Virulence Of Organism Research Articles

Distribution and seasonality of viral respiratory infections in pediatric oncologic patients in a tropical country

Abstract Background Comprehension of the distribution and seasonality viral respiratory infections pediatric oncologic patients in tropical localities remains poor. Due to lack mandatory report of all viruses and limitations in access to molecular biology techniques there is currently few data published on this topic. We report the distribution and seasonality of pediatric viral respiratory infections at a tertiary care oncological hospital in Guayaquil, Ecuador. Methods The laboratory database with results of real time PCR panel (Biofire® respiratory panel 2.1) from January to December 2021 was reviewed to collect the relative frequency and seasonality of identifiable viral infections in pediatric (ages < 18 years) patients. The frequency of infection according to sample characteristics including sex and age was compared using chi square test with a significance of 0.05. The frequency of detection was organized per month within the year. Percentage of coinfections was also determined. Results Out of 217 included PCR results, 143 (65.8%) tested positive for detectable viral organisms. Mean age was 7.3 years SD 4.8 years. The infection frequency for girl samples was higher than that of boy samples, X2 (1, N=217) = 4.16, p 0.041. The relation between age groups and the frequency of infection was not statistically significant X2 (2, N=217) = 5.24, p 0.073. The most prevalent viruses included Rhinovirus/Enterovirus 53 cases (relative frequency 37.0%), Parainfluenza virus 3 42 cases (relative frequency 29.3%) and Respiratory syncytial virus 19 cases (relative frequency 13.2%). Rhinovirus/Enterovirus, Paramyxovirus and Adenovirus respiratory infections occurred throughout the year while other respiratory viruses including Coronaviruses showed a higher detection rate during the rainy season (December – April). A total of 13 (5.9%) patients were coinfected with two up to four viruses. The most common combination was Parainfluenza virus 3 with Rhinovirus/Enterovirus. No bacterial targets were detected. Conclusion Larger and more complex studies are necessary to understand the burden of viral respiratory infections in pediatric oncological patients. In Ecuador, detection was higher in girls and school aged children. Rhinovirus/Enterovirus, Paramyxovirus and Adenovirus were present all year long while seasonality was common for other viruses following dry/rainy season pattern in the region. More understanding of seasonality will aim for preventive measures during more difficult months and to distribute resources accordingly. Figure 1. Seasonality of respiratory viral infections.

Listeria monocytogenes in the seafood industry: Exploring contamination sources, outbreaks, antibiotic susceptibility and genetic diversity.

Fish and seafood are rich sources of protein, vitamins, and minerals, significantly contributing to individual health. A global increase in consumption has been observed. Listeria monocytogenes is a known problem in food processing environments and is found in various seafood forms, including raw, smoked, salted, and ready-to-eat. Without heat treatment and given L. monocytogenes' ability to multiply under refrigerated conditions, consuming seafood poses a substantial health hazard, particularly to immunocompromised individuals. Numerous global outbreaks of listeriosis have been linked to various fish products, underscoring the importance of studying L. monocytogenes. Different strains exhibit varying disease-causing abilities, making it crucial to understand and monitor the organism's virulence and resistance aspects for food safety. This paper aims to highlight the genetic diversity of L. monocytogenes found in fish products globally and to enhance understanding of contamination routes from raw fish to the final product.

Microbiology of catheter-associated bloodstream infection: differences according to catheter type

ObjectivesCatheter-associated bloodstream infections (CABSI) cause preventable morbidity. We compared the microbiological etiology of CABSI across different types of central and peripherally-inserted catheters. MethodsWe analyzed prospectively collected CABSI data in a 2100-bed hospital network in Switzerland between 2016 and 2022. We included: short-term non-tunneled central venous catheters (CVC); long-term catheters (tunneled, or peripherally-inserted central catheters); arterial catheters; dialysis catheters; and peripheral venous catheters (PVC). We used multivariable logistic regression models to describe the risk of Staphylococcus aureus and Gram-negative pathogens according to catheter type. ResultsOverall, 416 CABSI episodes were included, including 60 episodes of S. aureus and 92 episodes of Gram-negative CABSI. Microbiological profiles differed between catheter types. Together, PVC and dialysis catheters accounted for 43/60 (72%) of all S. aureus CABSI. After adjusting for age, sex, and hematology/oncology care, the odds of S. aureus were higher for hemodialysis catheters (odds ratio [OR] 17.3, 95% confidence interval [CI] 5.75-52.2, P <0.01) and PVC (OR 2.96, 95% CI 1.22-7.20, P = 0.02) compared to short-term non-tunneled CVC. Odds of Gram-negative organism as the cause of CABSI were higher in long-term catheters versus short-term non-tunneled CVC (OR 2.70, 95% CI 1.37-5.24). ConclusionsCABSI in catheters other than short-term non-tunneled CVC is more commonly caused by virulent organisms including S. aureus and Gram-negative bacteria. Catheter type should be considered when selecting empirical antimicrobial therapies.

