Isolation and detection of bacterial species on mitis salivarius bacitracin agar from the plaque samples of caries active children.

BACKGROUND: In cariology studies, mitis-salivarius-bacitracin (MSB) agar has been commonly considered as the selective medium for Streptococcusmutans growth. The present study was the part of a funded project (a noninferiority randomized controlled trial) which compared the efficacy of a plant extract-based mouth rinse with that of a fluoride mouth rinse on the S.mutans counts of the children. AIM: This study aimed to identify the frequency of detection of S.mutans and nonstreptococcal bacterial species from the dental plaque of caries active children using a combined technique of anaerobic culture and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. SETTINGS AND METHODS: Caries-active children (8-12 years old) were enrolled from a pediatric dental outpatient department at a tertiary care hospital. From each participant, dental plaque samples were collected from carious surfaces under sterilized conditions and then subjected to anaerobic culture. After 48 h of incubation, the bacterial colonies were isolated by sub-culture and identified by the MALDI-TOF. RESULTS: In all, 13 different bacterial species were isolated from the MSB agar medium. Other than S.mutans species, colonies of bacterial species such as Veillonelladispar,Streptococcusanginosus, Veillonellaparvula, and Streptococcusgordonii were also frequently observed from the medium. CONCLUSIONS: The study concluded that several bacterial strains, both streptococcal and nonstreptococcal, could be isolated from the MSB agar medium; hence, this medium should no longer be considered selective medium for the culture of S.mutans in clinical and epidemiological studies.

Epidemiological, clinical, and laboratory characteristics of human granulocytic anaplasmosis in North India.

Human granulocytic anaplasmosis (HGA) is an emerging, rickettsial tick-borne disease caused by Anaplasma phagocytophilum. Sero-epidemiological data demonstrate that this pathogen has a worldwide distribution. The diagnosis of HGA requires a high index of clinical suspicion, even in endemic areas. In recent years, HGA has increasingly been reported from Asia and described in China, Japan, and Korea. We serologically and molecularly screened 467 patients with clinical suspicion of Anaplasmosis. The present study describes the epidemiology, clinical, and laboratory details of 6 confirmed and 43 probable cases of human granulocytic anaplasmosis. One of the HGA patients developed secondary invasive opportunistic Aspergillus fumigatus and Acinetobacter baumanii infection during the illness, which resulted in a fatal infection. The HGA patients without severe complications had excellent treatment responses to doxycycline. The emergence of this newly recognized tick-borne zoonotic HGA in North India is a significant concern for public health and is likely underdiagnosed, underreported, and untreated. Hence, it is also essential to establish a well-coordinated system for actively conducting tick surveillance, especially in the forested areas of the country.IMPORTANCEThe results of the present study show the clinical and laboratory evidence of autochthonous cases of Anaplasma phagocytophilum in North India. The results suggest the possibility of underdiagnosis of HGA in this geographical area. One of the HGA patients developed secondary invasive opportunistic Aspergillus fumigatus and Acinetobacter baumanii infection during the illness, which resulted in a fatal infection.

Breakthrough infection among healthcare personnel following exposure to COVID-19: Experience after one year of the world's largest vaccination drive.

Context: The COVID-19 vaccination drive globally was supposedly a game-changing event. However, the emerging variants of the virus and waning immunity over time posed new challenges for breakthrough infections. Standing at the frontline of defense against COVID-19, healthcare personnel (HCP) were vulnerable to such infections. Aims: This study estimates i) the vaccine breakthrough infections (VBI) among HCP following exposure to COVID-19 cases, and ii) the mean interval between the second dose of vaccine and laboratory-confirmed SARS-CoV-2 infection. Materials and Methods: A cross-sectional study was conducted including 385 HCP with a history of exposure to COVID-19 cases during January and February 2022. Demographic details and clinical and vaccination history were collected from the test forms and the Web-based hospital management system. Laboratory testing of COVID-19 was carried out by real-time RT-PCR test. Results: The majority of the HCP were males (262; 68.05%) and nurses (180; 46.75%) by occupation. Two doses of vaccines were received by 278 (87.7%) HCP. VBI was confirmed in 185 (66.55%) HCP. No significant difference in VBI between the COVAXIN and COVISHIELD recipients (P = 0.69) was observed. The interval between the second dose and confirmed SARS-CoV-2 infection was significantly higher (P < 0.00001) in COVAXIN recipients (median 228 days) than in COVISHIELD recipients (median 95 days). Conclusions: The incidence of VBI was very high among the HCP, but not statistically different among the COVAXIN and COVISHIELD-recipients. Waning immunity over time suggests boosting immunity with a third dose because of emerging variants.

