Harnessing the probiotic properties and immunomodulatory effects of fermented food-derived Limosilactobacillus fermentum strains: implications for environmental enteropathy

IntroductionEnvironmental enteropathy (EE), a chronic small intestine disease characterized by gut inflammation, is widely prevalent in low-income countries and is hypothesized to be caused by continuous exposure to fecal contamination. Targeted nutritional interventions using potential probiotic strains from fermented foods can be an effective strategy to inhibit enteric pathogens and prevent chronic gut inflammation.MethodsWe isolated potential strains from fermented rice water and lemon pickle and investigated their cell surface properties, antagonistic properties, adhesion to HT-29 cells, and inhibition of pathogen adherence to HT-29 cells. Bacteriocin-like inhibitory substances (BLIS) were purified, and in vivo, survival studies in Caenorhabditis elegans infected with Salmonella enterica MW116733 were performed. We further checked the expression pattern of pro and anti-inflammatory cytokines (IL-6, IL8, and IL-10) in HT-29 cells supplemented with strains.ResultsThe strains isolated from rice water (RS) and lemon pickle (T1) were identified as Limosilactobacillus fermentum MN410703 and MN410702, respectively. Strains showed probiotic properties like tolerance to low pH (pH 3.0), bile salts up to 0.5%, simulated gastric juice at low pH, and binding to extracellular matrix molecules. Auto-aggregation of T1 was in the range of 85% and significantly co-aggregated with Klebsiella pneumoniae, S. enterica, and Escherichia coli at 48, 79, and 65%, respectively. Both strains had a higher binding affinity to gelatin and heparin compared to Bacillus clausii. Susceptibility to most aminoglycoside, cephalosporin, and macrolide classes of antibiotics was also observed. RS showed BLIS activity against K. pneumoniae, S. aureus, and S. enterica at 60, 48, and 30%, respectively, and the protective effects of BLIS from RS in the C. elegans infection model demonstrated a 70% survival rate of the worms infected with S. enterica. RS and T1 demonstrated binding efficiency to HT-29 cell lines in the 38–46% range, and both strains inhibited the adhesion of E. coli MDR and S. enterica. Upregulation of IL-6 and IL-10 and the downregulation of IL-8 were observed when HT-29 cells were treated with RS, indicating the immunomodulatory effects of the strain.DiscussionThe potential strains identified could effectively inhibit enteric pathogens and prevent environmental enteropathy

Increasing Trends of Leptospirosis in Northern India: A Clinico-Epidemiological Study

Leptospirosis is often not suspected by physicians in patients with acute febrile illnesses reporting from supposedly “non-endemic areas,” including north India. Clinical manifestations are protean, and complications can affect most organ systems, including liver, kidneys, lungs, and the central nervous system. Timely diagnosis and specific therapy can reduce severity of illness and, in turn, mortality. In this study conducted at a tertiary care center in north India, we find how a much-neglected disease entity has emerged as a major cause of acute febrile illness in a so called “non-endemic area.” Incidence is increasing yearly. The majority of patients were from a rural background, and were farmers or farm labourers. Poor hygiene, contact with animals, rat infestation of houses, and contact with stagnant dirty water are the major determinants of disease. Apart from the usual symptoms of intermittent fever with chill and rigor, hepatosplenomegaly, renal decompensation, muscle pain and tenderness, and conjunctival suffusion, signs and symptoms indicating involvement of the respiratory and central nervous systems were also commonly observed. Severe complications resulting in mortality do occur and is especially due to late suspicion among primary level physicians, and the resulting inappropriate therapy

Evaluation of polymerase chain reaction using protein b primers for rapid diagnosis of tuberculous meningitis

Background: Rapid and specific diagnosis of tubercular meningitis (TBM) is of utmost importance. Aim: To evaluate polymerase chain reaction (PCR) using protein b primers directed against M. tuberculosis for the diagnosis of tuberculous meningitis. Materials and Methods: PCR using protein b primers was performed in ten patients with confirmed TBM (culture positive), 60 patients with clinically suspected TBM and 40 patients with no TBM (control group). Results: Protein b PCR had a sensitivity of 90% and a specificity of 100% in patients with confirmed TBM. In 60 clinically diagnosed TBM patients, protein b PCR was positive in 49 (81.7%) patients. The overall sensitivity of microscopy, culture and PCR using protein b primers was 1.4%, 14.3%, and 82.8% and specificity was 100%, 100%, and 100% respectively. Conclusion: Protein b PCR is valuable in rapid diagnosis of TBM

Evaluation of PCR using MPB64 Primers for Rapid Diagnosis of Tuberculous Meningitis

TBM control group and its comparison with conventional techniques like microscopy and culture. Materials and Methods A total of 130 CSF samples received for AFB smear and culture in laboratory of tertiary care hospital of India, between September 2008 and December 2009 were evaluated. Patient's age ranged from 12-90 years. The relevant history and other details of the patients were noted from the case records. The patients were divided into 3 groups: Group I: TBM (n=90): (a) confirmed TBM-culture/smear positive (n=9), and (b) suspected TBM: smear/culture negative, clinical and laboratory features suggestive of TBM and response to anti-tuberculosis therapy Processing of CSF sample All the 130 CSF samples were subjected to three microbiological tests: Ziehl-Neelsen staining (ZN), culture on Lowenstein-Jensen medium and PCR with MPB-64. The CSF samples of the subjects were Keywords: PCR; Tubercular meningitis Introduction Tuberculosis is a major cause of morbidity and mortality worldwide. The World Health Organization has noted that the global incidence of TB is increasing by 0.4% per annum The development of rapid, sensitive and specific test for detection of mycobacterium has been a long standing need. A number of mycobacterial antigen Nucleic Acid Amplification Techniques (NAAT) such as Polymerase Chain Reaction (PCR) has been reported to be more sensitive and specific. Several Mycobacterium tuberculosis specific sequences like IS6110, Protein antigen b [8], MPB64 and 65 kDa have been evaluated Abstract Purpose: Diagnosis of Tuberculosis (TB) is largely based on microscopy and culture, which either lack of sensitivity or time consuming. In the present study a PCR test based on DNA sequence coding for MPB64, specific for Mycobacterium tuberculosis was compared with Ziehl-Neelson (ZN) stained AFB smear examination, culture based on conventional LJ medium for diagnosis of tuberculosis using clinical samples obtained from Cerebro-Spinal Fluid (CSF) samples from TBM patients. Methods: PCR using MPB64 primers was performed on 9 TBM confirmed (culture was positive), 81TBM suspected and 40 Non TBM (control group) patients. Results: MPB64 PCR had sensitivity of 88.8% and specificity of 100% for confirmed TBM cases, where as in 81 clinically diagnosed but unconfirmed TBM cases MPB64 PCR was positive in 81.48% cases respectively. The overall sensitivity of microscopy, culture and PCR using MPB64 were 1.11%, 10%, 82.22% and specificity was 100%, 100% and 100% respectively