scholarly journals Characterization of PaxA and Its Operon: a Cohemolytic RTX Toxin Determinant from Pathogenic Pasteurella aerogenes

2000 ◽  
Vol 68 (1) ◽  
pp. 6-12 ◽  
Author(s):  
Peter Kuhnert ◽  
Bénédicte Heyberger-Meyer ◽  
Jacques Nicolet ◽  
Joachim Frey
Keyword(s):  
Sequence Similarity ◽  
Opportunistic Pathogen ◽  
Toxin Gene ◽  
Rrna Gene ◽  
Commensal Bacterium ◽  
Pathogenic Potential ◽  
Newborn Piglets ◽  
Toxin Genes ◽  
Rtx Toxin

ABSTRACT Pasteurella aerogenes is known as a commensal bacterium or as an opportunistic pathogen, as well as a primary pathogen found to be involved in abortion cases of humans, swine, and other mammals. Using broad-range DNA probes for bacterial RTX toxin genes, we cloned and subsequently sequenced a new operon namedpaxCABD encoding the RTX toxin PaxA in P. aerogenes. The pax operon is organized analogous to the classical RTX operons containing the activator genepaxC upstream of the structural toxin genepaxA, which is followed by the secretion protein genespaxB and paxD. The highest sequence similarity of paxA with known RTX toxin genes is found withapxIIIA (82%). PaxA is structurally similar to ApxIIIA and also shows functional analogy to ApxIIIA, since it shows cohemolytic activity with the sphingomyelinase of Staphylococcus aureus, known as the CAMP effect, but is devoid of direct hemolytic activity. In addition, it shows to some extent immunological cross-reactions with ApxIIIA. P. aerogenes isolated from various specimens showed that the pax operon was present in about one-third of the strains. All of the pax-positive strains were specifically related to swine abortion cases or septicemia of newborn piglets. These strains were also shown to produce the PaxA toxin as determined by the CAMP phenomenon, whereas none of thepax-negative strains did. This indicated that the PaxA toxin is involved in the pathogenic potential of P. aerogenes. The examined P. aerogenes isolates were phylogenetically analyzed by 16S rRNA gene (rrs) sequencing in order to confirm their species. Only a small heterogeneity (<0.5%) was observed between the rrs genes of the strains originating from geographically distant farms and isolated at different times.

scholarly journals Identification of Novel Toxin Genes from the Stinging Nettle Caterpillar Parasa lepida (Cramer, 1799): Insights into the Evolution of Lepidoptera Toxins

Insects ◽  
2021 ◽  
Vol 12 (5) ◽  
pp. 396
Author(s):  
Natrada Mitpuangchon ◽  
Kwan Nualcharoen ◽  
Singtoe Boonrotpong ◽  
Patamarerk Engsontia
Keyword(s):  
Protease Inhibitors ◽  
Proteolytic Enzymes ◽  
Gene Annotation ◽  
Sequence Similarity ◽  
New Drugs ◽  
Toxin Gene ◽  
Cone Snail ◽  
Stinging Nettle ◽  
Toxin Genes ◽  
Nettle Caterpillar

Many animal species can produce venom for defense, predation, and competition. The venom usually contains diverse peptide and protein toxins, including neurotoxins, proteolytic enzymes, protease inhibitors, and allergens. Some drugs for cancer, neurological disorders, and analgesics were developed based on animal toxin structures and functions. Several caterpillar species possess venoms that cause varying effects on humans both locally and systemically. However, toxins from only a few species have been investigated, limiting the full understanding of the Lepidoptera toxin diversity and evolution. We used the RNA-seq technique to identify toxin genes from the stinging nettle caterpillar, Parasa lepida (Cramer, 1799). We constructed a transcriptome from caterpillar urticating hairs and reported 34,968 unique transcripts. Using our toxin gene annotation pipeline, we identified 168 candidate toxin genes, including protease inhibitors, proteolytic enzymes, and allergens. The 21 P. lepida novel Knottin-like peptides, which do not show sequence similarity to any known peptide, have predicted 3D structures similar to tarantula, scorpion, and cone snail neurotoxins. We highlighted the importance of convergent evolution in the Lepidoptera toxin evolution and the possible mechanisms. This study opens a new path to understanding the hidden diversity of Lepidoptera toxins, which could be a fruitful source for developing new drugs.

scholarly journals Taxogenomics of the Genus Cyclobacterium: Cyclobacterium xiamenense and Cyclobacterium halophilum as Synonyms and Description of Cyclobacterium plantarum sp. nov.

