structuring natural communities been supported by experimental evidence. Most studies have focused on interactions between a pair of species at a single life-history stage. In this study positive interactions between a woody nitrogen-fixing shrub (Myrica pensylvanica) and two herbaceous sand dune species (Solidago sempervirens, Ammophila breviligulata) which frequently grow beneath shrub canopies are examined throughout the life cycles of the herbaceous species. Comparisons of S. sempervirens and A. breviligulata growing beneath and outside M. pensylvanica shrubs show that plants growing in association with shrubs are larger, are more likely to flower, produce greater numbers of flowers and seeds, have higher midday xylem water potentials, have higher tissue nitrogen concentrations, and have higher photosynthetic efficiencies. Measurements of environmental conditions show that areas beneath shrubs are more shaded, have lower soil temperatures, and have higher soil nitrogen levels. The results from experimental manipulations designed to test the effects of Myrica shrubs on understory species suggest that the observed differences in plant performance are strongly influenced by canopy shading and soil nutrient enrichment associated with the shrubs. The results demonstrate that M. pensylvanica facilitates growth, reproduction, and recruitment of S. sempervirens and A. breviligulata growing beneath it. This study, one of the few to examine positive interactions at different life-history stages, supports previous predictions that positive interactions may be particularly important in plant communities characterized by physiologically stressful conditions. Received: 21 July 1999 / Accepted: 18 January 2000     

Isolation and characterization of novel Serratia marcescens (AY927692) for pentachlorophenol degradation from pulp and paper mill waste

Seven aerobic bacterial strains were isolated from pulp paper mill waste and screened for pentachlorophenol (PCP) tolerance on PCP containing mineral salt agar medium (MSM). The organism was characterized by 16S rDNA sequencing which showed 99.7% sequence similarity with Serratia marcescens. PCP degradation was routinely monitored with spectrophotometric analysis and further confirmed by HPLC analysis. Among seven strains, ITRC S₇ was found to degrade up to 90.33% of 1.127 mM (300 mg/l) of PCP and simultaneous release of chloride ion (2.435 mM) emphasized the bacterial dechlorination in the medium in presence of glucose as an additional carbon and energy source under optimized condition within 168 h incubation. In absence of glucose bacterium was unable to utilize PCP indicating the phenomenon of co-metabolism. Bacterium was identified as S. marcescens (AY927692), was a novel and potential aerobic bacterial strain capable of degrading PCP in axenic condition. Further, this strain may be used for bioremediation of PCP containing pulp paper mill waste in the environment.     

Biological comparisons between arrhenotokous and thelytokous biotypes ofMesochorus nigripes [Hym.: Ichneumonidae]

The host preferences and sex ratios ofMesochorus biotypes from Sweden and Colorado (U.S.A.) were studied in the field and laboratory. TheseMesochorus are secondary parasites ofBathyplectes spp., parasites ofHypera postica (Gyllenhal). The Colorado biotype is 100% thelytokous, and clearly prefersBathyplectes stenostigma (Thomson) as a host. Only an arrhenotokous biotype was positively detected in Sweden, and it appeared to preferB. curculionis (Thomson). However, some of the data suggest that the thelytokous biotype was probably also present in Sweden, at least inB. stenostigma. The behavioral differences and reproductive isolation of the thelytokous and arrhenotokous biotypes indicate that they are separate sibling species, even though they cannot yet be separated morphologically, and isozyme analyses were inconclusive (possibly due to a mixture of biotypes in Sweden).      

Genetic studies of the ribosomal proteins inEscherichia coli

Summary Episomes ofE. coli K12, which coverthrleu region of the chromosome, were transferred toSerratia marcescens. Ribosomal proteins from these hybrid strains were analyzed with phosphocellulose column chromatography. TwoE. coli 30S ribosomal proteins, S2 and S20, could be detected in the ribosome of the hybrid strain in addition to all ribosomal proteins ofS. marcescens.