scholarly journals Differentiation of Bacillus anthracis and other Bacillus cereus group bacterial strains using multilocus sequence typing method

2020 ◽  
Vol 16 ◽  
pp. 12-21
Author(s):  
Grzegorz Graniak ◽  
Alina Olender ◽  
Katarzyna Naylor
Keyword(s):  
Bacillus Cereus ◽  
Phylogenetic Relationship ◽  
Multilocus Sequence Typing ◽  
Sequence Data ◽  
Animal Health ◽  
Housekeeping Genes ◽  
Negative Control ◽  
Bacterial Strains ◽  
Bacillus Cereus Group ◽  
Typing Method

The study describes the preparation of the phylogenetic differentiation of Bacillus cereus strains. The Bacillus cereus group of bacteria is very important for human and animal health. The multilocus sequence typing scheme has been used to present this group of bacteria’s phylogenetic relationship and structure. The MLST system was established using 60 isolates of B. anthracis, B. cereus sensu stricto, B. thuringiensis, and transitional environment strains of Bacillus spp. As a negative control, five strains of B. subtilis and B. megaterium were used. Primers for amplification and sequencing were designed to target highly conserved internal fragment of seven housekeeping genes: glpF, gmk, ilvD, pta, pur, pycA, and tpi. A total of 22 different sequence types (STs) were distinguished. Analysis of the sequence data showed that all of the Bacillus cereus strains are very closely related. The MLST scheme exhibited a high level of resolution that can be used as an excellent tool for studying the phylogenetic relationship, epidemiology, and population structure of the Bacillus cereus group strains. The MLST method additionally allows us to define the phylogenetic relationship between very closely related strains based on a combination of the sequences of all seven alleles fragments and each of them separately. Thus, this genetic investigation tool is very useful in epidemiological investigation of potential military/ bioterrorist use of B. anthracis.

scholarly journals Multilocus Sequence Typing Scheme for Bacteria of the Bacillus cereus Group

2004 ◽  
Vol 70 (1) ◽  
pp. 191-201 ◽  
Author(s):  
Erlendur Helgason ◽  
Nicolas J. Tourasse ◽  
Roger Meisal ◽  
Dominique A. Caugant ◽  
Anne-Brit Kolstø
Keyword(s):  
Population Structure ◽  
Bacillus Cereus ◽  
Multilocus Sequence Typing ◽  
Sequence Data ◽  
Housekeeping Genes ◽  
Industrial Applications ◽  
Enzyme Electrophoresis ◽  
Mlst Scheme ◽  
High Level ◽  
Sequence Types

ABSTRACT In this study we developed a multilocus sequence typing (MLST) scheme for bacteria of the Bacillus cereus group. This group, which includes the species B. cereus, B. thuringiensis, B. weihenstephanensis, and B. anthracis, is known to be genetically very diverse. It is also very important because it comprises pathogenic organisms as well as bacteria with industrial applications. The MLST system was established by using 77 strains having various origins, including humans, animals, food, and soil. A total of 67 of these strains had been analyzed previously by multilocus enzyme electrophoresis, and they were selected to represent the genetic diversity of this group of bacteria. Primers were designed for conserved regions of housekeeping genes, and 330- to 504-bp internal fragments of seven such genes, adk, ccpA, ftsA, glpT, pyrE, recF, and sucC, were sequenced for all strains. The number of alleles at individual loci ranged from 25 to 40, and a total of 53 allelic profiles or sequence types (STs) were distinguished. Analysis of the sequence data showed that the population structure of the B. cereus group is weakly clonal. In particular, all five B. anthracis isolates analyzed had the same ST. The MLST scheme which we developed has a high level of resolution and should be an excellent tool for studying the population structure and epidemiology of the B. cereus group.

scholarly journals Draft Genome Sequences of 18 Psychrotolerant and 2 Thermotolerant Strains Representative of Particular Ecotypes in the Bacillus cereus Group

2017 ◽  
Vol 5 (5) ◽  
Author(s):  
Marie-Hélène Guinebretière ◽  
Valentin Loux ◽  
Véronique Martin ◽  
Pierre Nicolas ◽  
Vincent Sanchis ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Draft Genome ◽  
Bacterial Strains ◽  
Genome Sequences ◽  
Bacillus Cereus Group ◽  
Content Type ◽  
Physiological Diversity

ABSTRACT Bacteria from the Bacillus cereus group exhibit genetic and physiological diversity through different ecotypes. Here, we present the draft genome sequences of 20 bacterial strains belonging to the contrasted psychrotolerant and thermotolerant ecotypes.

