The resulting peptides were extracted from GSI-IX clinical trial the gel plug with 0.1% (v/v) trifluoroacetic acid/50% (v/v) acetonitrile. Digests were spotted on a MALDI target using α-cyano-4-hydroxycinnamic acid as a matrix. Spectra were acquired on a 4800 MALDI TOF/TOF analyser (Applied Biosystems, Foster City, CA). Data

analysis and MS database searching were performed using GPS Explorer™ and mascot software. Total RNA was isolated from P. gingivalis cells grown to mid-exponential phase (OD600 nm of c. 1.0) by using an RNeasy Mini kit (Qiagen Science). DNA was removed with RNase-Free DNase. cDNA was generated in a reaction mixture containing a random primer (Promega), dNTP mixture, RNase inhibitor, DTT, Superscript III Reverse Transcriptase (Invitrogen) and

DEPC-treated water. Real-time qPCR was performed using Brilliant SYBR Green II QPCR Master Mix (Stratagene) with an Mx3005P™ Real-Time PCR System (Stratagene) according to the manufacturer’s instructions. Primers for the real-time qPCR are listed in Table S1 and were designed using the primer3 program. Real-time qPCR conditions were as follows: one cycle at 95 °C for 10 min, and 35 cycles of 95 °C for 30 s and 60 °C for 1 min. At each cycle, accumulation of PCR products was detected by the reporter dye from the dsDNA-binding SYBR Green. To confirm that a single PCR product was amplified, after the PCR, a dissociation curve (melting curve) was constructed in the range 55–95 °C. All data were analysed using Mx3005P software. The expression level of each targeted Ivacaftor mw gene was normalized to that of the 16S rRNA gene, which was used as a reference. All

PCR reactions were carried out in triplicate. The efficiency of primer binding was determined by linear regression by plotting the cycle threshold (CT) value versus the log of the cDNA dilution. Relative quantification of transcript was determined www.selleck.co.jp/products/Decitabine.html using the comparative CT method () calibrated to 16S rRNA gene. qPCR experiments were performed multiple times independently, yielding comparable results. We constructed an rgpA rgpB kgp porK mutant from an rgpA rgpB kgp strain and compared secreted proteins between the rgpA rgpB kgp and rgpA rgpB kgp porK strains to avoid degradation of secreted and surface proteins by gingipains as the wild-type strain secreted gingipains that had the ability to process both secreted and surface proteins, while the porK mutant secreted no gingipains. 2D-PAGE of the particle-free (membrane-free) culture supernatants from the kgp rgpA rgpB and kgp rgpA rgpB porK mutants was performed. As a control, three protein spots in each 2D gel, which exhibited the same amounts of proteins with the same molecular masses and isoelectric points, were subjected to MALDI-TOF mass analysis, resulting in the same proteins (PGN_0916, PGN_1367 and PGN_1587; Fig. 1). Their molecular masses and isoelectric points calculated from their amino acid sequences were 69 044 and 4.88 for PGN_0916, 49 199 and 5.

The views and conclusions contained hereon are those of the authors and should not be interpreted as necessarily representing

the official policies or endorsements, either expressed or Selleck Nutlin 3 implied, of IARPA, DOI, or the US Government. Abbreviations ACh acetylcholine BF basal forebrain Glu glutamate LGN lateral geniculate nucleus mAChRs muscarinic acetylcholine receptors PFC prefrontal cortex TRN thalamic reticular nucleus V1 primary visual cortex “
“miR-96 is a microRNA, a non-coding RNA gene which regulates a wide array of downstream genes. The miR-96 mouse mutant diminuendo exhibits deafness and arrested hair cell functional and morphological differentiation. We have previously shown that several genes are markedly downregulated in the diminuendo organ of Corti; one of these is Ptprq, a gene known to be important for maturation and maintenance of hair cells.

