Burkholderia pseudomallei, the causative agent for melliodiosis, is an opportunistic pathogen found in soil in tropic and subtropic regions worldwide.  B. pseudomallei is classified as a category B, Tier 1 Select Agent in the United States due to a high mortality rate when untreated, and the potential for its use as a biowarfare agent through aerosol transmission. Most strains of B. pseudomallei require handling under Biosafety level 3 (BSL 3). B. pseudomallei, strain Bp82 (deltapurM) has a 109 base pair deletion within the purM open reading frame, rendering the strain avirulent in mouse and hamster models.  B. pseudomallei, strain Bp82 (deltapurM) is now an Excluded Strain from the HHS and USDA Select Agents and Toxins ­Exclusions list and can be handled under BSL 2 conditions. Strain Bp82 (deltapurM) has been proposed as a vaccine candidate and has been used in antibiotic resistance studies.  This strain was graciously deposited into BEI Resources by Dr. Herbert Schweizer, and is available as item number NR-51280.

Reference:

Propst, K. L., et al. “A Burkholderia pseudomallei deltapurM Mutant is Avirulent in Immunocompetent and Immunodeficient Animals: Candidate Strain for Exclusion from Select-Agent Lists.” Infect. Immun. 78 (2010): 3136-3143. PubMed: 20404077.

Image:  Burkholderia pseudomallei culture on sheep blood agar (CDC/ Larry Stauffer)

BEI Resources continues to expand the catalog of luciferase-expressing parasitic protists. Recent additions include Leishmania mexicana, strain MNYC/BZ/62/M379 transfected with the luciferase gene from the sea pansy Renilla reniformis (NR-51210) and Trypanosoma cruzi, strain CL Brener expressing the thermostable red-shifted luciferase gene mutant, PpyRE9 (NR-49161). These models allow the high-throughput screening of large numbers of candidate compounds during infection in vitro and the possibility of tracking parasite distribution in vivo in laboratory mice. The expression of red-shifted luciferase in T. cruzi, in particular, results in a reporter system that is more sensitive than other bioluminescence systems previously reported. The availability of these new transgenic strains provides the researcher rapid quantification of parasite growth and simplification of the methodology for scoring inhibitor assays.

Reference:

Lewis, M.D., et al. “A New Experimental Model for Assessing Drug Efficacy Against Trypanosoma cruzi Infection Based on Highly Sensitive in vivo Imaging.” J. Biomol. Screen. 20 (2015): 36-43. PubMed: 25296657.

Image:  Trypanosoma cruzi CL Brener PpyRE9 (NR-49161) epimastigotes stained with GIEMSA (BEI Resources)

Now available are two well-characterized reference reagents for Zika virus (ZIKV) to facilitate evaluation of existing nucleic acid testing (NAT) assays and development of novel ZIKV assays. These reagents were produced by The Center for Biologics Evaluation and Research/U.S. Food and Drug Administration (CBER/FDA) from heat-inactivated (HI) ZIKV culture supernatant stock from two strains, PRVABC59 and FSS13025, which were diluted in dialyzed, defibrinated human plasma, and lyophilized. These reagents add to the expansive catalog of Zika virus materials now available from BEI Resources.

Image:  Female Aedes aegypti mosquito (CDC/James Gathany)

Coccidioidomycosis, also known as Valley fever, is an invasive fungal infection caused by the dimorphic fungi Coccidioides immitis and Coccidioides posadasii. These fungi are endemic to arid or semi-arid regions of the southwestern United Sates, Mexico, and Central and South America. Infection is acquired through inhalation of air-dispersed fungal spores termed arthroconidia. The initial or acute form of coccidioidomycosis is often mild with symptoms such as fever, cough, chest pain, and red, spotty rash. If infection is not controlled, it may progress to chronic pneumonia or even disseminate to other parts of the body including the brain, especially in immunocompromised patients. BEI Resources houses a well-characterized collection of Coccidioides strains from various origins, which have been used in studies of the ecology and distribution of different genotypes of the pathogen. Such studies are important for monitoring outbreaks, determining changes in virulence, and predicting disease progression. These strains were generously contributed by Dr. Bridget Barker, Montana State University.

