Distribution of virulence genes and biofilm characterization of human isolates of Streptococcus agalactiae: A pilot study.

This study included 21 newly isolated clinical samples of Streptococcus agalactiae (Group B Streptococcus) screened in patients (six male, fifteen female) from various states of India with different infections (urinary tract infections, blood, pus and eye infections). All isolates were identified as Group B Streptococcus (GBS) using hemolytic properties, serogrouping and MALDI-TOF-MS analysis. Six virulence genes, cfb (100%), cylE (90.4%), lmp (85.7%), bca (71.4%), rib (38%) and bac (4.7%) were detected via polymerase chain reaction (PCR). Distribution studies of these six genes revealed five isolates containing five virulence genes (23.8%), followed by ten isolates containing four virulence genes (47.6%). The twenty GBS isolates selected on the glass surface included non-biofilm producers (n = 6, 30%), weak (n = 11, 55%) and moderate biofilm producers (n = 3, 15%). On the polystyrene surface, weak (n = 4, 20%), moderate (n = 2, 10%) and strong (n = 14, 70%) biofilm producers were detected. Live-dead cell staining revealed that more viable cells accumulated in the S. ag 7420 isolate than in the AH1 isolate. Scanning electron microscope (SEM) biofilm analysis showed S. ag AH1 cells appeared as chain-like structures, whereas the S. ag 7420 isolate biofilm cells appeared as fork-like structures on the glass surface. Biofilm elements were analyzed using Energy Dispersive X-Ray Analysis (EDAX) for both isolates and 13 elements with different orders of composition were found. Thus, virulence gene detection, distribution and biofilm formation by these new clinical isolates suggested the virulent nature of these pathogens, which might cause different levels of disease severity in humans.

Intestinal carriage of colistin resistant Enterobacteriaceae in hospitalized patients from an Indian center.

There is limited data on the gut colonization rate of colistin resistant (Col-R) bacteria in patients and healthy volunteers in India. Aim of this study was to investigate the stool carriage rate of Col-R in hospitalized patients. Stool samples were inoculated in Eosin Methylene Blue agar plates supplemented with colistin. Colistin minimum inhibitory concentrations (MICs) were determined by the broth microdilution method. PCR for the mcr-1 was performed on Col-R Enterobacteriaceae isolates. Mutations in the mgrB gene were analyzed in K. pneumoniae isolates. Mcr-1 positive E. coli was subjected to whole-genome sequencing. Out of 65 stool samples screened, 33 (51%) samples carried Col-R bacteria. Majority (76.7%) of the isolates were sensitive to carbapenem.

Talaromyces marneffei Outside Endemic Areas in India: an Emerging Infection with Atypical Clinical Presentations and Review of Published Reports from India.

Talaromycosis is a disseminated disease caused by Talaromyces (Penicillium) marneffei, mainly seen in acquired immunodeficiency syndrome (AIDS) patients. Its distribution is restricted to southeast Asian countries; a small pocket of endemicity exists in the northeast Indian state of Manipur. Here, we present a series of five cases presenting to our tertiary care hospital, originating from non-endemic states neighboring Manipur. In addition to the geographical distinction, a variety of unique features were noted in our cases, including human immunodeficiency virus (HIV)-negative hosts, the absence of typical skin lesions, presentation as pneumonia and generalized lymphadenopathy. Our series highlights the importance of distinguishing this disease from histoplasmosis and tuberculosis, both endemic in India.

Assessment of core capacities for antimicrobial stewardship practices in indian hospitals: Report from a multicentric initiative of global health security agenda.

Introduction: Antimicrobial-resistant HAI (Healthcare associated infection) are a global challenge due to their impact on patient outcome. Implementation of antimicrobial stewardship programmes (AMSP) is needed at institutional and national levels. Assessment of core capacities for AMSP is an important starting point to initiate nationwide AMSP. We conducted an assessment of the core capacities for AMSP in a network of Indian hospitals, which are part of the Global Health Security Agenda-funded work on capacity building for AMR-HAIs. Subjects and Methods: The Centers for Disease Control and Prevention's core assessment checklist was modified as per inputs received from the Indian network. The assessment tool was filled by twenty hospitals as a self-administered questionnaire. The results were entered into a database. The cumulative score for each question was generated as average percentage. The scores generated by the database were then used for analysis. Results and Conclusion: The hospitals included a mix of public and private sector hospitals. The network average of positive responses for leadership support was 45%, for accountability; the score was 53% and for key support for AMSP, 58%. Policies to support optimal antibiotic use were present in 59% of respondents, policies for procurement were present in 79% and broad interventions to improve antibiotic use were scored as 33%. A score of 52% was generated for prescription-specific interventions to improve antibiotic use. Written policies for antibiotic use for hospitalised patients and outpatients were present on an average in 72% and 48% conditions, respectively. Presence of process measures and outcome measures was scored at 40% and 49%, respectively, and feedback and education got a score of 53% and 40%, respectively. Thus, Indian hospitals can start with low-hanging fruits such as developing prescription policies, restricting the usage of high antibiotics, enforcing education and ultimately providing the much-needed leadership support.

