Social and biological evaluation of antimicrobial resistance (SOBEAR) in rural India: a study protocol.

Background: Antimicrobial resistance (AMR) has been one of the biggest global health threats in recent years, mostly in low- and middle-income countries, which requires urgent research using a multidisciplinary research approach. The use of large quantities of antimicrobial drugs inappropriately for humans, poultry and agriculture has been recognized as a leading cause of antibiotic resistance and the predominance of drug-resistance pathogens in the environment. This protocol aims to describe the use/misuse of antibiotics (ABs) in the community and evaluate clinical samples from healthcare settings to detect genes associated with antimicrobial resistance. Methods: We will conduct a community-level survey in different villages of the Tigiria block to assess knowledge and awareness on ABs and AMR. We will conduct in-depth interviews (IDIs) with doctors, pharmacists, nurses and drug sellers, as well as focus group discussions (FGDs) with ASHA and ANM workers who are involved in antibiotic supplies to the community. Quantitative data from the community survey and qualitative data of IDIs and FGDs will be linked and analyzed using statistical modeling and iterative thematic content analysis. Specimens (stool, urine, blood and wound/pus) will be collected from clinically diagnosed patients of different healthcare centers of Tigiria block. The samples will be cultured for bacterial isolation and antibiotic sensitivity testing. Genomic DNA will be isolated from positive bacterial cultures and sequenced using PCR to evaluate high-threat multi-drug resistance organisms (MDROs), screening of plasmid-mediated quinolone resistance (PMQR) genes, antimicrobial genes responsible for MDR and quinolone resistance-determining regions (QRDRs). Conclusion: This is the community-based protocol to evaluate the knowledge, attitudes, awareness and practices regarding ABs and AMR. The study protocol establishes a foundation for evaluating population-based prevalence and risk factors for AMR and MDROs in rural areas of the Odisha state, India.

Sequential outbreaks due to mixed ctxB alleles of Vibrio cholerae O1 in Mayurbhanj district of Odisha, India.

INTRODUCTION: Cholera is a significant threat causing outbreaks/epidemics with high morbidity and mortality in coastal and tribal districts of Odisha. A sequential cholera outbreak reported from four places in Mayurbhanj district of Odisha during June to July 2009 was investigated. METHODOLOGY: Rectal swabs from diarrhea patients were analyzed for the identification, antibiogram profiles and detection of ctxB genotypes by double mismatch amplification mutation (DMAMA) polymerase chain reaction (PCR) assays and sequenced. The different virulent and drug resistant genes were detected by multiplex PCR assays. The clonality analysis on selected strains was done by pulse field gel electrophoresis (PFGE). RESULTS: Bacteriological analysis of rectal swabs revealed the presence of V. cholerae O1 Ogawa biotype El Tor which were resistant to co-trimoxazole, chloramphenicol, streptomycin, ampicillin, nalidixic acid, erythromycin, furazolidone and polymyxin B. DMAMA-PCR assay revealed that the cholera outbreak in Mayurbhanj district was due to both ctxB1 and ctxB7 alleles of V. cholerae O1 El Tor strains. All the V. cholerae O1 strains were positive for all virulence genes. The multiplex PCR assay on V. cholerae O1 strains revealed the presence of antibiotic resistance genes like dfrA1 (100%), intSXT (100%), sulII (62.5%) and StrB (62.5%). PFGE results on V. cholerae O1 strains exhibited two different pulsotypes with 92% similarity. CONCLUSIONS: This outbreak was a transition phase where both ctxB genotypes were prevalent after which the ctxB7 genotype gradually became dominant in Odisha. Therefore, close monitoring and continuous surveillance on diarrheal disorders is essential to prevent the future diarrheal outbreaks in this region.

Clinicodermoscopic and immunopathological profile of non-infectious non-eczematous inflammatory tattoo reactions: A retrospective study from a tertiary care centre of East India.

