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We evaluated use of maribavir (MBV) for treatment of 15 episodes of refractory/resistant cytomegalovirus infection in 13 solid organ transplant recipients. Treatment failure due to treatment-emergent MBV resistance or early virological recurrence after MBV discontinuation occurred in 7 (47%) episodes. Sustained viral clearance was achieved in 6 (40%) episodes.
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BACKGROUND: Better access to direct-acting antiviral (DAA) therapy has broadened the utilization of hepatitis C virus (HCV) nucleic acid testing (NAT) positive organs with excellent outcomes. However, DAA therapy has been associated with hepatitis B virus (HBV) reactivation. AIM: To determine the risk of HBV transmission or reactivation with utilization of HBV core antibody positive (HBcAb+) and HCV NAT positive (HCV+) organs, which presumably required DAA therapy. METHODS: The number of HBcAb+ donors with delineated HCV NAT status was obtained from the Organ Procurement and Transplantation Network (OPTN) database. The number of unexpected HBV infections from transplanted organs adjudicated as "proven" or "probable" transmission was obtained from the OPTN Ad Hoc Disease Transmission Advisory Committee database. A chart review of the donors of "proven" or "probable" cases was conducted. RESULTS: From January 1, 2016, to December 31, 2021, 7735 organs were procured from 3767 HBcAb+ donors and transplanted into 7469 recipients; 545 (14.5%) donors were also HCV+. HBV transmission or reactivation occurred in seven recipients. The rate is not significantly different between recipients of HCV+ (0.18%, 2/1115) and the HCV NAT negative (HCV-) organs (0.08%, 5/6354) (p = 0.28) or between recipients of HCV+ and HCV- livers as well as non-liver organs. HBV transmission or reactivation occurred within a median of 319 (range, 41-1117) days post-transplant in the setting of missing, inadequate, or truncated prophylaxis. CONCLUSION: HBV reactivation associated with DAA therapy for HBcAb+ HCV+ organs is less frequent than reported in the non-transplant population, possibly due to the common use of HBV prophylaxis in the at-risk transplant population.
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This study identified 26 late invasive primary surgical site infection (IP-SSI) within 4-12 months of transplantation among 2073 SOT recipients at Duke University Hospital over the period 2015-2019. Thoracic organ transplants accounted for 25 late IP-SSI. Surveillance for late IP-SSI should be maintained for at least one year following transplant.
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Cytomegalovirus (CMV) is a common viral pathogen in the transplant population and is associated with significant morbidity and mortality. CMV prevention is paramount; however, selecting the best preventive strategy depends on many factors including donor-recipient CMV serostatus, transplant-specific risks, antiviral toxicities and cost. Novel CMV therapeutics such as letermovir (LTV) are desperately needed to optimize CMV management. Uniquely among CMV antiviral therapies, LTV inhibits the viral terminase complex in the CMV DNA synthesis pathway and disrupts viral genome packaging. Further, it lacks side effects frequently associated with other CMV antiviral therapies and evades common mechanisms of resistance. LTV is approved by the US Food and Drug Administration for CMV prevention in adult CMV-seropositive hematopoietic cell transplant recipients but is increasingly applied off-label for prophylaxis and treatment. This review summarizes important concepts of CMV management in transplantation, with a specific focus on LTV pharmacology and clinical experience to date alongside future prospects for its application.
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Infecções por Citomegalovirus , Transplante de Células-Tronco Hematopoéticas , Adulto , Humanos , Citomegalovirus/genética , Infecções por Citomegalovirus/tratamento farmacológico , Infecções por Citomegalovirus/prevenção & controle , Infecções por Citomegalovirus/epidemiologia , Transplantados , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Antivirais/uso terapêuticoRESUMO
Purpose of Review: Histoplasmosis remains a challenging infection in solid organ transplantation. This review provides a topic update with emphasis on the changing Histoplasma epidemiology, along with new diagnostic and treatment innovations. Recent Findings: Recent years have observed expanding Histoplasma geographic distribution due to climate change, environmental disruption, and host factors. Early clinical experience also suggests a relationship between COVID-19 infection and histoplasmosis, particularly among immunocompromised individuals. Advances in diagnostic methods, such as newer enzyme immunoassays and molecular techniques, have broadened the capability for expedient and highly specific pathogen identification. Novel drug innovations, including the development of new formulations of existing antifungal agents, extended-spectrum azoles and new antifungal drug classes have expanded therapeutic options. Summary: Advances in organ transplantation have largely outpaced those for histoplasmosis. However, these emerging insights enhance our understanding of this pathogen and management of clinical infection, particularly for transplant recipients with a higher incidence and severity of disease.
