scholarly journals Detection and Identification of Bacillus anthracis: From Conventional to Molecular Microbiology Methods

2020 ◽  
Vol 8 (1) ◽  
pp. 125 ◽  
Author(s):  
Aleksandra A. Zasada
Keyword(s):  
Bacillus Anthracis ◽  
Special Focus ◽  
High Similarity ◽  
First Line ◽  
Closely Related Species ◽  
Molecular Microbiology ◽  
Detection And Identification ◽  
Deliberate Release ◽  
Anthrax Spores ◽  
Rapid Tests

Rapid and reliable identification of Bacillus anthracis is of great importance, especially in the event of suspected deliberate release of anthrax spores. However, the identification of B. anthracis is challenging due to its high similarity to closely related species. Since Amerithrax in 2001, a lot of effort has been made to develop rapid methods for detection and identification of this microorganism with special focus on easy-to-perform rapid tests for first-line responders. This article presents an overview of the evolution of B. anthracis identification methods from the time of the first description of the microorganism until the present day.

Multiplex polymerase chain reaction for simultaneous detection and identification of Bursaphelenchus xylophilus, B. mucronatus and B. fraudulentus – three closely related species within the xylophilus group

Nematology ◽  
2017 ◽  
Vol 19 (9) ◽  
pp. 1107-1116 ◽  
Author(s):  
Anna Filipiak ◽  
Przemysław Wieczorek ◽  
Marek Tomalak
Keyword(s):  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Morphological Characters ◽  
Bursaphelenchus Xylophilus ◽  
Chain Reaction ◽  
Closely Related Species ◽  
Detection And Identification ◽  
Different Populations ◽  
Polymerase Chain ◽  
Simultaneous Identification

Differentiation between Bursaphelenchus xylophilus and other related, non-pathogenic species can be ambiguous when based exclusively on morphological characters. The morphology of B. mucronatus and B. fraudulentus most closely resembles that of B. xylophilus. Moreover, all of these nematodes are found in both Asia and Europe and can colonise various species of pine. Therefore, for phytosanitary purposes it is necessary to identify the three species precisely and rapidly. We report the results of a multiplex PCR that utilises four primers to identify and discriminate the three Bursaphelenchus species simultaneously. The multiplex PCR yielded DNA fragments of 767, 305 and 132 bp, for B. xylophilus, B. mucronatus and B. fraudulentus, respectively. This primer combination has produced reliable results in multiplex PCR assays with a number of different populations of the listed species, and no cross-reactions were observed with other Bursaphelenchus species. The described approach is simple, reliable and cheaper than other molecular methods presently used for simultaneous identification of the above three species within the xylophilus group.

scholarly journals An Extracytoplasmic Function Sigma Factor Controls β-Lactamase Gene Expression in Bacillus anthracis and Other Bacillus cereus Group Species

2009 ◽  
Vol 191 (21) ◽  
pp. 6683-6693 ◽  
Author(s):  
Cana L. Ross ◽  
Kerrie S. Thomason ◽  
Theresa M. Koehler
Keyword(s):  
Bacillus Cereus ◽  
Bacillus Anthracis ◽  
Resistant Strain ◽  
Sigma Factor ◽  
Susceptible Strain ◽  
Closely Related Species ◽  
Naturally Occurring ◽  
Lactamase Gene ◽  
Extracytoplasmic Function Sigma Factor ◽  
Group Species