Image:  Colonies of Coccidioides immitis, Strain RMSCC 3476 (NR-48939) growing at 37°C on Emmons' Modification of Sabouraud agar.

Parechovirus A (formerly Human parechovirus or HPeV) is an emerging infection of public health concern among pediatric populations in North America. HPeV belongs to the family Picornaviridae and is currently divided into 19 genotypes. Parechovirus A type 3 (HPeV-3) is clinically the most important genotype as it has been associated with more severe clinical manifestations in the form of sepsis-like and central nervous system (CNS) illnesses, particularly in neonates and infants. HPeV3 was initially isolated in 1999 from a patient in Japan and has been detected with increasing frequency over the last decade in Asia, Europe, Canada, South America, and more recently, in the United States. HPeV3 is the most common single cause of aseptic meningitis/ meningoencephalitis in infants less than 90 days old in North America. BEI Resources now offers two HPeV3 isolates from the United States, including one from the 2014 outbreak in Missouri.
Image:  Parechovirus (ViperDB)

Chagas’ disease caused by the protozoan parasite Trypanosoma cruzi (T. cruzi) is widespread in Central and South America infecting an estimated 12 to 14 million people, with infection often leading to chronic myocarditis resulting in high morbidity and mortality. In addition to our catalog of T. cruzi parasites and triatomine vectors, BEI Resources now offers monoclonal antibodies raised against parasite stage-specific surface antigens, which play important roles during infection. These antigens include the trypomastigote-specific antigen Ssp-1 (BEI Resources NR-50891) and the amastigote-specific surface antigen Ssp-4 (BEI Resources NR-50892). Studies have shown the presence of Ssp-4 in tissue culture supernatants of T. cruzi-infected cells and the presence of anti-Ssp4 antibodies in sera from chagasic patients. These monoclonal antibodies were generously contributed by Dr. Norma Andrews, University of Maryland.  Dr. Andrews generated these monoclonal antibodies while at the laboratory of Dr. Ruth Nussenzweig, New York University.
Image:  Immunofluorescence staining of amastigotes of Trypanosoma cruzi, strain TcVT-1 (BEI Resources NR-36630) with Monoclonal Anti-Trypanosoma cruzi, NR-50892. A, Light microscopy image of T. cruzi-infected kidney epithelial cells (ATCC^® CCL-26™). B, Immunostaining of intracellular amastigotes with NR-50892 labeled with a FITC-conjugated rabbit anti-mouse antibody. Host cell and parasite nuclei are stained with DAPI.

References:

1. Andrews, N. W., et al. “Stage-Specific Surface Antigens Expressed during the Morphogenesis of Vertebrate Forms of Trypanosoma cruzi.”  Exp. Parasitol. 64 (1987):  474-484.  PubMed:  3315736.
2. Andrews, N. W., et al. “Developmentally Regulated, Phospholipase C-Mediated Release of the Major Surface Glycoprotein of Amastigotes of Trypanosoma cruzi.”  167 (1988):  300-314.  PubMed:  3279152.

Enterobacter cloacae is part of the normal human flora and not considered a primary pathogen. Unfortunately, there is increasing incidence of nosocomial infections with E. cloacae, which frequently involve intravenous and other medical and hospital devices as well as medical solutions and disinfectants used routinely in hospitals and patient care facilities. The situation is further complicated by the ability of these bacteria to acquire resistance to antibiotics, notably carbapenems. BEI Resources has obtained a set of 15 isolates of the E. cloacae complex with defined susceptibility patterns against 16 antibiotics. This valuable set is now available to aid your studies on carbapenem-resistant Enterobacteriaceae (CRE).