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The cooperation between the actin and microtubule (MT) cytoskeletons is important for cellular processes such as cell migration and muscle cell development. However, a full understanding of how this cooperation occurs has yet to be sufficiently developed. The MT plus-end tracking protein CLIP-170 has been implicated in this actin-MT coordination by associating with the actin-binding signaling protein IQGAP1 and by promoting actin polymerization through binding with formins. Thus far, the interactions of CLIP-170 with actin were assumed to be indirect. Here, we demonstrate using high-speed cosedimentation assays that CLIP-170 can bind to filamentous actin (F-actin) directly. We found that the affinity of this binding is relatively weak but strong enough to be significant in the actin-rich cortex, where actin concentrations can be extremely high. Using CLIP-170 fragments and mutants, we show that the direct CLIP-170-F-actin interaction is independent of the FEED domain, the region that mediates formin-dependent actin polymerization, and that the CLIP-170 F-actin-binding region overlaps with the MT-binding region. Consistent with these observations, in vitro competition assays indicate that CLIP-170-F-actin and CLIP-170-MT interactions are mutually exclusive. Taken together, these observations lead us to speculate that direct CLIP-170-F-actin interactions may function to reduce the stability of MTs in actin-rich regions of the cell, as previously proposed for MT end-binding protein 1.
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Actinas , Microtúbulos , Actinas/metabolismo , Forminas , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo , Proteínas Associadas aos Microtúbulos/metabolismo , Microtúbulos/metabolismo , Proteínas de Neoplasias/metabolismoRESUMO
BACKGROUND: Solid organ transplant recipients (SOTR) have diminished humoral immune responses to COVID-19 vaccination and higher rates of COVID-19 vaccine breakthrough infection than the general population. Little is known about COVID-19 disease severity in SOTR with COVID-19 vaccine breakthrough infections. METHODS: Between 4/7/21 and 6/21/21, we requested case reports via the Emerging Infections Network (EIN) listserv of SARS-CoV-2 infection following COVID-19 vaccination in SOTR. Online data collection included patient demographics, dates of COVID-19 vaccine administration, and clinical data related to COVID-19. We performed a descriptive analysis of patient factors and evaluated variables contributing to critical disease or need for hospitalization. RESULTS: Sixty-six cases of SARS-CoV-2 infection after vaccination in SOTR were collected. COVID-19 occurred after the second vaccine dose in 52 (78.8%) cases, of which 43 (82.7%) occurred ≥14 days post-vaccination. There were six deaths, three occurring in fully vaccinated individuals (7.0%, n = 3/43). There was no difference in the percentage of patients who recovered from COVID-19 (70.7% vs. 72.2%, p = .90) among fully and partially vaccinated individuals. We did not identify any differences in hospitalization (60.5% vs. 55.6%, p = .72) or critical disease (20.9% vs. 33.3%, p = .30) among those who were fully versus partially vaccinated. CONCLUSIONS: SOTR vaccinated against COVID-19 can still develop severe, and even critical, COVID-19 disease. Two doses of mRNA COVID-19 vaccine may be insufficient to protect against severe disease and mortality in SOTR. Future studies to define correlates of protection in SOTR are needed.
