Analysis of epidemic characteristics of anthrax in China from 2017 to 2019 and molecular typing of Bacillus anthracis / 中华预防医学杂志

Objective: To analyze the epidemiological characteristics of anthrax in China from 2017 to 2019 and molecular typing of Bacillus anthracis isolated from some provinces (autonomous regions). Methods: Surveillance data of anthrax cases reported from 2017 to 2019 in the Infectious Disease Surveillance information System of China Disease Prevention and Control and the Public Health Emergency Reporting and Management Information System were collected, and descriptive epidemiological methods were used to analyze the epidemic characteristics, including the temporal, geographic and demographic distribution of this disease. A total of 47 strains of Bacillus anthracis isolated from 2017 to 2019 were analyzed by canSNP and MLVA15. Results: A total of 951 cases of anthrax were reported from 2017 to 2019, of which 938 were cutaneous anthrax, representing 98.63% of the total number reported. It was mainly distributed in the west and northeast of China, and the three provinces with the highest number of cases were Gansu (215), Sichuan (202) and Qinghai (191). Cases had been reported throughout the year, more cases occurred in the summer and autumn, and August was the month with the most cases,66.35% (211/318), 72.32% (243/336) and 68.01% (202/297) of cases were reported during June to September. The age distribution was mainly between 20 and 59 years old, accounting for more than 80% of all cases. The number of male cases was significantly higher than that of female cases, the ratio of male to female was about 3∶1. The occupations were mainly herdsmen and farmers, accounting for 49.70% to 58.18% and 31.45% to 36.70%, respectively. Public health events occurred every year, and 29 events had been reported from 2017 to 2019. canSNP analysis showed that 37 of the 47 strains belonged to the A.Br.001/002 subgroup and 10 belonged to the A.Br.Ames subgroup. MLVA15 analysis showed that there were 17 genotypes, of which 10 genotypes contained only one strain. Conclusion: Cutaneous anthrax was the predominant clinical type in China from 2017 to 2019.The seasonal, geographic and demographic distribution characteristics were evident.Molecular typing methods such as canSNP and MLVA15 can be used to trace the source of infectious diseases and provide technical support for anthrax prevention and control.

Seguridad ocupacional contra Bacillus anthracis en la industria de curtiembre peruana / Occupational safety against Bacillus anthracis in the Peruvian tannery industry

El carbunco es una zoonosis que afecta a animales herbívoros, causada por el microorganismo Bacillus anthracis; que se encuentra en su forma vegetativa en el organismo infectado, y una vez en contacto con el aire forma esporas que infectan al huésped, liberando toxinas que inducen edema, septicemia y necrosis tisular. En humanos, la transmisión se produce por cortes, pinchazos o contacto directo de piel lesionada con esporas o tejidos de animales infectados. En Perú, para la década de 1980, se reportaron 400 casos anuales en áreas rurales agrícolas, adquiridos por manipulación del ganado vacuno, muerto por ántrax. Es por ello, que la piel y el cuero procedente de animales infectados representa un riesgo de infección para los trabajadores de industrias curtiembres. Se realizó un estudio transversal para evaluar el riesgo biológico ocupacional y las medidas de bioseguridad relacionadas con el Bacillus anthracis en pequeñas, medianas y grandes empresas curtiembres peruanas. Se aplicó un instrumento a los trabajadores para cuantificar las variables, componentes de riesgo biológico y conocer las medidas higiénicas adoptadas. Posteriormente se calculó e interpretó el nivel de riesgo biológico. Se encontró que el nivel de riesgo supera el límite de exposición biológica para el Bacillus anthracis en todas las empresas evaluadas y los trabajadores de las pequeñas empresas tienen poco conocimiento acerca del riesgo biológico al que están expuestos, siendo necesaria la capacitación de los trabajadores acerca del Bacillus anthracis y adopción de medidas de bioseguridad en empresas curtiembres, que garanticen la salud de los trabajadores(AU)

