Investigating the Clinico-Demographic Characteristics of Dengue Fever and Its Seroprevalence at a Tertiary Care Hospital in Northern India.

Background and objective Dengue virus (DENV) is a major global health threat, causing over 50,000 deaths annually. The state of Uttar Pradesh (UP) in India faces significant challenges due to the increasing number of dengue cases detected. This study aimed to assess DENV seropositivity in the Raebareli district of UP, to offer crucial insights into the region's effective control and management strategies. Materials and methods This study, after obtaining approval from the ethics committee, analyzed blood samples of individuals suspected of having dengue at a teaching hospital in rural UP between January and December 2022. To determine the disease's seroprevalence, both dengue NS1 antigen ELISA and dengue IgM Microlisa were conducted. Furthermore, RT-PCR was performed on NS1-positive samples to confirm the serotypes. The collected data were analyzed using Epi Info 7.0. Results Of the 589 suspected dengue cases, 86 (14.60%) tested positive for dengue NS1 and/or IgM. Our findings showed that males (n=330, 56.03%) and adolescents and young adults (n=301, 51.1%) from rural areas (n=523, 88.4%) were predominantly affected. Cases peaked post-monsoon, and platelet levels were notably low in NS1-positive cases. Dengue serotype 2 (DEN-2) was found in all RT-PCR-positive samples. Our results revealed a dengue seroprevalence of 14.60% (n=86), which peaked in post-monsoon months. The higher incidence among males and young adults from rural areas attending the outpatient department highlights the importance of targeted interventions and community surveillance. RT-PCR confirmed the circulation of a single serotype in the region. Conclusions This study contributes crucial insights into dengue's epidemiology and clinical profile and its findings are all the more significant now as India prepares for phase 3 trials of a quadrivalent dengue-virus vaccine in 2024. Adolescent and young adult males have an increased likelihood of acquiring the virus, and this demographic can be prioritized for vaccine trials.

Actinomycetoma by Nocardia asteroides: A Case Report of a Unique Neglected Tropical Disease From North India.

Nocardial mycetoma is a neglected tropical disease reported worldwide, especially in tropical and subtropical regions. It is ubiquitous in nature and is a soil-borne, gram-positive, filamentous, aerobic bacteria with acute angle branching. Traumatic inoculation in endemic areas is the primary mode of infection of this debilitating disease. The clinical triad of tumefaction, draining sinus, and pus discharge with granules is very much characteristic and specific for clinching the diagnosis of mycetoma. However, the painless nature of the primary skin lesion often makes the patient present late to the clinician, often in the advanced stages of the disease. Here, we present a very intriguing case report of a young female patient who presented with a single neck nodule but was later diagnosed as a case of nocardial mycetoma. Timely diagnosis and initiation of therapy proved to be a boon for the patient with almost complete recovery within a few weeks in the form of healed skin lesions and insignificant scarring.

Commencement and Monitoring: Microbiological Surveillance in Operation Theaters at a Tertiary Care Center in North India.