In order to study the contribution that downregulation of Ptprq makes to the diminuendo phenotype, we carried out microarrays, scanning electron microscopy and single hair cell electrophysiology to compare diminuendo mutants (heterozygous and homozygous) with mice homozygous for a functional null allele of Ptprq. In terms of both morphology and electrophysiology, the auditory phenotype of mice lacking Ptprq resembles that of diminuendo heterozygotes, while diminuendo homozygotes are more severely affected. A comparison of transcriptomes indicates there is a broad similarity between diminuendo homozygotes Methocarbamol and Ptprq-null mice. The reduction in Ptprq observed in diminuendo see more mice appears to be a major contributor to the morphological, transcriptional and electrophysiological phenotype, but does not account for the complete diminuendo phenotype. “
“The dopaminergic projections to the basal ganglia have long been implicated in reward-guided behavior and decision-making, yet little is known about the role of the posterior pedunculopontine nucleus (pPPN), a major source of excitatory input to the mesolimbic dopamine

system. Here we studied the contributions of the pPPN to decision-making under risk, using excitoxic lesions and reversible inactivation in rats. Rats could choose between two options – a small but certain reward on one lever; or a large but uncertain reward on the other lever. The overall payoff associated with each choice is the same, but the reward variance (risk) associated with the risky choice is much higher. In Experiment 1, we showed that excitotoxic lesions of the pPPN before training did not affect acquisition of lever pressing. But whereas the controls strongly preferred the safe choice, the lesioned rats did not. In Experiment 2, we found that muscimol inactivation of the pPPN also produced similar effects, but reversibly. These results show that permanent lesions or reversible inactivation of the pPPN both abolish risk aversion in decision-making.

The glycopeptide antibiotic gene clusters reported for balhimycin, chloroeremomycin, A40926, A47934 and teicoplanin do not contain Fd or FdR genes (Donadio et al., 2005). Only the biosynthetic gene cluster for the related natural product complestatin contains a single Fd gene (comK) (Chiu et al., 2001). To advance in vitro studies of the cross-linking steps in glycopeptide antibiotic biosynthesis, we present efforts to identify and characterize Fd genes in the balhimycin producer A. balhimycina. The sequencing and annotation

of the entire genome is currently underway. We describe in silico analyses, which reveal 11 different Fd genes in A. balhimycina. Furthermore, we demonstrate the production and KPT-330 concentration purification of two of the newly identified Fds, as well as their ability to participate in electron transfers to OxyB from both A. balhimycina and A. orientalis. The A. balhimycina DSM5908 genome was analyzed using a blast search carried out with six Fd sequences (NP-631171, NP-629284,

NP-631715, NP-625075, NP-628054 and NP-625924) from Streptomyces coelicolor A3(2) (Lamb et al., 2002; Chun et al., 2007), putidaredoxin (P00259) from Pseudomonas putida (Peterson et al., 1990; Pochapsky et al., 1994; Sevrioukova et al., 2003) and the putative ferredoxin comFd (AAK81833) from the complestatin producer Streptomyces lavendulae (Chiu et al., 2001). Eleven putative Fds, named balFd-I to balFd-XI, containing expect (E)-values