Management of periprosthetic joint infection of the shoulder: A narrative review

Evidence for management of shoulder periprosthetic joint infection (PJI) has traditionally originated from the hip and knee literature. The differing microbiome, anatomy and implants used in the shoulder mean this evidence is not always directly transferrable. The 2018 Philadelphia International Consensus Meeting for the first-time produced evidence-based guidelines and diagnostic criteria relating specifically to PJI of the shoulder. These guidelines and criteria recognize the pathogenicity of lower virulence organisms in the shoulder which often means clinical presentation is less obvious than other joints.The role of Cutibacterium acnes in shoulder PJI is the subject of increasing basic science and clinical research and advances in microbiological research may help to understand the pathology behind shoulder infections. There is new evidence that outcomes after revision shoulder arthroplasty are dependent on the virulence of the causative organism. An individualised approach to treatment considering host factors, organism, soft tissues and bone stock is recommended.Debate continues in the literature regarding the indications of one- or two-stage revision and the latest evidence is discussed and synthesized in this review article. We advocate careful multidisciplinary team decision making for cases of shoulder PJI and recognize a limited role for debridement and implant retention in acute shoulder PJI (<6 weeks). There appears to be a role for one-stage revision in lower risk cases with low virulence organisms but caution against its’ universal adoption. In higher risk or complex cases, there remains a clear role for two-stage revision arthroplasty, and we detail the specifics of this protocol and procedure from our tertiary shoulder and elbow unit.

Hypoalbuminemia is Highly Prevalent in Patients with Periprosthetic Joint Infection and Strongly Associated with Treatment Failure.

The role of hypoalbuminemia throughout the course of chronic periprosthetic joint infection (PJI) remains poorly understood. This study aimed to determine the prevalence and risk factors of hypoalbuminemia in periprosthetic joint infection (PJI) patients and to explore the association between hypoalbuminemia and treatment outcomes. This retrospective cohort study included 387 PJI cases who underwent two-stage exchange arthroplasty between January 2007 and August 2020, of which 342 were reimplanted. The mean follow-up period was 7.9 years. Multivariate logistic regression analyses were performed to identify risk factors for hypoalbuminemia and to assess the effect of hypoalbuminemia at 1st- and 2nd-stage exchange on the treatment outcome. Furthermore, the impact of dynamic changes in hypoalbuminemia was investigated. The prevalence of hypoalbuminemia at 1st- and 2nd-stage exchange was 22.2% and 4.7%, respectively. Patients with age ≥ 68 years and those with isolation of Staphylococcus aureus, Streptococcus, or Gram-negative bacteria exhibited a higher risk of hypoalbuminemia. Hypoalbuminemia at 1st-stage was significantly related to treatment failure (OR = 3.3), while hypoalbuminemia at 2nd-stage raised the OR to 10.0. Patients with persistent hypoalbuminemia at both the 1st- and 2nd-stage exchanges had a significantly higher rate of treatment failure than patients with hypoalbuminemia at the 1st-stage but normal albumin levels at the 2nd-stage exchange (55.6% vs 20.0%, p = 0.036). One in five patients with chronic PJI exhibits hypoalbuminemia. Hypoalbuminemia is more likely to develop in patients of advanced age and those infected by specific highly virulent organisms. Also, our results highlight the close association between hypoalbuminemia and treatment outcomes.

Characterising the allergic fungal rhinosinusitis microenvironment using full-length 16S rRNA gene amplicon sequencing and fungal ITS sequencing.