INTRODUCTION: Preserving sterility and safety in hospital operation theaters (OTs) is vital. We have implemented a comprehensive microbiological surveillance program for OTs, encompassing both commencement and ongoing monitoring. This study assesses the prevalence of microorganisms, identifies their types, and detects contamination on surfaces and in the air. METHODS: Commencement and monitoring samples were collected from October 2021 to July 2023, from nine OTs. OTs were cleaned with soap and water, disinfected, and fogged with quaternary ammonium compounds. After sealing the OTs overnight, samples were collected aseptically. Air was sampled using the settle plate method, and surfaces were swabbed. Six surfaces, namely, the floor, wall, table, light, anesthesia workstation, and door handle, were swabbed. Samples were transported immediately to the institution's microbiology laboratory. RESULTS: During OT commencement, 247 swabs from nine OTs yielded 19 (7.29%) positives for bacterial growth. These microorganisms were primarily non-pathogenic, including aerobic spore-forming bacilli and Micrococcus, with an average bioload of 9.5 colony-forming units (CFU)/m3 of air. During OT monitoring, swab positivity was 10.79% (23/213). The General Surgery OT and Obstetrics and Gynecology OT showed the highest bacterial growth (5/23). Surface sampling revealed prevalent methicillin-resistant coagulase-negative staphylococci (MRCoNS) (9/23), followed by methicillin-sensitive Staphylococcus aureus (MSSA) (4/23) and methicillin-sensitive coagulase-negative staphylococci (MSCoNS) and aerobic spore-forming bacilli (ASB) (3/10). The General Surgery, Obstetrics and Gynecology, and ENT OTs displayed elevated air bioloads of 53, 49, and 47 CFU/m3, respectively. CONCLUSION: In newly constructed non-operational OTs, non-pathogenic organisms prevailed. However, as the OTs became functional, pathogenic organisms became more prevalent. Sampling emphasized contamination in areas with high patient loads, such as General Surgery, Obstetrics and Gynecology, and ENT OTs. Notably, OT tables and OT walls exhibited higher pathogenic microorganism presence. By combining both initial commencement and ongoing monitoring, the institution has effectively managed the microbial environment within its OTs.

Recent trends in targeted delivery of smart nanocarrier-based microbial enzymes for therapeutic applications.

Smart carrier-based immobilization has widened the use of enzymes for the treatment of several disorders. Large surface areas, tunable morphology, and surface modification ability aid the targeted and controlled release of therapeutic enzymes from such formulations. Smart nanocarriers, such as polymeric carriers, liposomes, and silica have also increased the stability, half-life, and permeability of these enzymes. In this review, summarize recent advances in the smart immobilization of microbial enzymes and their development as precision nanomedicine for the treatment of cancer, thrombosis, phenylketonuria (PKU), and wound healing. We also discuss the challenges and measures to be adopted for the successful clinical translation of these formulations.

Monkeypox: another pandemic in the making?

Monkeypox is a zoonotic disease caused by the monkeypox virus, which is a member of the Poxviridae family of viruses. It is transmitted through direct or indirect contact with fluid secretions. Initial symptoms include fever, chills, headache, and malaise, followed by a maculopapular rash that starts on the face and progresses centrifugally. Polymerase chain reaction is the preferred laboratory test for the diagnosis, and management is mostly supportive. The clinical presentation of monkeypox is quite similar to that of another member of the Poxviridae family: smallpox, which wreaked havoc in the 20th century, before being eradicated with the help of the vaccinia virus vaccine in 1977. This vaccine protects not only against smallpox but also monkeypox; therefore, when use of this vaccine was discontinued, monkeypox had a new susceptible population to infect and way to proliferate and evolve. Initially the disease spread in Africa, but now the more evolved monkeypox is quickly spreading to other countries. On July 23, 2022, the World Health Organization declared this multicountry outbreak a public health emergency of international concern. Given its mutating ability and high transmissibility, we need to quickly devise measures to control this virus before it turns into a pandemic.

Possible Long-Term Cardiovascular Effects of COVID-19.

Coronavirus Disease 2019 is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and has become a worldwide pandemic. Since 2019, the virus has mutated into multiple variants that have made it harder to eradicate and have increased the rate of infection. This virus can affect the structure and the function of the heart and can lead to cardiovascular symptoms that can have long-lasting effects despite recovery from COVID-19. These symptoms include chest pain, palpitations, fatigue, shortness of breath, rapid heartbeat, arrhythmias, cough and hypotension. These symptoms may persist due to myocardial injury, cardiac inflammation or systemic damage that may have been caused during infection. If these symptoms persist, the patient should visit their cardiologist for diagnosis and treatment plan for any type of cardiovascular disease that may have developed Post-COVID 19.

Preidentification of high-risk pregnancies to improve triaging at the time of admission and management of complications in labour room: a quality improvement initiative.