Experience with ß-D-Glucan Assay in the Management of Critically ill Patients with High Risk of Invasive Candidiasis: An Observational Study.

BACKGROUND: The (1,3)-ß-D-glucan assay (BDG) is recommended for the early diagnosis of invasive candidiasis (IC). METHODS: Records of 154 critically ill adults with suspected IC, on whom BDG was done, were analyzed. Patients were divided into three groups: Group A (confirmed IC), Group B (alternative diagnosis or cause of severe sepsis), and Group C (high candidal score and positive BDG [>80 pg/mL] but without a confirmed diagnosis of IC). RESULTS: Mean BDG levels were significantly higher in Group A (n = 32) as compared to Group B (n = 60) and Group C (n = 62) (448.75 ± 88.30 vs. 144.46 ± 82.49 vs. 292.90 ± 137.0 pg/mL; P < 0.001). Discontinuation of empiric antifungal therapy based on a value <80 resulted in cost savings of 14,000 INR per day per patient. CONCLUSION: A BDG value of <80 pg/ml facilitates early discontinuation of empirical antifungal therapy, with considerable cost savings.

Carbapenem-resistant enterobacteriaceae screening: A core infection control measure for critical care unit in India?

BACKGROUND: Infection/colonization due to carbapenem-resistant enterobacteriaceae (CRE) are emerging as an important challenge, particularly in high risk patients due to widespread use of Carbapenems. Therefore, preventing both CRE infections and their transmission has become an important infection control objective. AIMS AND OBJECTIVE: Determine the proportion of asymptomatic carriers of CRE among patients admitted to our critical care unit (CCU) from the community and other health care facilities. Enumerate risk factors and guide implementation of infection control interventions. METHODS: This prospective surveillance study was done in a 24 bed CCU of a tertiary care hospital, at Chennai, India between August2017 through December 2017. Patients were screened based on a composed questionnaire framed from Centers for Diseases Control and Prevention CRE tool-kit. Two rectal swabs were collected from each patient. They were processed in microbiology laboratory. RESULTS: A total of 102 patients were included. CRE colonization were identified in 8 (7.8%) of the total samples. Among 8 CRE colonized patients 3 (37.5%) patients developed systemic infection. Patients who were exposed to high end antibiotic and past history of surgery had significant association with CRE colonization of (P = 0.0029) and (P = 0.0167) respectively. CONCLUSION: Overall CRE colonization rates among our CCU patients were found to be low. Risk factors associated with CRE colonization were high end antibiotic exposure and surgery in past 90 days. Hence rectal screening should be a risk factor-based active surveillance. Association of systemic infection among CRE colonizers was more significant. This study led us to modify our infection control practices in CCU.

Melioidosis: An Emerging Infection with Fatal Outcomes.

According to the Centers for Disease Control and Prevention, from being nonendemic for melioidosis, India has now become endemic for the disease since 2012. Until then, melioidosis cases were being reported sporadically from India. There have been isolated case reports from few states across the country for the past few years. Most of the times, Burkholderia pseudomallei may be misreported as Pseudomonas species, especially in resource-poor laboratories. Due to its varied clinical presentation, the specific clinical diagnosis can be difficult, thereby making laboratory diagnosis mandatory. This could make a huge impact on patient care as this organism has a different treatment protocol as well as virulence determinants which influence the course of management. Although known for its endemicity in Australia, Thailand, and other Southeast Asian countries, B. pseudomallei has emerged in new areas such as India, Southern China, Brazil, and Malawi. We present a rare case of melioidosis with rapid disease progression to fatal outcome from Chennai, South India.

Antimicrobial susceptibility of Salmonella enterica serovars in a tertiary care hospital in southern India.