Allergic fungal rhinosinusitis (AFRS) is a severe phenotype of chronic rhinosinusitis with nasal polyposis (CRSwNP), characterised by localised and exaggerated type 2 inflammation. While fungal antigenic stimulation of unregulated Th2-mediated inflammation is the core pathophysiological mechanism, the direct and synergistic role of bacteria in disease modification is a pervasive hypothesis. We set out to define the microenvironment of AFRS to elucidate virulent organisms that may be implicated in the pathophysiology of AFRS. We undertook a cross-sectional study of AFRS patients and non-fungal CRSwNP patients. Demographics, disease severity, culture and microbiome sequences were analysed. Multimodality microbiome sequencing included short-read next-generation sequencing (NGS) on the Illumina Miseq (16S rRNA and ITS) and full-length 16S rRNA sequencing on the Oxford Nanopore Technologies GridION (ONT). Thirty-two AFRS and 29 non-fungal CRSwNP patients (NF) were included in this study. Staphylococcus aureus was the dominant organism cultured and sequenced in both AFRS and NF groups (AFRS 27.54%; NF 18.04%; p = .07). Streptococcus pneumoniae (AFRS 12.31%; NF 0.98%; p = .03) and Haemophilus influenzae (AFRS 15.03%; NF 0.24%; p = .005) were significantly more abundant in AFRS. Bacterial diversity (Shannon's index) was considerably lower in AFRS relative to NF (AFRS 0.6; NF 1.0, p = .008). Aspergillus was the most cultured fungus in AFRS (10/32, 31.3%). The AFRS sequenced mycobiome was predominantly represented by Malassezia (43.6%), Curvularia (18.5%) and Aspergillus (16.8%), while the NF mycobiome was nearly exclusively Malassezia (84.2%) with an absence of Aspergillus or dematiaceous fungi. A low diversity, dysbiotic microenvironment dominated by Staphylococcus aureus, Streptococcus pneumoniae and Haemophilus influenzae characterised the bacterial microbiome of AFRS, with a mycobiome abundant in Malassezia, Aspergillus and Curvularia. While Staphylococcus aureus has been previously implicated in AFRS through enterotoxin superantigen potential, Streptococcus pneumoniae and Haemophilus influenzae are novel findings that may represent alternate cross-kingdom pathophysiological mechanisms.

EFFECT OF LEAD AND CADMIUM ON IMMUNITY OF TWO VACCINES

This study was performed to investigate the effect of lead exposure on resistance of mice immunized against haemorrhagic septicaemia and challenged with virulent organisms. Lead treated mice showed greater susceptibility to challenge with P. multucida than control especially if lead nitrate is administered orally. this result showed that treatment with lead initials resistance of mice and might interferes with the vaccination programs. Cadmium chloride (cd) injected into rabbits at various time intervals in relation to tetanus toxoid injection was found to have some effect on the immune response. A reduction in the primary immune response was detected in case if (cd) is given two weeks prior to toxoid injection. On the other hand, enhancement of secondary antibody titers was detected. Cadmium reduced both primary and secondary antibody response if it is given seven days before immunization.

Exploiting CRISPR/Cas system in controlling antibiotic resistance of bacteria

The emergence and spread of antimicrobial resistance is a threat to human health, therefore there is a need to explore new strategies to combat antibiotic-resistant bacteria. Antimicrobial resistance genes are mainly transmitted through horizontal gene transfer (HGT) mediated by transposable elements, phages, and plasmids, which leads to the emergence of multidrug-resistant bacteria. Clustered Regularly interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated proteins (Cas) protect archaebacteria and eubacteria from potentially harmful foreign DNA such as plasmids, transposable elements, and phages. Several studies have introduced the CRISPR/Cas system as an innovative and effective strategy to trigger antibiotic resistance and virulent genes in bacteria. Therefore, this study aimed to introduce the CRISPR-Cas system in controlling antibiotic resistance of bacteria. We hope exploiting CRISPR/Cas could provide a bright future for the treatment of disastrous infections with resistant and virulent organisms.

Virulence phenotypes differ between toxigenic Vibrio parahaemolyticus isolated from western coasts of Europe