Complications can occur anytime during pregnancy and childbirth. Pregnancies associated with high-risk factors have a higher-than-normal risk for fetomaternal complications. Bhagwan Mahavir hospital is a public sector hospital catering to low-risk and high-risk pregnant women (PW) in the labour room (LR)). The obstetrics and gynaecology team observed that at times the LR team failed to identify high-risk pregnancy (HRP) during admission in LR and to manage complications timely and efficiently. Therefore, the team started a quality improvement (QI) project in January 2019 with the aim to admit preidentified HRP in LR from existing 0% to 80% in 3 months.The QI team followed the point-of-care quality improvement methodology to conduct this improvement process. They identified HRP in the outpatient department (OPD) during their antenatal care (ANC) visits, mentioned an HRP number on their ANC cards, and did risk stratification with yellow and red stickers into moderate and severe HRP respectively. Preidentified HRP were attended, admitted and managed on priority in the LR. The team achieved its aim in the ninth week of the QI initiative and sustaining to date. The team also measured and analysed the type of HRP identified in OPD, complications occurring around the process of childbirth in LR, maternal near-miss, maternal death and PW referred out from LR. They observed a 6.5%-point reduction (68.93%) in the median complication rate of major life-threatening complications following this improvement process.This new intervention facilitated the team in early initiation of management of HRP in OPD, their triaging in LR, preparedness towards managing complications, involvement of support staff, PW and their relatives in the patient care, and redistribution of human resources according to priority area. The lessons learnt are generalisable and can be used in other facilities with similar settings.

Improvement in Pain and Quality of Life After Ultrasound-Guided Saphenous Nerve Block in Patients With Knee Osteoarthritis.

Introduction Osteoarthritis (OA) of the knee is a common complaint in the elderly population and results in considerable disability in advanced stages. Though many pharmacological, electrotherapeutic, and interventional options are available for the effective treatment of knee OA in the early stages, these modalities fail to provide effective and long-term relief in some cases where peripheral nerve blocks may prove beneficial. Hence, this study was conducted to assess the efficacy of the saphenous nerve block in knee pain due to OA. Objective To evaluate improvement in pain and quality of life after ultrasound-guided saphenous nerve block in patients with knee OA. Material and methods An interventional prospective study in patients with knee OA, with medial compartment knee pain, was conducted from March 2016 to March 2017. All patients were evaluated prior to the procedure, and then at one week, one month, three months, and six months. The pain was evaluated using the visual analog scale (VAS) and functional improvement using the Knee injury and Osteoarthritis Outcome Score (KOOS). Results Forty patients with unilateral knee OA underwent saphenous nerve block. Fifty percent of the patients reported pain relief within one week, whereas 58%, 33%, and 23% exhibited relief at subsequent follow-ups at one, three, and six months. A statistically significant difference (p < 0.0001) was observed in pain (VAS and KOOS pain) and functional scales (KOOS symptom, quality of life (QOL), and activities of daily living (ADL)) at follow-up evaluations. Conclusion Ultrasound-guided saphenous nerve block results in a significant improvement in pain and QOL in patients with knee OA.

Private Health Sector in India-Ready and Willing, Yet Underutilized in the Covid-19 Pandemic: A Cross-Sectional Study.

Background: The private medical sector is a resource that must be estimated for efficient inclusion into public healthcare during pandemics. Methods: A survey was conducted among private healthcare workers to ascertain their views on the potential resources that can be accessed from the private sector and methods to do the same. Results: There were 213 respondents, 80% of them being doctors. Nearly half (47.4%) felt that the contribution from the private medical sector has been suboptimal. Areas suggested for improved contributions by the private sector related to patient care (71.8%) and provision of equipment (62.4%), with fewer expectations (39.9%) on the research front. Another area of deemed support was maintaining continuity of care for non-COVID patients using virtual consultation services (77.4%), tele-consultation being the preferred option (60%). 58.2% felt that the Government had not involved the private sector adequately; and 45.1% felt they should be part of policy-making. Conclusion: A streamlined pathway to facilitate the private sector to join hands with the public sector for a national cause is the need of the hour. Through our study, we have identified gaps in the current contribution by the private sector and identified areas in which they could contribute, by their own admission.