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COVID-19 , Transplante de Órgãos , COVID-19/prevenção & controle , Vacinas contra COVID-19 , Humanos , Transplante de Órgãos/efeitos adversos , SARS-CoV-2 , Transplantados , VacinaçãoAssuntos
COVID-19/epidemiologia , Transplante de Órgãos/efeitos adversos , Transplantados/estatística & dados numéricos , Adulto , COVID-19/imunologia , COVID-19/virologia , Vacinas contra COVID-19/imunologia , Centers for Disease Control and Prevention, U.S./estatística & dados numéricos , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/prevenção & controle , Humanos , Imunossupressores/administração & dosagem , Imunossupressores/efeitos adversos , Vacinação em Massa/estatística & dados numéricos , Medição de Risco/estatística & dados numéricos , SARS-CoV-2/imunologia , Estados Unidos/epidemiologiaRESUMO
The purpose of this investigation was to determine the effects of varying facemask reinforcement and visor tint on peripheral visuomotor abilities in collegiate football players. Division I NCAA football players (n = 14) completed two peripheral visuomotor experiments: (1) Varying facemask reinforcement, (2) Varying visor tinting. In experiment 1, participants were tested under the following conditions: baseline (no helmet; BL), helmet + light (HL), helmet + medium (HM), helmet + heavy (HH), and helmet + extra heavy (HXH) reinforced facemasks. In experiment 2, participants were tested under the following conditions: baseline (no helmet; BL), helmet only (HO), helmet + clear (HCV), helmet + smoke-tinted (HSV), and helmet + mirror-tinted (HMV) visors. For each condition, a 60 s peripheral visuomotor test was completed on a Dynavision D2 visuomotor board. For experiment 1, the BL peripheral reaction time (PRT) was faster than all facemask conditions (p < 0.05). Furthermore, PRT was impaired with the HXH compared to HL (p < 0.001), HM (p < 0.001), and HH (p = 0.001). Both HH and HXH resulted in the potentiation of PRT impairments in the outermost and inferior peripheral visual areas (p < 0.05). In experiment 2, BL PRT was faster than all helmeted conditions (p < 0.05). Additionally, PRT was slower in HSV (p = 0.013) and HMV (p < 0.001) conditions compared to HO. HMV resulted in slower PRT in all peripheral areas (p < 0.05) while PRT was impaired only in outer areas for HSV (p < 0.05). Wearing protective football headgear impairs peripheral visuomotor ability. Lighter reinforced facemasks and clear visors do not appear to exacerbate impairment. However, heavier reinforced facemasks and tinted visors further decrease visuomotor performance in outer and inferior visual areas, indicating a potential need for considerations of on-field player performance and safety.
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Portable and inexpensive analytical tools are required to monitor pharmaceutical quality in technology limited settings including low- and middle-income countries (LMICs). Whole cell yeast biosensors have the potential to help meet this need. However, most of the readouts for yeast biosensors require expensive equipment or reagents. To overcome this challenge, we have designed a yeast biosensor that produces a unique scent as a readout. This inducible scent biosensor, or "scentsor", does not require the user to administer additional reagents for reporter development and utilizes only the user's nose to be "read". In this Letter, we describe a scentsor that is responsive to the hormone estradiol (E2). The best estimate threshold (BET) for E2 detection with a panel of human volunteers (n = 49) is 39 nM E2 (15 nM when "non-smellers" are excluded). This concentration of E2 is sensitive enough to detect levels of E2 that would be found in dosage forms. This paper provides evidence that scent has the potential for use in portable yeast biosensors as a readout, particularly for use in LMICs.
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Técnicas Biossensoriais , Preparações Farmacêuticas , Humanos , Saccharomyces cerevisiae/genéticaRESUMO
PURPOSE OF REVIEW: Nocardia is a ubiquitous pathogen associated with life-threatening opportunistic infections. Organ transplant recipients are uniquely predisposed to Nocardia infections due to their iatrogenic cell-mediated immune deficit necessary to maintain allograft function. This review aims to address recent updates in the epidemiology, clinical presentation, diagnostics, treatment, and outcomes of Nocardia infections in solid-organ transplant recipients. RECENT FINDINGS: The incidence of Nocardia infection depends on multiple patient and environmental factors. Among transplant recipients, lung recipients are most commonly affected. Species identification and antimicrobial susceptibility testing are critical for optimizing therapy as substantial variation occurs among and within Nocardia spp. This has been increasingly accomplished through advances in molecular methods leading to improved accuracy and wider accessibility to testing. There are emerging data applying novel therapeutics and short course therapy that may offer alternative management approaches for transplant associated nocardiosis to minimize drug toxicity and intolerance. SUMMARY: Further prospective, multicenter studies are needed to better characterize the epidemiology of Nocardia in transplant recipients, as well as evaluate the impact of diagnostic advancements and new treatment strategies.
