Virulence Of Organism Research Articles

A study on characterisation of antimicrobial resistance and antibiotic susceptibility pattern in late preterm and term neonates in a tertiary hospital, Imphal

Background: Neonatal sepsis is defined as a clinical syndrome of bacteremia with systemic signs and symptoms of infection in the first four weeks of life. It may further be divided into two main classes: early onset sepsis, which presents within the first 72 hours of birth and late onset sepsis, which usually presents after 72 hours after birth. The pattern of organisms causing neonatal sepsis has been constantly changing and the indiscriminate use of antibiotics had resulted in the emergence of multidrug resistant and virulent organisms. This study aimed to evaluate neonatal infections and the antibiotic susceptibility patterns.Methods: An institution based cross sectional study in a NICU of a tertiary care hospital. Cases enrolled were both intra and extramural who got admitted during the study period. Informed consent was obtained from the parents/guardians.Results: There were a total of 138 participants in the study. All of them tested positive for sepsis screen. Neonatal sepsis was more common in late pre-terms (57.2%) than terms (42.8%). Gram negative bacteria were the main causative agents with E. coli (9.4%) being the most common isolate followed by Acinetobacter baumanii (8%) and Klebsiella (8%). The most common gram-positive organism isolated was budding yeast cells (Candida spp.). Among gram positive bacteria the most common isolates were methicillin-resistant Staphylococcus aureus (MRSA) (5.8%) and coagulase-negative Staphylococci (CONS) (3.6%). Tigecycline and colistin were most sensitive antibiotics showing the least resistance to all organisms.Conclusions: The antibiotic sensitivity patterns have changed and the morbidity load is higher in pre-terms compared to terms.

The contribution of the intestinal microbiome to immune recovery after HCT.

Allogenic hematopoietic stem-cell transplantation (allo-HCT) is a curative-intent immunotherapy for high-risk hematological malignancies and immune deficiencies. Allo-HCT carries a high risk of treatment-related mortality (TRM), largely due to infection or graft-versus-host disease (GVHD). Robust immune recovery is essential for optimal patient outcomes, given the immunologic graft-versus-leukemia effect prevents relapse, and functional innate and adaptive immunity are both needed for the prevention and control of infection. Most simply, we measure immune recovery by enumerating donor lymphocyte subsets in circulation. In functional terms, ideal immune recovery is more difficult to define, and current lab techniques are limited to the measurement of specific vaccine-responses or mitogens ex vivo. Clinically, poor immune function manifests as problematic infection with viral, bacterial and fungal organisms. Furthermore, the ideal recovering immune system is capable of exerting graft-versus-tumor effects to prevent relapse, and does not induce graft-versus-host disease. Large clinical observational studies have linked loss of diversity within the gut microbiome with adverse transplant outcomes including decreased overall survival and increased acute and chronic GVHD. Furthermore, the correlation between intestinal microbial communities and numeric lymphocyte recovery has now been reported using a number of approaches. Large sets of clinically available white blood cell count data, clinical flow cytometry of lymphocyte subsets and bespoke flow cytometry analyses designed to capture microbiota-specific T cells (e.g. Mucosal-associated invariant T cells, subsets of the gd T cells) have all been leveraged in an attempt to understand links between the microbiota and the recovering immune system in HCT patients. Additionally, preclinical studies suggest an immunomodulatory role for bacterial metabolites (including butyrate, secondary bile acids, and indole derivatives from tryptophan metabolism) in transplant outcomes, though further studies are needed to unravel mechanisms relevant to the post-HCT setting. An understanding of mechanistic relationships between the intestinal microbiome and post-transplant outcomes is necessary for reduction of risk associated with transplant, to inform prophylactic procedures, and ensure optimal immune reconstitution without alloreactivity. Here, we summarize the current understanding of the complex relationship between bacterial communities, their individual members, and the metabolites they produce with immune function in both the allo-HCT and steady-state setting.