Vibrio parahaemolyticus is the leading bacterial cause of gastroenteritis associated with seafood consumption worldwide. Not all members of the species are thought to be pathogenic, thus identification of virulent organisms is essential to protect public health and the seafood industry. Correlations of human disease and known genetic markers (e.g. thermostable direct hemolysin (TDH), TDH-related hemolysin (TRH)) appear complex. Some isolates recovered from patients lack these factors, while their presence has become increasingly noted in isolates recovered from the environment. Here, we used whole-genome sequencing in combination with mammalian and insect models of infection to assess the pathogenic potential of V. parahaemolyticus isolated from European Atlantic shellfish production areas. We found environmental V. parahaemolyticus isolates harboured multiple virulence-associated genes, including TDH and/or TRH. However, carriage of these factors did not necessarily reflect virulence in the mammalian intestine, as an isolate containing TDH and the genes coding for a type 3 secretion system (T3SS) 2α virulence determinant, appeared avirulent. Moreover, environmental V. parahaemolyticus lacking TDH or TRH could be assigned to groups causing low and high levels of mortality in insect larvae, with experiments using defined bacterial mutants showing that a functional T3SS1 contributed to larval death. When taken together, our findings highlight the genetic diversity of V. parahaemolyticus isolates found in the environment, their potential to cause disease and the need for a more systematic evaluation of virulence in diverse V. parahaemolyticus to allow better genetic markers.

In vitro analysis of various antibiotic and cement combinations against S. epidermidis and S. lugdunensis for treatment of periprosthetic shoulder infection

BackgroundPeriprosthetic joint infections (PJIs) are devastating potential complications after arthroplasty surgery with significant therapeutic challenges. Many authors advocate for two-stage revision arthroplasty for PJI of the shoulder which includes explanation with antibiotic spacer placement. The optimal antibiotic/cement combination for PJI in the shoulder is not known. The purpose of this study is to analyze various cement and antibiotic concentrations against Coagulase negative staphylococcus species, S. epidermidis and S. lugdunensis. MethodsFive strains of coagulase negative Staph were taken from clinically documented orthopedic infections Both Simplex and Palacos cement were used to create antibiotic laden beads and mixed with either ertapenem, vancomycin, gentamicin, or a combination of ertapenem + vancomycin or ertapenem + gentamicin. The bacteria were plated on lysogeny broth agar plates and 3 beads were added per plate. Samples were analyzed for zone of inhibition at 24 hours, 72 hours, and 1 week. After 1 week, beads were transferred to new plates with bacteria and the process was repeated for 6 weeks. ResultsResults showed that ertapenem beads with both Simplex and Palacos cement showed the largest zones of inhibition for all samples. Vancomycin in Palacos cement and vancomycin in combination with ertapenem in Simplex and Palacos cement showed consistent zones of inhibition for the duration of the experiments. Ertapenem in combination with either vancomycin or gentamicin may allow for a powerful initial burst of killing followed by consistent antibiotic elution as opposed to gentamicin alone. ConclusionWhile many premade spacers on the market are infused with gentamicin, our in vitro models demonstrate more efficacious bacterial eradication for antibiotics such as ertapenem and vancomycin specifically for certain low virulence organisms that are commonly found in PJI around the shoulder.

Immunological and molecular diagnostic techniques in fish health: present and future prospectus.

Fish health management is critical to aquaculture and fisheries as it directly affects sustainability and productivity. Fish disease diagnosis has taken a massive stride because of advances in immunological and molecular diagnostic tools which provide a sensitive, quick, and accurate means of identifying diseases. This review presents an overview of the main molecular and immunological diagnostic methods for determining the health of fish. The immunological techniques help to diagnose different fish diseases by detecting specific antigens and antibodies. The application of immunological techniques to vaccine development is also examined in this review. The genetic identification of pathogens is made possible by molecular diagnostic techniques that enable the precise identification of bacterial, viral, and parasitic organisms in addition to evaluating host reactions and genetic variation associated with resistance to disease. The combination of molecular and immunological methods has resulted in the creation of novel techniques for thorough evaluation of fish health. These developments improve treatment measures, pathogen identification and provide new information about the variables affecting fish health, such as genetic predispositions and environmental stresses. In the framework of sustainable fish farming and fisheries management, this paper focuses on the importance of these diagnostic techniques that play a crucial role in protecting fish populations and the aquatic habitats. This review also examines the present and potential future directions in immunological and molecular diagnostic techniques in fish health.