2020 ◽  
Vol 8 (4) ◽  
pp. 610 ◽  
Author(s):  
Azadeh Shahinpei ◽  
Mohammad Ali Amoozegar ◽  
Leila Mirfeizi ◽  
Mahdi Moshtaghi Nikou ◽  
Antonio Ventosa ◽  
...  
Keyword(s):  
Sequence Similarity ◽  
Threshold Value ◽  
Amino Acid Identity ◽  
Rrna Gene ◽  
Unidentified Phospholipid ◽  
Average Amino Acid Identity ◽  
Halophytic Plant ◽  
Taxonomic Characterization ◽  
Current Species

The genus Cyclobacterium belongs to the phylum Bacteroidetes and includes eight species. Our study, based on the genomic parameters in silico DNA–DNA hybridization (GGDC), average nucleotide identity (OrthoANI), and average amino acid identity (AAI), confirmed that all current species of Cyclobacterium belong to this genus and constitute a coherent phylogenomic group, but with species forming two separate branches. In addition, the genome-based analyses revealed that Cyclobacterium xiamenense and Cyclobacterium halophilum are members of the same species. Besides, we carried out a taxonomic characterization of the new strain GBPx2T, isolated from the halophytic plant Salicornia sp. Analysis of its 16S rRNA gene sequence showed the highest sequence similarity (97.5%) to Cyclobacterium lianum HY9T. Percentages of GGDC and OrthoANI between strain GBPx2T and species of the genus Cyclobacterium were lower than the threshold value for species delineation. The DNA G+C content was 43.0 mol%. The polar lipids included phosphatidylethanolamine as well as one unidentified phospholipid and four unidentified lipids, and its major cellular fatty acids were iso-C15:0 and summed feature 3 (C16:1ω7c and/or iso-C15:0 2-OH). The only quinone present was menaquinone 7. Based on a combination of phenotypic, chemotaxonomic, and phylogenomic features, the GBPx2T strain represents a novel species of the genus Cyclobacterium, for which the name Cyclobacterium plantarum sp. nov. is proposed. The type strain of Cyclobacterium plantarum is GBPx2T (= IBRC-M 10634T = LMG 28551T).

Isolation and Characterization of Soil Myxobacteria from Nepal

2019 ◽  
Vol 24 (2) ◽  
pp. 7-16
Author(s):  
Nabin Rana ◽  
Saraswoti Khadka ◽  
Bishnu Prasad Marasini ◽  
Bishnu Joshi ◽  
Pramod Poudel ◽  
...  
Keyword(s):  
Sequence Similarity ◽  
Antimicrobial Properties ◽  
Antimicrobial Activities ◽  
Rrna Gene ◽  
Isolation And Characterization ◽  
The Family ◽  
Global Concern ◽  
Bacteria And Fungi ◽  
Antimicrobial Metabolites

 Realizing myxobacteria as a potential source of antimicrobial metabolites, we pursued research to isolate myxobacteria showing antimicrobial properties. We have successfully isolated three strains (NR-1, NR-2, NR-3) using the Escherichia coli baiting technique. These isolates showed typical myxobacterial growth characteristics. Phylogenetic analysis showed that all the strains (NR-1, NR-2, NR-3) belong to the family Archangiaceae, suborder Cystobacterineae, and order Myxococcales. Furthermore, 16S rRNA gene sequence similarity searched through BLAST revealed that strain NR-1 showed the closest similarity (91.8 %) to the type strain Vitiosangium cumulatum (NR-156939), NR-2 showed (98.8 %) to the type of Cystobacter badius (NR-043940), and NR-3 showed the closest similarity (83.5 %) to the type of strain Cystobacter fuscus (KP-306730). All isolates showed better growth in 0.5-1 % NaCl and pH around 7.0, whereas no growth was observed at pH 9.0 and below 5.0. All strains showed better growth at 32° C and hydrolyzed starch, whereas casein was efficiently hydrolyzed by NR-1 and NR-2. Besides, preliminary antimicrobial tests from crude extracts showed activities against Gram-positive, Gram-negative bacteria, and fungi. Our findings suggest that the arcane soil habitats of Nepal harbor myxobacteria with the capability to produce diverse antimicrobial activities that may be explored to overcome the rapidly rising global concern about antibiotic resistance.

scholarly journals Mycobacterium celeriflavum sp. nov., a rapidly growing scotochromogenic bacterium isolated from clinical specimens