<10−6, were identified in the whole genome of A. balhimycina (Table 1). Assignment of the iron–sulfur cluster type was achieved through a blast search of the nonredundant R428 purchase database and a comparison with known Fds. The sequences of the ferredoxins balFd-I to balFd-XI have been deposited in the EMBL database under the accession numbers FN594523-FN594533. The genomic DNA of A. balhimycina DSM5908 was used as a PCR template to amplify the genes coding for the putative [3Fe–4S] ferredoxins balFd-V and balFd-VII. The primers used are shown below (restriction sites underlined): 5′-balFd-V: 5′-TAGACCATATGAAGGTTGTTGTCGACG-3′ 3′-balFd-V: 5′-ATCTACTCGAGGGCCTCTTCGAGGGC-3′ selleckchem 5′-balFd-VII: 5′-TAGACCATATGAAGGTCACCGTGGACG-3′ 3′-balFd-VII: 5′-ATCTACTCGAGCGCGTCCTCGCTCACCG-3 After digestion with NdeI and XhoI, the fragments were cloned between the NdeI/XhoI sites of plasmid pET22b (Novagen) for expression as C-terminal His6-tagged fusion proteins. The nucleotide sequence of each insert was confirmed by sequencing. For production in Escherichia coli Rosetta2(DE3)pLysS (balFd-V) or Origami2(DE3) (balFd-VII), terrific broth medium (400 mL) supplemented with antibiotics and FeSO4 (0.5 mM) was induced with isopropyl-β-d-thiogalactopyranoside (0.2 mM), and shaking at 30 °C for 6 h. Cell pellets in buffer-A (25 mL, 50 mM sodium phosphate pH 7.4, 300 mM KCl, 1 mM dithiothreitol, 1 mM phenylmethylsulfonyl fluoride, 1 mM benzamidine, 0.

1) and 100% 16S rRNA gene sequence identity, Selleck Gefitinib supporting their close affiliation.

The mean sequence identity for the concatenated five protein-coding loci was 98.8% between strains DY05T and 47666-1 and 94.4% between these strains and the relatives V. harveyi, V. campbellii and V. rotiferianus. Discrimination between these species on the basis of phenotypic and 16S rRNA gene analyses is difficult and additional molecular methods such as MLSA have become important tools for correct species delineation and identification (Sawabe et al., 2007; Thompson et al., 2007). Phylogenetic trees generated for concatenated sequences of the five protein-coding loci using NJ, MP and ML methods confirmed the clustering of strains DY05T and 47666-1 (bootstrap values of 100%, 100% and 95%, respectively) and their distinction to close species (Fig. 2, Fig. S1a and b). An extended phylogenetic analysis was performed to detect public database sequences that could potentially belong to the same species as strains DY05T and 47666-1. Using database sequences for the pyrH, topA and mreB loci, Vibrio sp. CAIM 994 clustered with DY05T and 47666-1 in single-gene phylogenetic analyses. Thus, we acquired this strain, isolated from snapper (Lutjanus guttatus) in the northwest coast of Mexico, and determined its 16S rRNA and rpoA gene sequences. Strain CAIM 994 was initially identified as V. rotiferianus, but described as a

possible buy Cabozantinib intermediate strain according to MLSA (Thompson et al., 2007). Phylogenies based on 16S rRNA gene sequences (Fig. 1) and concatenated sequences of five protein-coding loci (Fig. 2) confirmed that CAIM 994, 47666-1 ZD1839 in vitro and DY05T formed a monophyletic group with bootstrap support values

of 99–100%. CAIM 994 shared 99.9% (16S rRNA gene) and 98.3% (five protein-coding loci) gene sequence identities with DY05T and 47666-1. These are greater than the identities shared between CAIM 994 and V. rotiferianus LMG 21460T (99.4% for 16S rRNA gene and 93.2% for five protein-coding loci). Therefore, 16S rRNA gene and MLSA support the notion that CAIM 994 was previously misidentified. Further studies based on phenotypic and genotypic characterization would be required to clarify the relatedness of this and other strains clustering with the Vibrio owensii sp. nov. proposed here. Strains DY05T and 47666-1 showed 76% DNA–DNA hybridization values with each other and 44–55% with V. harveyi LMG 4044T, V. campbellii LMG 11216T and V. rotiferianus LMG 21460T (Table S2). As a DNA–DNA hybridization value of 70% is generally accepted as the limit for species delineation (Wayne et al., 1987), it can be concluded that strains DY05T and 47666-1 belong to a single novel species. The DNA mol% G+C content of DY05T (45.3 mol%) and 47666-1 (45.9 mol%) support their affiliation with Vibrio (Baumann & Schubert, 1983). It can be concluded that strains DY05T and 47666-1 are closely related to V. harveyi, V. campbelli and V.