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Nocardiose/epidemiologia , Transplante de Órgãos/estatística & dados numéricos , Humanos , Nocardia/isolamento & purificação , Nocardiose/diagnóstico , Nocardiose/imunologia , Infecções Oportunistas/diagnóstico , Infecções Oportunistas/epidemiologia , Infecções Oportunistas/imunologia , Transplante de Órgãos/efeitos adversosRESUMO
BACKGROUND: Short duration, antimicrobial prophylaxis that includes antistaphylococcal activity is recommended at the time of left ventricular assist device (LVAD) implantation to reduce infection-related complications. There continues to be wide variability in surgical infection prophylaxis (SIP) regimens among implantation centers. The aim of this study is to characterize current SIP regimens at different LVAD centers. METHODS: A survey study was conducted from 26 September 2017 to 25 October 2017. Surveys were distributed electronically to LVAD coordinators and infectious diseases specialists at 75 US medical centers identified as having an LVAD program. Data collection included information about antimicrobial selection, duration, Staphylococcus aureus screening, and decolonization procedures. RESULTS: We received 29 survey responses. The majority of surveys were completed by infectious diseases physicians (72.4% [21 out of 29]). Most responding centers reported LVAD programs established for greater than 10 years (20 out of 29 [69%]). Cardiac transplantation was performed in 28 out of 29 (96%) centers. Of centers reporting a defined SIP regimen for non-penicillin allergic patients (96% [28 out of 29]), 17.9% (5 out of 28) reported a four-drug regimen, 35.7% (10 out of 28) reported a three-drug regimen, and 46.4% (13 out of 28) reported a two-drug regimen, while no centers reported a single-drug regimen. Empiric fluconazole was common (50% [14 out of 28]) and 96.4% (27 out of 28) of regimens included vancomycin. Duration of antimicrobial prophylaxis (24 hours to 5 days), S. aureus screening, decolonization procedures, and alterations due to drug allergies varied across participating centers. CONCLUSIONS: Our survey results indicate wide variation in SIP regimens among participating LVAD centers. These results highlight the need for studies evaluating the implications of SIP regimens, and whether clinical factors that prolong antimicrobial duration impact postoperative infection rates.
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Antibacterianos/administração & dosagem , Antibioticoprofilaxia , Coração Auxiliar/efeitos adversos , Implantação de Prótese/efeitos adversos , Infecções Relacionadas à Prótese/prevenção & controle , Infecção da Ferida Cirúrgica/prevenção & controle , Inquéritos e Questionários , Aztreonam/administração & dosagem , Cefalosporinas/administração & dosagem , Estudos Transversais , Quimioterapia Combinada , Fluconazol/administração & dosagem , Humanos , Levofloxacino/administração & dosagem , Infecções Relacionadas à Prótese/etiologia , Rifampina/administração & dosagem , Infecção da Ferida Cirúrgica/etiologia , Vancomicina/administração & dosagemAssuntos
Aspergilose Pulmonar Invasiva , Aspergilose Pulmonar , Antifúngicos/uso terapêutico , Humanos , Aspergilose Pulmonar Invasiva/diagnóstico , Aspergilose Pulmonar Invasiva/prevenção & controle , Pulmão , Aspergilose Pulmonar/diagnóstico , Aspergilose Pulmonar/tratamento farmacológico , Aspergilose Pulmonar/prevenção & controle , TransplantadosRESUMO
BACKGROUND: Respiratory syncytial virus (RSV) infection in lung transplant recipients (LTRs) causes mortality rates of 10%-20% despite antiviral therapy. Ribavirin (RBV) has been used to treat RSV-infected LTRs with limited data. METHODS: A retrospective study including all LTRs at Duke Hospital during January 2013-May 2017 with positive RSV polymerase chain reaction respiratory specimens was performed. RESULTS: Fifty-six of 70 patients in the oral RBV group and 29 of 32 in the inhaled RBV group had symptomatic RSV infection. One patient receiving oral RBV had to prematurely stop drug due to significant nausea and vomiting. While unadjusted all-cause 1-year mortality was significantly higher in the inhaled RBV group (24.1% versus 7.1% [oral RBV], P = 0.03), adjusted hazard ratio (HR) for death and oral RBV use (compared to inhaled RBV), accounting for oxygen requirement and need for mechanical ventilation, showed the HR for death and oral RBV use was 0.38 ([0.10, 1.46], P = 0.38). The HR for death in patients with supplemental oxygen >2 L/min at diagnosis was 6.18 ([1.33, 26.83], P = 0.02). Kaplan-Meier curves showed patients with forced expiratory volume in 1 second decline ≥5% and ≥10% at 90 days post-RSV infection had a higher 1-year mortality (P = 0.004 and P = 0.001, respectively). CONCLUSIONS: Oral and inhaled RBV appear to be well tolerated in LTRs, and our data support the use of oral RBV as a safe alternative to inhaled ribavirin in LTRs. Oxygen requirement >2 L/min at diagnosis and forced expiratory volume in 1 second decline ≥5% postinfection may be markers for increased mortality.