Introduction Tattoo-associated complications are on the rise due to the popularity of decorative tattoos in recent years. The exact pathogeneses of various tattoo reaction patterns are still unclear, and their dermoscopic details are sparsely reported. Aim We aimed to retrospectively study the clinical, dermoscopic and immunopathological details of patients with non-infectious, non-eczematous inflammatory tattoo reaction patterns in a tertiary care centre of East India. Method The clinical, dermoscopic and pathological details of all the patients who had non-infectious, non-eczematous inflammatory tattoo reactions were collected. In all the cases, immunohistochemistry was done for CD1a, CD3, CD4, CD8, FoxP3, CD20 and CD56. Results A total of five patients of skin phototypes IV and V and six tattoo reactions were analysed. Five lesions had reactions at the site of a black tattoo, and one at the site of red tattoo. Clinically, the patients presented with erythematous or blue-grey flat-topped to verrucous papules and plaques. Dermoscopic features were dominated by a central white to pink-white structureless area, a peripheral grey-white to bluish-white structureless area, white scales, comedo-like opening with keratotic plugging, milia-like cysts and shiny white structures. Pathologically, except for one lesion that only showed a lichenoid reaction pattern in the red tattoo, all had a combination of reaction patterns. Immunohistochemistry showed increased epidermal and dermal Langerhans cells, predominantly CD8 positive T cells in the epidermis and dermis, sparse dermal B cells and CD4 positive T cells, reduced T regulatory cells and a complete absence of CD56 positive NK cells. Limitations Small sample size was the limitation of the study. Conclusion The clinical morphology and dermoscopy may not differentiate between various types of non-infectious non-eczematous inflammatory tattoo reactions. The immunological profile supports a delayed hypersensitivity reaction due to contact sensitisation to tattoo pigment, and CD8 positive T cells play a central role in executing various pathological reaction patterns, both in the epidermis and dermis.

Clinico-Dermoscopic-Pathological Features of a Rare Case of Locally Invasive Multifocal Primary Cutaneous Diffuse Large B Cell Lymphoma-Leg Type Over the Face and Scalp.

Primary cutaneous diffuse large B-cell lymphoma-leg type (PCDLBCL-LT) is characterized by diffuse monotonous proliferation of centroblasts and immunoblasts. It commonly presents as erythematous to violaceous nodules on one or both the legs and has a poor prognosis. We report the clinico-dermoscopic-pathological features and therapeutic response of a rare case of PCDLBCL-LT in a 62-year-old diabetic man, who presented with multifocal plaques, one lobulated and two arcuate-shaped, on the face and scalp. During the investigation, one of the plaques had eroded the underlying bone without any evidence of malignant cells in the cerebrospinal fluid. He was successfully treated with R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) along with intrathecal methotrexate.

Adsorption of As(III) and As(V) from aqueous solution by magnetic biosorbents derived from chemical carbonization of pea peel waste biomass: Isotherm, kinetic, thermodynamic and breakthrough curve modeling studies.

The purpose of this research was to investigate the adsorption of arsenic (As) from aqueous solutions using MPAC-500 and MPAC-600 (magnetic-activated carbons synthesized from the peel of Pisum sativum (pea) pyrolyzed at 500 °C and 600 °C temperatures, respectively). The potential of both biosorbents for As adsorption was determined in batch and column mode. The characterization of both biosorbents was performed by energy dispersive spectroscopy, scanning electron microscope, pHZPC, particle size distribution, X-ray diffraction, zeta potential and Fourier-transform infrared spectroscopy. It was found that the efficiency of MPAC-600 was better than MPAC-500 for the adsorption of As(III) and As(V) ions. The adsorption capacities of MPAC-500 and MPAC-600 in removing As(III) were 0.7297 mg/g and 1.3335 mg/g, respectively, while the values of Qmax for As(V) on MPAC-500 and MPAC-600 were 0.4930 mg/g and 0.9451 mg/g, respectively. The Langmuir isotherm model was found to be the best fit for adsorption of As(III) by MPAC-500 and MPAC-600, as well as adsorption of As(V) by MPAC-500. The Freundlich isotherm model, on the other hand, was optimal for As(V) removal with MPAC-600. With R2 values close to unity, the pseudo-second-order kinetics were best fitted to the adsorption process of both As species. The Thomas model was used to estimate the breakthrough curves. The effects of coexisting oxyanions and regeneration studies were also carried out to examine the influence of oxyanions on As adsorption and reusability of biosorbents.