Mannitol Anaphylaxis in the Setting of Septic Emboli-Induced Intracranial Hemorrhage

Neurological complications are a significant problem in bacterial endocarditis. Cerebral embolism is the most frequent concern. Acute embolic disease may trigger focal seizures or mycotic aneurysms. Miliary infection is also common, and lumbar puncture can guide in determining the infective organism. Purulent cerebrospinal fluid (CSF) consists often of Staphylococcus aureus, a virulent organism, whereas non-virulent organisms (i.e., viridans streptococci) have normal CSF formulae. Microscopic abscesses suggest the potential for aneurysm from bacterial endocarditis amplifying the risk of intracranial hemorrhage. Mannitol and hypertonic (3%) saline are intravenous medications used as a rescue treatment for brain hemorrhage. A patient diagnosed with mycoplasma pneumonia and septic shock secondary to tricuspid endocarditis with extensive pulmonary emboli and metastatic infection to his spine was initiated on antibiotics. He developed a massive intracranial bleed from the rupture of mycotic septic emboli and was given mannitol to decrease intracranial pressure, which caused anaphylaxis.

A scoping review of penile implant biofilms-what do we know and what remains unknown?

BackgroundPenile prosthesis (PP) is a gold standard for treatment of erectile dysfunction given its reliability and efficacy. Infection remains the most feared complication of prosthetic surgery, which usually results in device removal, and places a significant economic burden on the healthcare system. While biofilms have shown to support the persistence of microorganisms, the degree by which this matrix is truly pathogenic remains unknown given its high prevalence even in asymptomatic patients. We aim to review and summarize the current literature pertaining to biofilm formation in the setting of PP surgeries in clinically infected and non-infected cases.MethodsSearches were performed in the MEDLINE online database through PubMed using a combination of keywords “penile prosthetic” OR “penile prosthesis” OR “penile implant” AND “biofilm” OR “revision” OR “removal” OR “infection” OR “explant”. Eleven articles met inclusion criteria. There were only three studies that explicitly listed the number of biofilms identified in their cohort, but we also included eight articles that mentioned swabbing and culturing of any bacterial biofilm during revision procedures for both clinically infected and non-infected implants.ResultsInfected PP yielded a 11–100% rate of biofilm presence, while non-infected PP yielded a 3–70% rate of biofilm presence. Time to reoperation from initial PP placement were also largely variable, ranging from 2 weeks to over 2 years. Coagulase-negative staphylococcus (i.e., Staphylococcus epidermidis) were the most commonly reported organisms among non-infected implants, however, newer studies have identified a change towards more virulent organisms.ConclusionsSince the advent of PP surgery, diabetes control, revision washout protocols and antibiotic-impregnated devices have led to an overall decrease in biofilm formation and infectious complications. There is an overall paradigm shift in microbial profiles with more virulent organisms, such as Escherichia coli, Pseudomonas aeruginosa, Enterococcus species, and even fungal species beginning to replace the more common coagulase-negative staphylococcal species, especially in clinically infected implants. Additional studies are necessary to define the significance of bacterial presence in biofilms using impactful technologies such as next-generation sequencing. Currently, preliminary and experimental biofilm-control strategies are also underway to further address this clinical issue.

Role of TlyA in the Biology of Uncultivable Mycobacteria.

TlyA proteins are related to distinct functions in a diverse spectrum of bacterial pathogens, including mycobacterial spp. There are several annotated proteins that function as hemolysin or pore-forming molecules that play an important role in the virulence of pathogenic organisms. Many studies reported the dual activity of mycobacterial TlyA as 'hemolysin' and 'Sadenosylmethionine dependent rRNA methylase'. To act as a hemolysin, a sequence must have a signal sequence and transmembrane segment, which helps the protein enter the extracellular environment. Interestingly, the mycobacterial tlyA has neither traditional signal sequences of general/ sec/tat pathways nor any transmembrane segments. Still, it can reach the extracellular milieu with the help of non-classical signal mechanisms. Also, retention of tlyA in cultivable mycobacterial pathogens (such as Mycobacterium tuberculosis and M. marinum) as well as uncultivated mycobacterial pathogens despite their extreme reductive evolution (such as M. leprae, M. lepromatosis and M. uberis) suggests its crucial role in the evolutionary biology of pathogenic mycobacteria. Numerous virulence factors have been characterised by the uncultivable mycobacteria, but the information of TlyA protein is still limited in terms of molecular and structural characterisation. The genomic insights offered by comparative analysis of TlyA sequences and their conserved domains reveal pore-forming activity, which further confirms its role as a virulence protein, particularly in uncultivable mycobacteria. Therefore, this review presents a comparative analysis of the mycobacterial TlyA family by sequence homology and alignment to improve our understanding of this unconventional hemolysin and RNA methyltransferase TlyA of uncultivable mycobacteria.