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Antivirais/administração & dosagem , Transplante de Pulmão/efeitos adversos , Complicações Pós-Operatórias/terapia , Infecções por Vírus Respiratório Sincicial/terapia , Ribavirina/administração & dosagem , Administração por Inalação , Administração Oral , Adulto , Idoso , Feminino , Volume Expiratório Forçado , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Oxigênio/administração & dosagem , Complicações Pós-Operatórias/imunologia , Complicações Pós-Operatórias/mortalidade , Complicações Pós-Operatórias/virologia , RNA Viral/isolamento & purificação , Respiração Artificial/estatística & dados numéricos , Infecções por Vírus Respiratório Sincicial/imunologia , Infecções por Vírus Respiratório Sincicial/mortalidade , Infecções por Vírus Respiratório Sincicial/virologia , Vírus Sinciciais Respiratórios/genética , Vírus Sinciciais Respiratórios/imunologia , Vírus Sinciciais Respiratórios/isolamento & purificação , Estudos Retrospectivos , Transplantados/estatística & dados numéricos , Resultado do TratamentoRESUMO
The purpose of this study was to examine the effects of protective football headgear on peripheral vision reaction time and visual target detection. Twenty-five Division I NCAA football players (age = 20.5 yrs ± 0.9, height = 185.9 cm ± 6.8, body mass = 99.2 kg ± 19.2, BMI = 29.6 ± 4.5) participated. In a crossover counterbalanced study design, subjects participated in one visit with three conditions: Baseline (BL) without headgear, helmet only (HO), helmet with an eye shield (HE). Subjects completed a 1-min peripheral vision reaction time test for each condition separated by 3-min recovery periods. Tests were administered using a 64 light Dynavision D2 Visuomotor board. Target detection (total hit score) was higher during BL than HO (p < 0.001) and HE (p < 0.001). Average (p < 0.001), peak (p < 0.001), minimum (p < 0.001), and median (p < 0.001) peripheral reaction times were faster during BL than HO and HE. No significant differences were observed for any measures between HO and HE conditions (p > 0.05). Findings indicate that protective football headgear impaired reaction time to peripheral visual stimuli. The addition of an eye shield to the helmet had a small non-significant effect on reaction time and target detection. These results may hold important implications in helmet design and player safety.
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The unexpected transmission of donor-derived infection through organ transplantation is a rare event with current donor screening practices. In this case report we describe a probable donor-derived transmission of Herpes Simplex Virus (HSV)-2 via deceased donor kidney transplantation resulting in HSV hepatitis in the recipient. This manifested as acute liver failure which resolved with appropriate anti-viral therapy. Following recovery from the acute liver insult, the patient developed fibrotic liver morphology and portal hypertension, an unusual departure from the typical course.