ABSTRACT The susceptibility of most Bacillus anthracis strains to β-lactam antibiotics is intriguing considering that the closely related species Bacillus cereus and Bacillus thuringiensis typically produce β-lactamases and the B. anthracis genome harbors two β-lactamase genes, bla1 and bla2. We show that β-lactamase activity associated with B. anthracis is affected by two genes, sigP (BA2502) and rsiP (BA2503), predicted to encode an extracytoplasmic function sigma factor and an anti-sigma factor, respectively. Deletion of the sigP-rsiP locus abolished β-lactamase activity in a naturally occurring penicillin-resistant strain and had no effect on β-lactamase activity in a prototypical penicillin-susceptible strain. Complementation with sigP and rsiP from the penicillin-resistant strain, but not with sigP and rsiP from the penicillin-susceptible strain, conferred constitutive β-lactamase activity in both mutants. These results are attributed to a nucleotide deletion near the 5′ end of rsiP in the penicillin-resistant strain that is predicted to result in a nonfunctional protein. B. cereus and B. thuringiensis sigP and rsiP homologues are required for inducible penicillin resistance in these species. Expression of the B. cereus or B. thuringiensis sigP and rsiP genes in a B. anthracis sigP-rsiP-null mutant confers inducible production of β-lactamase activity, suggesting that while B. anthracis contains the genes necessary for sensing β-lactam antibiotics, the B. anthracis sigP and rsiP gene products are not sufficient for bla induction.

scholarly journals Attenuated Nontoxinogenic and Nonencapsulated Recombinant Bacillus anthracis Spore Vaccines Protect against Anthrax

2000 ◽  
Vol 68 (8) ◽  
pp. 4549-4558 ◽  
Author(s):  
S. Cohen ◽  
I. Mendelson ◽  
Z. Altboum ◽  
D. Kobiler ◽  
E. Elhanany ◽  
...  
Keyword(s):  
Bacillus Anthracis ◽  
Protective Immunity ◽  
Protective Antigen ◽  
Cell Vaccine ◽  
Lethal Challenge ◽  
Electrospray Mass Spectroscopy ◽  
Immunological Tests ◽  
Anthrax Spores ◽  
Biochemical Analyses ◽  
Recombinant Protective Antigen

ABSTRACT Several highly attenuated spore-forming nontoxinogenic and nonencapsulated Bacillus anthracis vaccines differing in levels of expression of recombinant protective antigen (rPA) were constructed. Biochemical analyses (including electrospray mass spectroscopy and N terminus amino acid sequencing) as well as biological and immunological tests demonstrated that the rPA retains the characteristics of native PA. A single immunization of guinea pigs with 5 × 107 spores of one of these recombinant strains, MASC-10, expressing high levels of rPA (≥100 μg/ml) from a constitutive heterologous promoter induced high titers of neutralizing anti-PA antibodies. This immune response was long lasting (at least 12 months) and provided protection against a lethal challenge of virulent (Vollum) anthrax spores. The recombinant B. anthracis spore vaccine appears to be more efficacious than the vegetative cell vaccine. Furthermore, while results clearly suggest a direct correlation between the level of expression of PA and the potency of the vaccine, they also suggest that some B. anthracisspore-associated antigen(s) may contribute in a significant manner to protective immunity.

scholarly journals Determination of effi ciency of the methods for isolating Bacillus anthracis pathogen spores from soil

2019 ◽  
pp. 29-35
Author(s):  
I. Rublenko ◽  
S. Rublenko
Keyword(s):  
Bacillus Anthracis ◽  
Causative Agent ◽  
Tween 80 ◽  
Economic Losses ◽  
Effi Ciency ◽  
Higher Spin ◽  
Anthrax Spores ◽  
Livestock Products ◽  
Favorable Conditions ◽  
Soil Distribution