Incidence and Risk Factors for Hepatitis C Virus and Hepatitis B Virus Seroconversion in End-Stage Renal Failure Patients on Maintenance Hemodialysis.

BACKGROUND: Renal replacement therapy in the form of either dialysis or transplantation is the only option for end-stage renal disease (ESRD). Blood-borne infections such as hepatitis B virus (HBV) and hepatitis C virus (HCV) are of special concern in these patients because of their high incidence. Although there are sufficient data from the developed world, there is scarcity of data from developing countries such as India. METHODS: All newly diagnosed ESRD patients initiated on hemodialysis after attending the Department of Nephrology, PGIMER, Chandigarh between January 2015 and October 2015 were included in the study. All the subjects were initially screened for HCV and HBV serology status and subsequent HCV and HBV status on follow-up at the end of 6 months and evaluated by standardized precoded questionnaires and biochemical examinations. Univariate and multivariate analyses were done to identify the risk factors for seroconversion. RESULTS: A total of 196 patients were recruited for the study after confirming seronegative status. At the end of 6 months, 61 patients lost to follow-up. Anti-HCV antibody had shown moderate association to HCV RNA testing at the end of 6 months by kappa test. Out of 135, 16.3% seroconverted to HCV RNA positive and 0.7% patient became hepatitis B surface antigen positive. Isolation of dialysis machine and nursing staff was associated with lower seroconversion. CONCLUSION: In a real-life scenario, HCV seroconversion is observed in 15% of the patients initiated on hemodialysis. Isolation of both dialysis machine and personnel was associated with lower seroconversion.

Fever of Unknown Origin in a Renal Transplant Recipient: Lactate Dehydrogenase as an Important Clue to Diagnosis.

Histoplasmosis is a rare disease in nonendemic areas. We report a case of a 23-year-old male patient who presented with fever of unknown origin, cytopenias, organomegaly, and allograft dysfunction 4 months after renal transplant with father as donor. Bone marrow examination showed intracellular budding yeast cells, which was confirmed as histoplasmosis by culture of bone marrow biopsy sample. The patient was treated with intravenous liposomal amphotericin and responded well.

Multiple cerebral abscesses in a renal transplant recipient: Two swords in one scabbard!

Although rare, both Cladophialophora bantiana (C. bantiana) and Toxoplasma gondii have been known to be associated with brain abscess in renal transplant recipients (RTRs), however co-infection has never been reported till date. In the present case, 40 years old renal transplant recipient on curtailed immunosuppressive therapy presented with progressive headache and altered sensorium. The computed tomography of head showed multiple ring-enhancing discrete lesions in the left frontal lobe, with moderate perilesional oedema. Left frontal craniotomy and aspiration revealed thick yellowish brown pus, which on culture showed the growth of dematiaceous fungal hyphae "C. bantiana" and co-infection with "Toxoplasma" was confirmed by PCR as well as serology (both IgM and IgG - Toxoplasma) positivity. Stereotactic aspiration/open craniotomy and drainage is imperative to arrive at microbiological diagnosis and provide timely therapy to the patient.

First report of peritonitis caused by the vancomycin-resistant coccus Pediococcus pentosaceus in a patient on continuous ambulatory peritoneal dialysis.