Symptom profiling for infectious intestinal disease (IID): Do symptom profiles alter with age?

Symptom profiles have previously been identified for infectious intestinal disease (IID) which distinguish bacterial from viral organisms. However, there is evidence that the seasonality, severity, and duration of IID may differ between children, adults and elderly. A secondary data analysis was undertaken to explore whether symptom profiles for bacterial and viral IID vary across different age groups. Data from 844 cases of IID were divided into three age categories: <16 years, 16–65 years and >65 years. Multivariable logistic regression modelling was used to compare the significance of different symptoms across the three age groups. The odds of bacterial IID in children were increased by onset in the summer, diarrhoea in the absence of vomiting and fever. These symptoms were also associated with lower odds of a viral pathogen. In adults, diarrhoea but no vomiting, bloody diarrhoea and diarrhoea lasting more than 3 days were associated with increased odds of a bacterial organism, whilst onset in the winter or spring and a loss of appetite were associated with viral IID. In the elderly, diarrhoea in the absence of vomiting and diarrhoea lasting more than 3 days were associated with higher odds of bacterial IID and lower odds of a viral cause. Only diarrhoea in the absence of vomiting emerged as a key symptom across all three age groups. Variation in symptom profiles by age has implications for clinicians, public health specialists and epidemiologists who use symptoms to guide presumptive diagnoses in the absence of microbiological confirmation.

Lower Respiratory Tract Coinfection in the ICU: Prevalence and Clinical Significance of Coinfection Detected via Microbiological Analysis of Bronchoalveolar Lavage Fluid With a Comparison of Invasive Methodologies.

OBJECTIVES:Pneumonia remains a significant cause of morbidity and mortality, with increasing interest in the detection and clinical significance of coinfection. Further investigation into the impact of bronchoalveolar lavage (BAL) sampling methodology and efficient clinical utilization of microbiological analyses is needed to guide the management of lower respiratory tract infection in the ICU.DESIGN:Retrospective observational study.SETTING:ICUs at a single center between August 1, 2012, and January 1, 2018.PATIENTS:Mechanically ventilated adult patients who underwent BAL testing during an ICU admission were included.INTERVENTIONS:None.MEASUREMENTS AND MAIN RESULTS:BAL methodology (bronchoscopic vs nonbronchoscopic), microbiological diagnostic testing, and clinical outcomes measures were obtained. Chi-square or Fisher exact tests assessed associations between categorical variables, whereas Kruskal-Wallis tests analyzed differences in distributions of measures. BAL samples from 803 patients met inclusion criteria. Coinfection was detected more frequently via bronchoscopic BAL compared with nonbronchoscopic BAL (26% vs 9%; p < 0.001). Viruses were detected more frequently in bronchoscopic (42% vs 13%; p < 0.001) and bacteria in nonbronchoscopic (42% vs 33%; p = 0.011) BALs. A positive correlation between mortality and the number of organisms isolated was identified, with 43%, 48%, and 58% 30-day mortality among those with 0, 1, and more than 2 organisms, respectively (p = 0.003). Viral organism detection was associated with increased 30-day mortality (56% vs 46%; p = 0.033).CONCLUSIONS:Even in the setting of standardized institutional techniques, retrospective evaluation of bronchoscopic and nonbronchoscopic BAL methodologies did not reveal similar microbiologic yield in critically ill patients, though bronchoscopic BAL overall yielded more organisms, and occurrence of multiple organisms in BAL was associated with worse outcome. Prospective data are needed for direct comparison of both methods to develop more standardized approaches for use in different patient groups.

