A closer look at the link between cycle threshold, clinical features and biomarkers: An observational study in COVID-19 patients.

Background: Symptoms for severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) appear 2-3 days after exposure to the virus. Being a virus, detection is primarily by polymerase chain reaction as this offers superior sensitivity and specificity. There was a misconception that patients with low cycle threshold (Ct) have severe coronavirus disease (COVID), and for individuals with higher Ct, it is the other way around. The prognosis for COVID was derived from various biomarkers and physicians heavily relied on them. Materials and Methods: A cross-sectional study spanning a duration of 2 years was conducted at a tertiary care centre in western India. A total of 201 individuals were included and the correlation between Ct, clinical features and biomarkers was studied. Results: In the E-gene, 43.28% had lower Ct values and 40.79% had low Ct values in the RdRp gene. 50% of all patients had diabetes, with 60% being between the ages of 61 and 80. 54.1% of hypertension patients belonged to ages between 61 and 80. 90.54% of COVID-positive individuals had lactose dehydrogenase levels ranging from 440 to 760. 79% of patients had a procalcitonin value of more than one but less than six. 79.1% of patients had an erythrocyte sedimentation rate between 36 and 90. Conclusion: Ct value though has a research value; it is a poor prognostic marker when compared to the various biomarkers that have been studied earlier. We cannot conclusively state that all our findings are accurate due to a lack of data but further research into the prognostic value of Ct should be conducted which will help in the ongoing scenario.

Seroprevalence of SARS-CoV-2 IgG in blood donors in a teaching institute from Western part of Maharashtra.

Introduction: COVID-19 is a disease caused by the severe acute respiratory syndrome coronavirus 2 that has appeared as a global pandemic in recent times. Currently, the transmission rate has slowed down significantly, but the definite pathological reason behind this is still unknown. Therefore, the prevalence of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibody must be studied to establish the relation between the rate of transmission and antibody presence. Materials and Methods: A clinical assessment was performed to evaluate the seroprevalence of SARS-CoV-2 Immunoglobulin G (IgG) antibodies among 299 healthy volunteers in the period of February to May 2021. Serum samples were analyzed using chemiluminescent microparticle immunoassay (CMIA) technology to detect the presence of IgG antibodies. Result: It was observed that 21% of the participants were seropositive, and 78% of the population was seronegative across the different genders. This confirmed that the generation of antibodies is independent of gender. Simultaneously, a t-test was performed that further suggested no statistical correlation between gender and seroprevalence. Moreover, a comprehensive analysis was performed to establish the relation between age and blood group with the seroprevalence. However, there was no statistical relationship found among these parameters. Conclusion: This study assisted in examining the underlying causes of high or low seroprevalence among healthy volunteers.

The microbial revolution: Unveiling the benefits of vaginal probiotics and prebiotics.

Vaginal health is essential to a woman's overall well-being, as abnormalities in vaginal health can lead to a variety of gynaecological disorders, such as urinary tract infections, yeast infections, and bacterial vaginosis. The vaginal microbiome is essential for the prevention of these infections. Disruptions in this microbial ecosystem can significantly impact vaginal health. The concept of utilizing probiotics and prebiotics to stimulate the growth of protective vaginal microbiota has gathered substantial interest in recent years. Probiotics are live micro-organisms that strengthen and restore vaginal microbial balance by lowering pH levels, production of bacteriocins, biofilm disruption, modulation of immune response, and production of hydrogen peroxide (H2O2), consequently combating the development of pathogens. Prebiotics are oligosaccharides that encourage the development of probiotics such as lactobacilli species. Probiotics and prebiotics also have some broader implications for vaginal health, including their role in minimizing the incidence of premature birth, optimizing fertility, managing menopausal symptoms, and preventing vaginal infections. Synbiotics are a combination of probiotics and prebiotics that deliver additional benefits by encouraging the development and activity of beneficial microbes. Furthermore, postbiotics are bioactive compounds derived from probiotic bacteria during fermentation that have immunomodulatory actions and provide an additional layer of protection against vaginal infections. The present study highlights the most prevalent vaginal infections and limitations of existing therapies that influence the vaginal microbiota. The profound consequences of probiotics and prebiotics in women's health, including their role in minimizing the prevalence of vaginal infections and promoting overall vaginal health, as well as advanced therapeutic strategies such as synbiotics and postbiotics, are also discussed. The literature offers significant insights into the mechanism, efficacy, and safety of probiotics and prebiotics to healthcare providers and researchers.

Commonly associated aerobic microbial pathogens and their antibiotic susceptibility profile in diabetic foot ulcers in tertiary care centre in Western Maharashtra.