Anthrax is a zoonosis that affects herbivorous animals, caused by the microorganism Bacillus anthracis; which is found in its vegetative form in the infected organism, and once in contact with the air it forms spores that infect the host, releasing toxins that induce edema, septicemia and tissue necrosis. In humans, transmission occurs through cuts, punctures, or direct contact of broken skin with spores or tissues of infected animals. In Peru, for the 1980s, 400 annual cases were reported in rural agricultural areas, acquired by handling cattle, which died of anthrax. For this reason, the skin and leather from infected animals represent a risk of infection for workers in tanneries. A cross-sectional study was carried out to evaluate the occupational biological risk and biosafety measures related to Bacillus anthracis in small, medium and large Peruvian tanneries. An instrument was applied to the workers to quantify the variables, components of biological risk and to know the hygienic measures adopted. Subsequently, the level of biological risk was calculated and interpreted. It was found that the level of risk exceeds the biological exposure limit for Bacillus anthracis in all the companies evaluated and the workers of the small companies have little knowledge about the biological risk to which they are exposed, being necessary the training of the workers about the Bacillus anthracis and adoption of biosafety measures in tanneries, which guarantee the health of workers(AU)

Calidad microbiológica del aire en seis áreas de la microestación biológica- Zoocriadero de la Universidad Nacional Agraria de la Selva ­ Tingo María ­ Perú / Microbiological quality of air in six areas of the biological micro station - Zoocriadero of the Universidad Nacional Agraria de la Selva - Tingo Maria ­ Peru

El presente trabajo de investigación tiene como objetivo evaluar la calidad microbiológica del aire en seis áreas de la Microestación Biológica-Zoocriadero de la Universidad Nacional Agraria de la Selva-Tingo María-Perú; procediéndose de la siguiente manera: Se seleccionaron seis áreas de muestreo (A1: Boletería, A2: Caseta gallito de las rocas, A3: Zona de las tortugas, A4: Caseta del otorongo, A5: Caseta de los monos y A6: Cocina), de los cuales para la determinación de los parámetros físicos (PAS, temperatura, humedad relativa, precipitación y rosa de viento) se obtuvo información de la estación meteorológica de la UNAS y de lecturas directas en campo, para la determinación de las PAS se realizó por el método pasivo obteniéndose la mayor concentración de PAS en el área 1 "Boletería" (12.61 t/km2/mes, 11.08 t/km2/mes y 8.4 t/km2/mes) y la más baja en el área 5 "caseta de los monos" (2.80 t/km2/mes, 2.80 t/km2/mes y 3.2 t/km2/mes); para el análisis microbiológico se realizó por el método de borboteo en líquidos, identificándose 27 géneros entre bacterias y hongos, siendo las bacterias más frecuentes Bacillus sp., Staphylococcus sp., Enterobacter agglomerans, Klebsiella pneumoniae, Enterobacter hafniae y algunas bacterias patógenas para el hombre como Streptococcus sp., Clostridium perfringens, Bacillus anthracis. Los hongos de mayor frecuencia son Penicillium sp., Aspergillius sp., Geotrichum sp., algunos hongos patógenos identificados: Microsporum sp., Epirophyton sp. Por otro lado, para la percepción del público como trabajadores se aplicó encuestas, donde indicaron que la calidad microbiológica del aire en la Microestación Biológica-Zoocriadero es regular(AU)

The objective of this research paper is to assess the microbiological quality of the air in six areas of the Biologial Micro-station and animal breeding farm of the Peruvian Univeristy "Universidad Nacional Agraria de la Selva in Tingo María, Perú", proceeding as follows: Six sampling areas were selected (A1: Ticket Office, A2: Cock-of-the-Rock hut, A3: Turtle area, A4: Otorongo hut, A5: Monkey hut, A6: Kitchen), of wich for the determination of the phisical parameters (SBP, temperature, relative humidity, precipitation and wind rose) information was obtained from the UNAS meteorological station and direct readings in the field. For the determination of SBP was performed by the passive method obtaining the high concentration of SBP in area 1 "Ticket Office" (12.61 t/km2/month, 11.08 t/km2/month and 8.4 t/km2/month) and the lowest in area 5 "monkey house" (2.80 t/km2/month, 2.80 t/km2/month and 3.2 t/km2/month); microbiological analysis was performed by the liquid bubbling method, identifying 27 genera incluiding bacteria and fungi, with the most frequent bacteria being Bacillus sp., Staphylococcus sp., Enterobacter agglomerans, Klebsiella pneumoniae, Enterobacter hafniae and some bacteria pathogenic for humans such as Streptococcus sp., Clostridium perfringens and Bacillus anthracis. The most frequent fungi are Penicillium sp., Aspergillius sp. and Geotrichum sp.; some pathogenic fungi identified: Microsporum sp., Epirophyton sp. On the other hand,regarding the perception of the public and workers, surveys were applied, wich indicated that the microbiologial quality of the air in the Biological Micro-station and animal breeding farm is regular(AU)