2015 ◽  
Vol 65 (Pt_2) ◽  
pp. 510-515 ◽  
Author(s):  
Abdolrazagh Hashemi Shahraki ◽  
Cengiz Çavuşoğlu ◽  
Emanuele Borroni ◽  
Parvin Heidarieh ◽  
Orhan Kaya Koksalan ◽  
...  
Keyword(s):  
Type Strain ◽  
Type Species ◽  
Sequence Similarity ◽  
Housekeeping Genes ◽  
Mycolic Acid ◽  
Rrna Gene ◽  
Content Type ◽  
Link Type ◽  
Polyphasic Characterization

Six strains of a rapidly growing scotochromogenic mycobacterium were isolated from pulmonary specimens of independent patients. Biochemical and cultural tests were not suitable for their identification. The mycolic acid pattern analysed by HPLC was different from that of any other mycobacterium. Genotypic characterization, targeting seven housekeeping genes, revealed the presence of microheterogeneity in all of them. Different species were more closely related to the test strains in various regions: the type strain of Mycobacterium moriokaense showed 99.0 % 16S rRNA gene sequence similarity, and 91.5–96.5 % similarity for the remaining six regions. The whole genome sequences of the proposed type strain and that of M. moriokaense presented an average nucleotide identity (ANI) of 82.9 %. Phylogenetic analysis produced poorly robust trees in most genes with the exception of rpoB and sodA where Mycobacterium flavescens and Mycobacterium novocastrense were the closest species. This phylogenetic relatedness was confirmed by the tree inferred from five concatenated genes, which was very robust. The polyphasic characterization of the test strains, supported by the ANI value, demonstrates that they belong to a previously unreported species, for which the name Mycobacterium celeriflavum sp. nov. is proposed. The type strain is AFPC-000207T ( = DSM 46765T = JCM 18439T).

scholarly journals Identification and Characterization of Hemolysin-Like Proteins Similar to RTX Toxin in Pasteurella pneumotropica

2009 ◽  
Vol 191 (11) ◽  
pp. 3698-3705 ◽  
Author(s):  
Hiraku Sasaki ◽  
Eiichi Kawamoto ◽  
Yoshikazu Tanaka ◽  
Takuo Sawada ◽  
Satoshi Kunita ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Opportunistic Pathogen ◽  
Pcr Analysis ◽  
Type I ◽  
Secretion Systems ◽  
Rtx Toxin ◽  
Rtx Toxins ◽  
Genes Encoding ◽  
Encoding Genes

ABSTRACT Pasteurella pneumotropica is an opportunistic pathogen that causes lethal pneumonia in immunodeficient rodents. The virulence factors of this bacterium remain unknown. In this study, we identified the genes encoding two RTX toxins, designated as pnxI and pnxII, from the genomic DNA of P. pneumotropica ATCC 35149 and characterized with respect to hemolysis. The pnxI operon was organized according to the manner in which the genes encoded the structural RTX toxin (pnxIA), the type I secretion systems (pnxIB and pnxID), and the unknown orf. The pnxII gene was involved only with the pnxIIA that coded for a structural RTX toxin. Both the structural RTX toxins of deduced PnxIA and PnxIIA were involved in seven of the RTX repeat and repeat-like sequences. By quantitative PCR analysis of the structural RTX toxin-encoding genes in P. pneumotropica ATCC 35149, the gene expression of pnxIA was found to have increased from the early log phase, while that of pnxIIA increased from the late log to the early stationary phase. As expressed in Escherichia coli, both the recombinant proteins of PnxIA and PnxIIA showed weak hemolytic activity in both sheep and murine erythrocytes. On the basis of the results of the Southern blotting analysis, the pnxIA gene was detected in 82% of the isolates, while the pnxIIA gene was detected in 39%. These results indicate that the products of both pnxIA and pnxIIA were putative associations of virulence factors in the rodent pathogen P. pneumotropica.

scholarly journals Molecular diagnosis of Kingella kingae osteoarticular infections by specific real-time PCR assay

2009 ◽  
Vol 58 (1) ◽  
pp. 65-68 ◽  
Author(s):  
Abdessalam Cherkaoui ◽  
Dimitri Ceroni ◽  
Stéphane Emonet ◽  
Yan Lefevre ◽  
Jacques Schrenzel
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Cell Types ◽  
Cross Reactivity ◽  
Toxin Gene ◽  
Rrna Gene ◽  
Putative Virulence Factor ◽  
Pcr Assay ◽  
Kingella Kingae ◽  
Rtx Toxin