PURPOSE: Diabetes mellitus (DM-II) is a metabolic disorder either due to reduced insulin production or reduced insulin sensitivity. Diabetic foot ulcer (DFU) is one of the most devastating complications of DM-II. This study was performed to assess commonly isolated micro-organisms and their anti-microbial sensitivity pattern in diabetic foot ulcers in a tertiary care centre in Western Maharashtra. METHODS: Adult patients with a known case of DM-II with foot lesions, suspected to be a Diabetic Foot Infections (DFIs) at the tertiary care hospital from Aug 2022 to Sept 2022 were included in the study. After obtaining informed written consent, pus sample was collected with sterile swab from lesion's base and submitted to Microbiology Laboratory for aerobic culture and sensitivity. RESULTS: Out of 56 enrolled patients, 47 (83.9%) patients tested positive for bacteriological growth and there was 'no growth' in 9 (16.07%) patients at the end of 48 h of aerobic incubation. There was male preponderance and patients were in age group of 35-85 years. The most commonly isolated micro-organisms were P. aeruginosa (17.8%), followed by S. aureus (14.2%), K. pneumonia and P. mirabilis (12.5% each). The resistance markers observed was ESBL producer, AmpC producer, MBL producer, Methicillin resistance and Inducible Clindamycin Resistance (ICR). CONCLUSION: Due to the injudicious use of antibiotics, antibiotic resistance has been increased in all types of soft tissue infections. The empirical formula for the treatment of DFIs should be decided for given geographical reasons according to antimicrobial susceptibility profile from particular geographical area or health care institute.

Development of a machine learning model to predict risk of development of COVID-19-associated mucormycosis.

Aim: The study aimed to identify quantitative parameters that increase the risk of rhino-orbito-cerebral mucormycosis, and subsequently developed a machine learning model that can anticipate susceptibility to developing this condition. Methods: Clinicopathological data from 124 patients were used to quantify their association with COVID-19-associated mucormycosis (CAM) and subsequently develop a machine learning model to predict its likelihood. Results: Diabetes mellitus, noninvasive ventilation and hypertension were found to have statistically significant associations with radiologically confirmed CAM cases. Conclusion: Machine learning models can be used to accurately predict the likelihood of development of CAM, and this methodology can be used in creating prediction algorithms of a wide variety of infections and complications.

Fungal infections caused by the Mucorales order of fungi usually target patients with a weakened immune system. They are usually also associated with abnormal blood sugar states, such as in diabetic patients. Recent work during the COVID-19 outbreak suggested that excessive steroid use and diabetes may be behind the rise in fungal infections caused by Mucorales, known as mucormycosis, in India, but little work has been done to see whether we can predict the risk of mucormycosis. This study found that these fungal infections need not necessarily be caused by Mucorales' species, but by a wide variety of fungi that target patients with weak immune systems. Secondly, we found that diabetes, breathing-assisting devices and high blood pressure states had associations with COVID-19-associated fungal infections. Finally, we were able to develop a machine learning model that showed high accuracy when predicting the risk of development of these fungal infections.

A post-transplant infection by Nocardia cyriacigeorgica.

Nocardia are Gram-positive, acid-fast, filamentous bacteria that cause opportunistic infections in susceptible populations. We describe a case of post-transplant infection of pulmonary nocardiosis caused by the rare strain Nocardia cyriacigeorgica and the challenges faced in reaching a definitive diagnosis. This case report emphasizes on keeping nocardiosis as a differential diagnosis in transplant recipients, as this disease is largely underdiagnosed and underreported.

Rhino-orbital cerebral mycosis: a case series of non-mucorales in COVID patients.

Introduction: Rhino-Orbito-cerebral mycoses are not only caused by Aspergillus spp. and Zygomycetes spp. but also can be associated with other rare species such as Neurospora spp., Cladosporium spp. and Fusarium spp. Mucormycosis is associated causatively with immunocompromised states, for example patients with comorbidities such as diabetes mellitus. Clinical symptoms of coronavirus disease (COVID) and mucormycosis in tandem are critical and relentless, frequently with no life-saving treatment. Case series: We report three patients with COVID-19 infection, who during the course of treatment developed rhino-orbital-cerebral mycosis including oral cavity involvement. Rhinocerebral mycosis along with oral cavity involvement was diagnosed by radiological investigations and preliminary screening for fungal infection (KOH mount) in all three cases. Empirical treatment was started but patients did not respond to treatment. All patients died even after debridement and maxillectomy. On culture, rare species of fungi were isolated in all three cases which, with the help of a reference laboratory, were identified as Neurospora, Cladosporium and Fusarium. Neurospora is considered nonpathogenic to humans. Cladosporium is a dematiaceous fungus found in soil in all climates, associated with disseminated or cerebral infections; and Fusarium, though considered a saprophytic colonizer of skin and respiratory mucosa along with other bacteria, is a common cause of mycotic keratitis worldwide. Conclusion: Immune system modifications due to COVID-19 with/without other risk factors can result in fungal co-infections that prove to be fatal for the patients. It is vital to be aware that COVID-19 patients, particularly those who are critically ill, may acquire secondary fungal infections and early detection is critical.