Perception and determinants leading to antimicrobial (mis)use: A knowledge, attitude, and practices study in the rural communities of Odisha, India.

Background: Antimicrobial resistance (AMR) has emerged as one of the major public health issues globally. This cross-sectional study determined knowledge, attitudes and practices (KAP) regarding antimicrobial use and AMR among rural communities of Tigiria (Odisha), India. Methods: A semi-structured questionnaire based on socio-demographic characteristics, antibiotics usage, awareness of antimicrobial resistance, healthcare utilization and quality of life were asked to the participants using an electronic device with Open Data Kit. Descriptive statistics, independent t-test and ANOVA were performed to analyze the variables. Results: A total of 1,003 participants were surveyed in the study from 25 villages of Tigiria. About 44.47% (95% CI: 41.36-47.60) of study participants have heard about antimicrobial medicines and 14.75% (95% CI: 12.65-17.13) of participants were involved in buying antibiotics without prescription over the counter. Around 20.14% (95% CI: 17.72-22.78) of participants, stopped taking antibiotics before completing the full course. The physical domain was the most affected with low scores compared to other domains of quality of life (QOL). The QOL scores were found significant (p < 0.05) across age, gender, education and ethnicity. Conclusion: The study documented a significant level of KAP regarding antimicrobial (mis)use in the study. It is essential that antimicrobial stewardship programs for various stakeholders and educational programmes must be initiated to increase awareness of people on antimicrobial resistance.

Genomic diversities of ctxB, tcpA and rstR alleles of Vibrio cholerae O139 strains isolated from Odisha, India.

The genome of Vibrio cholerae O139 strains has undergone cryptic changes since its first emergence in 1992 in South India. This study aimed to determine the presence of genotypic changes marked in ctxB, tcpA and rstR genes located within the CTX prophages among the strains of V. cholerae O139 isolated from 1999 to 2017 in Odisha. Antibiotic susceptibility test was conducted on 59 V. cholerae O139 strains. A conventional PCR assay was done for ctxB gene typing followed by sequencing along with identification of rstR and tcpA gene. Pulsed-field gel electrophoresis (PFGE) was carried out to reveal clonal variations among the V. cholerae O139 strains. Among V. cholerae O139 isolates more than 60% showed resistance to ampicillin, co-trimoxazole, furazolidone, streptomycin, neomycin and nalidixic acid. The ctxB sequencing and rstR allele-specific PCR assay revealed the presence of three genotypes 1, 3 and 4 with at least one copy of CTX Calc φ in addition to CTX ET and CTX Cl prophages in V. cholerae O139 isolates. PFGE analysis revealed 13 pulsotypes with two clades having 60% similarity among V. cholerae O139 strains. The circulating V. cholerae O139 strains in Odisha showed variation in genotypes with multiple clonal expansions over the years.

Variants of ctxB alleles of Vibrio cholerae O1 caused sequential cholera outbreaks in the tribal areas of Odisha, India.

Cholera localized outbreaks/epidemics accounting for high morbidity and mortality have been reported in different years both from the coastal and tribal districts of Odisha. In the present study, the emergence and spread of two sequential cholera outbreaks reported in July to October 2012 from Rayagada and Kalahandi districts of Odisha was investigated. Environmental water samples from different sources and rectal swabs from diarrhoea patients were analysed for identification, antibiogram profiles and molecular studies using DMAMA-PCR assays. The pulsed field gel electrophoresis (PFGE) was done on some selected Vibrio cholerae O1 strains isolated from these cholera outbreak areas. Results showed 42% of rectal swabs and 2.3% of water samples collected from both the districts were positive for Vibrio cholerae O1 Ogawa biotype El Tor carrying both ctxB1 and ctxB7 genotypes. The common resistance profile of V. cholerae O1 strains was ampicillin, nalidixic acid, furazolidone and co-trimoxazole. The PFGE analysis on selected V. cholerae O1 strains of ctxB1 and ctxB7 genotypes showed three pulsotypes with 96% similarity matrix exhibiting the relationship with their respective water sources. Hence, continuous surveillance is highly essential to monitor the antibiogram profile and changing pattern of ctxB genotypes of V. cholerae O1 in this region.