A Novel Potential Treatment for Diabetic Foot Ulcers and Non-Healing Ulcers - Case Series.

INTRODUCTION: Appropriate care and treatment of a wound is the need of the hour whether it is an infected or a non-infected wound. If wound healing is delayed for some reason, it leads to serious complications and further increases the hospital stay and cost of treatment. Herein, we describe a novel antimicrobial wound dressing formulation (VG111), with an objective to generate the preliminary data showing the distinct advantages in various types of wounds. METHOD: This case series involved the treatment of acute cases of wounds or chronic wounds that did not respond well to conventional wound healing treatments with VG111 in patients with different etiologies. Thirteen cases of patients that included patients with diabetes, pressure ulcers, burns, trauma, and others treated with VG111 showed rapid wound healing in all the cases even obviating the need for a graft when complete skin regeneration occurred. RESULT: This was illustrated by clearing of the wound infections, reduction/disappearance of the exudate, appearance of intense granulation, epithelialization, and anti-biofilm activity followed by complete wound closure. This VG111 precludes the need for systemic antimicrobial agents in localized infections and therefore, this single agent is an attempt to address the limitations and the drawbacks of the available products. CONCLUSION: Despite patients belonging to the old age group and having comorbidities like diabetes, still VG111 showed effective rapid wound healing, and that too without any scar formation in hardto- heal, infected, and non-infected wounds.

Relevance of Conventional Microscopy in the Diagnosis of Mucormycosis during COVID-19 Pandemic.

Introduction: Mucormycosis is a lethal disease which bewildered the health-care community of India during the ongoing second wave of the COVID-19 pandemic. The diagnosis is challenging considering the poor isolation in culture. Aims: The aim of the study was to emphasize the utility of potassium hydroxide (KOH) mount examination using conventional light microscopy for early diagnosis of mucormycosis in resource-limited settings. Materials and Methods: A retrospective analysis of results for all the samples including tissue biopsies, swabs, and pus received in the laboratory for KOH microscopy was done, and results were recorded. The clinical and demographic details of the patients were collected from the hospital information system. Results: A total of 75 samples from 50 patients were received in the laboratory. Out of these, 43 samples from 35 patients showed fungal hyphae (38 patients with only nonseptate hyaline hyphae, 2 with septate hyaline hyphae, and 3 samples with mixed infections). All patients except one were positive for severe acute respiratory syndrome coronavirus 2 infection. The most common age group was 45-59 years (40%), followed by 30-44 years (34.28%) with a male predominance. There was a significant difference in hemoglobin A1C (P = 0.005) and ferritin (P = 0.017) levels between laboratory-confirmed mucormycosis patients and clinically suspected mucormycosis patients without confirmation. Conclusion: Early diagnosis and initiation of targeted therapy is the cornerstone for treating mucormycosis patients. Hence, a rapid and reliable mode of diagnosis is the need of the hour. Conventional microscopy is such a tool that may be used, especially in resource-limited settings.

Serological profile of patients suspected with non-scrub typhus rickettsioses.