scholarly journals Estimating Recombinational Parameters in Streptococcus pneumoniae From Multilocus Sequence Typing Data

Genetics ◽  
2000 ◽  
Vol 154 (4) ◽  
pp. 1439-1450 ◽  
Author(s):  
Edward J Feil ◽  
John Maynard Smith ◽  
Mark C Enright ◽  
Brian G Spratt
Keyword(s):  
Streptococcus Pneumoniae ◽  
Multilocus Sequence Typing ◽  
Clonal Complex ◽  
Average Length ◽  
Housekeeping Genes ◽  
Single Locus ◽  
Recent Common Ancestor ◽  
Typing Method ◽  
Molecular Events ◽  
Molecular Typing Method

Abstract Multilocus sequence typing (MLST) is a highly discriminatory molecular typing method that defines isolates of bacterial pathogens using the sequences of ~450-bp internal fragments of seven housekeeping genes. This technique has been applied to 575 isolates of Streptococcus pneumoniae and identifies a number of discrete clonal complexes. These clonal complexes are typically represented by a single group of isolates sharing identical alleles at all seven loci, plus single-locus variants that differ from this group at only one out of the seven loci. As MLST is highly discriminatory, the members of each clonal complex can be assumed to have a recent common ancestor, and the molecular events that give rise to the single-locus variants can be used to estimate the relative contributions of recombination and mutation to clonal divergence. By comparing the sequences of the variant alleles within each clonal complex with the allele typically found within that clonal complex, we estimate that recombination has generated new alleles at a frequency ~10-fold higher than mutation, and that a single nucleotide site is ~50 times more likely to change through recombination than mutation. We also demonstrate how to estimate the average length of recombinational replacements from MLST data.

scholarly journals Multilocus Sequence Type System for the Plant Pathogen Xylella fastidiosa and Relative Contributions of Recombination and Point Mutation to Clonal Diversity

2005 ◽  
Vol 71 (12) ◽  
pp. 8491-8499 ◽  
Author(s):  
Mark Scally ◽  
Erin L. Schuenzel ◽  
Richard Stouthamer ◽  
Leonard Nunney
Keyword(s):  
Point Mutation ◽  
Plant Pathogen ◽  
Recombination Rate ◽  
Sequence Data ◽  
Xylella Fastidiosa ◽  
Bacterial Species ◽  
Type System ◽  
Clonal Diversity ◽  
Housekeeping Genes ◽  
Bacterial Strains

ABSTRACT Multilocus sequence typing (MLST) identifies and groups bacterial strains based on DNA sequence data from (typically) seven housekeeping genes. MLST has also been employed to estimate the relative contributions of recombination and point mutation to clonal divergence. We applied MLST to the plant pathogen Xylella fastidiosa using an initial set of sequences for 10 loci (9.3 kb) of 25 strains from five different host plants, grapevine (PD strains), oleander (OLS strains), oak (OAK strains), almond (ALS strains), and peach (PP strains). An eBURST analysis identified six clonal complexes using the grouping criterion that each member must be identical to at least one other member at 7 or more of the 10 loci. These clonal complexes corresponded to previously identified phylogenetic clades; clonal complex 1 (CC1) (all PD strains plus two ALS strains) and CC2 (OLS strains) defined the X. fastidiosa subsp. fastidiosa and X. fastidiosa subsp. sandyi clades, while CC3 (ALS strains), CC4 (OAK strains), and CC5 (PP strains) were subclades of X. fastidiosa subsp. multiplex. CC6 (ALS strains) identified an X. fastidiosa subsp. multiplex-like group characterized by a high frequency of intersubspecific recombination. Compared to the recombination rate in other bacterial species, the recombination rate in X. fastidiosa is relatively low. Recombination between different alleles was estimated to give rise to 76% of the nucleotide changes and 31% of the allelic changes observed. The housekeeping loci holC, nuoL, leuA, gltT, cysG, petC, and lacF were chosen to form the basis of a public database for typing X. fastidiosa (www.mlst.net ). These loci identified the same six clonal complexes using the strain grouping criterion of identity at five or more loci with at least one other member.

scholarly journals 179 Using a 100% Dried Distillers Grains Cube as a Supplement for Steers Grazing Mixed Grass Prairie in Oklahoma