Multidrug-Resistant and Virulent Organisms Trauma Infections: Trauma Infectious Disease Outcomes Study Initiative.

During the wars in Iraq and Afghanistan, increased incidence of multidrug-resistant (MDR) organisms, as well as polymicrobial wounds and infections, complicated the management of combat trauma-related infections. Multidrug resistance and wound microbiology are a research focus of the Trauma Infectious Disease Outcomes Study (TIDOS), an Infectious Disease Clinical Research Program, Uniformed Services University, research protocol. To conduct comprehensive microbiological research with the goal of improving the understanding of the complicated etiology of wound infections, the TIDOS MDR and Virulent Organisms Trauma Infections Initiative (MDR/VO Initiative) was established as a collaborative effort with the Brooke Army Medical Center, Naval Medical Research Center, U.S. Army Institute of Surgical Research, and Walter Reed Army Institute of Research. We provide a review of the TIDOS MDR/VO Initiative and summarize published findings. Antagonism and biofilm formation of commonly isolated wound bacteria (e.g., ESKAPE pathogens-Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.), antimicrobial susceptibility patterns, and clinical outcomes are being examined. Isolates collected from admission surveillance swabs, as part of infection control policy, and clinical infection workups were retained in the TIDOS Microbiological Repository and associated clinical data in the TIDOS database. Over the TIDOS study period (June 2009 to December 2014), more than 8,300 colonizing and infecting isolates were collected from military personnel injured with nearly one-third of isolates classified as MDR. At admission to participating U.S. military hospitals, 12% of wounded warriors were colonized with MDR Gram-negative bacilli. Furthermore, 27% of 913 combat casualties with ≥1 infection during their trauma hospitalization had MDR Gram-negative bacterial infections. Among 335 confirmed combat-related extremity wound infections (2009-2012), 61% were polymicrobial and comprised various combinations of Gram-negative and Gram-positive bacteria, yeast, fungi, and anaerobes. Escherichia coli was the most common Gram-negative bacilli isolated from clinical workups, as well as the most common colonizing MDR secondary to extended-spectrum β-lactamase resistance. Assessment of 479 E. coli isolates collected from wounded warriors found 188 pulsed-field types (PFTs) from colonizing isolates and 54 PFTs from infecting isolates without significant overlap across combat theaters, military hospitals, and study years. A minority of patients with colonizing E. coli isolates developed subsequent infections with the same E. coli strain. Enterococcus spp. were most commonly isolated from polymicrobial wound infections (53% of 204 polymicrobial cultures). Patients with Enterococcus infections were severely injured with a high proportion of lower extremity amputations and genitourinary injuries. Approximately 65% of polymicrobial Enterococcus infections had other ESKAPE organisms isolated. As biofilms have been suggested as a cause of delayed wound healing, wound infections with persistent recovery of bacteria (isolates of same organism collected ≥14 days apart) and nonrecurrent bacterial isolates were assessed. Biofilm production was significantly associated with recurrent bacteria isolation (97% vs. 59% with nonrecurrent isolates; P < 0.001); however, further analysis is needed to confirm biofilm formation as a predictor of persistent wound infections. The TIDOS MDR/VO Initiative provides comprehensive and detailed data of major microbial threats associated with combat-related wound infections to further the understanding of wound etiology and potentially identify infectious disease countermeasures, which may lead to improvements in combat casualty care.

Efficacy of single pass UVC air treatment for the inactivation of coronavirus, MS2 coliphage and Staphylococcus aureus bioaerosols