BACKGROUND: Rickettsial pathogens are Gram-negative, obligate intracellular bacteria. They are transmitted by arthropods and are responsible for a wide variety of disease, from minor to life-threatening, which have a global effect on human health. Limited data are available on the prevalence of rickettsial diseases from India, and the disease epidemiology is not fully described. This study aimed to diagnose non-scrub typhus rickettsioses including spotted fever and typhus group of Rickettsia in clinically suspected patients by using standard serological tests and recognition of common epidemiological conditions and clinical manifestations. METHODS: During the study period, a total of 700 patients of all ages with acute febrile illness were enrolled. Patients were screened for rickettsial infection using IgM Enzyme-linked immunosorbent assay (ELISA) and Immunofluorescence assay (IFA) was performed to confirm the ELISA positive results. The relevant demographic, clinical, and laboratory details of patients were documented and analyzed. RESULTS: Of 700 samples tested, 141 (20.2%) were found to be positive for IgM antibodies against rickettsioses using ELISA and IFA. SFGR was positive in 15 (2.2%), TGR was positive in 112 (16%) and 14 (2%) samples were positive for both groups. 20 (14.2%) patients required admission to the intensive care unit (ICU), and 24 (17%) in-hospital deaths occurred. CONCLUSIONS: The prevalence of rickettsioses in India appears to be underestimated; therefore, increased awareness and improved diagnostic testing could facilitate early detection of cases, pathogen-targeted appropriate treatment, and improved outcomes for patients. Despite the fact that Rickettsiae can be isolated or detected using molecular techniques in clinical specimens, serology still remains the most commonly used diagnostic method for rickettsioses around the world. Our study helps bridge the gap of limited data on Rickettsia in north India and could be useful for future epidemiological investigation of rickettsial diseases and outbreaks.

Case report of SARS-CoV-2 with secondary Clostridioides difficile infection.

There has been an overuse of antibiotics in most patients suffering from COVID-19 which predisposes patients to hospital acquired C. difficile Infection (CDI). Also, COVID infection of the gastrointestinal tract also predisposes the patient to CDI. We here present a fatal case of SARS-CoV-2 coinfection with CDI.

Bacteriophages as therapeutic & disinfectant agents to tackle multidrug-resistant Acinetobacter baumannii.

Background & objectives: Multidrug-resistant (MDR) Acinetobacter baumannii is a serious threat for human health worldwide. The studies on agents targeting A. baumannii are imperative due to identified A. baumannii co-infections in COVID-19. Bacteriophages are promising antibacterial agents against drug-resistant bacteria. This study intended to isolate bacteriophages against MDR A. baumannii from the water of river Ganga, to be used potentially as therapeutic and disinfectant particles. Methods: Acinetobacter phages were isolated from the Ganga water collected from Kanpur and further tested on 50 MDR A. baumannii isolates to determine host range. The phages were morphologically characterized by transmission electron microscopy. The disinfectant property of the isolated phages was tested by spraying of bacteriophage cocktail on MDR A. baumannii contaminated plastic surface, analyzed by colony-forming unit (CFU) and bioluminescence assay (adenosine triphosphate monitoring). Results: A total of seven bacteriophages were isolated against MDR A. baumannii. The bacteriophages lysed three MDR A. baumannii isolates out of 50 tested, showing narrow host range. Electron microscopy revealed hexagonal heads and long tails of bacteriophages, belonging to order Caudovirales. The bacteriophage cocktail reduced the MDR A. baumannii load efficiently on plastic surface, evidenced by reduction in CFUs and bioluminescence. Interpretation & conclusions: The findings of this study suggest that the isolated bacteriophages are potential lytic agents for MDR A. baumannii clinical isolates, and may be used as potential therapeutic agents as well as disinfectant to combat MDR A. baumannii with due consideration to phage host specificity, with further characterization.

Phylogeny and evolution of SARS-CoV-2 during Delta and Omicron variant waves in India.

SARS-CoV-2 evolution has continued to generate variants, responsible for new pandemic waves locally and globally. Varying disease presentation and severity has been ascribed to inherent variant characteristics and vaccine immunity. This study analyzed genomic data from 305 whole genome sequences from SARS-CoV-2 patients before and through the third wave in India. Delta variant was reported in patients without comorbidity (97%), while Omicron BA.2 was reported in patients with comorbidity (77%). Tissue adaptation studies brought forth higher propensity of Omicron variants to bronchial tissue than lung, contrary to observation in Delta variants from Delhi. Study of codon usage pattern distinguished the prevalent variants, clustering them separately, Omicron BA.2 isolated in February grouped away from December strains, and all BA.2 after December acquired a new mutation S959P in ORF1b (44.3% of BA.2 in the study) indicating ongoing evolution. Loss of critical spike mutations in Omicron BA.2 and gain of immune evasion mutations including G142D, reported in Delta but absent in BA.1, and S371F instead of S371L in BA.1 could explain very brief period of BA.1 in December 2021, followed by complete replacement by BA.2. Higher propensity of Omicron variants to bronchial tissue, probably ensured increased transmission while Omicron BA.2 became the prevalent variant possibly due to evolutionary trade-off. Virus evolution continues to shape the epidemic and its culmination.Communicated by Ramaswamy H. Sarma.