INTRODUCTION: Worldwide, about one-tenth of end-stage renal disease (ESRD) patients are on peritoneal dialysis (PD). Peritonitis is a major cause of PD failure and change of therapy to haemodialysis. An update on peritoneal dialysis-related infections has recommended the use of a first generation cephalosporin or vancomycin as an empirical therapy for Gram-positive organisms. Pediococcus spp. is a Gram-positive environmental cocci that have been increasingly reported from various nosocomial infections but very rarely from peritoneal dialysis infections. It is intrinsically resistant to Vancomycin but sensitive to ampicillin. So, diagnosis of this bacteria is important if isolated from PD infections. CASE PRESENTATION: An elderly female patient of ESRD on continuous ambulatory peritoneal dialysis (CAPD) was admitted with complaints of high fever and cloudy PD effluent for 2 days. She was started with vancomycin and imipenem empirically but did not improve even after 4 days. Pus cells were seen when PD fluid was examined microscopically. BACTEC culture of PD fluid isolated growth of Gram-positive cocci, which was confirmed as Pediococcus pentosaceus . It was resistant to vancomycin. The antibiotic of the patient was changed to ciprofloxacin IV. The patient responded in 2 days and was discharged after 7 days. CONCLUSION: This is the first case report of Pediococcus pentosaceus peritonitis in an ESRD patient on CAPD. Accurate diagnosis and antibiotic sensitivity test of the bacteria is important especially if isolated in critical patients as it is intrinsically resistant to vancomycin.

Procalcitonin as a diagnostic biomarker of sepsis: A tertiary care centre experience.

INTRODUCTION: Despite the advancement in diagnostic modalities of sepsis, it is still a leading cause of morbidity and mortality. Differentiation between sepsis and non-infectious disease states remains a diagnostic challenge. Procalcitonin (PCT) is useful for the diagnosis of sepsis but it varies in cut-off ranges at different clinical settings. The aim of this study was to correlate serum PCT levels with cultures and to evaluate the best cut-off values with high sensitivity and specificity for PCT. METHODOLOGY: This prospective study included 305 patients from different medical wards; the patients were classified into group I: controls (n=46), group II: culture-negative sepsis (n=76) and group III: culture-positive sepsis (n=196). Mean p value <0.05 was considered significant. RESULTS: PCT levels were significantly higher in group II and group III as compared with group I. In group II, the best cut-off point for PCT was 1.3ng/ml with 87.30% sensitivity and 78.26% specificity (area under curve 0.86). In group III, the best cut-off value of 2.20ng/ml with 98.47% sensitivity and 89.13% specificity was found (AUC 0.96). CONCLUSION: Procalcitonin can accurately differentiate culture-negative and culture-positive sepsis from non-infectious diseases, thus making it a promising biomarker in diagnosis of bacterial sepsis.

Novel variant NDM-11 and other NDM-1 variants in multidrug-resistant Escherichia coli from South India.

OBJECTIVES: Carbapenem resistance mediated by New Delhi metallo-ß-lactamase 1 (NDM-1) and its variants has caused a major public-health concern worldwide. Here we report for the first time an Escherichia coli isolate positive for a novel variant (NDM-11). METHODS: blaNDM genes were investigated in E. coli by PCR and sequencing, and blaNDM variants were further characterised. The susceptibility pattern of novel blaNDM-11 towards different antimicrobials was compared with blaNDM-1 by cloning and expression in E. coli TOP10. RESULTS: A total of 33 carbapenem-resistant E. coli isolates were screened by PCR for the presence of blaNDM, of which 15 (45.5%) were positive. Sequencing of the PCR products revealed 10 isolates with NDM-1 and 5 isolates with NDM variants (one each of NDM-4, NDM-8 and NDM-11 and two NDM-5). Other resistance genes, including blaTEM-1, blaCTX-M-15, blaVIM, plasmid-encoded AmpC blaCMY-2 and 16S methyltransferases (rmtB and rmtC), were also associated with NDM variants in different combinations. The blaNDM variants were located on a transferable IncF-type plasmid of >100kb. Pulsed-field gel electrophoresis (PFGE) showed that all five E. coli isolates were unrelated, and multilocus sequence typing (MLST) revealed that they all belonged to ST131. Expression of the blaNDM-1 and blaNDM-11 genes in E. coli TOP10 showed no significant difference in MICs to various ß-lactams, including carbapenems. CONCLUSIONS: This study underlines the spread of NDM variants with other antimicrobial resistance genes in E. coli in South India. It also describes a novel NDM variant (blaNDM-11) having an antimicrobial resistance pattern similar to blaNDM-1.