Demographic Study to Evaluate the Morbidity Status of Children in Jodhpur City

Every nation's foundation is its children, and that nation's top priority is to ensure their health. Their early years of life are characterized by their development and growth. The first few years of life are the most important time because of the fast growth and development that is common at this age and necessitates ongoing observation. During this time, roughly 40% of physical growth and 80% of mental development take place. Any obstacle they face at this age could severely restrict their ability to develop and grow. A child who lacks access to health care during these formative years is denied the chance to develop into a healthy adult, and the harm caused in the early years may be permanent or in the long run. Children under the age of five continue to die from ailments that are treatable or preventable. Despite the significant progress in immunization to strengthen the immune system, medical research has not yet developed a fully effective defense against virulent organisms, and occasionally we fail in the struggle for survival. To understand the primary causes of death and the carers' patterns of seeking medical attention before the deaths of such children, a cross-section study encompassing the area of Jodhpur, Rajasthan, was done. The survey was conducted on 1000 children living in the surroundings of the Post Graduate Institute of Ayurved, Dr. S. R. Rajasthan Ayurved University, Jodhpur. Demographic studies show that Respiratory tract Infection is the main cause of morbidity in children. In Respiratory tract Infection recurrent cough, recurrent running nose, recurrent nasal obstruction, recurrent sore throat, and breathlessness are included. Out of which maximum number of children suffered from recurrent running nose.

Epidemiological etiology of Erysiphe sp. and putative viral and phytoplasma-like symptoms in Ayocote bean (Phaseolus coccineus)

Introduction/Objective. Ayocote bean (Phaseolus coccineus) has potential as a source of resistance in breeding programs because it exhibits greater tolerance to plant pathogens than P. vulgaris. However, its sanitary characterization is insipient; therefore, the purpose of this work was to carry out an etiological-epidemiological diagnosis, with emphasis on presumptive symptoms of viral and phytoplasmic organisms, and a typical fungal signs of powdery mildew. Materials and Methods. A plot (50 x 62 m) of flowering Ayocote bean was selected. It was divided into 80 (8 x 10) quadrats (6 x 6 m) and 720 subquadrats (2 x 2 m). From 25 plants with powdery-mildew-type leaf symptoms, mycelium was collected with adhesive tape for light microscopy observation and taxonomic identification. Length-width measurements were made on 60 conidia. Pure mycelium collected in situ and ex situ from 1-5 leaflets/plant was used for genomic analysis by PCR with universal primers ITS1 and ITS4. Samples were sequenced in Macrogen Inc. Korea. A total of 63 plants and 121 trifoliate leaves with viral and phytoplasmic symptoms were collected by direct sampling. In 88/121 samples, genomic analysis was performed by PCR with universal primers for Potyvirus (1), Begomovirus (2), and Phytoplasmas (1). Sequence editing and analysis were performed in SeqAssem and BLASTn/GenBank. Phylogenetic constructions were developed in Mega 11 with MUSCLE, Maximum Likelihood (ML), and HKY substitution model (1000-Bootstrap). Putative powdery mildew severity (%), flower damage (%), Macrodactylus sp. adult density, and plant vigor (%) were evaluated in 80 quadrats (3subquadrats/quadrat) with App-Monitor®v1.1 configured with a 5-class scale. In GoldenSurfer® v10, Kriging geostatistical analysis was performed to determine the spatial interrelationship between these variables. Results. Erysiphe vignae was identified as associated with powdery mildew of P. coccineus. The fungus, with hyaline, ovoid to ellipsoid conidia measuring 31.74 ± 0.3419 μm x 15.11 ± 0.1579 μm, without the presence of fibrosin bodies, had 100% genomic homology. This is the first report in Mexico. With average July-August temperature and relative humidity of 16.3 °C (±5.8) and 92.8 % (±10.7), respectively, powdery mildew leaf incidence and severity were 65.3 and 22.7 % (±16.9, range: 0 - 66.5 %), respectively. The most inductive focus (60-80 % severity) had an aggregate e 4-quadrat pattern (96 m2, lag = 4 and σ2-s = 450). Inoculum dispersal was significantly associated with dominant North-South winds and plant vigor (lag = 4 and σ2-s = 470). Flower damage was inconclusive in its spatial association with powdery mildew and Macrodactylus sp. suggesting uncorrelated events. No Potyvirus, Begomovirus, or Phytoplasmas were detected associated with yellowing, leaf distortion, mosaic, internode shortening, and other symptoms observed in situ. This confirms the relative tolerance/resistance reported for P. coccineus. Conclusion. E. vignae (Erysiphales: Erysiphaceae) associated with P. coccineus is reported for the first time in Mexico with moderate to intense epidemic level, which indicates its susceptible condition to this fungus. However, negative results for Potyvirus, Begomovirus, and Phytoplasmas, validate the apparent tolerance/resistance of P. coccineus to these organisms.

Centrifugal microfluidic system for colorimetric sample-to-answer detection of viral pathogens.