There is strong evidence that SARS-CoV-2 is spread predominantly by airborne transmission, with high viral loads released into the air as respiratory droplets and aerosols from the infected subject. The spread and persistence of SARS-CoV-2 in diverse indoor environments reinforces the urgent need to supplement distancing and PPE based approaches with effective engineering measures for microbial decontamination – thereby addressing the significant risk posed by aerosols. We hypothesized that a portable, single-pass UVC air treatment device (air flow 1254 L/min) could effectively inactivate bioaerosols containing bacterial and viral indicator organisms, and coronavirus without reliance on filtration technology, at reasonable scale. Robust experiments demonstrated UVC dose dependent inactivation of Staphylococcus aureus (UV rate constant (k) = 0.098 m2/J) and bacteriophage MS2, with up to 6-log MS2 reduction achieved in a single pass through the system (k = 0.119 m2/J). The inclusion of a PTFE diffuse reflector increased the effective UVC dose by up to 34% in comparison to a standard Al foil reflector (with identical lamp output), resulting in significant additional pathogen inactivation (1-log S. aureus and MS2, p < 0.001). Complete inactivation of bovine coronavirus bioaerosols was demonstrated through tissue culture infectivity (2.4-log reduction) and RT-qPCR analysis – confirming single pass UVC treatment to effectively deactivate coronavirus to the limit of detection of the culture-based method. Scenario-based modelling was used to investigate the reduction in risk of airborne person-to-person transmission based upon a single infected subject within the small room. Use of the system providing 5 air changes per hour was shown to significantly reduce airborne viral load and maintain low numbers of RNA copies when the infected subject remained in the room, reducing the risk of airborne pathogen transmission to other room users. We conclude that the application of single-pass UVC systems (without reliance on HEPA filtration) could play a critical role in reducing the risk of airborne pathogen transfer, including SARS-CoV2, in locations where adequate fresh air ventilation cannot be implemented.

Antiviral Activity of Approved Antibacterial, Antifungal, Antiprotozoal and Anthelmintic Drugs: Chances for Drug Repurposing for Antiviral Drug Discovery.

Drug repurposing process aims to identify new uses for the existing drugs to overcome traditional de novo drug discovery and development challenges. At the same time, as viral infections became a serious threat to humans and the viral organism itself has a high ability to mutate genetically, and due to serious adverse effects that result from antiviral drugs, there are crucial needs for the discovery of new antiviral drugs, and to identify new antiviral effects for the exciting approved drugs towards different types of viral infections depending on the observed antiviral activity in preclinical studies or clinical findings is one of the approaches to counter the viral infections problems. This narrative review article summarized mainly the published preclinical studies that evaluated the antiviral activity of drugs that are approved and used mainly as antibacterial, antifungal, antiprotozoal, and anthelmintic drugs, and the preclinical studies included the in silico, in vitro, and in vivo findings, additionally some clinical observations were also included while trying to relate them to the preclinical findings. Finally, the structure used for writing about the antiviral activity of the drugs was according to the families of the viruses used in the studies to form a better image for the target of antiviral activity of different drugs in the different kinds of viruses and to relate between the antiviral activity of the drugs against different strains of viruses within the same viral family.

A Pain in the Neck: An Adolescent with Neck Pain.

A Pain in the Neck: An Adolescent with Neck Pain.

Quantification of Helicobacter pylori and its oncoproteins in the oral cavity: A cross-sectional study.

To identify Helicobacter pylori (H.pylori) and related oncogenic and virulent proteins (CagA and VacA) in patients with gingivitis, periodontitis, oral cancer and gastric cancer. Subgingival plaque samples were collected from 90 individuals with either gingivitis/periodontitis (group A, n=30), oral cancer (group B, n=30) and gastric cancer (group C, n=30). H.pylori was identified by real-time polymerase chain reaction (RT-PCR). The virulent organisms were detected by identification of proteins CagA and VacA through Enzyme Linked Immuno-Sorbent Assay (ELISA). We identified the presence of H.pylori in subgingival plaque samples among a large majority (76/90) of our study cohort. The proportions of CagA and VacA identified among H.pylori individuals with periodontal inflammation and oral cancer were lower than those diagnosed with gastric cancer. Furthermore, the relative risk of oral cancer based on the presence of the organism was no different to those with gingivitis/periodontitis. The findings of our study do not indicate significant association between the organism and oral cancer, but preludes that the oral cavity could act as a potential niche for H.pylori. The possibility for CagA and VacA proteins to be pathogenic in oral cavity is highly possible and must be researched extensively.