Sporadic human cutaneous anthrax outbreak in Shaanxi Province, China: report of two cases from 2018

ABSTRACT China's compulsory annual livestock anthrax vaccination policy has remarkably reduced but not completely eradicated human anthrax infections. Herein we describe a sporadic human cutaneous anthrax outbreak involving two cases in 2018 in Shaanxi Province, both involving herdsman who dealt with unvaccinated and potentially sick cattle. Both patients showed Bacillus anthracis-positive blister smear and blood culture. Treatment with penicillin was followed by uneventful recovery for both. The prompt performance of the prophylactic measures successfully interrupted the further transmission of this sporadic human cutaneous anthrax outbreak.

Detection of Bacillus anthracis and Bacillus anthracis-like spores in soil from state of Rio de Janeiro, Brazil

BACKGROUND Bacillus anthracis is the aetiologic agent of anthrax, a re-emerging, septicaemic, haemorrhagic and lethal disease that affects humans, domestic ruminants and wildlife. Plasmids pXO1 and pXO2 are attributes that confer pathogenicity to B. anthracis strains. This bacterium was used as biological weapon in the World Wars and in the biological attack in the United States of America at 2001. B. anthracis is classified as a Tier 1 bioterrorism agent by the Centers for Diseases Control and Prevention. Anthrax is recognised as a re-emerging disease. Several studies concerning the dynamics of B. anthracis cycle in soil revealed that nonpathogenic B. anthracis strains due to lack of pXO2 plasmid are commonly found in some types of soil. OBJECTIVES This study aimed isolation and identification of B. anthracis spores in soil samples of the state of Rio de Janeiro, Brazil. METHODS Phenotypic and genotypic approaches were used to identify isolates including MALDI-TOF/MS, motility test, susceptibility to gamma phage and penicillin, survey for pag and cap genes as surrogates of pXO1 and pXO2 plasmids, respectively, and sequencing of 16SrRNA-encoding gene. Physicochemical analysis of the soil samples were carried out to describe soil characteristics. FINDINGS We observed the presence of one B. anthracis pXO1+ and pXO2- isolated from clay loam soil; one B. anthracis-like strain pXO1+ and pXO2-isolated from loamy sand; and 10 Bacillus spp. strains sensitive to phage-gamma that need better characterisation to define which their species were recovered from loamy sand. MAIN CONCLUSIONS This work showed promising results and it was the first study to report results from an active surveillance for B. anthracis in Brazil.

Evaluation of in vitro antimicrobial susceptibility of Bacillus anthracis strains isolated during anthrax outbreaks in Italy from 1984 to 2017