Serology is useful since this kind of patient has not had any previous contact with the fungus. All traveler patients diagnosed in our laboratory had a positive immunodiffusion test. RT-PCR was positive in only five of the nine patients studied, probably due to the limited amount of DNA circulating in immunocompetent

selleck chemicals llc patients. Respiratory samples provided better results than sera or blood samples. For most patients, only sera samples were available for reaching diagnosis, a fact which could explain the low sensitivity of RT-PCR in the case of travelers. More studies should be performed on this kind of patient. Finally, the fungi were never cultured. In immigrant cases, we found mainly disseminated histoplasmosis in immunosuppressed patients. Histoplasmosis

occurred as a result of the reactivation MDV3100 chemical structure of a latent focus of infection acquired years earlier.30 A total of 29 out of 30 immigrants had AIDS as an underlying disease. This figure matches previously reported studies.31 Patients with disseminated histoplasmosis present fever, weight loss, anorexia, cough, vomiting, diarrhea, and abdominal pain.6 Only 8 patients out of 20 had a positive result in a serological test. In 73% (22/30) of cases the fungus was isolated. Cultures showed good sensitivity in detecting H capsulatum; however, the average time needed to obtain positive cultures was 15 days. RT-PCR showed good sensitivity (89%). The technique

was performed in 27 patients and was positive in 24. Respiratory samples and biopsies were the most useful samples, with 100% sensitivity. Blood samples appeared to have lower sensitivity than sera samples (37.5% vs 69%); however, we obtained a positive result for sera sample and a negative result for blood only in patients 15 and 11 (Table 4). In these cases there may be a partial inhibition which was reflected in a slightly lower melting curve for the internal control. In the other cases, sera and blood samples were either both negative (Table 3, patient 9; and Table 4, patients 1, 18, and 20) or both positive (Table 2, patients 19 and 21). These results may correlate with the clinical status of each patient. More blood samples should therefore C-X-C chemokine receptor type 7 (CXCR-7) be analyzed to reach a conclusion. PCM in non-endemic areas is rarely suspected because of the extremely long silent period of this disease.9 Diagnosis was delayed in four of the six cases diagnosed in our laboratory; we have no data on the other two cases. In all cases described in this paper characteristic yeasts were visualized at the hospitals. The fungus was cultured in only one case (patient 5) and growth was very slow. Serology proved to be useful since it was positive in all patients. RT-PCR showed good sensitivity as we obtained positive results for all patients. Respiratory and biopsy samples proved more suitable than blood samples.

g. malate and glyoxylate), because a variety of acetate assimilation pathways convert acetate into 20s Proteasome activity these compounds (e.g. the glyoxylate shunt of the tricarboxylic acid cycle, the ethylmalonyl-CoA pathway, the citramalate cycle, and the methylaspartate cycle). In this review, we summarize the history of facultative methanotrophy, describe scenarios for the basis

of facultative methanotrophy, and pose several topics for future research in this area. Aerobic methanotrophs are widely distributed in the environment, found wherever methane : air interfaces develop, including in wetlands, bogs, agricultural, forest and urban soils, rice paddies, groundwater, landfill cover soils, among many other locations (Semrau et al., 2010). These cells play a critical role in the global carbon cycle by utilizing methane as a source of carbon and energy – it is estimated that in soils, aerobic methanotrophs consume ∼30 Tg methane year−1 (Kolb, 2009). It was initially widely believed

that aerobic methanotrophs were obligate, i.e., that these microorganisms could only grow utilizing methane or methanol, and in some cases, other C1 compounds such as formaldehyde, formate, and methylamine, but not compounds with carbon–carbon bonds (Bowman, 2006). The cause for obligate methanotrophy is still unresolved (Wood et al., 2004), and, interestingly, many reports have recently been published of methanotrophs that also able