Diseases of animals and people with anthrax continues to appear in traditionally endemic regions, indicating a real threat of disease in any territory: in Europe, Asia and other regions of the globe. Spores of the causative agent of anthrax are very stable in the environment. It is known that they remain viable for up to 300 years. Under favorable conditions, activation (mobilization of metabolic processes) of spores occurs within 1–2 minutes, followed by initiation and germination (20–30 minutes) and growth into a vegetative cell (60–90 minutes). Zoonotic anthrax disease is relevant not only for Ukraine, but also for most countries in Europe and around the world, as the incidence remains high, thousands of deaths are registered annually, large sums of citizens and states are spent on the treatment of patients, and producers of livestock products bear considerable economic losses due to the death of animals and carrying out anti-epizootic, anti-epidemiological and preventive measures. The article presents the results of determining the eff ectiveness of methods for isolating the spores of the causative agent of the strain Bacillus anthracis UA–07 from the soil. It was found that using the method using Tween-80, 1 % serum albumin in phosphate-buff ered saline and PLET agar, it was possible to increase the excretion of pathogen spores from the soil by 16.8 % (p <0.001). At the same time, it should be noted that with a slight contamination of the soil or material under study, there is probably a problem of obtaining an unreliable study result. According to our advanced methodology, only 2.5 g of soil is needed for the study, whereas according to the method № 1 – 60 g, according to the method № 2 and № 3 – 95, № 4 – 10 g. higher spin speed, uses 100% ethyl alcohol, which destroys the soil's vegetative and some spore like microfl ora (except for spores of Bas. anthracis). Key words: anthrax, spores, Bacillus anthracis, animals, soil, distribution, methodology.

scholarly journals Use of Long-Range Repetitive Element Polymorphism-PCR To Differentiate Bacillus anthracisStrains

2001 ◽  
Vol 67 (7) ◽  
pp. 3021-3028 ◽  
Author(s):  
Michael J. Brumlik ◽  
Urszula Szymajda ◽  
Dorota Zakowska ◽  
Xudong Liang ◽  
Rajendra J. Redkar ◽  
...  
Keyword(s):  
Bacillus Anthracis ◽  
Long Range ◽  
Repetitive Dna ◽  
Related Species ◽  
Molecular Level ◽  
Repetitive Element ◽  
Bacterial Genome ◽  
Chromosomal Marker ◽  
Closely Related Species ◽  
Pcr Method

ABSTRACT The genome of Bacillus anthracis is extremely monomorphic, and thus individual strains have often proven to be recalcitrant to differentiation at the molecular level. Long-range repetitive element polymorphism-PCR (LR REP-PCR) was used to differentiate various B. anthracis strains. A single PCR primer derived from a repetitive DNA element was able to amplify variable segments of a bacterial genome as large as 10 kb. We were able to characterize five genetically distinct groups by examining 105B. anthracis strains of diverse geographical origins. AllB. anthracis strains produced fingerprints comprising seven to eight bands, referred to as “skeleton” bands, while one to three “diagnostic” bands differentiated between B. anthracisstrains. LR REP-PCR fingerprints of B. anthracis strains showed very little in common with those of other closely related species such as B. cereus, B. thuringiensis, and B. mycoides, suggesting relative heterogeneity among the non-B. anthracis strains. Fingerprints from transitional non-B. anthracis strains, which possessed the B. anthracis chromosomal marker Ba813, scarcely resembled those observed for any of the five distinct B. anthracis groups that we have identified. The LR REP-PCR method described in this report provides a simple means of differentiating B. anthracisstrains.

scholarly journals Information on which to base assessments of risk from environments contaminated with anthrax spores

1994 ◽  
Vol 113 (3) ◽  
pp. 479-490 ◽  
Author(s):  
A. Watson ◽  
D. Keir
Keyword(s):  
Risk Assessment ◽  
Bacillus Anthracis ◽  
Causative Agent ◽  
Scientific Literature ◽  
Health Hazards ◽  
Environmental Materials ◽  
Anthrax Spores ◽  
Available Information

SUMMARYAlthough there has been a considerable amount of research conducted intoBacillus anthracis, the causative agent of anthrax, the data are widely disseminated in the scientific literature and are therefore not always easy to assimilate. In view of continuing concern about potential anthrax contamination in environmental materials and sites, this review brings together the currently available information relating to the health hazards fromB. anthracis.The relevance of the available information for risk assessment purposes is assessed.