A case series and literature review of infections due to Myroides spp.: identification of contributing factors and emerging antibiotic susceptibility trends.

Introduction: Infections forby Myroides spp. can lead to significant morbidity and mortality, particularly in immunocompromised patients with underlying co-morbidities. Recent reports have highlighted its intrinsic and acquired drug resistance, making it a particularly challenging infectious agent to combat. Methods: Myroides spp. isolated and reported in clinically significant urine samples were considered for the study. Identification of the organism was done via the VITEK 2C system. Antibiotic susceptibility testing was done using both manual and automated methods following Clinical and Laboratory Standards Institute (CLSI) guidelines. Existing literature was searched on MEDLINE using PubMed. Results: We present a series of five catheter-associated urinary tract infections due to Myroides odoratimimus , with sensitivity to only minocycline. This is the first case from Western India, and the third case in the existing literature that shows Myroides sensitivity only to minocycline. Our literature review is the first to systematically describe contributory factors to infection, allowing us to devise a clinically relevant tool that delineates contributory factors and efficacious drugs in Myroides spp. infection. Conclusion: Myroides spp. infections, previously considered rare and opportunistic, need cognizance and diagnostic suspicion especially in particular associated conditions.

Lest we forget spinal tuberculosis (Potts's spine): Case series with unusual presentation.

Pott's disease, also known as TB spondylitis, is a very uncommon extrapulmonary infection caused by Mycobacterium tuberculosis. As its prevalence is not high it can easily be underdiagnosed. Magnetic resonance imaging (MRI), computed tomographic (CT) guided needle aspiration, or biopsy are known to be the best techniques for early histopathological diagnosis along with confirmation by microbiological results. Ziehl Neelson stain (ZN) can detect Mycobacterium infections when clinically suspected samples are adequate and optimally stained. No single method or simple guideline can diagnose spinal tuberculosis. Early diagnosis and prompt treatment are necessary to prevent permanent neurological disability and to minimize spinal deformity. We are reporting three cases of Potts disease which could have been easily missed if we would have relied on one single investigation.

Lymphopenia with Altered T Cell Subsets in Hospitalized COVID-19 Patients in Pune, India.

The cellular immune cell subsets affecting COVID-19 disease severity are being studied by researchers from many countries. The current study was carried out to investigate the alteration of peripheral blood mononuclear cells (PBMCs) and their subsets in hospitalized COVID-19 patients in a tertiary care center in Pune, India. The PBMCs were isolated from enrolled study participants, and flow cytometry analysis was done to assess peripheral white blood cell alterations. The lymphocyte subsets of naive, effector, central memory, and effector memory CD4+ or CD8+ T cells were then evaluated in COVID-19 patients with different disease categories and compared to healthy controls. The immunophenotypic characterization of the immune cell subset was done for 139 COVID-19 patients and 21 healthy controls. These data were evaluated based on the disease severity. A total of 139 COVID-19 patients were classified as mild (n = 30), moderate (n = 57), or severe (n = 52) cases. The decreased percentages of total lymphocytes, CD3+ T cells, CD4+ T cells, naive T cells, central memory T cells, and Natural Killer (NK) cytotoxic cells were found, and there was increase in effector T (TEf) cells and effector memory T cells in patients with severe COVID-19 compared to healthy controls. The severity of SARS-CoV-2 infection has an effect on lymphocyte subsets, resulting in reduced T memory cells and NK cells but increased TEf cells in severe cases. Clinical Trial Registration: CTRI ID-CTRI/2021/03/032028.

Role of GeneXpert MTB/RIF assay for the diagnosis of cervical lymph node tuberculosis and rifampicin resistance.