Anthrax, caused by Bacillus anthracis, is a non-contagious infectious disease that affects a wide range of animal species (primarily ruminants) including humans. Due to the often-fatal outcome in humans, quick administration of definitely effective antimicrobials is crucial either as prophylaxis or as a clinical case therapy. In this study, 110 B. anthracis strains, temporally, geographically, and genetically different, isolated during anthrax outbreaks in Italy from 1984 to 2017, were screened using a broth microdilution method to determine their susceptibility to 16 clinically relevant antimicrobial agents. The strains were isolated from various matrices (human, animal, and environmental samples) and were representative of thirty distinct genotypes previously identified by 15-loci multiple-locus variable-number of tandem repeats analysis. The antimicrobials tested were gentamicin, ceftriaxone, streptomycin, penicillin G, clindamycin, chloramphenicol, vancomycin, linezolid, cefotaxime, tetracycline, erythromycin, rifampin, amoxicillin, ciprofloxacin, doxycycline, and trimethoprim. All isolates were susceptible to most of the tested antimicrobials, with the exception of trimethoprim for which all of them showed high minimal inhibitory concentration values. An intermediate level of susceptibility was recorded for ceftriaxone and cefotaxime. Although the Centers for Disease Control and Prevention recommend the use of doxycycline, ciprofloxacin, penicillin G, and amoxicillin for treatment of human cases and for post-exposure prophylaxis to anthrax spores, this study shows a high degree of in vitro susceptibility of B. anthracis to many other antimicrobials, suggesting the possibility of an alternative choice for prophylaxis and therapy.

Emergency Response for a Laboratory Biosafety Incident / 生物医学与环境科学（英文）

On December 14, 2017, a faculty member of a university in Hunan Province reported that an anthrax vaccine strain might have recovered virulence during an undergraduate experiment and potential exposure could not be ruled out for the students involved. Upon receiving the case report, the CDC, health bureaus, and local governments at the county, prefectural, and provincial levels promptly organized experts in different fields (including epidemiologists, biosafety experts, and laboratory testing experts) for case investigation, evaluation, and response. As the investigation results showed, no virulence recovery was identified in the involved anthrax vaccine strain; and no contamination of Bacillus anthracis was detected at the involved areas. Thus, the university returned to normal functioning.

Ántrax cutáneo, último brote diagnosticado en Chile / Cutaneous anthrax, the last outbreak diagnosed in Chile

Resumen El ántrax, es una zoonosis causada por una bacteria generadora de esporas, llamada Bacillus anthracis. En forma natural tiene una distribución global, con una predilección en zonas agrícolas con pocas normativas de sanidad pública veterinaria. El contagio humano ocurre por el consumo de carnes de animales enfermos, por contacto a través de una puerta de entrada en la piel o por la inhalación de esporas de productos derivados del animal afectado (lana, cuero, huesos). La infección en los seres humanos compromete con mayor frecuencia la piel, seguido por el tracto gastrointestinal y los pulmones. El control de la enfermedad se basa en la prevención, de allí la importancia de la vigilancia en la detección de casos y brotes. Presentamos el último brote de ántrax cutáneo diagnosticado en Chile con descripción de dos primeros casos clínicos del brote.

Anthrax is a zoonosis caused by a spore-forming bacterium, called Bacillus anthracis. Naturally it is of global distribution, with a predilection in agricultural zones with few norms of public veterinary health. Human contagion occurs through the consumption of diseased animal's meat or through a doorway into the skin or through the spores inhalation of products derived from the affected animal (wool, leather, bones). The most frequent infection in humans occurs in the skin, followed by the gastrointestinal tract and lungs. We present the last outbreak of cutaneous anthrax diagnosed in Chile with a description of the first two clinical cases of the outbreak. Control disease is based on prevention, hence the importance of surveillance in detecting cases and outbreaks.

Genome sequencing of two Bacillus anthracis strains: a virulent strain and a vaccinal strain

ABSTRACT Bacillus anthracis strain SPV842_15 was isolated from bovine fetus, while B. anthracis strain Brazilian vaccinal was recovered from a commercial vaccine. We report here the genome sequences of both strains. The SPV842_15 genome is composed of a single circular chromosome with a length of 5,228,664 base pairs, and comprises 5911 coding sequences. In turn, the Brazilian vaccinal genome remains in 201 contigs with 5733 coding sequences. Both genomes have an overall C + G content of 35.4%, and 11 genes encoding the ribosomal RNAs (rRNAs) 5S, 16S and 23S. Only the plasmid pX01 sequence, which carries genes for toxins synthesis, was detected and completely assembled for both strains. These plasmids have a length of 181,684 base pairs and a C + G content of 32.5%. These genomic data generate insights about vaccinal B. anthracis virulence.