PCR amplification and DNA sequence analysis of parasitic intestinal protozoa in specimens stained with Chlorazol Black E

2013 ◽  
Vol 58 (2) ◽  
Author(s):  
Norihito Morimoto ◽  
Masataka Korenaga ◽  
Yoshie Nishida ◽  
Hiroaki Takeuchi ◽  
Yoshitaka Kumon ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Pcr Amplification ◽  
Giardia Intestinalis ◽  
High Similarity ◽  
Amplification Product ◽  
Intestinal Protozoa ◽  
Detection And Identification ◽  
Nuclear Extracts ◽  
Storage Term

AbstractChlorazol Black E (CBE) stain has been used for the detection and identification of intestinal parasitic protozoa. In recent years, genotyping of protozoa has been performed to examine pathogenicity and for epidemiologic analysis. In this study, protozoan DNA was amplified from preserved human fecal specimens stained with CBE that were positive for Giardia intestinalis (syn. G. lamblia and G. duodenalis), Chilomastix mesnili, Pentatrichomonas hominis, and Entamoeba histolytica. DNA was amplified from 11 of the 12 (91.6%) samples examined. DNA from CBE-stained smears of G. intestinalis, E. histolytica, and P. hominis was amplified, whereas any amplification product could not be obtained from one of three smears of C. mesnili. Storage term and protozoan number had no association with results of PCR amplification. In genotyping of G. intestinalis, four out of six (66.7%) samples were of genotype AI, while the remaining two (33.3%) samples were of genotype B. The amplified DNA sequences showed high similarity (>99%) with that of G. intestinalis in the GenBank database. These results suggest that DNA remains stable in CBE-stained smears for long term. The present study demonstrates that nuclear extracts from specimens stained with CBE can be amplified by PCR and suggests that specimens stored for extended periods could be applied to genetic and prospective epidemiologic analyses.

scholarly journals Is There Still a Place for Tyrosine Kinase Inhibitors for the Treatment of Hepatocellular Carcinoma at the Time of Immunotherapies? A Focus on Lenvatinib

Cancers ◽  
2021 ◽  
Vol 13 (24) ◽  
pp. 6310
Author(s):  
Marie Decraecker ◽  
Caroline Toulouse ◽  
Jean-Frédéric Blanc
Keyword(s):  
Hepatocellular Carcinoma ◽  
Tyrosine Kinase ◽  
Tyrosine Kinase Inhibitors ◽  
Kinase Inhibitors ◽  
Standard Of Care ◽  
Special Focus ◽  
Second Line ◽  
First Line ◽  
First Line Therapy ◽  
Therapeutic Algorithms

The systemic treatment of hepatocellular carcinoma is changing rapidly. Three main classes of treatment are now available. Historically, multi-targeted tyrosine kinase inhibitors (TKIs) (sorafenib and lenvatinib as first-line; regorafenib and cabozantinib as second-line) were the first to show an improvement in overall survival (OS). Anti-vascular endothelial growth factor (anti-VEGF) antibodies can be used in first-line (bevacizumab) or second-line (ramucirumab) combination therapy. More recently, immuno-oncology (IO) has profoundly changed therapeutic algorithms, and the combination of atezolizumab-bevacizumab is now the first-line standard of care. Therefore, the place of TKIs needs to be redefined. The objective of this review was to define the place of TKIs in the therapeutic algorithm at the time of IO treatment in first-line therapy, with a special focus on lenvatinib that exhibits one of the higher anti-tumoral activity among TKI in HCC. We will discuss the place of lenvatinib in first line (especially if there is a contra-indication to IO) but also after failure of atezolizumab and bevacizumab. New opportunities for lenvatinib will also be presented, including the use at an earlier stage of the disease and combination with IOs.

scholarly journals Isolation Bac. anthracis at different pH and soil humus content

2018 ◽  
Vol 20 (88) ◽  
pp. 147-151
Author(s):  
I. Rublenko ◽  
S. Rublenko
Keyword(s):  
Bacillus Anthracis ◽  
Humus Content ◽  
Diagnostic Techniques ◽  
Sterile Soil ◽  
Soil Humus ◽  
Bacillus Anthracis Spores ◽  
Anthrax Spores ◽  
Ancient Times ◽  
Ph Range ◽  
Great Damage