We describe a microfluidic system for conducting thermal lysis, polymerase chain reaction (PCR) amplification, hybridization, and colorimetric detection of foodborne viral organisms in a sample-to-answer format. The on-chip protocol entails 24 steps which are conducted by a centrifugal platform that allows for actuating liquids pneumatically during rotation and so facilitates automation of the workflow. The microfluidic cartridge is fabricated from transparent thermoplastic polymers and accommodates assay components along with an embedded micropillar array for detection and read-out. A panel of oligonucleotide primers and probes has been developed to perform PCR and hybridization assays that allows for identification of five different viruses, including pathogens such as norovirus and hepatitis A virus (HAV) in a multiplexed format using digoxigenin-labelled amplicons and immunoenzymatic conversion of a chromogenic substrate. Using endpoint detection, we demonstrate that the system can accurately and repetitively (n = 3) discriminate positive and negative signals for HAV at 350 genome copies per μL. As part of the characterization and optimization process, we show that the implementation of multiple (e.g., seven) micropillar arrays in a narrow fluidic pathway can lead to variation (up to 50% or more) in the distribution of colorimetric signal deriving from the assay. Numerical modeling of flow behaviour was used to substantiate these findings. The technology-by virtue of automation-can provide a pathway toward rapid detection of viral pathogens, shortening response time in food safety surveillance, compliance, and enforcement as well as outbreak investigations.

Development of a Simple and Effective Water Disinfection System Using an Electrochemical Mixed Oxidants-Generating System to Remove Waterborne Pathogens in Developing Countries

Despite tremendous efforts from various international agencies such as World Health Organization (WHO), waterborne diseases are a still major cause of morbidity and mortality in the world – with more than 1 billion incidences and more than 2 million deaths per year. As an effort to mitigate this global burden, we developed a water disinfection system using electrochemical mixed oxidants-generating system. This system is simple, inexpensive, and easy to use. In this study, we determined the effectiveness of the new water disinfection system against waterborne pathogens using &lt;i&gt;Escherichia coli&lt;/i&gt; CN13, bacteriophage MS2 and &lt;i&gt;Bacillus subtilis&lt;/i&gt; spores as bacterial, viral, and protozoa indicator organisms, respectively. The results of this study showed that the new water disinfection system is very effective against &lt;i&gt;E. coli&lt;/i&gt; CN13 (~3 log&lt;sub&gt;10&lt;/sub&gt; inactivation within 3 minutes with 0.3 mg/L of free chlorine) and coliphage MS2 (&gt; 4 log&lt;sub&gt;10&lt;/sub&gt; inactivation within 3 minutes with 0.5 mg/L of free chlorine) Although the inactivation of &lt;i&gt;B. subtilis&lt;/i&gt; spores by the new disinfection system was somewhat slow (~0.5 log&lt;sub&gt;10&lt;/sub&gt; inactivation in 60 minutes with 10 mg/L of free chlorine), the result is similar to previous studies with free chlorine. Overall, the results of this study indicate that the new water disinfection system using an electrochemical mixed oxidants-generating system is easy to use, convenient to carry, and also very effective against most waterborne pathogens, so it could be a sustainable solution for providing safe drinking water to the people in developing countries.

Neonatal Sepsis: Insight into Incidence, Classification, Risk Factors, Causative Organisms, Pathophysiology, Prognosis, Clinical Manifestations, Complications, Systemic Examination, and Treatment