ÁNTRAX: enfermedad aún vigente / ANTHRAX: disease still current

El ántrax es una zoonosis producida por Bacillus anthracis, único miembro del género Bacillus que es capaz de causar enfermedad epidémica en humanos y otros mamíferos. Afecta principalmente a los animales herbívoros domésticos y silvestres. Los humanos son hospederos accidentales y se infectan por contacto directo o indirecto con animales o productos contaminados. Las esporas pueden vivir en el suelo por años y los humanos pueden contraer el ántrax al tener contacto con animales infectados, productos provenientes de estos que al consumir carne infectada; esto se presenta principalmente en países poco desarrollados donde los niveles de vacunación animal contra esta enfermedad son bajos. Este escrito tiene por objetivo presentar una revisión sobre el tema, especialmente sobre aspectos como el modo de infección, manifestaciones clínicas, diagnóstico y tratamiento de la enfermedad.

Anthrax is a zoonosis produced by Bacillus anthracis, the only member of the genus Bacillus that is capable of causing epidemic disease in humans and other mammals. It mainly affects wild and domestic animals. Humans are accidental hosts and are infected through direct or indirect contact with animals or contaminated animal products. B. anthracisspores can live in the soil for many years and humans can become infected with anthraxby contact with infected animals or products contaminated from eating meat infected. This disease occurs mainly in developing countries where vaccine levels are low.The objective of this paper is to present a review on thesubject, especially on aspects such as mode ofinfection, clinical features diagnostic assessment and treatment of thedisease.

Expresión y renaturalización del antígeno protector de Bacillus anthracis: un modelo para evaluar el replegado de proteínas antigénicas recombinantes a gran escala / Expression and refolding of the protective antigen of Bacillus anthracis: A model for high-throughput screening of antigenic recombinant protein refolding

Bacillus anthracis protective antigen (PA) is a well known and relevant immunogenic protein that is the basis for both anthrax vaccines and diagnostic methods. Properly folded antigenic PA is necessary for these applications. In this study a high level of PA was obtained in recombinant Escherichia coli. The protein was initially accumulated in inclusion bodies, which facilitated its efficient purification by simple washing steps; however, it could not be recognized by specific antibodies. Refolding conditions were subsequently analyzed in a high-throughput manner that enabled nearly a hundred different conditions to be tested simultaneously. The recovery of the ability of PA to be recognized by antibodies was screened by dot blot using a coefficient that provided a measure of properly refolded protein levels with a high degree of discrimination. The best refolding conditions resulted in a tenfold increase in the intensity of the dot blot compared to the control. The only refolding additive that consistently yielded good results was L-arginine. The statistical analysis identified both cooperative and negative interactions between the different refolding additives. The high-throughput approach described in this study that enabled overproduction, purification and refolding of PA in a simple and straightforward manner, can be potentially useful for the rapid screening of adequate refolding conditions for other overexpressed antigenic proteins.

El antígeno protector de Bacillus anthracis (protective antigen, PA) es una importante proteína inmunogénica, en la que se basan tanto las vacunas contra el ántrax/carbunclo como varios métodos diagnósticos. Para estas aplicaciones es esencial que el PA mantenga sus propiedades antigénicas, para lo cual debe estar correctamente plegado. En este estudio se obtuvieron altos niveles del PA en Escherichia coli recombinante. Inicialmente, la proteína se acumuló desnaturalizada en cuerpos de inclusión, lo que facilitó su eficiente purificación en simples pasos de lavado, pero no fue reconocida por anticuerpos específicos. Se analizaron las condiciones de replegado con un sistema de alto rendimiento, evaluando simultáneamente casi un centenar de condiciones diferentes. La recuperación de la capacidad del PA de ser reconocido por los anticuerpos se evaluó por dot blot utilizando un coeficiente que proporcionó una medida de los niveles de proteína correctamente plegada, con un alto grado de discriminación. Las mejores condiciones de renaturalización permitieron un aumento de diez veces en la intensidad de los dot blots con respecto del control. El único aditivo que produjo buenos resultados de forma constante fue la L-arginina. El análisis estadístico de las interacciones entre los diferentes aditivos de replegado permitió identificar tanto interacciones cooperativas como negativas. El enfoque de alto rendimiento descripto en este trabajo, que permitió la sobreproducción, purificación y plegado del PA de una manera sencilla y directa, puede ser potencialmente útil para el rápido screening de las condiciones adecuadas de replegado cuando se sobreexpresan otras proteínas antigénicas.