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Hepatite Viral Humana/virologia , Herpes Simples/virologia , Hipertensão Portal/etiologia , Transplante de Rim/efeitos adversos , Cirrose Hepática/virologia , Falência Hepática Aguda/etiologia , Aciclovir/uso terapêutico , Adulto , Aloenxertos/virologia , Antivirais/uso terapêutico , Biópsia , Feminino , Hepatite Viral Humana/diagnóstico , Hepatite Viral Humana/patologia , Hepatite Viral Humana/transmissão , Herpes Simples/diagnóstico , Herpes Simples/patologia , Herpes Simples/transmissão , Herpesvirus Humano 2/isolamento & purificação , Humanos , Hipertensão Portal/patologia , Hipertensão Portal/terapia , Rim/virologia , Fígado/patologia , Fígado/virologia , Cirrose Hepática/complicações , Cirrose Hepática/patologia , Cirrose Hepática/terapia , Falência Hepática Aguda/patologia , Falência Hepática Aguda/terapia , Resultado do TratamentoRESUMO
The Salmonella cytolethal distending toxin (S-CDT), first described as the "typhoid toxin" in Salmonella enterica subsp. enterica serotype Typhi, induces DNA damage in eukaryotic cells. Recent studies have shown that more than 40 nontyphoidal Salmonella (NTS) serotypes carry genes that encode S-CDT, yet very little is known about the activity, function, and role of S-CDT in NTS. Here we show that deletion of genes encoding the binding subunit (pltB) and a bacteriophage muramidase predicted to play a role in toxin export (ttsA) does not abolish toxin activity in the S-CDT-positive NTS Salmonella enterica subsp. enterica serotype Javiana. However, S. Javiana strains harboring deletions of both pltB and its homolog artB, had a complete loss of S-CDT activity, suggesting that S. Javiana carries genes encoding two variants of the binding subunit. S-CDT-mediated DNA damage, as determined by phosphorylation of histone 2AX (H2AX), producing phosphorylated H2AX (γH2AX), was restricted to epithelial cells in S and G2/M phases of the cell cycle and did not result in apoptosis or cell death. Compared to mice infected with a ΔcdtB strain, mice infected with wild-type S. Javiana had significantly higher levels of S. Javiana in the liver, but not in the spleen, ileum, or cecum. Overall, we show that production of active S-CDT by NTS serotype S. Javiana requires different genes (cdtB, pltA, and either pltB or artB) for expression of biologically active toxin than those reported for S-CDT production by S. Typhi (cdtB, pltA, pltB, and ttsA). However, as in S. Typhi, NTS S-CDT influences the outcome of infection both in vitro and in vivoIMPORTANCE Nontyphoidal Salmonella (NTS) are a major cause of bacterial food-borne illness worldwide; however, our understanding of virulence mechanisms that determine the outcome and severity of nontyphoidal salmonellosis is incompletely understood. Here we show that S-CDT produced by NTS plays a significant role in the outcome of infection both in vitro and in vivo, highlighting S-CDT as an important virulence factor for nontyphoidal Salmonella serotypes. Our data also contribute novel information about the function of S-CDT, as S-CDT-mediated DNA damage occurs only during certain phases of the cell cycle, and the resulting damage does not induce cell death as assessed using a propidium iodide exclusion assay. Importantly, our data support that, despite having genetically similar S-CDT operons, NTS serotype S. Javiana has different genetic requirements than S. Typhi, for the production and export of active S-CDT.