Prevalence and Molecular Characterization of New Delhi Metallo-Beta-Lactamases in Multidrug-Resistant Pseudomonas aeruginosa and Acinetobacter baumannii from India.

New Delhi metallo-beta-lactamase (NDM)-mediated carbapenem resistance in Pseudomonas aeruginosa and Acinetobacter baumannii is a major concern. We investigated the presence of NDM and its variants in P. aeruginosa and A. baumannii at a tertiary hospital in North India. A total of 236 isolates (130 P. aeruginosa and 106 A. baumannii) were included; 38 (29.23%) P. aeruginosa and 20 A. baumannii isolates (18.8%) were resistant to carbapenems and all of them were blaNDM positive. All 38 carbapenem-resistant P. aeruginosa harbored blaNDM-1, while 12 (60%) of 20 A. baumannii harbored blaNDM-2. Pulsed-field gel electrophoresis showed that all 58 isolates were clonally unrelated. By Southern blot analysis, blaNDM-2 was located on chromosome. The blaNDM-2-positive isolates were more frequently recovered from tracheal aspirate (67% vs.16%; p = 0.02) and intensive care unit (67% vs. 20%; p = 0.001) than blaNDM-1. Among other carbapenemases, VIM was significantly associated with blaNDM-1 than blaNDM-2 (61% vs. 17%; p = 0.006). Mortality between blaNDM-1- and blaNDM-2-infected patients was comparable. When expressed in Escherichia coli, blaNDM-2 transformant conferred one doubling dilution higher MIC value for cefotaxime, piperacillin/tazobactam than blaNDM-1. The study shows the emergence of blaNDM-mediated resistance among P. aeruginosa and A. baumannii and rapid evolution of blaNDM-2 in A. baumannii with its chromosomal localization.

Proximal femoral focal deficiency associated with fibular duplication and diplopodia and complete agenesis of the tibia: a case report.

A 14-year-old boy reported with congenital deformity of the left lower limb, with gross shortening, hip and knee flexion deformities, ankle with equinus deformity, and polydactyly of the left foot. Radiologic examination showed proximal femoral focal deficiency, double fibula, and duplication of the tarsal bones, and a diagnosis of 'proximal femoral focal deficiency associated with fibular duplication and diplopodia with complete agenesis of tibia' was made. Such association of deformities is very rare and poses difficulties in rehabilitation of the case. This patient was managed with elective knee disarticulation, early prosthetic fitment, and gait training.

Microbiology of Non-Tunnelled Catheter-Related Infections.

INTRODUCTION: Aerobic bacterial infections often complicate vascular access in patients receiving haemodialysis, leading to Catheter-Related Blood Stream Infections (CRBSI). Various studies report Gram - positive bacteria, Staphylococcus aureus (S. aureus) in particular, as the most common aetiologic agent. Studies on microbiological analysis in this subset of population from India are very few. AIM: To examine clinical and bacteriological profiles of haemodialysis patients developing CRBSI, the antibiotic susceptibility of the bacteria isolated from these patients and determine nasal carriage of S. aureus in the study subjects. MATERIALS AND METHODS: Using a prospective observational design 127 patients receiving haemodialysis (84 males; 43 females) from October 2011 to March 2013 were enrolled in this study. At each dialysis session, catheters were examined for any evidence of infection. In case of suspicion for infection, pus swab, blood culture and the catheter tips were sent to microbiology laboratory for site specific investigations. Vancomycin injection was empirically administered to these patients pending culture results. Data obtained was examined for relationship of CRBSI with clinical and socio-demographic risk factors. RESULTS: Out of 127 patients, 19 developed CRBSI, 10 developed exit-site infections and 33 patients were noted to have colonization of their catheters. The most common organisms included S. aureus in 24 (45.2%) catheter tips, followed by Pseudomonas aeruginosa in 9 (17%), Acinetobacter spp. in 5 (9%), Enterobacter spp. in 4 (7.5%) and Klebsiella pneumoniae in 3 (5.6%) catheter tips. Bacteraemia was found in 19 (20.7%) patients and P. aeruginosa was the most commonly isolated organism amongst them (38.8%). Staphylococcal nasal carriage was seen in 60 (69%) patients and 36 (41.4%) of these isolates were methicillin-resistant. Significant factors associated with CRBSI included history of bacteraemia, presence of diabetes mellitus, long duration (>15 days) of catheterization and antibiotic use within three months (p<0.05 for all). CONCLUSION: Although S. aureus was the most common colonizer of non-tunnelled central access catheters among haemodialysis patients, CRBSI was most frequently caused by P. aeruginosa, which may have a bearing on our current antibiotic policy.