Origin and Dissemination of Altered El Tor Vibrio cholerae O1 Causing Cholera in Odisha, India: Two and Half Decade's View.

The origin, spread and molecular epidemiology of altered El Tor Vibrio cholerae O1 strains isolated from cholera outbreaks/surveillance studies between 1995 and 2019 from different district of Odisha were analyzed. The stock cultures of V. cholerae O1 strains from 1995 to 2019 were analyzed through molecular analysis using different PCR assays and pulse field gel electrophoresis (PFGE) analysis. The spread map (month, year and place) was constructed to locate the dissemination of altered El Tor variants of V. cholerae O1 in this region. A total of 13 cholera outbreaks were caused by V. cholerae O1 Ogawa biotype El Tor carrying ctxB1 and ctxB7 genotypes. The ctxB1 alleles of V. cholerae O1 mostly confined to the coastal areas, whereas the ctxB7 genotypes, though originating in the coastal region of Odisha, concentrated more in the tribal areas. The positive correlation between virulence-associated genes (VAGs) was found through Pearson's correlation model, indicative of a stronger association between the VAGs. The clonal relationship through PFGE between ctxB1 and ctxB7 genotypes of V. cholerae O1 strains exhibited 80% similarity indicating single- or multi-clonal evolution. It is evident from this study that the spread of multidrug-resistant V. cholerae O1-altered El Tor was dominant over the prototype El Tor strains in this region. The origin of altered El Tor variants of V. cholerae O1 occurred in the East Coast of Odisha established that the origin of cholera happened in the Gangetic belts of Bay of Bengal where all new variants of V. cholerae O1 might have originated from the Asian countries.

Dermoscopic features of acute cutaneous lupus erythematosus: A retrospective analysis from a tertiary care centre of East India.

BACKGROUND: The cutaneous features of acute cutaneous lupus erythematosus (ACLE) can be confused with other dermatoses. The dermoscopic features of ACLE have rarely been reported. AIMS: We aimed to study the dermoscopic features of ACLE, malar rash, and generalized ACLE. MATERIALS AND METHODS: The dermoscopic features of all biopsy and direct immunofluorescence confirmed ACLE cases were analysed retrospectively between May 2019 and May 2020. RESULTS: A total of 21 patients with skin phototypes III, IV, or V were included in the analysis. A pinkish-white to reddish-white homogenous area (94.1%), brown dots, globules and peppering (76.4%), and keratotic follicular plugging (64.7%) were the common dermoscopic features observed in the case of generalized ACLE. Similarly, in the case of malar rash, a pinkish-white to reddish-white homogenous area (77.8%) was the most common dermoscopic feature, followed by keratotic follicular plugging (66.7%), scales (44.5%), and brown dots, globules, and peppering (44.5%). In both variants, vascular structures were dominated by linear vessels with or without branching. CONCLUSIONS: A multicomponent pattern comprising white scales, homogenous pinkish-white to the reddish-white area, brown dots/globules/peppering, keratotic follicular plugging, and linear vessels with or without branching were the common dermoscopic features observed in our study and may indicate the diagnosis of both localized and generalized ACLE. In addition, a paucity of dotted vessels may serve as an additional clue.

Emergence and spread of different ctxB alleles of Vibrio cholerae O1 in Odisha, India.

This study reports variants of the ctxB allele of Vibrio cholerae O1 isolated between 1995 and 2019 in Odisha, India. ctxB1 genotypes dominated from 1995 to 2016. The Haitian variant and El Tor ctxB3 genotypes of V. cholerae O1 emerged in 1999, and were most common in 2018-2019 and 2005-2011, respectively. The ctxB7 genotype of the Haitian variant of V. cholerae O1 was quiescent from 2000 to 2006, but further spread was noted from 2007 to 2019.