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Toxinas Bacterianas/metabolismo , Toxinas Bacterianas/farmacologia , Dano ao DNA/efeitos dos fármacos , Salmonella/metabolismo , Salmonella/patogenicidade , Animais , Ciclo Celular/efeitos dos fármacos , Ciclo Celular/genética , Divisão Celular/efeitos dos fármacos , Divisão Celular/genética , Linhagem Celular , Dano ao DNA/genética , Fase G2/efeitos dos fármacos , Fase G2/genética , Células HeLa , Humanos , Fígado/microbiologia , Camundongos , Camundongos Endogâmicos C57BL , Fase S/efeitos dos fármacos , Fase S/genética , Salmonella enterica/metabolismo , Salmonella enterica/patogenicidade , Virulência/efeitos dos fármacos , Virulência/genéticaRESUMO
While some species in the Bacillus cereus group are well-characterized human pathogens (e.g., B. anthracis and B. cereus sensu stricto), the pathogenicity of other species (e.g., B. pseudomycoides) either has not been characterized or is presently not well understood. To provide an updated characterization of the pathogenic potential of species in the B. cereus group, we classified a set of 52 isolates, including 8 type strains and 44 isolates from dairy-associated sources, into 7 phylogenetic clades and characterized them for (i) the presence of toxin genes, (ii) phenotypic characteristics used for identification, and (iii) cytotoxicity to human epithelial cells. Overall, we found that B. cereus toxin genes are broadly distributed but are not consistently present within individual species and/or clades. After growth at 37°C, isolates within a clade did not typically show a consistent cytotoxicity phenotype, except for isolates in clade VI (B. weihenstephanensis/B. mycoides), where none of the isolates were cytotoxic, and isolates in clade I (B. pseudomycoides), which consistently displayed cytotoxic activity. Importantly, our study highlights that B. pseudomycoides is cytotoxic toward human cells. Our results indicate that the detection of toxin genes does not provide a reliable approach to predict the pathogenic potential of B. cereus group isolates, as the presence of toxin genes is not always consistent with cytotoxicity phenotype. Overall, our results suggest that isolates from multiple B. cereus group clades have the potential to cause foodborne illness, although cytotoxicity is not always consistently found among isolates within each clade.IMPORTANCE Despite the importance of the Bacillus cereus group as a foodborne pathogen, characterizations of the pathogenic potential of all B. cereus group species were lacking. We show here that B. pseudomycoides (clade I), which has been considered a harmless environmental microorganism, produces toxins and exhibits a phenotype consistent with the production of pore-forming toxins. Furthermore, B. mycoides/B. weihenstephanensis isolates (clade VI) did not show cytotoxicity when grown at 37°C, despite carrying multiple toxin genes. Overall, we show that the current standard methods to characterize B. cereus group isolates and to detect the presence of toxin genes are not reliable indicators of species, phylogenetic clades, or an isolate's cytotoxic capacity, suggesting that novel methods are still needed for differentiating pathogenic from nonpathogenic species within the B. cereus group. Our results also contribute data that are necessary to facilitate risk assessments and a better understanding as to which B. cereus group species are likely to cause foodborne illness.
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Bacillus/patogenicidade , Toxinas Bacterianas/metabolismo , Bacillus/química , Bacillus/genética , Bacillus cereus/química , Bacillus cereus/genética , Bacillus cereus/patogenicidade , Genoma Bacteriano , FilogeniaRESUMO
The Bacillus cereus group comprises nine species, several of which are pathogenic. Differentiating between isolates that may cause disease and those that do not is a matter of public health and economic importance, but it can be particularly challenging due to the high genomic similarity within the group. To this end, we have developed BTyper, a computational tool that employs a combination of (i) virulence gene-based typing, (ii) multilocus sequence typing (MLST), (iii) panC clade typing, and (iv) rpoB allelic typing to rapidly classify B. cereus group isolates using nucleotide sequencing data. BTyper was applied to a set of 662 B. cereus group genome assemblies to (i) identify anthrax-associated genes in non-B. anthracis members of the B. cereus group, and (ii) identify assemblies from B. cereus group strains with emetic potential. With BTyper, the anthrax toxin genes cya, lef, and pagA were detected in 8 genomes classified by the NCBI as B. cereus that clustered into two distinct groups using k-medoids clustering, while either the B. anthracis poly-γ-d-glutamate capsule biosynthesis genes capABCDE or the hyaluronic acid capsule hasA gene was detected in an additional 16 assemblies classified as either B. cereus or Bacillus thuringiensis isolated from clinical, environmental, and food sources. The emetic toxin genes cesABCD were detected in 24 assemblies belonging to panC clades III and VI that had been isolated from food, clinical, and environmental settings. The command line version of BTyper is available at https://github.com/lmc297/BTyper In addition, BMiner, a companion application for analyzing multiple BTyper output files in aggregate, can be found at https://github.com/lmc297/BMinerIMPORTANCE Bacillus cereus is a foodborne pathogen that is estimated to cause tens of thousands of illnesses each year in the United States alone. Even with molecular methods, it can be difficult to distinguish nonpathogenic B. cereus group isolates from their pathogenic counterparts, including the human pathogen Bacillus anthracis, which is responsible for anthrax, as well as the insect pathogen B. thuringiensis By using the variety of typing schemes employed by BTyper, users can rapidly classify, characterize, and assess the virulence potential of any isolate using its nucleotide sequencing data.