Anthrax is one of the diseases that cause great damage to livestock and at the same time is a dangerous disease for humans. This disease under different names is known from ancient times. The source of the pathogen is diseased animals. However, an important factor in the transmission of the pathogen is the burial place of animals that died from anthrax. The pathogen can be stored in the soil for more than 100 years. Anthrax is one of the diseases that cause great damage to livestock and at the same time is a dangerous disease for humans. This disease under different names is known from ancient times. The source of the pathogen is diseased animals. However, an important factor in the transmission of the pathogen is the burial place of animals that died from anthrax. In Ukraine there is burial of dead animals from anthrax. Most of the Siberian burial ground is registered in Vinnytsia, Kharkiv, Chernihiv, Ternopil, Lugansk and Kirovograd regions of Ukraine. In this regard, there is a need to study the issues of epizootology and improve diagnostic techniques, measures for general and specific anthrax prevention. The pathogen can be stored in the soil for more than 100 years. The purpose of these studies was to select the Bacillus anthracis spores for the influence of pH and different humus content in the soil. The pH and soil humus content were determined, which was used in experiments at the Laboratory of Agrochemistry of the Institute of Bioenergetic Cultures and Sugar Beet at the National Academy of Agrarian Sciences of Ukraine. The work was performed on the basis of the State Scientific-Control Institute of Biotechnology and strains of microorganisms. Used methodology in which soil sample – 2.5 grams grow. In studies, spores of the avirulence strain Bacillus anthracis were introduced into a sterile soil. Registered methods of separating the argument of anthracite from the soil are obsolete. The method allows distinguishing anthrax spores from soils with different pH and humus content. when 21.76 ± 0.94 spores were applied to sterile soil (which was 17.41 ± 0.75 s/gram), the amount of spores selected was 3.33 ± 0.27 spores/grams (19.13%). The amount of controversy in the experimental soil was lower (17.41 ± 0.75), compared with the amount of controversy submitted to the soil with a pH of 6.5 (P < 0.05), 6.7 (P < 0.05), 7.5 and 7.8, however, the number of allocated – higher – 3.33 ± 0.27 (19.1%). The introduction of 18.54 ± 1.29 spores/grams of soil (3.67% of humus content) amounting to 18% of the deductions. Isolation Bac. anthracis with different pH indicates that the most spores of the pathogen are released within the pH range of 6.7–7.5. The content of humus in the soil affects the number of selected spores. With a higher content of humus, fewer amounts of spores is released anthracis, optimum pH for spore selection – 7.0.

scholarly journals Assessing the zoonotic potential ofAscaris suumandTrichuris suis: looking to the future from an analysis of the past

2012 ◽  
Vol 86 (2) ◽  
pp. 148-155 ◽  
Author(s):  
P. Nejsum ◽  
M. Betson ◽  
R.P. Bendall ◽  
S.M. Thamsborg ◽  
J.R. Stothard
Keyword(s):  
Production Systems ◽  
Pig Manure ◽  
Future Research ◽  
Human Consumption ◽  
Special Focus ◽  
Zoonotic Potential ◽  
Closely Related Species ◽  
The Past ◽  
Open Questions ◽  
Close Proximity

AbstractThe two geohelminths,Ascaris lumbricoidesandTrichuris trichiura, infect more than a billion people worldwide but are only reported sporadically in the developed part of the world. In contrast, the closely related speciesA. suumandT. suisin pigs have a truly global distribution, with infected pigs found in most production systems. In areas where pigs and humans live in close proximity or where pig manure is used as fertilizer on vegetables for human consumption, there is a potential risk of cross-infections. We therefore review this relationship betweenAscarisandTrichurisin the human and pig host, with special focus on recent evidence concerning the zoonotic potential of these parasites, and identify some open questions for future research.

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