Lateral throat form re-classified using a customized gauge: A clinical study.

BACKGROUND: A common problem faced by prosthodontists is achieving adequate retention and stability in the mandibular dentures. Recording the lateral throat form (LTF) correctly can aid in the retention and stability. Till date, Neil's classification has been considered as the gold standard in measuring the depth of the LTF. This is a subjective classification and varies among different operators. In this study, a customized tool was used to measure the depth of the LTF, and a classification was proposed according to the measured depths. OBJECTIVES: The objective of this study is to measure the exact depth of LTF using customized gauge and to propose a classification based on the measured depth. MATERIALS AND METHODS: A customized gauge was made to measure the depth of the LTF. Two different observers classified the LTFs according to Neil's classification and according to the proposed classification in a total group of 50 patients. The customized gauge was inserted into the alveolo-lingual sulcus to measure the depth. The Pearson's correlation statistics was carried out to observe the inter-observer relationships of sulcus depth using this customized gauge. ANOVA test was used to compare the mean depth of the sulcus as measured by observers 1 and 2. RESULTS: There was more inter-observer variability when Neil's classification was used as compared to the one with the proposed classification using the gauge. The inter-observer agreement for the proposed new classification was assessed by Cohen's kappa value, with P < 0.001. The mean depth of the sulcus as calculated by observers 1 and 2 was compared with ANOVA test and found to be significant with P < 0.001. CONCLUSION: The proposed new classification for LTF gave consistent results and was easier to use with less variability when compared to the Neil's classification.

High-density linkage map construction and mapping of seed trait QTLs in chickpea (Cicer arietinum L.) using Genotyping-by-Sequencing (GBS).

This study reports the use of Genotyping-by-Sequencing (GBS) for large-scale SNP discovery and simultaneous genotyping of recombinant inbred lines (RILs) of an intra-specific mapping population of chickpea contrasting for seed traits. A total of 119,672 raw SNPs were discovered, which after stringent filtering revealed 3,977 high quality SNPs of which 39.5% were present in genic regions. Comparative analysis using physically mapped marker loci revealed a higher degree of synteny with Medicago in comparison to soybean. The SNP genotyping data was utilized to construct one of the most saturated intra-specific genetic linkage maps of chickpea having 3,363 mapped positions including 3,228 SNPs on 8 linkage groups spanning 1006.98 cM at an average inter marker distance of 0.33 cM. The map was utilized to identify 20 quantitative trait loci (QTLs) associated with seed traits accounting for phenotypic variations ranging from 9.97% to 29.71%. Analysis of the genomic sequence corresponding to five robust QTLs led to the identification of 684 putative candidate genes whose expression profiling revealed that 101 genes exhibited seed specific expression. The integrated approach utilizing the identified QTLs along with the available genome and transcriptome could serve as a platform for candidate gene identification for molecular breeding of chickpea.