to utilize Enzalutamide multicarbon PFKL compounds as sole growth substrates (Semrau et al., 2010). Hence, it appears that facultative methanotrophy may be more common than originally thought. In this review, the history and basis of facultative methanotrophy is summarized, as well as the implications and applications of such metabolism. The defining characteristic of a methanotroph is its ability to utilize methane as its sole carbon and energy source, and there are at least two forms of the key enzyme involved in the initial oxidation of methane to methanol, the methane monooxygenase (MMO). Most but not all methanotrophs express a membrane-bound or particulate methane monooxygenase (pMMO), while some can either express in addition, or as the unique form, a cytoplasmic, or soluble methane monooxygenase (sMMO). Phylogenetically, aerobic methanotrophs belong primarily to the Alpha- and Gammaproteobacteria, although recently aerobic methanotrophs have also been found that belong to the Verrucomicrobia phylum (Op den Camp et al., 2009; Semrau et al., 2010). The alphaproteobacterial methanotrophs can be further divided in the Beijerinckiaceae and Methylocystaceae families, while the gammaproteobacterial methanotrophs belong to the Methylococcaceae family.

To construct plasmid pENA9, a 0.25-kb DNA fragment containing the phaR promoter region was amplified by PCR. Cyclopamine After digestion with EcoRI and HindIII, the DNA fragment was inserted into pLC4 (Ray et al., 1985), which carries the xylE reporter gene. To construct plasmid pENA10, a 2-bp mutation was introduced into the −35 region of the putative σA-like promoter of phaR using a two-step PCR method (Higuchi et al., 1988). The resulting DNA fragment was digested with EcoRI plus HindIII and cloned into pLC4. Disruptions of the chromosomal phaC, sigB, sigH, spo0A, spo0F, or sigF gene of B. thuringiensis by integrations of plasmid pRN5101-derived pENA1, pENA2, pENA3, pENA4, pENA5, or pENA6, respectively,

through a Campbell-like single-crossover recombination were performed as described previously (Fedhila et al., 2002). The correct integrations were verified by PCR. Construction of the abrB deletion mutant BNA7, in which the abrB gene was replaced with the kanamycin resistance gene kan via a two-step gene replacement event, was performed according to the method described previously (Arnaud et al., 2004). The correct replacement of the chromosomal abrB sequence was verified by PCR. To construct the abrB spo0A double mutant BNA8, the pRN5101-derived plasmid pENA4 was introduced into

the abrB deletion mutant BNA7. Chromosomal integration of pENA4 through a single-crossover recombination was carried out as described above. The correct integration was verified click here by PCR. The PHB contents of B. thuringiensis cells were determined by GC as described Celastrol previously (Tseng et al., 2006) as well as using the method involving spectrophotometric determination of crotonic acid generated from digestion of PHB with concentrated sulfuric acid (Law & Slepecky, 1961). The sample preparation was carried out according to the method described in the literature (Tian et al., 2005), but with a slight modification as described previously (Chen et

al., 2009). Thin sections were examined on a JEM 2000EXII TEM (JEDL Inc.). Transformation of B. thuringiensis cells by electroporation was carried out as described previously(Bone & Ellar, 1989). Total RNA was isolated from B. thuringiensis according to the previously described method (Zuber & Losick, 1983). Northern blotting and primer extension analysis were performed using standard methods (Sambrook & Russell, 2001). An established method was used for spectrophotometric measurement of XylE (catechol 2,3-dioxygenase) activity (Ray et al., 1985). Protein concentrations were determined using the BCA protein assay method according to the instructions of the manufacturer (Pierce Biotechnology Inc.). Measurement of the PHB contents of B. thuringiensis revealed that this bacterium gradually accumulated PHB in the stationary phase after growth in LB medium (Fig. 1a). To demonstrate that the B. thuringiensis homolog of the B.