Mayaro virus (MAYV) is a New World alphavirus and the etiologic agent of Mayaro fever, an acute febrile illness sometimes accompanied by severe and persistent arthritis similar to Chikungunya fever. Its transmission cycle occurs mainly through mosquito vectors, especially those of the genus Haemagogus, but Aedes spp. mosquitoes may also be competent vectors. The enzootic transmission cycle of MAYV is not fully understood, but the occurrence of relatively large outbreaks of Mayaro fever and the competence of Aedes mosquitoes for the transmission of MAYV suggest the potential for an urban human-mosquito-human transmission cycle to emerge. MAYV was first isolated in Trinidad in 1954, with sporadic outbreaks of Mayaro fever in Bolivia, Brazil, and other regions throughout Central and South America. Recently, MAYV was found in a DENV co-infected patient in Haiti¹, the first report of MAYV in the Caribbean nation, underscoring the need for MAYV inclusion in surveillance activities in these regions in addition to Dengue, Chikungunya, and Zika viruses.

Una virus (UNAV) is the closest genetic relative of MAYV that also causes a similar febrile illness. UNAV was first identified in northern Brazil in 1959. Serosurveys have identified antibodies against UNAV in birds, horses, and humans; however, the extent of viral distribution and human disease risk is unknown.

References:1. eid2211.161015

Plasmid maintenance by bacteria can be unpredictable without selective pressure. This is of particular concern during in vivo studies. To circumvent this in Staphylococcus species, the Bose Lab at The University of Kansas Medical Center has provided plasmids to BEI Resources that remain stable inStaphylococcus cells without the requirement of antibiotics during both in vitro and in vivo experiments¹. These E. coli-S. aureus shuttle vectors are based on the LAC-p01 plasmid.

Plasmid pKK22 (Accession KX085042) is designed to be used with S. aureus USA300 strains that contain LAC-p01 and will render the strains isogenic. Plasmid pKK30 (Accession KX085043) is a variant of pKK22, in which the predicted open reading frames (ORFs) that are not needed for plasmid maintenance have been deleted, and is intended for use inStaphylococcus cells that do not contain LAC-p01. Both plasmids contain a single trimethoprim resistance cassette that is functional in both E. coliand Staphylococcus. In addition, they contain the E. coli R6Kγ origin of replication and require pir+ cells for replication. DH5αλpir and the pir+dam- dcm- strain GM2163λpir are provided as a host strains, courtesy of Dr. Eric Stabb (University of Georgia)². These plasmids and host strains are now supplied individually or as a kit through BEI Resources.

References:

1. Krute, C. N., K. K. Krausz, M. A. Markiewicz, J. A. Joyner, S. Pokhrel, P. R. Hall, and J. L. Bose. 2016. "Generation of a Stable Plasmid for in vitro and in vivostudies of Staphylococcus." Applied and Environmental Microbiology 82:6859-6869. PMID 27637878

2. Dunn A. K., M. O. Martin, and E. V. Stabb. 2005. "Characterization of pES213, a Small Mobilizable Plasmid from Vibrio fischeri." Plasmid 54:114-134. PMID: 16122560

Clostridium difficile research is easier with strains from BEI Resources

For the benefit of researchers requiring nucleic acids from pathogens, BEI Resources offer more than 1,000 nucleic acids derived from pathogens. These reagents can be used at lower containment levels for molecular studies; to ensure their biosafety, the preparations have been safety tested for the absence of residual live organisms. Please check our website to see the wide variety viral nucleic acids extracted from various BSL-2 through BSL-4 pathogens. Notable new additions to this category of reagents include Genomic RNA from Enterovirus D68 and human metapneumovirus.