Kingella kingae is an emerging pathogen that is recognized as a causative agent of septic arthritis and osteomyelitis, primarily in infants and children. The bacterium is best detected by rapid inoculation in blood culture systems or by real-time PCR assays. Pathogenesis of the agent was linked recently to the production of a potent cytotoxin, known as RTX, which is toxic to a variety of human cell types. The locus encoding the RTX toxin is thought to be a putative virulence factor, and is, apparently, essential for inducing cytotoxic effects on respiratory epithelial, synovial and macrophage-like cells. Herein, we describe a novel real-time PCR assay that targets the RTX toxin gene and illustrate its use in two clinical cases. The assay exhibited a sensitivity of 30 c.f.u., which is 10-fold more sensitive than a previously published semi-nested broad-range 16S rRNA gene PCR, and showed no cross-reactivity with several related species and common osteoarticular pathogens.

scholarly journals Geobacillus lituanicus sp. nov.

2004 ◽  
Vol 54 (6) ◽  
pp. 1991-1995 ◽  
Author(s):  
Nomeda Kuisiene ◽  
Juozas Raugalas ◽  
Donaldas Chitavichius
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Novel Species ◽  
Sequence Similarity ◽  
Rrna Gene ◽  
The Novel ◽  
Gene Sequence Analysis ◽  
Pcr Rflp ◽  
Group 5

Obligately thermophilic, aerobic, proteolytic, endospore-forming strain N-3T was isolated from a high-temperature oilfield in Lithuania. 16S rRNA gene sequence analysis placed this strain in genetic group 5 of the endospore formers. Geobacillus thermoleovorans appeared to be the closest phylogenetic neighbour (99·4 % sequence similarity). The G+C content of strain N-3T was 52·5 mol% and matched the range established for the genus Geobacillus. Studies of DNA–DNA relatedness and morphological and physiological analyses enabled strain N-3T to be described as a member of the genus Geobacillus, but could not assign this strain to any other known species of this genus. Results of this polyphasic study allowed characterization of strain N-3T as a novel species in the genus Geobacillus – Geobacillus lituanicus sp. nov. This species can be distinguished from G. thermoleovorans and Geobacillus stearothermophilus on the basis of 16S rRNA gene PCR-RFLP assays with the restriction endonucleases AluI, HaeIII and TaqI. The type strain of the novel species is N-3T (=DSM 15325T=VKM B-2294T).

scholarly journals Six novel species of the obligate marine actinobacterium Salinispora, Salinispora cortesiana sp. nov., Salinispora fenicalii sp. nov., Salinispora goodfellowii sp. nov., Salinispora mooreana sp. nov., Salinispora oceanensis sp. nov. and Salinispora vitiensis sp. nov., and emended description of the genus Salinispora

2020 ◽  
Vol 70 (8) ◽  
pp. 4668-4682 ◽  
Author(s):  
Brenda Román-Ponce ◽  
Natalie Millán-Aguiñaga ◽  
Dulce Guillen-Matus ◽  
Alexander B. Chase ◽  
Joape G.M. Ginigini ◽  
...  
Keyword(s):  
Sequence Similarity ◽  
Taxonomic Status ◽  
Phenotypic Characterization ◽  
Comparative Genomic ◽  
Rrna Gene ◽  
Content Type ◽  
Link Type ◽  
Physiological And Biochemical Characteristics ◽  
Genome Phylogeny

Ten representative actinobacterial strains isolated from marine sediments collected worldwide were studied to determine their taxonomic status. The strains were previously identified as members of the genus Salinispora and shared >99 % 16S rRNA gene sequence similarity to the three currently recognized Salinispora species. Comparative genomic analyses resulted in the delineation of six new species based on average nucleotide identity and digital DNA–DNA hybridization values below 95 and 70 %, respectively. The species status of the six new groups was supported by a core-genome phylogeny reconstructed from 2106 orthologs detected in 118 publicly available Salinispora genomes. Chemotaxonomic and physiological studies were used to complete the phenotypic characterization of the strains. The fatty acid profiles contained the major components iso-C16 : 0, C15 : 0, iso-17 : 0 and anteiso C17 : 0. Galactose and xylose were common in all whole-sugar patterns but differences were found between the six groups of strains. Polar lipid compositions were also unique for each species. Distinguishable physiological and biochemical characteristics were also recorded. The names proposed are Salinispora cortesiana sp. nov., CNY-202T (=DSM 108615T=CECT 9739T); Salinispora fenicalii sp. nov., CNT-569T (=DSM 108614T=CECT 9740T); Salinispora goodfellowii sp. nov., CNY-666T (=DSM 108616T=CECT 9738T); Salinispora mooreana sp. nov., CNT-150T (=DSM 45549T=CECT 9741T); Salinispora oceanensis sp. nov., CNT-138T (=DSM 45547T=CECT 9742T); and Salinispora vitiensis sp. nov., CNT-148T (=DSM 45548T=CECT 9743T).