Background: Neonatal sepsis is a clinical syndrome of bacteremia with systemic signs and is the most important cause of neonatal death in developing country. Mortality increase with delay in diagnosis and initiation of antibiotics. So, improvements in diagnosis and management of sepsis can significantly decrease the sepsis complications and improve its outcome. Objectives: The current review aims to high light on the incidence of neonatal sepsis, classification, risk factors, causative organisms, pathophysiology, prognosis, clinical manifestations, complications, systemic examination, and treatment. The incidence of neonatal sepsis varies among the different geographic areas, the highest being registered in Africa and Asia and the lowest in the U.S.A and Australia. The sepsis has been classified as early-onset sepsis (EOS) and the late-onset sepsis. Late-onset infections onset after 1 month of life, which occurs particularly in VLBW preterm infants or term infants requiring prolonged neonatal intensive care for other chronic problems. Neonatal sepsis includes maternal risk factors and neonatal risk factors. Septic or traumatic risk factors delivery, maternal poverty, pre-eclampsia, cardiac disease and diabetes mellitus are significant risk factors. Prolonged rupture of fetal membranes before the onset of labour, chorio-amnionitis, and maternal genitourinary tract infections have been significantly associated with a wide range of adverse perinatal and maternal outcomes, including miscarriage, stillbirth, preterm birth, fetal growth restriction, neonatal and puerperal sepsis, neonatal encephalopathy and neonatal and maternal mortality. Improvement in the socio-economic status of the population and availability of affordable antenatal care would reduce the incidence of neonatal septicemia. Amniocentesis, cervical cerclage, transcervical chorionic villus sampling, or percutaneous blood sampling, can permit entry of skin or vaginal organisms, causing amnionitis and secondary fetal infection. Infection of skin abrasions after use of obstetric forceps increases the risk of infection in neonates. Infant birth weight is inversely related to risk of early- onset sepsis. Prematurity and low birth weights are the most important neonatal factors predisposing to infection. Endotracheal intubation, insertion of an umbilical vessel catheter, or both, is associated with an increased risk of bacterial infection. Prolonged intravascular access and mechanical ventilation, the use of intravenous intralipids, and the duration of total parenteral nutrition are among the proposed risk factors associated with NICU-acquired coagulase- negative staphylococcus infections. Males have an approximately two-fold higher incidence of sepsis than females. Systemic antibiotics potentiate the overgrowth of certain organisms as Staphylococcus epidermidis. Corticosteroid treatment increases the risk of invasive candidiasis among premature infants. Hospital Acquired Infections are one of the leading causes of mortality and morbidity in (NICU). Group B Streptococcus (GBS) and E. coli as the dominant early onset sepsis (EOS) pathogens and coagulase- negative staphylococci (CONS) as the dominant late onset sepsis (LOS) pathogen followed by GBS and Staph aureus. In developing countries, Gram negative organisms are more common and are mainly represented by Klebsiella, E.coli and Pseudomonas of the Gram positive organisms, Staph aureus, CONS, Streptococcus pneumoniae, and Streptococcus pyogenes are most commonly isolated. Agents that commonly cause nosocomial infection are coagulase-negative staphylococci, gram-negative bacilli, Enterococci, and S. aureus. Causative organisms are bacterial, fungal, viral, and protozoal organism's infections. Mode of infection is prenatal, natal, and postnatal infections. Soon after birth, the newborns are exposed to the "hostile world" of bacteria, viruses, fungi and parasites, so they must immediately defend themselves. Necotizing enterocolitis, Neonatal thrombocytopenia, endophthalmitis, abdominal aortic-iliac thrombosis, fatal complete atrioventricular block, and wide range of signs and symptoms that may be observed in neonates with septicemia frequently suggest a number of alternative diagnoses. Identification of infection may be made by isolating the etiologic agent from a sterile body fluid (blood, CSF, and urine), or by demonstrating endotoxin or bacterial antigen in a body fluid. Investigations are including isolation of the microorganism, blood culture, urine culture, tracheal aspirate culture, isolation of organisms from CSF, complete blood cell count, cerebrospinal fluid analysis, C-Reactive protein, and tissue release of procalcitonin. Purpose of supportive care is to normalize the temperature, stabilize the cardiopulmonary status, correct hypoglycemia, and prevent bleeding tendency. Treatment of sepsis includes antibiotics, antiviral, antifungal, intravenous immunoglobulin, granulocyte transfusion, .......

DIFFICULTY IN THE DIFFERENTIAL DIAGNOSIS OF BRODIE’S ABSCESS OF PROXIMAL TIBIA

Brodie’s abscess, a subtype of subacute osteomyelitis, is very rare, accounting for 2.4% of subacute osteomyelitis. It is caused by a lower virulence organism, so it is not accompanied by systemic symptoms such as fever, and local symptoms appear insidiously. The disease when it is found through radiologic exams, is likely to be confused with bone tumors, resulting in delayed diagnosis and treatment. The authors experienced a brodie abscess of the proximal tibia, which was difficult to diagnose due to the inconspicuous symptoms.

A Rare Case of HACEK Endocarditis in the Context of a Permanent Pacemaker

Introduction: Infective endocarditis (IE) is a serious medical condition that can lead to high morbidity and mortality rates. The prevalence of healthcare-associated IE is increasing due to the use of therapeutic devices, such as intravascular catheters and cardiac devices. The HACEK group of bacteria, are commonly found in human respiratory and genitourinary tracts, are low virulence and fastidious organisms that are associated with blood culture-negative endocarditis. Proper diagnosis requires a combination of clinical, microbiological, and imaging results. The following study presents a case of atypical device-associated IE caused by Haemophilus influenzae in a 56-year-old man with a permanent pacemaker. Methodology: description of a clinical problem, its diagnosis, management, and outcome, accompanied with a literature review. Conclusion: This study highlights the importance of considering the HACEK group of bacteria in culture-negative IE cases and the use of advanced imaging techniques in diagnosis to prevent further complications and improve patient outcomes.