BACKGROUND & OBJECTIVES: Salmonella enterica serovars Typhi and Paratyphi are predominantly known to cause enteric fever. Multidrug resistance in S. Tphi and S. Paratyphi has emerged as a cause of concern. This study was done to evaluate status in antimicrobial susceptibility patterns of Salmonella enterica serovar Typhi (S. Typhi) and S. Paratyphi obtained from blood culture in a tertiary care hospital in south India. METHODS: Blood isolates of Salmonella species over a two year period between May 2009 and June 2011 were studied. A total of 322 isolates of Salmonella species were tested for antimicrobial susceptibility by Kirby-Bauer disc diffusion method. The MIC of ciprofloxacin was obtained by E-test, and azithromycin MIC was confirmed by agar dilution method for a limited number of isolates. RESULTS: Of the total of 322 isolates studied, 186 (57.8%) were S. Typhi, 134 (41.6%) were S. Paratyphi A, and two were S. Paratyphi B. Of these, 44(13.66%) were resistant to ciprofloxacin (MIC <0.50 µg/ml) and 296 (91.9%) were nalidixic acid resistant. Of these 296 nalidixic acid resistant isolates, 278 (94%) were susceptible to ciprofloxacin by MIC criteria (<0.5 µg/ml). Of the 262 isolates tested for azithromycin sensitivity, only 120 (46%) were susceptible, whereas 81 (31%) were resistant and 55 (21%) showed intermediate susceptibility. Of the isolates, 322 (90%) were susceptible to ampicillin and (95%) were susceptible to co-trimoxazole. However, all the isolates were susceptible to chloramphenicol and ceftriaxone. INTERPRETATION & CONCLUSIONS: Nalidixic acid resistance screening is not a reliable surrogate indicator of ciprofloxacin resistance. Ciprofloxacin MIC should to be routinely done. Azithromycin resistance appears to be emerging. However, isolates showed a high degree of susceptibility to ampicillin, co-trimoxazole and chloramphenicol. Thus, antibiotics like ampicillin and co-trimoxazole may once again be useful for the management of enteric fever in southern India.

Melioidosis :an emerging infection in India.

BACKGROUND: Melioidosis caused by the gram-negative bacterium Burkholderia pseudomallei is endemic in Southeast Asia but may be under-diagnosed and under-reported in the Indian subcontinent. This study was undertaken to analyse the clinical presentation and epidemiological risk factors for melioidosis in India. METHODS: We carried out a retrospective study of 32 culture proven cases of melioidosis at a tertiary care hospital in South India between 2005 and 2010. RESULTS: Thirty two culture confirmed cases of melioidosis were included in the study. Patient age varied from 4 to 60 years with a median age of 42.5 years. Males constituted 75% of cases and 78.12% of cases were from rural areas. Three-fourth (24 of 32) had at least one risk factor that predisposed to melioidosis: diabetes (43.75%) followed by alcoholism (21.87%) were the commonest. Fever was the most common symptom (68.75%) and mean duration of symptoms was 2.34 months before diagnosis. More than half of the cases (56.25%) presented as disseminated disease with the remainder having localised disease, usually septic arthritis or abscesses. Three fourth of patients (75%) were treated successfully on follow-up, with a regimen of parenteral ceftazidime followed by oral doxycycline and cotrimoxazole. CONCLUSION: Melioidosis is an emerging infection in India especially in males from rural areas, with diabetes and alcoholism being the commonest risk factors. Both sepsis with bacteraemia and localised disease involving joints or focal abscess were common presentations. Diagnosis is readily made by culturing the organism from appropriate clinical specimens and identifying non-fermenting Gram negative bacteria to the species level. As there was an excellent response in 75% of patients, early suspicion, culture confirmation and therapy is warranted in India.

Mucormycosis in patients without cancer: a case series from A tertiary care hospital in South India.

BACKGROUND: Mucormycosis (Zygomycosis) is a life-threatening infection. We attempted to analyse clinical features and risk factors of Mucormycosis cases in a tertiary care referral institution in India, in patients without underlying malignancy. METHODS: We retrospectively analyzed data of patients diagnosed as having Mucormycosis over a 10 year period of 2000-2010. Patients with a histopathology report and/or a Microbiology report of Zygomycetes or Mucor from a biopsy specimen were included in the study. RESULTS: Out of the 27 cases, rhino-orbital/rhino-cerebral involvement occurred in 12 (44.4%) patients, pulmonary involvement in 3 (11.1%) cases, soft tissue involvement in 11 (40.7%) cases and gastrointestinal involvement in one patient (3.7%). Diabetes mellitus is the main risk factor, followed by renal failure and trauma. Mean ESR value of these patients was 118 mm/1 hour. Mean WBC count was 20 x 10(9)/L, and neutrophil count 82%. The mean absolute neutrophil count (ANC) was 16.8 x 10(9)/L. CONCLUSION: The interesting finding in our study was the presence of neutrophilic leucocytosis and high ESR in most of the patients. In a predisposed individual, especially in a diabetic, in countries with high environmental fungal burden; presence of high ESR and neutrophilic leucocytosis with a compatible clinical presentation should raise suspicion of Mucormycosis.