Development and comparative evaluation of up-converting phosphor technology based lateral flow assay for rapid detection of Yersinia pestis, Bacillus anthracis spore and Brucella spp / 中华预防医学杂志

<p><b>OBJECTIVE</b>To develop an up-converting phosphor technology based lateral flow (UPT-LF) assay for rapid and quantitative detection of Yersinia pestis, Bacillus anthracis spore and Brucella spp.and make the comparison with BioThreat Alert (BTA) test strips (Tetracore Inc., USA).</p><p><b>METHODS</b>Using up-converting phosphor nano-particles (UCP-NPs) as the bio-marker, three double-antibody-sandwich model based UPT-LF strips including Plague-UPT-LF, Anthrax-UPT-LF, Brucella-UPT-LF were prepared and its sensitivity, accuracy, linearity and specificity were determined by detecting 10(10), 10(9), 10(8), 10(7), 10(6), 10(5) and 0 CFU/ml series of concentrations of Y.pestis, B.anthracis, Brucella standards and other 27 kinds of 10(9) CFU/ml series of contrations of bacteria strains.Furthermore, the speed, sensitivity and accuracy of bacteria standards and simulated sample detection were compared between UPT-LF and BTA system.</p><p><b>RESULTS</b>The detection limit of Plague-UPT-LF, Anthrax-UPT-LF and Brucella-LF was 10(5) CFU/ml. The CV of series of bacteria concentrations was ≤ 15%, and the r between lg (T/C-cut-off) and lg (concentration) was 0.996,0.998 and 0.999 (F values were 1 647.57, 743.51 and 1 822.17. All the P values were <0.001), respectively. The specificity of Plague-UPT-LF and Brucella-LF were excellent, while that of Anthrax-UPT-LF was a little bit regretful because of non-specific reaction with two isolates of B. subtilis and one B.cereus. On-site evaluation showed the detection time of UPT-LF for all Y.pestis, B.anthracis spore and Brucella spp.was 33, 36 and 37 min, while BTA was 115, 115 and 111 min, which revealed the higher detection speed and sensitivity of UPT-LF comparing with BTA. The negative rate of two methods for blank standard was both 5/5, the sensitivity of UPT-LF for Y.pestis,B.anthracis spore and Brucella spp. was all 10(5) CFU/ml, then BTA was 10(6), 10(6) and 10(5) CFU/ml, respectively. The detection rate of UPT-LF for all three bacteria analog positive samples was 16/16, while BTA for B.anthracis was 7/16 only.</p><p><b>CONCLUSION</b>The good performance including rapidness, simplicity and high sensitivity will bring the bright future of UPT-LF to be broadly used on-site as first response to bio-terrorism.</p>

The Effect of Polyphenols Isolated from Cynanchi wilfordii Radix with Anti-inflammatory, Antioxidant, and Anti-bacterial Activity

Recently, Cynanchi wilfordii Radix has gained wide use in Asian countries as a functional food effective for relieving fatigue, osteoporosis, and constipation, particularly in menopausal disorders. However, its anti-inflammatory and anti-microbial activities have not been explored in detail to date. The anti-inflammatory, antioxidant, and anti-bacterial properties of the Cynanchi wilfordii Radix extracts obtained with water, methanol, ethanol, and acetone were compared. All 4 polyphenol-containing extracts exhibited anti-inflammatory and antioxidant effects. The ethanol extract was found to elicit the most potent reduction of nitric oxide (NO), prostaglandin E2 (PGE2), and cytokine (IL-1beta, IL-6, IL-10, and TNF-alpha) levels, as well as inhibit the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in a concentration-dependent manner. The evaluation of antioxidant activity also revealed the ethanol extract to have the highest free radical scavenging activity, measured as 85.3+/-0.4%, which is equivalent to 99.9% of the activity of alpha -tocopherol. In the assessment of anti-bacterial activity, only ethanol extract was found to inhibit the growth of the Bacillus species Bacillus cereus and Bacillus anthracis. These results show that polyphenols of Cynanchi wilfordii Radix have anti-inflammatory, antioxidant, and anti-bacterial properties that can be exploited and further improved for use as a supplementary functional food, in cosmetics, and for pharmaceutical purposes.