Tick-tock, beat the clock: comparative analysis of disc diffusion testing with 6-, 10-, and 24-h growth for accelerated antimicrobial susceptibility testing and antimicrobial stewardship.

Disc diffusion testing by Kirby-Bauer technique is the most used method for determining antimicrobial susceptibility in microbiological laboratories. The current guidelines by The Clinical and Laboratory Standards Institute (CLSI) 2022 specify using an 18- to 24-h growth for testing by disc diffusion. We aim to determine if using an early growth (6 h and 10 h) would produce comparable results, thus ultimately leading to reduced turnaround time. Six-hour, 10-h, and 24-h growths of 20 quality control strains and 6-h and 24-h growths of 48 clinical samples were used to perform disc diffusion testing using a panel of appropriate antimicrobial agents. Disc diffusion zone sizes were interpreted for all and comparative analyses were performed to determine categorical agreement, minor errors (mE), major errors (ME), and very major errors (VME) according to CLSI guidelines. On comparing with the standard 24 h of incubation, disc diffusion from 6-h and 10-h growths of quality control strains showed 94.38% categorical agreement, 5.10% mE, 0.69% MEs, and no VMEs. Disc diffusion testing for the additional 40 clinical samples yielded a similarly high level of categorical agreement (98.15%) and mE, ME, and VME of 1.29%, 1.22%, and 0% respectively. Disc diffusion testing using early growth is a simple and accurate method for susceptibility testing that can reduce turnaround time and may prove to be critical for timely patient management.

Low prevalence of Clostridioides difficile infection in acute severe ulcerative colitis: A retrospective cohort study from northern India.

BACKGROUND: The incidence of Clostridioides difficile infection (CDI) is high in ulcerative colitis and is associated with disease flares and adverse outcomes. However, the data on the dynamics of CDI in patients with acute severe ulcerative colitis (ASUC) is rather scarce. We evaluated the prevalence of CDI in patients with ASUC. METHODS: This retrospective analysis of a prospectively maintained cohort admitted to the All India Institute of Medical Sciences, India, from May 2016 to December 2021, included patients with ASUC (as per Truelove and Witts criteria) who were tested for CDI. CDI testing was performed using enzyme-linked immunoassay for toxins A and B. Risk factors for developing CDI were analyzed along with short-term outcomes of ASUC. Steroid failure was defined as the need for medical rescue therapy or colectomy. RESULTS: Total 153 patients with ASUC were included (mean age 34.92 ± 12.24 years; males 56.2%; disease duration 36 (IQR: 16-55.5) months, pancolitis 67.3%). Ninety-eight (63.4%), 72 (47%) and 10 (6.5%) patients, respectively, had received steroids, azathioprine and biologics in the past. Forty patients (26.14%) had a prior history of ASUC. Among risk factors for CDI, 14% of the patients had prior admission within 30 days, 22.2% had a recent history of antibiotics and 3.9% had long-term non-steroidal anti-inflammatory drug intake. Only one sample was positive for Clostridioides difficile toxin assay. Tissue Cytomegalovirus DNA-PCR positivity was noted in 57 patients (37.3%). Fifty-seven patients (37.3%) had steroid failure, 35 required medical rescue therapy and 30 (19.6%) required colectomy (eight after medical rescue therapy failure). CONCLUSION: Despite antecedent risk factors for CDI, the overall prevalence of CDI in ASUC was low and the outcomes were determined by underlying disease severity.

A pattern of microbiological colonization of orthodontic miniscrew implants.