Comparison of Illumina and Oxford Nanopore Sequencing Technologies for Pathogen Detection from Clinical Matrices Using Molecular Inversion Probes

Next-generation sequencing is rapidly finding footholds in numerous microbiological fields, including infectious disease diagnostics. Here, we describe a molecular inversion probe panel for the identification of bacterial, viral, and parasitic pathogens. We describe the ability of Illumina and Oxford Nanopore Technologies (ONT) to sequence small amplicons originating from this panel for the identification of pathogens in complex matrices. The panel correctly classified 31 bacterial pathogens directly from positive blood culture bottles with a genus-level concordance of 96.7% and 90.3% on the Illumina and ONT platforms, respectively. Both sequencing platforms detected 18 viral and parasitic organisms directly from mock clinical samples of plasma and whole blood at concentrations of 104 PFU/mL with few exceptions. In general, Illumina sequencing exhibited greater read counts with lower percent mapped reads; however, this resulted in no effect on limits of detection compared with ONT sequencing. Mock clinical evaluation of the probe panel on the Illumina and ONT platforms resulted in positive predictive values of 0.91 and 0.88 and negative predictive values of 1 and 1 from de-identified human chikungunya virus samples compared with gold standard quantitative RT-PCR. Overall, these data show that molecular inversion probes are an adaptable technology capable of pathogen detection from complex sample matrices on current next-generation sequencing platforms.

Association of High-risk Human Papillomavirus Titer and Pathogenic Co-infections with Cervical Tissue Cytopathology

Human papillomaviruses (HPV) are one of the first viral organisms acknowledged to causing carcinogenesis. Among gynecologic cancers, Pap smear represents a gold standard diagnostic procedure for precancerous cervical lesions. It is efficiently interpreted through a standardized reporting system; The Bethesda System, which aids in distinguishing squamous categories from other entities. Co-infections with other sexually transmitted infections (STIs) could exacerbate cervical lesion severity caused by initial HPV infection as co-infections can lead to distinct reprogramming of host cells and genome integrity. The intricate pathways and effect of the unique cellular microenvironment that HPV and co-infecting STIs create that cause local inflammation and eventually cervical lesion progression will be reviewed in this manuscript. Besides, it is also crucial to consider HPV viral load and distinguish its correlation with cervical lesion severity. Varying amounts of viral titer and its impact on cervical lesions could indicate a mutagenic transformation of the human host cells and HPV. Thus, this review aims to discuss the correlation of co-infections and viral titer on cervical lesion severity and its progression to cancer. Based on these factors, clear clinical reasoning with more effective treatment plans and specific diagnostics can be achieved. Keywords: Cervical cancer, Cervical lesions, Co-infection, Human papillomavirus, Viral titer

Future Applications of Metagenomic Next-Generation Sequencing for Infectious Diseases Diagnostics.

Metagenomic next-generation sequencing (mNGS) has the theoretical capacity to detect any microbe present in a host. mNGS also has the potential to infer a pathogen's phenotypic characteristics, including the ability to colonize humans, cause disease, and resist treatment. Concurrent host nucleic acid sequencing can assess the infected individual's physiological state, including characterization and appropriateness of the immune response. When the pathogen cannot be identified, host RNA sequencing may help infer the organism's nature. While the full promise of mNGS remains far from realization, the potential ability to identify all microbes in a complex clinical sample, assess each organism's virulence and antibiotic susceptibility traits, and simultaneously characterize the host's response to infection provide opportunities for mNGS to supplant existing technologies and become the primary method of infectious diseases diagnostics.

THE CLINICAL PROFILE OF DENGUE PATIENTS IN CHILDREN

Background: To study the clinical profile of dengue in children&#x0D; Methods: The hospital based study was conducted on patients presenting to paediatric hospital, who fulfilled inclusion and exclusion criteria.&#x0D; Results: Based on the symptoms, the most common symptoms noticed were fever 94.00% followed by myalgia 85.00% decreased appetite 83%, retroorbital pain in 84.0% and vomiting 81.00%&#x0D; Conclusion: It concluded that common symptoms observed were fever, myalgia, decreased appetite and headache The common complications presented were hepatic dysfunction and shock with no mortality indicating the presence of less virulent organisms..&#x0D; Keywords: Dengue, Complication, Shock

Dynamics of Antibacterial Drone Establishment in Staphylococcus aureus: Unexpected Effects of Antibiotic Resistance Genes.