2020 ◽  
Vol 98 (Supplement_2) ◽  
pp. 46-46
Author(s):  
Zane N Grigsby ◽  
Paul A Beck ◽  
Stacey A Gunter
Keyword(s):  
Animal Health ◽  
Late Summer ◽  
Estradiol Benzoate ◽  
Positive Control ◽  
Negative Control ◽  
Early Summer ◽  
Trenbolone Acetate ◽  
Mixed Grass Prairie ◽  
Mixed Grass ◽  
Main Plot

Abstract This research was conducted to determine effects of supplementation and implanting on BW gain by steers grazing mixed grass prairie (n = 12 pastures, 19.9 ± 0.7 ha) in northwest Oklahoma. Three main plot treatments were: 1) Negative Control (NC), no supplementation, 2) Positive Control (PC), supplemented with DDGS cubes, 1.8 kg/steer on alternate days in late summer, 3) High Supplement (HS), 1/3 increase in stocking rate with 0.75% BW supplemental DDGS cubes all season. Steers (n = 125, BW = 223.1 ± 23.2 kg) were stocked at 2.2 ha/steer for PC and NC, 1.3 ha/steer for HS. Grazing was from May 17 – September 27 (132 d). All steers were implanted with 200 mg progesterone and 20 mg estradiol benzoate (SYN, Synonvex S, Zoetis Animal Health) on May 17. On July 18 three reimplant treatments were applied: 1) no reimplant; 2) SYN; or 3) 40 mg trenbolone acetate and 8 mg estradiol (Revalor G, Merck Animal Health). Data were analyzed using the PROC MIXED in SAS as a split-plot experimental design. In early summer HS had 0.26 kg greater (P < 0.01) ADG than NC and PC. Late summer gains of PC were 0.33 kg/d more (P ≤ 0.01) than NC; and HS gained 0.49 and 0.16 kg/day more (P ≤ 0.04) than NC and PC, respectively. Gain per hectare for PC (46 kg/ha) were greater (P < 0.01) than NC (35 kg/ha) and more than doubled (P < 0.01) with HS (89 kg/ha). Reimplanting had no effect on ADG (P ≥ 0.28). Late season supplementation with PC resulted in supplemental efficiency of 2.7 kg supplement/kg added gain compared with NC. Increased stocking rates with season long supplementation in HS resulted in supplemental efficiency of 3.8 kg supplement/kg added gain per hectare. Based on these data, a 100% DDGS cube is an effective supplement option to increase BW gain during the late summer or increase carrying capacity and gain during the summer grazing period in northwestern Oklahoma.

Phenotypic properties and genotyping analysis of Bacillus cereus group isolates from dairy and potato products

LWT ◽  
2021 ◽  
Vol 140 ◽  
pp. 110853
Author(s):  
Yiying Huang ◽  
Steve H. Flint ◽  
Shubo Yu ◽  
Yu Ding ◽  
Jon S. Palmer
Keyword(s):  
Bacillus Cereus ◽  
Bacillus Cereus Group ◽  
Phenotypic Properties ◽  
Potato Products

Development of Rapid Real-Time PCR and Most-Probable-Number Real-Time PCR Assays To Quantify Enterotoxigenic Strains of the Species in the Bacillus cereus Group

2007 ◽  
Vol 70 (12) ◽  
pp. 2774-2781 ◽  
Author(s):  
I-CHEN YANG ◽  
DANIEL YANG-CHIH SHIH ◽  
JAN-YI WANG ◽  
TZU-MING PAN
Keyword(s):  
Bacillus Cereus ◽  
Real Time ◽  
Real Time Pcr ◽  
Food Samples ◽  
Most Probable Number ◽  
Pcr Assay ◽  
Bacillus Cereus Group ◽  
Probable Number ◽  
Cooked Rice ◽  
Group Detection

Members of the Bacillus cereus group may produce diarrheal enterotoxins and could be potential hazards if they enter the food chain. Therefore, a method capable of detecting all the species in the B. cereus group rather than B. cereus alone is important. We selected nhe as the target and developed a real-time PCR assay to quantify enterotoxigenic strains of the B. cereus group. The real-time PCR assay was evaluated with 60 B. cereus group strains and 28 others. The assay was also used to construct calibration curves for different food matrices and feces. The assay has an excellent quantification capacity, as proved by its linearity (R2 > 0.993), wide dynamic quantification range (102 to 107 CFU/g for cooked rice and chicken, 103 to 107 CFU/ml for milk, and 104 to 107 CFU/g for feces), and adequate relative accuracy (85.5 to 101.1%). For the low-level contaminations, a most-probable-number real-time PCR assay was developed that could detect as low as 100 CFU/ml. Both assays were tested with real food samples and shown to be considerably appropriate for B. cereus group detection and quantification.

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