INTRODUCTION: Current orthodontic literature reveals a lack of studies on bacterial colonization of orthodontic miniscrew implants (MSI) and their role in the stability of MSI. This study aimed to determine the pattern of microbiological colonization of miniscrew implants in 2 major age groups, to compare it with the microbial flora of gingival sulci in the same group of patients and to compare microbial flora in successful and failed miniscrews. METHODS: The study involved 102 MSI placed in 32 orthodontic subjects in 2 age groups: (1) aged ≤14 years and (2) aged >14 years. Gingival and peri-mini implant crevicular fluid samples were collected using sterile paper points (International Organization for Standardization no. 35) >3 months and processed by conventional microbiologic culture and biochemical techniques. A microbiologist characterized and identified the bacteria, and the results were subjected to statistical analysis. RESULTS: Initial colonization was reported within 24 hours, with Streptococci being the dominant colonizer. The relative proportion of anaerobic bacteria over aerobic bacteria increased over time in peri-mini implant crevicular fluid. Group 1 had greater Citrobacter (P = 0.036) and Parvimonas micra (P = 0.016) colonizing MSI than group 2. Failed MSI showed a significantly higher presence of Parvimonas micra (P = 0.008) in group 1 and Staphylococci (P = 0.008), Enterococci (P = 0.011), and Parvimonas micra (P <0.001) in group 2. CONCLUSIONS: Microbial colonization around MSI is established within 24 hours. Compared to gingival crevicular fluid, peri-mini implant crevicular fluid is colonized by a higher proportion of Staphylococci, facultative enteric commensals and anaerobic cocci. The failed miniscrews showed a higher proportion of Staphylococci, Enterobacter, and Parvimonas micra, suggesting their possible role in the stability of MSI. The bacterial profile of MSI varies with age.

Aerobic and anerobic contamination of mobile phones of health personnels with probable transfer of bacterial microbes to their hands at a tertiary care hospital of India.

Background and Aims: Mobile phone (MP) contamination of health personnels (HPs) in hospitals is a potential health hazard to the patients and the HP themselves. However, transfer of microbes from MPs of HP to their hands has not been demonstrated before, which would make potential threat into an actual peril. The primary objective was to determine aerobic and anerobic bacterial contamination of MP and hands of HP. The secondary objective was to determine probable transfer of bacterial microbes from MP to hands of tested HP. Material and Methods: Three swabs each were taken from 374 HP first from their MP, second from their dominant hand and third from their dominant hand after cleaning with disinfectant followed by a mock phone call of one minute (DHM). Aerobic and anerobic bacterial microbes were identified with standard methods. Results: Three hundred twenty-two HPs were recruited. Bacterial contamination was seen in 92% MP, 85% dominant hands, and 68% DHM of tested HP. Of these, contamination with potentially pathogenic bacterias (PPB) was 50% in MP, 25.6% in hands, and 31% in DHM. Anerobic contamination (1.6%) was present on MP but not in hands or DHM. In 54.7% HP, there was presence of similar bacterial microbes in MP and DHM of which 30% were PPB. When disinfectant was used in non-protocolized way in DHM, decrease in aerobic spore forming bacteria (ASB) was seen but not of gram-positive and gram-negative bacterial microbes. Conclusion: There is significant aerobic bacterial contamination, including PPB, seen in MP, hands, and DHM of HP in a tertiary care hospital of India; however, anerobic bacterias are found only in MP. Similar bacterial microbes in MP and DHM point to probable transfer of aerobic bacterias from MP to hands of HP which does not decrease when hand disinfectants are used in non-protocolized way, which is a point of concern.

Role of a probiotic strain in the modulation of gut microbiota and cytokines in inflammatory bowel disease.