ABSTRACTThe antibacterial drone (ABD) system is based on repurposing the phage-inducible staphylococcal pathogenicity islands (SaPIs) for use as antibacterial agents that are indifferent to antibiotic resistance. The ABDs were constructed by inserting tetM for tetracycline resistance (Tcr) selection, replacing the SaPI virulence genes with bactericidal or bacteriostatic genes such as CRISPR/cas9/agrA, whose expression kills by double-strand cleavage of agrA, or CRISPR/dcas9/agrP2P3, whose expression blocks the target organism’s virulence. ABD DNA is packaged in phage-like particles that attack their staphylococcal targets in vivo as well as in vitro. We determine ABD titers by transfer frequency, enumerate surviving cells as a function of multiplicity, and analyze the fate of ABD DNA with green fluorescent protein. An initial study revealed surprisingly that many more cells were killed by the ABD than were measured by transduction. Our study of this phenomenon has revealed several important features of the ABD system: (i) a significant number of entering ABD DNA molecules do not go on to establish stable transductants (i.e., are abortive); (ii) ABD cargo genes are expressed immediately following entry, even by the abortive ABDs; (iii) immediate plating on Tc-containing agar seriously underestimates particle numbers, partly owing to Tc inhibition of protein synthesis; (iv) replacement of tetM with cadA (conferring resistance to CdCl2) provides more accurate particle enumeration; (v) ABDs expressing CRISPR/cas9/agrA kill ∼99.99% of infected cells and provide the most accurate measurement of particle numbers as well as proof of principle for the system; and (vi) surprisingly, TetM interferes with stable establishment of ABD DNA independently of Tcr.

Isolation And Characterization Of Biosurfactant Producing Bacteria From Different Environmental Soil Samples

Isolation And Characterization Of Biosurfactant Producing Bacteria From Different Environmental Soil Samples

A site assessment tool for inpatient controlled human infection models for enteric disease pathogens.

The use of the controlled human infection model to facilitate product development and to advance understanding of host-pathogen interactions is of increasing interest. While administering a virulent (or infective) organism to a susceptible host necessitates an ongoing evaluation of safety and ethical considerations, a central theme in conducting these studies in a safe and ethical manner that yields actionable data is their conduct in facilities well-suited to address their unique attributes. To that end, we have developed a framework for evaluating potential sites in which to conduct inpatient enteric controlled human infection model to ensure consistency and increase the likelihood of success.

Scalable Nucleic Acid Storage and Retrieval Using Barcoded Microcapsules.

Rapid advances in nucleic acid sequencing and synthesis technologies have spurred a major need to collect, store, and sequence the DNA and RNA from viral, bacterial, and mammalian sources and organisms. However, current approaches to storing nucleic acids rely on a low-temperature environment and require robotics for access, posing challenges for scalable and low-cost nucleic acid storage. Here, we present an alternative method for storing nucleic acids, termed Preservation and Access of Nucleic aciDs using barcOded micRocApsules (PANDORA). Nucleic acids spanning kilobases to gigabases and from different sources, including animals, bacteria, and viruses, are encapsulated into silica microcapsules to protect them from environmental denaturants at room temperature. Molecular barcodes attached to each microcapsule enable sample pooling and subsequent identification and retrieval using fluorescence-activated sorting. We demonstrate quantitative storage and rapid access to targeted nucleic acids from a pool emulating standard retrieval operations implemented in conventional storage systems, including recovery of 100,000-200,000 samples and Boolean logic selection using four unique barcodes. Quantitative polymerase chain reaction and short-read sequencing of the retrieved samples validated the sorting experiments and the integrity of the released nucleic acids. Our proposed approach offers a scalable long-term, room-temperature storage and retrieval of nucleic acids with high sample fidelity.