Comparison of double disc and three dimensional methods to screen for ESBL producers in a tertiary care hospital.

Extended spectrum beta lactamases (ESBLs) continue to be a major problem in clinical setups world over, conferring resistance to the expanded spectrum cephalosporins. An attempt was made to study ESBL production among Enterobacteriaceae members from a tertiary care center in Chennai. A total of seventy randomly collected isolates of the family Enterobacteriaceae from a tertiary care center was studied for their susceptibility patterns to various antibiotics and detection of ESBL producers by double disc synergy (DDS) test and three dimensional test (TDT). Eighty percent of the isolates were multidrug resistant (MDR) and 20% were ESBL producers. TDT detected 85.7% whereas only 14.2% were detected by DDS. In the present study, a large number of isolates were found to be MDR and ESBL producers. TDTs were found to be better than DDS in the detection of ESBLs. Continued monitoring of drug resistance is necessary in clinical settings for proper disease management.

The vertical displacement of the center of mass during walking: a comparison of four measurement methods.

Measuring the vertical displacement of the center of mass (COM) of the body during walking may provide useful information about the energy required to walk. Four methods of varying complexity to estimate the vertical displacement of the COM were compared in 25 able-bodied, female subjects. The first method, the sacral marker method, utilized an external marker on the sacrum as representative of the COM of the body. The second method, the reconstructed pelvis method, which also utilized a marker over the sacrum, theoretically controlled for pelvic tilt motion. The third method, the segmental analysis method, involved measuring motion of the trunk and limb segments. The fourth method, the forceplate method, involved estimating the COM displacement from ground reaction force measurements. A two-tailed paired t-test within an ANOVA showed no statistically significant difference between the sacral marker and the reconstructed pelvis methods (p = 0.839). There was also no statistically significant difference between the sacral marker and the segmental analysis method (p = 0.119) or between the reconstructed pelvis and the segmental analysis method (p = 0.174). It follows that the first method, which is the most simple, can provide essentially the same estimate of the vertical displacement of the COM as the more complicated second and third measures. The forceplate method produced data with a lower range and a different distribution than the other three methods. There was a statistically significant difference between the forceplate method and the other methods (p < 0.01 for each of the three comparisons). The forceplate method provides information that is statistically significantly different from the results of the kinematic methods. The magnitude of the difference is large enough to be physiologically significant and further studies to define the sources of the differences and the relative validity of the two approaches are warranted.

A tool to assess biomechanical gait efficiency; a preliminary clinical study.

A goal of many physiatric interventions is to improve biomechanical walking efficiency. Thus, a tool that helps assess this efficiency, independent of cardiac, pulmonary, psychologic, or other nonbiomechanical factors, would be useful. Currently used methods to measure efficiency, including comfortable walking speed, are not specific to biomechanical variables. A potential tool, the biomechanical efficiency quotient (BEQ), which uses three variables--average stride length, vertical displacement of the trunk during walking, and sacral height during standing--is proposed and preliminarily tested. This quotient is based on Saunders, Inman, and Eberhart's theories and on a prior study in able-bodied subjects. The BEQ was computed in 20 consecutive patients with neurologically based gait disability referred for gait laboratory evaluation who subjectively reported that one or two ankle-foot-orthoses (AFOs) reduced the effort necessary to walk. The quotient was calculated with and without the AFO(s) by dividing the average vertical displacement of the sacrum, which was measured with an optoelectronic system, and by a predicted displacement, which was based on the patient's sacral height and average stride length. The mean BEQ with the AFO(s) (6.3 +/- 4.4) was significantly less than the mean BEQ without the AFO(s) (9.7 +/- 7.1); P = 0.005. Furthermore, the BEQ was less with the AFO(s) compared with trials without the AFO(s) in all subjects. Percent change in BEQ with the AFO(s) (26.8 +/- 19.6) correlated with percent change in comfortable walking velocity (24.8 +/- 31.8), r = 0.73, P<0.001, across all subjects. The BEQ may be useful in specifically assessing the effect on biomechanical efficiency of physiatric interventions, despite variable nonbiomechnical factors. An instrument to measure vertical trunk displacement during walking outside of the gait laboratory would be extremely useful for further necessary longitudinal studies.