Anti-Inflammatory and Anti-Superbacterial Properties of Sulforaphane from Shepherd's Purse

Shepherd's purse, Capsella bursa-pastoris (L.) Medik., has been considered a health food for centuries in Asia and is known to contain the isothiocyanate compound sulforaphane. In this study, we evaluated the anti-inflammatory and antibacterial properties of a sulforaphane-containing solution (SCS) isolated from shepherd's purse. SCS had significant anti-inflammatory activity indicated by the decreased levels of nitric oxide (NO), cytokines (interleukin 1beta [IL-1beta], IL-6, and IL-10), and prostaglandin E2 (PGE2) in lipopolysaccharide-stimulated RAW 264.7 murine macrophages. In addition, SCS decreased the inducible NO synthase (iNOS) and cyclooxygenase 2 (COX-2) levels, which confirmed the anti-inflammatory activity of SCS. Further, SCS inhibited vancomycin-resistant enterococci (VRE) and Bacillus anthracis. The minimal inhibitory concentration was 250 microg/ml for VRE and 1,000 microg/ml for B. anthracis. Taken together, these data indicate that SCS has potential anti-inflammatory and anti-superbacterial properties, and thus it can be used as a functional food or pharmaceutical.

Anti-Inflammatory and Anti-Superbacterial Activity of Polyphenols Isolated from Black Raspberry

The fruit of the black raspberry (Rubus coreanus Miquel) has been employed in traditional medicine, and recent studies have demonstrated its measureable biological activities. However, the root of the black raspberry has not been studied. Therefore, in this study, we evaluated the anti-inflammatory and antibacterial properties of the root and unripe fruit polyphenols of the black raspberry. Both polyphenols proved to have anti-inflammatory activity as evidenced by the decreased nitric oxide (NO), cytokines (IL-1beta , IL-6, and IL-10) and prostaglandin E2 (PGE2) levels in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages. However, root polyphenols showed stronger anti-inflammatory activity than fruit polyphenols. LPS-induced mRNA and protein expressions of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 levels were also decreased, confirming the anti-inflammatory activity. Root polyphenols showed lethal activity against methicillin-resistant Staphylococcus aureus (MRSA), carbapenem-resistant Acinetobacter baumannii (CRAB), and Bacillus anthracis. In contrast, the black raspberry fruit did not demonstrate these properties. These data provide the first demonstration that black raspberry root has potential anti-inflammatory and anti-superbacterial properties that can be exploited as alternatives for use in the food and cosmetic industries and/or as pharmaceuticals.

Genetic diversity of Korean Bacillus anthracis isolates from soil evaluated with a single nucleotide repeat analysis

Bacillus (B.) anthracis, the etiological agent of anthrax, is one of the most genetically monomorphic bacteria species in the world. Due to the very limited genetic diversity of this species, classification of isolates of this bacterium requires methods with high discriminatory power. Single nucleotide repeat (SNR) analysis is a type of variable-number tandem repeat assay that evaluates regions with very high mutation rates. To subtype a collection of 21 isolates that were obtained during a B. anthracis outbreak in Korea, we analyzed four SNR marker loci using nucleotide sequencing analysis. These isolates were obtained from soil samples and the Korean Center for Disease Control and Prevention. The SNR analysis was able to detect 13 subgenotypes, which allowed a detailed evaluation of the Korean isolates. Our study demonstrated that the SNR analysis was able to discriminate between strains with the same multiple-locus variable-number tandem repeat analysis genotypes. In summary, we obtained SNR results for four SNR marker loci of newly acquired strains from Korea. Our findings will be helpful for creating marker systems and help identify markers that could be used for future forensic studies.