Isolation of Shiga Toxin Producing Escherichia coli 0157:H7 from Environmental and Clinical Samples in Dhaka City

Escherichia coli O157:H7, a Shiga toxin producing microbe was first acknowledged as a virulent organism in 1982 during an analysis of an outbreak of haemorrhagic colitis associated with consumption of hamburgers from a fast-food chain restaurant. Ability of Escherichia coli O157:H7 to induce injury in humans is a result of its ability to produce numerous virulence factors, most notably Shiga toxins Stx1 and Stx2, both of which constitute one of the most potent toxins known to man. Besides, Shiga toxin, Escherichia coli O157:H7 produces several other virulence factors, which include proteins which aid in the attachment and colonization of the bacteria in the intestinal wall and which can break down red blood cells and release iron to help support metabolism in Escherichia coli . Virulence factors facilitate this organism’s ability to cause intestinal and extra-intestinal diseases such as diarrhoea, haemorrhagic colitis (HC), haemolytic uremic syndrome (HUS), urinary tract infections (UTI), septicaemia and neonatal meningitis. In this study r, 7 samples from Dhaka city was collected, cultured in various media for enumeration, isolation and screening of Escherichia coli colonies which were further analysed to check for the presence of stx genes using PCR and gel electrophoresis. The seven samples collected were: Door knob swab, tea water, bhel puri, kitchen pipe swab, vegetable water, Lake water and Skin swab. The samples collected initially were enriched in enrichment media overnight, followed by a dilution series which were then used for spread plating on nutrient agar and MacConkey agar and EMB for confirmation with the observation of pink colonies and metallic sheen. The confirmed Escherichia coli isolates were later subjected to DNA extraction and amplification after which the bands for stx genes were observed and recorded. Out of the seven samples tested for stx1 and stx2 genes, two showed the presence of stx1 genes and one showed the presence of stx2 gene. The presence of the stx1 and stx2 genes in regular food and in the surrounding signifies how close is a large outbreak. Knowledge of processing such food or avoiding such environmental contacts or taking precautions when possible may prevent occurrence of diseases.

B-232 Versatile, Extraction-Free Nucleic Acid Sample Preparation in 15 Seconds Via Hyperbaric Heating

Abstract Nucleic acid extraction-based PCR sample prep is currently limited by long processing times, liquid transfers, kit cost, and lab storage requirements. Extraction-free approaches enable streamlined sample prep, but lack sensitivity compared to extraction-based methods. We studied several sample types to demonstrate the effectiveness of a novel 15-second Hyperbaric Heating (HBH) process to prepare nucleic acids for PCR. HBH exposes the sample to a short duration, high temperature environment to effectively lyse organisms and denature PCR inhibitors such as nucleases in seconds. Samples are introduced and sealed in a pressure-tight metallic vessel, which is then heated in seconds using magnetic induction. Within the sealed metallic vessel, the vaporization of the aqueous sample raises the internal pressure, enabling an internal temperature higher than 100°C to be achieved. Chelating agents and other proteins in HBH also protect the target nucleic acids from fragmentation. Post HBH, the sample is ready for direct PCR amplification without further dilution or extraction. Efficacy of HBH for efficient PCR amplification was tested against commercial kits. HBH was applied to viral, bacterial, eukaryotic, and fungal organisms. For SARS-CoV-2, HBH detected down to 250 viral copies/mL. Additionally, HBH outperformed commercial kits for PCR sample prep of Candida albicans and Bacillus subtilis. HBH is a generalizable sample prep technology with sensitivity comparable to that of current extraction kits. We have applied HBH to a multitude of organisms and have demonstrated that HBH can be employed as a rapid and comprehensive nucleic acid prep approach for applications that involve PCR amplification post sample prep. Future integration of HBH in devices suitable for point of care molecular testing are underway. We also envision HBH to be a useful, generalizable sample prep method for the molecular lab in applications where cost, simplicity, and speed are essential for preparing nucleic acids for amplification.