Objectives: Tuberculosis (TB) of lymph node (TB lymphadenitis) is one of the most common forms of extrapulmonary TB (EPTB) whose diagnosis is critically challenging. Although new diagnostic methods have been developed, especially in patients without a history of TB, the cervical tuberculous lymphadenitis diagnosis is still elusive. This study assessed the applicability of GeneXpert in early diagnosis of EPTB, especially cervical lymphadenopathy. Materials and Methods: The study was conducted in a tertiary care hospital from January 2018 to December 2020 at the department of microbiology. All the samples of cervical lymph node tissue and lymph node aspirate were followed as per the routine protocol for mycobacterial identification. The sample was divided into two parts: one was used for the new molecular-based GeneXpert MTB/RIF assay and the second one was tested by direct and concentrated acid-fast bacilli microscopy by Z-N staining and culture for the detection of MTB. Results: Among the 145 samples tested, the GeneXpert detected the DNA of MTB in 89 samples (61.37%), whereas the culture test was positive in 42 (28.93%) specimens. GeneXpert also detected 7 rifampicin resistance cases. GeneXpert sensitivity and specificity results were assessed according to culture results. The sensitivity and specificity of the GeneXpert assay were 85.71% and 48.54%, respectively. Conclusion: GeneXpert MTB/RIF should be used in conjunction with clinical presentation and other molecular investigation in nonrespiratory specimens.

Molecular Characterization Identifies Upstream Presence of IS Aba1 to OXA Carbapenemase Genes in Carbapenem-Resistant Acinetobacter baumannii.

Background Evaluating the expression pattern of oxacillinases (OXA) carbapenemases is essential to understand the prevalence and spread of carbapenem resistance Acinetobacter baumannii . Objectives The aim of the study is to evaluate the presence of OXA carbapenemase genes and IS Aba1 upstream to these genes in carbapenem-resistant A. baumannii clinical isolates. Materials and Methods A. baumannii isolated from clinical samples were phenotypically identified and antibiotics sensitivity was performed. Multiplex polymerase chain reaction (PCR) was used to detect OXA51-like gene, OXA carbapenemases genes (OXA-23-like, OXA-24-like, and OXA-58-like), and IS Aba1 in carbapenem-resistant isolates. Results Out of 55 Acinetobacter isolates, 54 were confirmed as A. baumannii by PCR. Bla OXA-23 -like gene was observed in 51 isolates of A. baumannii and none of the isolates showed the presence of bla OXA-24 -like and bla OXA-58 -like genes. Presence of IS Aba1 upstream to OXA-23-like gene, OXA-51-like gene, and both OXA-51-like/OXA-23-like genes was observed in 51, 7, and 4 A. baumannii isolates, respectively. Conclusion The genetic pattern of carbapenem-resistant A. baumannii isolated in this study was unique, which should be factored for clinical protocols to manage infections caused by emerging resistant strains of A. baumannii .

Isolation and identification of Acinetobacter species with special reference to antibiotic resistance.

BACKGROUND: Acinetobacter is clinically important pathogen with widespread resistance to various antibiotics. We assessed the incidence of Acinetobacter infection at a tertiary care hospital, analyze their resistance pattern and identify the production of extended spectrum ß-lactamases (ESBLs) and metallo ß-lactamases (MBLs). MATERIALS AND METHODS: The study was conducted in tertiary care hospital, India over a period of 2 years. Acinetobacter species were isolated from various clinical samples received in Department of Microbiology. After identification, Acinetobacter isolates were speciated and antibiotic susceptibility was determined by the standard disc diffusion method. ESBL and MBL production was detected by the double disc synergy test and combined disc diffusion test respectively. RESULTS: Of 3298 infected samples, 111 (3.36%) were found to be Acinetobacter. The most predominant species was Acinetobacter calcoaceticus-A. baumannii (Acb) complex (72%). High incidence of resistance was recorded for piperacillin (55%), followed by ceftriaxone (46%) and ceftazidime (46%). Isolation rate and antibiotic resistance was higher in the Intensive Care Units (ICUs) of the hospital. ESBL and MBL production was detected in 31.5% and 14.4% of the isolates respectively. DISCUSSION AND CONCLUSION: A high level of antibiotic resistance was observed in our study and maximum isolation rate of Acinetobacter was in the ICUs. Acb complex was the most predominant and most resistant species. The analysis of susceptibility pattern will be useful in understanding the epidemiology of this organism in our hospital setup, which will help in treating individual cases and controlling the spread of resistant isolates to other individuals.

Pencillium keratitis in an Immunocompetent Patient from Pune, Maharashtra, India.