Apoptosis, antimicrobial and antioxidant activities of phytochemicals from Garcinia malaccensis Hk.f

OBJECTIVE@#To study the chemical constituents of stembark of Garcinia malaccensis (G. malaccensis) together with apoptotic, antimicrobial and antioxidant activities.@*METHODS@#Purification and structure elucidation were carried out by chromatographic and spectroscopic techniques, respectively. MTT and trypan blue exclusion methods were performed to study the cytotoxic activity. Antibacterial activity was conducted by disc diffusion and microdilution methods, whereas antioxidant activities were done by ferric thiocyanate method and DPPH radical scavenging.@*RESULTS@#The phytochemical study led to the isolation of α,β-mangostin and cycloart-24-en-3β-ol. α-Mangostin exhibited cytotoxic activity against HSC-3 cells with an IC(50) of 0.33 μM. β- and α-mangostin showed activity against K562 cells with IC(50) of 0.40 μM and 0.48 μM, respectively. α-Mangostin was active against Gram-positive bacteria, Staphylococcus aureus (S. aureus) and Bacillus anthracis (B. anthracis) with inhibition zone and MIC value of (19 mm; 0.025 mg/mL) and (20 mm; 0.013 mg/mL), respectively. In antioxidant assay, α-mangostin exhibited activity as an inhibitor of lipid peroxidation.@*CONCLUSIONS@#G. malaccensis presence α- and β-mangostin and cycloart-24-en-3β-ol. β-Mangostin was found very active against HSC-3 cells and K562. The results suggest that mangostins derivatives have the potential to inhibit the growth of cancer cells by inducing apoptosis. In addition, α-and β-mangostin was found inhibit the growth of Gram-positive pathogenic bacteria and also showed the activity as an inhibitor of lipid peroxidation.

Genetic populations of Bacillus anthracis isolates from Korea

Bacillus (B.) anthracis is the pathogen that causes fatal anthrax. Strain-specific detection of this bacterium using molecular approaches has enhanced our knowledge of microbial population genetics. In the present study, we employed molecular approaches including multiple-locus variable-number tandem repeat analysis (MLVA) and canonical single-nucleotide polymorphism (canSNP) analysis to perform molecular typing of B. anthracis strains isolated in Korea. According to the MLVA, 17 B. anthracis isolates were classified into A3a, A3b, and B1 clusters. The canSNP analyses subdivided the B. anthracis isolates into two of the three previously recognized major lineages (A and B). B. anthracis isolates from Korea were found to belong to four canSNP sub-groups (B.Br.001/2, A.Br.005/006, A.Br.001/002, and A.Br.Ames). The A.Br.001/002 and A.Br.Ames sub-lineages are closely related genotypes frequently found in central Asia and most isolates were. On the other hand, B. anthracis CH isolates were analyzed that belonged to the B.Br.001/002 sub-group which found in southern Africa, Europe and California (USA). B.Br.001/002 genotype is new lineage of B. anthracis in Korea that was not found before. This discovery will be helpful for the creation of marker systems and might be the result of human activity through the development of agriculture and increased international trade in Korea.

Staphylococcus aureus Membrane Vesicles and Its Potential Role in Bacterial Pathogenesis

The production of extracellular vesicles is a ubiquitous process in both Gram-negative and Gram-positive bacteria. Gram-negative bacteria produce and secrete outer membrane vesicles during in vitro culture and in vivo infection and their contribution to bacterial pathogenesis has been well characterized. However, little is known about extracellular vesicles in Gram-positive bacteria. Until now, only few Gram-positive bacterial species, Staphylococcus aureus, Bacillus anthracis, B. cereus, and B. subtilis, have been found to produce membrane vesicles (MVs), but their contribution to bacterial pathogenesis has not been understood. Here, I discuss S. aureus MVs in terms of MV production, interaction of MVs with host cells, and immune response against MVs to understand its potential role in S. aureus pathogenesis.