, 2005). These include two ATP-binding cassette (ABC) transporters, TcyABC and TcyJKLMN, and a symporter TcyP (Burguiere et al., 2005). The TcyJKLMN and TcyP uptake systems are high-affinity transporters

while TcyABC is a low-affinity l-cystine transporter (Burguiere et al., 2005). The TcyJKLMN transporter, encoded within a large operon called the ytmI operon, was found to be the most sensitive to l-cystine starvation compared with other transporters in that its expression was repressed more than 200-fold in the presence of sulfate or l-cystine (Carlsson, 1970). In addition, the expression of the ytmI operon was induced during disulfide stress by the thiol oxidant diamide (Chapot-Chartier et al., 1993). TcyP and TcyABC l-cystine transporters have also been identified in Staphylococcus aureus and were shown to be negatively regulated by the CymRSA regulator, a global regulator that controls cysteine metabolism Selleck Olaparib in response to its availability (Coppee et al., 2001). Cysteine metabolism has not been extensively studied in S. mutans. However, Sperandio et al. 2010 recently characterized two LysR-type transcriptional regulators, CysR and HomR, which activate transcription of genes involved in cysteine metabolism and transport. These authors also identified two l-cystine importers, TcyABC and TcyDEFGH, whose expression was activated by CysR and HomR, respectively (Sperandio et al.,

2010). We sought to characterize the tcyABC tri-cistronic operon encoding the TcyABC transporter in S. mutans. Mutagenesis of tcyABC severely impaired the ability of

S. mutans to transport l-cystine and survive under cystine starvation conditions. BAY 80-6946 solubility dmso We also identified a novel Lys-type regulator of TcyABC which we termed TcyR. Unlike most Lys-type regulators, TcyR was found to repress transcription of the tcyABC operon. Streptococcus mutans strain UA159 was used to construct mutants. Unless otherwise specified, strains were routinely cultured in Todd-Hewitt yeast extract (THYE) medium (BD Biosciences) at 37 °C in air with 5% CO2 without agitation. Mutant strains were propagated in THYE agar plates supplemented with erythromycin at 10 μg mL−1. Optical density (OD) was measured using an Ultrospec 3000 UV/Visible Spectrophotometer (Fisher Scientific). Streptococcus mutans UA159 was Chlormezanone used as the wild-type strain. The S. mutans ΔtcyA (SmTcyA), ΔtcyB (SmTcyB), ΔtcyC (SmTcyC), ΔtcyABC (SmTcyABC), and ΔtcyR (SmTcyR) mutants were constructed in UA159 by a PCR ligation-based deletion strategy as described previously (Cvitkovitch et al., 1997). Briefly, an erythromycin resistance cassette was used to disrupt the tcyC, tcyABC, and tcyR coding regions in the S. mutans UA159 wild-type chromosome using the primer pairs listed below. To confirm successful integration of the erythromycin gene into these coding regions, chromosomal DNA was isolated from erythromycin-resistant transformants and subjected to validation using PCR and nucleotide sequence analysis.

Other exclusion criteria were a history of central nervous system (CNS) infection, stroke, serious head injury, or other neurologic event likely to affect cognition. Many patients were exposed to low doses of psychoactive drugs, either on a prescription or recreational basis. Given that this is a clinical reality in this population, we excluded only those in whom drug effects might be expected to substantially affect cognition. Of the patients originally referred selleck screening library for the study, only three

met one of these exclusion criteria (one with MoCA <20, one with a history of another CNS process, and one with intoxication at the time of testing). The protocol was approved by the ethics board of the McGill University Health Centre, and all participants provided informed consent. All tests Pirfenidone price were administered