The incidence of fungal keratitis is less common than bacterial and viral keratitis. However, it remains a diagnostic and therapeutic challenge. Delayed clinical diagnosis is common mainly because of lack of suspicion. Further slow growth of fungus increases the time for confirmed laboratory diagnosis. After accurate diagnosis, patient's management remains inadequate due to lack of availability of antifungal agents and its poor corneal penetration. Multitude of genera of molds and yeast have been identified in fungal keratitis. Due to their ubiquitous nature and easy isolation from the environment, their role in true pathogenesis is difficult to ascertain. Worldwide, incidence of fungal keratitis is rising at present. The predisposing factors comprises trauma, use of contact lenses and topical steroids. Filamentous fungi and dematiaceous fungi are the frequently encountered etiological agents of fungal keratitis. Dimorphic fungi are reported less frequently. Fungal keratitis tends to occur more frequently in young males and usually in winter and monsoon. Penicillium genera includes several species. By far Penicillium marneffei (P. marneffei) infection is most common, mainly associated with AIDS. A number of infections caused by species other than P. marneffei have been reported as well. Here we report a case of Penicillium keratitis in a young, HIV negative male farmer.

Tuberculous Otitis with Proteus mirabilis Co-Infection: An Unsuspected Presentation Encountered in Clinical Practice.

Tuberculosis, a contagious bacterial disease which is caused by Mycobacterium tuberculosis, primarily involves the lungs.Though Pulmonary tuberculosis (PTB) is the commonest clinical presentation, there is a need for alertness towards uncommon presentations which involve other organs. Tuberculous otitis media (TOM) is one such rare presentation seen in paediatric practice. It is characterized by painless otorrhoea which fails to respond to the routine antibacterial treatment. TOM usually occurs secondary to PTB. Here is a case of tuberculous otitis media with Proteus mirabilis co-infection, with no evidence of PTB. In the sample of ear discharge obtained from the patient, acid fast bacilli were demonstrated on direct microscopy after Ziehl-Neelsen staining. Culture done on Lowenstein-Jensen medium demonstrated slow-growing Mycobacterium. Bacteriological culture and identification helped in isolating Proteus mirabilis. PCR, followed by Line- Probe Assay for early identification and susceptibility testing to primary drugs, was done. Further, patient tested negative for the Mantoux test. Patient was enrolled in National Tuberculosis programme- RNTCP. This case emphasizes on one of the less common presentations of a common disease. A high clinical suspicion and laboratory confirmation are required for appropriate patient management.

The spectrum and aetiology of mycotic infections from a tertiary care hospital from Western part of India.

BACKGROUND: In the past few years, there has been an increase in infections caused by fungal aetiology. This is mainly due to increase in sizes of populations which are at risk. Also, fungi which were previously considered as non-pathogenic have been increasingly implicated. Hence, this study was taken up. AIM: To assess the magnitude of mycotic infections in this set up. To assess the spectrum of fungi which are involved in various infections. MATERIAL AND METHODS: Total 704 samples taken over two and half period were included. They consisted of various samples like sputum, blood, urine, sterile body fluids, corneal scrapings. These were processed by conventional mycological techniques. These included direct microscopy after Gram staining and KOH mount, culture on Sabouraud's dextrose agar (SDA), corn meal agar, brain heart infusion (BHI), brain heart infusion agar (BHIA) and others, as required. For non-albicans Candida, the automated Vitek 2 C system was used. Various moulds and yeasts were identified up to genus level and species level. RESULTS: Of 704 samples, 336 (48%) were positive for fungal culture. Of these, 244 isolates were yeast like fungi, 81 were moulds, 7 were yeasts and 4 were dimorphic. Among blood stream infections, non-albicans Candida were the commonest isolates. In the urine samples, C. albicans was the commonest isolate. From corneal scrapings, only moulds and dimorphic fungi were isolated. CONCLUSION: Incidence of non- albicans Candidal infections is increasing. Culture and identification of mycotic infections is essential for commencement of suitable antifungal therapy.

Primary splenic tubercular abscess in an immunocompromised patient-rapid diagnosis by line probe assay.

Diagnosing extra-pulmonary tuberculosis is a challenge that can confound even the most practiced clinicians as clinical manifestations are vague, non-specific and typical chest radiograph findings may not be evident till late in the disease. Conventional methods for mycobacteriological culture and drug susceptibility testing are slow and cumbersome. Novel technologies for rapid detection of Mycobacterium tuberculosis and its anti-TB drug resistance have therefore become a priority hence with the development of molecular line probe assays are most advanced. Herewith we are reporting a case of splenic tuberculosis in an immunocompromised patient for its rarity and to emphasis the fact that such patients can be diagnosed early for better treatment outcome to enhance the longevity if a health setup possesses all the modern diagnostic services.