Whooping cough is a disease caused by bacteria from the genus Bordetella. Currently under widespread resurgence, whooping cough ranks globally among the 10 leading causes of childhood mortality. BEI Resources offers several isolates of Bordetella pertussis, B. bronchiseptica, and B. holmesii, along with toxins, nucleic acids, and proteins for Bordetella research. To see the full list of Bordetella reagents and strains currently available from BEI resources, please click here

Image:  Bordetella holmesii NR-43499 grown on Bordet Gengou media

Toxoplasma gondii is an obligate intracellular protozoan parasite of the phylum Apicomplexa that is the causal agent of toxoplasmosis. BEI Resources houses a comprehensive assortment of T. gondii cultures that include GFP-expressing strains; reference genome strains; genetic crosses between type strains I, II, and III; and strains used as models for parasite transmission and cyst formation. For a complete list of our Toxoplasma catalog, please contact us at Contact@BEIResources.org.

¹ Available soon. Send inquiries to Contact@BEIResources.org for availability.

BEI Resources offers 42 distinct Escherichia coli-Staphylococcus cassette-based shuttle vectors that provide all the tools you need to complete your research. These shuttle vectors are deposited as part of the Network on Antimicrobial Resistance in Staphylococcus aureus (NARSA) reagents. Choose from staphylococcal low-copy-number, high-copynumber, thermosensitive, and theta-replicating plasmids. Select from erythromycin, tetracycline, chloramphenicol, kanamycin, or spectinomycin selectable markers in Staphylococcus, and ampicillin, erythromycin, kanamycin, and chloramphenicol resistance markers in E. coli. The modular design of the plasmid vector system offers flexibility and variety for expression analysis and provides the opportunity to address questions about gene dosage, complementation, and cis-trans effects. The shuttle vector system provides effective tools for studying gene regulation of staphylococci among ecosystems.

For further published details on these plasmids, please see Charpentier et al. Appl

Environ Microbiol 70(10): 6076-6085, 2004

Image:  NR-46158. Staphylococcus aureus strain 4221 containing shuttle vector pCN57, grown on TSA + 10 μg/ml erythromycin

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We have a number of expression plasmids for various Filoviral proteins for Zaire Ebolavirus and Marburgvirus. Many of the proteins have been tagged with HA or FLAG. Continue to check our catalog for new plasmid additions, such as those expressing GP, NP, VP24, VP30, VP35, and VP40 in the next few months for a complete list.

Registration for the BEI Resources program can be found at this link below. NARSA Strain requestors will need to register for Level 2 for continued access to the NARSA Strains. ** Please note if you are an existing NARSA registrant, BEI Resources will contact you after the Annual Meeting with specific registration instructions and paperwork to facilitate your registration.

http://www.beiresources.org/Register.aspx

Once registered with BEI Resources, researchers will have access to the full BEI Resources catalog (within the respective Biosafety Level). The catalog has over 12,000 research reagents representing more than 400 Genus and Species focusing on NIAID's Category A, B and C priority pathogen list and Emerging Infections.

Once registered with BEI Resources, ALL investigators will receive FREE shipping and handling on orders placed through BEI Resources.

Please feel free to contact BEI Resources by email: contact@beiresources.org or by phone: Toll-free telephone number: (800) 359-7370 (8:30 AM to 4:30 PM Eastern Time) with any questions or concerns.

General Information about the Schistosoma spp. Life Cycle Training Course

Biomedical Research Institute

12111 Parklawn Drive Rockville, MD.

May 21-23, 2014

This course is designed to acquaint investigators working in the field of schistosomiasis with basic techniques and strategies for maintaining Schistosoma mansoni, S. haematobium, and S. japonicum in the laboratory, and it offers directions into making one’s research more productive.  Among the topics covered will be procedures for setting up and maintaining the complete S. mansoni/Biomphalaria glabrata life cycle in the research laboratory, an overview of schistosomiasis, the pathology of the disease, collection of adult worms and eggs from tissues, exposure of snails to miracidia, shedding snails, exposure of mice and hamsters to cercariae, and an examination of all the intramolluscan life stages.  Hands-on exercises relating to some of these topics will also be presented (ex. perfusion of mice, exposing mice/hamsters, identification of sporocysts in snails).  Since we first began offering this basic course, over 120 scientists, technicians and students have benefited from completion of the course, and have expanded their own research agendas once back in their own laboratories.