in the same session by a trained research technician, in a quiet room, in the patient’s choice of either French or English. Clinical information was collected using a semi-structured interview at the time of testing, supplemented by clinic chart review. Patient age, sex, educational level, and mother tongue were recorded and evaluated for their impact on cognitive test performance. Age was coded into 5-year bins and educational level was coded as some vs. no education at the university level. Mother tongue was coded as English, French or other. Clinical characteristics deemed relevant to cognitive test performance and HIV-infection-related

variables were also recorded, including the presence of self-reported cognitive complaints (no/yes), and Selleckchem Sunitinib the presence of depressive symptoms as evaluated with the Beck Depression Inventory II (BDI-II; minimal, mild, moderate or severe). The MoCA was administered and scored according to the published instructions for this test (http://www.mocatest.org). Individual items of the MoCA test were coded dichotomously as failed or passed for each patient, with the exception of Serial 7s subtraction. For this item we used a polytomous scoring system of 0 to 5 based on the sum of correct responses over five consecutive subtractions. Participants performed seven tasks examining different aspects of frontal lobe function. Reversal learning. Participants learned to make response selections based on feedback. The score was the total number of correct selections [28]. Emotion recognition. Participants rated the degree of emotional expression in a series of faces and were scored based on the difference between ratings of emotional and neutral faces [29]. Letter 2-back task. In this working memory test, a series of letters was presented and participants were scored on their ability to detect letters matching the one presented two trials previously [30]. Stop-signal task.

The genes required for PGA synthesis in B. anthracis

are part of the cap operon (capBCADE) located on the pX02 plasmid (Makino et al., 1989; Uchida et al., 1993; Candela et al., 2005). The capD gene encodes a γ-glutamyltranspeptidase, or PGA depolymerase, which was found to be required for the covalent anchoring of PGA to the peptidoglycan in B. anthracis (Candela & Fouet, 2005). PGA producers that lack capD produce a loose slime layer instead of a covalently linked capsule (Candela & Fouet, 2005). While all Group I and II isolates in this study tested negative for the four cap genes tested (including capD), they were still able to produce a covalently linked PGA capsule. Homologs to several of the cap genes AC220 have been found in other bacteria, such as Bacillus subtilis, Bacillus licheniformis, and Staphylococcus epidermidis (Ashiuchi & Misono, 2002; Urushibata et al., 2002; Candela & Fouet, 2005, 2006; Candela et al., 2005; Kocianova Selleckchem Verteporfin et al., 2005). It is possible that the Group I and II isolates contain PGA synthesis genes that more closely resemble these homologs than those in the cap operon of B. anthracis and are thus too divergent to amplify using the cap PCR primers used in this study. A clear difference in colony morphology on bicarbonate

agar was evident for about half of the isolates. These isolates, as well as the B. cereus G9241-positive control, produced dull or dry colonies, whereas virulent strains of B. anthracis produce the shiny or the mucoid morphology. Thus, although commonly used for capsule expression and observation in B. anthracis, the colony morphology of other species on bicarbonate agar alone is not an accurate indicator of capsule production. The d-PGA capsule of B. anthracis is required for virulence, in addition to Phospholipase D1 the tripartite toxin encoded by the pX01 plasmid. The role in virulence, if any, of the B. anthracis-like PGA capsules produced by

Group I and II isolates, however, is unknown. We cannot be certain that these isolates were responsible for the infections, as Bacillus spp. are frequent contaminates of clinical samples, and no further epidemiological data were available (Farrar & Reboli, 2006). However, during the course of this study, 20 clinical isolates from CDC’s Special Bacteriology Reference Laboratory’s (SBRL) culture collection were identified as having a high degree of 16S rRNA gene sequence similarity (99.87–100%) to one or more of the Group I or II isolates, and were subsequently tested for capsule production. These isolates had been sent by various states or territories (AK, CA, CO, MO, OH, PA, PR, TN, VA, and VT) for identification from 2001 to 2008, and were ultimately identified by SBRL as either Bacillus spp. or Brevibacterium spp. Nineteen of the 20 isolates tested positive for capsule production, detected by both methods described above.