Dissemination of Vibrio cholerae O1 isolated from Odisha, India.

The present study reported the antimicrobial susceptibility trends, virulence genes, and drug resistance genes of Vibrio cholerae O1 strains isolated from outbreaks and epidemics over two and half decades (1995-2019) from Odisha, India. Antimicrobial susceptibility testing was performed by disc diffusion method. Virulence and drug resistance genes were detected by multiplex PCR assays. All V. cholerae O1 strains were sensitive to gentamicin, chloramphenicol, norfloxacin and ciprofloxacin while resistant to one or more antibiotics used. About 90% of the isolates of V. cholerae O1 carried antibiotic drug resistant genes (SulII, dfrA1 and strB) and SXT elements and the results correlated with the phenotypic antibiotic data obtained through disc diffusion assay. The tcpA Haitian variant V. cholerae O1 first appeared in 1999, gradually showing its increasing number upto 2019. TcpA El Tor strains only prevailed from 1995 to 2006; whereas the tcpA classical strains of V.choleraeO1 were found in less number from 1995 to 2016. Two multiplex PCR assays confirmed the presence of various toxigenic and virulence genes (toxR, ompU, ace, rtxC, ctxA, tcpA, rfbO1 and ompW) in all isolate of V. cholerae O1 strains. The present findings demonstrated the origin and spread of Haitian variants tcpA in V. cholerae O1 strains over two and half decades.

Dissemination of Polymyxin B Sensitivity in El Tor Vibrio cholerae O1 Strains in Odisha, India.

The Vibrio species undergo cryptic changes in their genetic material for better adaptability, which accounts for antibiotic resistance. In the present study, we investigated the emergence and spread of sensitivity to polymyxin B (PB) by El Tor V. cholerae O1 strains from 1995 to 2019 in Odisha, India. The results showed that out of 1200 V. cholerae O1 strains, 89.4% were resistant and the remaining 10.6% strains were sensitive to PB. The sensitivity to PB of V. cholerae O1 strains emerged from 2005 to 2019, except in 2015, clearly signifying the presence of classical biotype characteristics in the El Tor variant of V. cholerae O1 strains. The Etest assay revealed some interesting traits of PB susceptibility in the ctxB1 and ctxB7 genotypes of V. cholerae O1 strains. The minimum inhibitory concentration (MIC) of ctxB7 genotypes showed reduced MIC values of ≤ 4 µg/mL, whereas ctxB1 genotypes exhibited higher MIC values of 24 and 32 µg/mL.

Spread of Haitian Variant Vibrio cholerae O1 Causing Cholera Outbreaks in Odisha, India.

Cholera posed a significant threat causing outbreaks/epidemics with high morbidity and mortality in Odisha. This study envisages the characterisation of isolated pathogen from two cholera outbreaks reported in 2018 and 2019 from Bargarh and Rayagada districts of Odisha respectively. Vibrio cholerae O1 were isolated following standard techniques. The different virulent and drug resistant genes were detected by multiplex PCR assays; whereas the ctxB genotypes were characterised through double mismatch amplification mutation (DMAMA) PCR assay. The ctxB genes were further sequenced and pulse-field gel electrophoresis (PFGE) was done on some selected strains. The clinical and water isolates of Haitian variant (HCT) V. cholerae O1 Ogawa biotype El Tor with multi drug resistant strains were isolated from both the places. All the V. cholerae O1 strains were positive for virulence genes. The antibiotic resistant genes like dfrA1 (100%), strB (76.9%), intSXT (61.5%) were detected. The PFGE results on V. cholerae O1 strains exhibited two different pulsotypes. These cholera outbreaks were due to multidrug resistant HCT variant V. cholerae O1 strains which were circulating and caused the cholera outbreaks in Odisha. So continuous surveillance on diarrheal disorders is highly essential to prevent the future diarrheal outbreaks in this region.