The workshop incorporates aspects of all three Schistosoma spp. life cycles.  Attendees will be exposed to aspects of maintaining S. japonicum and S. haematobium life cycles that include general snails maintenance, collection of cercariae from infected snails, and exposing mice/hamsters.  It is our hope that incorporating the other two life cycles will encourage interest in maintaining these species in attendees’ laboratories.  The three-day course will also include a lecture on NIH-NIAID parasitology and grant programs. 

We anticipate that attendees will come away with a greater appreciation of the basic biology of Schistosoma spp. parasites, greater confidence in maintaining life cycles, and they will learn strategies for optimizing life cycles in their home laboratories.  Time will be available to ask questions about life cycle maintenance and other aspects of schistosomiasis research. There is no fee for the course.  Attendees are responsible for travel and lodging expenses, but travel assistance may be available. If you have any questions about the course, please contact me at mtucker@afbr-bri.com or (301) 881-3300 x31, or Mitzi Sereno at msereno@afbr-bri.com or (301) 881-3300 x39.

BEI Resources has the MERS-CoV virus (strain EMC/2012) available for distribution to qualified laboratories.  This virus will require a BEI Level 3 registration.  Middle East Respiratory Syndrome (MERS) is viral respiratory illness first reported in Saudi Arabia in 2012. It is caused by a coronavirus called MERS-CoV. Most people who have been confirmed to have MERS-CoV infection developed severe acute respiratory illness. They had fever, cough, and shortness of breath. About half of these people died.

The catalog number for the MERS-CoV virus is NR-44260. You must be logged in with a valid BEI Level 3 Account to place an order. Please email our customer service department if you have any questions about obtaining the MERS-CoV virus.

The much awaited H7N9 Influenza virus is now available for your research needs. NR-44078 is a 6:2 reassortant virus produced using plasmid-based reverse genetics to contain the hemagglutinin and neuraminidase genes from A/Shanghai/1/2013 (H7N9) and internal genes from A/Puerto Rico/8/1934 (H1N1).

Click here to see all H7N9 reagents.

As of Wednesday, August 20th we have updated the BEI Resources and MR4 websites to further optimize your online experience; making it easier, more accessible, and hopefully a bit more pleasurable to access your desired BEI and MR4 reagents. We have rolled out a new redesigned website that we anticipate will provide a more user-friendly navigation and shopping cart experience, easier searching and reading throughout the site, and an optimal viewing experience from essentially any web accessible platform whether it is desktop, laptop, mobile phone, or tablet.

This Responsive Web Design approach suggests that design and development should respond to the user’s behavior and environment based on screen size, platform, and orientation. As more and more of our customers in the scientific community are switching to the duality of laptops and mobile devices to conduct and support their research efforts, we at BEI Resources, would like to let our customers know… We’ve heard you… and would like to thank you for your continued feedback.

This redesigned website has been engineered to automatically respond to user preferences; switching to accommodate for resolution, image size and the scripting capabilities as dictated by the device being used. We can now make images and layouts more flexible. With this design, we have essentially eliminated the need to make individual adjustments in design and development for each new mobile device produced and made available on the market.

While Responsive Design is not a complete solution, we believe this solution provides more options for our users and a more optimal online experience overall when accessing the BEI Resources and MR4 sites; from any website accessible device; mobile or otherwise.

So feel free to access the BEI Resources and MR4 sites while in the office, lab, or on-the-go with your mobile device. An option to provide feedback is available through our Online User Feedback form, provided at the completion of your online order. Tell us what you think; we look forward to hearing from you.

At MR4 and BEI Resources, “We take care of the details - so you can focus on your research.”  

We have available for the first time in the BEI Resources catalog, two SARS Coronaviruses (SARS-CoV) derived from the Urbani strain. NR-18925 is produced from a recombinant infectious clone, icSARS-CoV. NR-15418 is the mouse-adapted strain, Urbani v2163, developed by Dr. Barnard as a lethal model for evaluating antiviral agents (PubMed:19853271). To view a complete listing of all coronaviruses in the catalog, click here.