scholarly journals Characterization of a Halotolerant Fungus from a Marine Sponge

2019 ◽  
Vol 2019 ◽  
pp. 1-9 ◽  
Author(s):  
Yitayal S. Anteneh ◽  
Melissa H. Brown ◽  
Christopher M. M. Franco
Keyword(s):  
Sequence Similarity ◽  
Marine Sponges ◽  
Rrna Gene ◽  
28S Rrna ◽  
Symbiotic Interaction ◽  
28S Rrna Gene ◽  
Symbiotic Interactions ◽  
Isolation And Characterization ◽  
Halotolerant Fungus

Introduction. Marine sponges have established symbiotic interactions with a large number of microorganisms including fungi. Most of the studies so far have focussed on the characterization of sponge-associated bacteria and archaea with only a few reports on sponge-associated fungi. During the isolation and characterization of bacteria from marine sponges of South Australia, we observed multiple types of fungi. One isolate in particular was selected for further investigation due to its unusually large size and being chromogenic. Here, we report on the investigations on the physical, morphological, chemical, and genotypic properties of this yeast-like fungus. Methods and Materials. Sponge samples were collected from South Australian marine environments, and microbes were isolated using different isolation media under various incubation conditions. Microbial isolates were identified on the basis of morphology, staining characteristics, and their 16S rRNA or ITS/28S rRNA gene sequences. Results. Twelve types of yeast and fungal isolates were detected together with other bacteria and one of these fungi measured up to 35 μm in diameter with a unique chromogen compared to other fungi. Depending on the medium type, this unique fungal isolate appeared as yeast-like fungi with different morphological forms. The isolate can ferment and assimilate nearly all of the tested carbohydrates. Furthermore, it tolerated a high concentration of salt (up to 25%) and a range of pH and temperature. ITS and 28S rRNA gene sequencing revealed a sequence similarity of 93% and 98%, respectively, with the closest genera of Eupenidiella, Hortaea, and Stenella. Conclusions. On the basis of its peculiar morphology, size, and genetic data, this yeast-like fungus possibly constitutes a new genus and the name Magnuscella marinae, gen nov., sp. nov., is proposed. This study is the first of its kind for the complete characterization of a yeast-like fungus from marine sponges. This novel isolate developed a symbiotic interaction with living hosts, which was not observed with other reported closest genera (they exist in a saprophytic relationship). The observed unique size and morphology may favour this new isolate to establish symbiotic interactions with living hosts.

Revisiting theAcanthamoebaspecies that form star-shaped cysts (genotypes T7, T8, T9, and T17): characterization of seven new Brazilian environmental isolates and phylogenetic inferences

Parasitology ◽  
2011 ◽  
Vol 139 (1) ◽  
pp. 45-52 ◽  
Author(s):  
ANA C. M. MAGLIANO ◽  
MARTA M. G. TEIXEIRA ◽  
SILVIA C. ALFIERI
Keyword(s):  
Opportunistic Infections ◽  
Phylogenetic Analyses ◽  
Sequence Divergence ◽  
Rrna Gene ◽  
Pathogenic Potential ◽  
Morphological Group ◽  
Isolation And Characterization ◽  
Group I ◽  
Group 2

SUMMARYFree-living amoebae of the genusAcanthamoebaare the agents of both opportunistic and non-opportunistic infections and are frequently isolated from the environment. Of the 17 genotypes (T1–T17) identified thus far, 4 (T7, T8, T9, and T17) accommodate the rarely investigated species of morphological group I, those that form large, star-shaped cysts. We report the isolation and characterization of 7 new Brazilian environmentalAcanthamoebaisolates, all assigned to group I. Phylogenetic analyses based on partial (∼1200 bp) SSU rRNA gene sequences placed the new isolates in the robustly supported clade composed of the species of morphological group I. One of the Brazilian isolates is closely related toA. comandoni(genotype T9), while the other 6, together with 2 isolates recently assigned to genotype T17, form a homogeneous, well-supported group (2 0% sequence divergence) that likely represents a newAcanthamoebaspecies. Thermotolerance, osmotolerance, and cytophatic effects, features often associated with pathogenic potential, were also examined. The results indicated that all 7 Brazilian isolates grow at temperatures up to 40°C, and resist under hyperosmotic conditions. Additionally, media conditioned by each of the newAcanthamoebaisolates induced the disruption of SIRC and HeLa cell monolayers.

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