OBJECTIVE: To assess the effect of a probiotic strain Bacillus clausii UBBC-07 on gut microbiota and cytokines in IBD patients. METHOD: Patients were randomly allocated to either placebo or probiotic Bacillus clausii UBBC-07 for four weeks along with the standard medical treatment (SMT). Enrolled patients were evaluated before and after intervention for presence of the given probiotic, change in gut microbiota, change in serum cytokines, serotonin and dopamine, symptoms of disease, physical, behavioral and psychological parameters. RESULTS: Probiotic strain Bacillus clausii UBBC-07 showed good survival in IBD patients in the treatment group (p < 0.01) without any reported adverse event. Metagenomic analysis showed that the given probiotic strain was able to modulate the gut microbiota in treated group. Phylum Firmicutes was increased and phylum Bacteroidetes was decreased in the probiotic treated group. A significant increase was observed in the abundance of anaerobic bacterial genera Lactobacillus, Bifidobacterium and Faecalibacterium in the probiotic treated group (p < 0.01) as compared to placebo group. Significant increase was observed in IL-10 (p < 0.05) and variable decrease in the secretion of IL-1ß, TNF- α, IL-6, IL -17 and IL -23 in probiotic treated group. In the treatment group a significant decrease in the symptoms of IBD and improvement in the psychological parameter to various degrees was noted. CONCLUSION: These results indicated that probiotic strain B clausii UBBC-07 affected the gut microbiota and cytokine secretion and shown efficacy in IBD patients.

A rare case of complicated pancreatitis due to leptospirosis.

The COVID-19 pandemic and the actions taken to combat it have greatly impacted the health infrastructure of all nations. Here we present a rare case of leptospirosis with severe acute pancreatitis, bilateral peripheral gangrene, disseminated intravascular coagulopathy and multiorgan failure. This is a rare presentation of leptospirosis wherein the patient had no history suggestive of acquisition of leptospires. The patient was started on doxycycline but still could not be saved due to the multisystem involvement.

Comparative Analysis of Clinical and Genomic Characteristics of Hypervirulent Klebsiella pneumoniae from Hospital and Community Settings: Experience from a Tertiary Healthcare Center in India.

Hypervirulent Klebsiella pneumoniae (hvKp) is a hypermucoviscous phenotype of classical Klebsiella pneumoniae (cKp) that causes serious infections in the community. The recent emergence of multidrug-resistant hvKp isolates (producing extended-spectrum beta-lactamases and carbapenemases) along with other virulence factors in health care settings has become a clinical crisis. Here, we aimed to compare the distribution of virulence determinants and antimicrobial resistance (AMR) genes in relation to various sequence types (STs) among the clinical hvKp isolates from both settings, to reinforce our understanding of their epidemiology and pathogenic potential. A total of 120 K. pneumoniae isolates confirmed by matrix-assisted laser desorption ionization-time of flight mass spectrometry were selected. hvKp was phenotypically identified by string test and genotypically confirmed by the presence of the iucA gene using PCR. Molecular characterization of hvKp isolates was done by whole-genome sequencing (WGS). Of the K. pneumoniae isolates, 11.6% (14/120) isolates were confirmed as hvKp by PCR (9.1% [11/120] string positive and 3.3% [4/120] positive by both methods); these were predominantly isolated from bloodstream infection (50%, 7/14), urinary tract infection (29%, 4/14), and respiratory tract infection (21%, 3/14). For all 14 hvKp infections, for 14.2% the source was in the community and for 85.7% the source was a health care setting. Two virulent plasmids were identified by WGS among the hvKp isolates from both settings. K64 was found to be the commonest capsular serotype (28.5%, 4/14), and ST2096 was the most common ST (28.5%, 4/14) by WGS. Two new STs were revealed: ST231 (reported to cause outbreaks) and ST43. The genome of one isolate was determined to be carrying AMR genes (blaCTX-M-15, blaNDM-1, blaNDM-5, blaOXA-181, blaOXA-232, etc.) in addition to virulence genes, highlighting the clonal spread of hvKp in both community and health care settings. IMPORTANCE To date, studies comparing the genomic characteristics of hospital- and community-acquired hvKp were very few in India. In this study, we analyzed the clinical and genomic characteristics of hvKp isolates from hospital and community settings. ST2096 was found as the most common ST along with novel STs ST231 and ST43. Our study also revealed the genome is simultaneously carrying AMR as well as virulence genes in isolates from both settings, highlighting the emergence of MDR hvKp STs integrated with virulence genes in both community and health care settings. Thus, hvKp may present a serious global threat, and essential steps are needed to prevent its further dissemination.