BEI Resources announces the addition of Sand Flies, Reduviids and Ticks for the research community.

CDC recommends handling naïve ticks from these species at ABSL2 containment in appropriate barrier protected facilities; pathogen research may require elevated ABSL containment. Check the BEI Vector Resources page for a list of other vectors and related reagents available through BEI Resources.

Chikungunya Viruses - Additional Strains Available

BEI Resources announces the addition of two novel BSL3 strains of Chikungunya virus (CHIKV), R-91142 and 182/25, to the catalog of available items. CHIKV is a mosquito-borne alphavirus that is mainly associated with acute febrile illness. Strains R-91142 and 182/25 CHIKV are of Asian origin. Strain 182/25 is the recently developed vaccine candidate and the only CHIKV strain to be tested in humans. Coming soon to the BEI catalogue is the CHIKV African prototype strain, S-27.

Recent acquisitions in BEI Resources aim to contribute to research focused on understanding the mechanisms of disease and the development of novel therapeutic drugs for Trypanosome research. Such acquisitions include SSGCID clones, proteins and reference parasite strains used in genome and virulence studies. For more information on the availability of these materials please contact BEI Resources customer service.

BEI Resources is working to expand the fungal resources so that researchers have a broader range of organisms and reagents for their work.  BEI Resources currently has eight genus and species represented with several more currently going through the production pipeline.  BEI Resources is actively encouraging depositors with medically relevant strains to deposit into the repository. If you are interested in depositing, please click here.

The ISO Guide 34 accreditation by the American Association for Laboratory Accreditation (A2LA) builds upon the earlier certification for compliance with the ISO 9001:2000 standard for quality management systems received by ATCC in 2007. Biological reference materials produced under an ISO Guide 34-accredited process have confirmed identity, well-defined characteristics and an established chain of custody — qualities essential to their effectiveness as biological standards in research and development.

BEI Resources is pleased to announce a new and improved website!  We heard your feedback concerning difficult searches and long checkout times…and, we invite you to explore and discover our new dynamic search package, state-of-the-art shopping cart experience and enhanced product information pages for your items on interest.

Our new search engine provides the community with a user friendly interface in which to research, search, and order reagents. We will now be using the same search and navigation features of many of the world's largest internet sites. Some of the major search features include dynamic filter searching, Intuitive Auto-Complete Text Searches, Suggested Search Terms, Dynamic Clustering and multiple sorting parameters for result sets.

Your ordering experience is also vastly improved with our new Web Services E-Commerce software.  No more waiting for your items to load to the shopping cart; with the new software, your checkout is fast and reliable. Look for all these updates today!

The agent of bubonic plague Yersinia pestisis a NIAID Category A Priority Pathogen and likely biothreat. In support of research and product development in this area BEI Resources released a Yersinia pestis Plasmid Detection Kit (NR-9562) in April 2008.   

This assay allows the researcher to quickly identify the presence of plasmid-encoded virulence factors pPCP1, pMT1 and pCD1 and is designed to be used for characterization purposes in conjunction with standard microbiological techniques.

The assay consists of primer sets designed to specifically detect the pPCP1, pMT1 and pCD1 plasmids using standard polymerase chain reactions and includes a species-specific positive control primer set and internal positive control template.

PCR amplification of genomic DNA from Yersinia pestis displaying a pPCP1+, pMT1+, pCD1– profile (lanes 2-4 respectively). Lanes 5 and 6 are the Yersinia pestis and internal positive controls. Lane 7 is a non-template control."

Panel A. Growth of Yersinia pestis on Congo Red media showing red/orange colonies typical of pgm+ organisms. Organisms that are confirmed pgm+ and pCD1+ are considered Select Agents.

Panel B. Bacteria lacking pgm showing distinctive variations in colony morphology and reduced coloring on Congo Red media as compared to pgm+ organisms.

For technical questions about these products or to learn more, please contact our Technical Services Department.