SEROVAR PROFILE AND DETECTION OF INVA VIRULENCE GENE AMONG NON-TYPHOIDAL SALMONELLAE SEROVARS ISOLATED FROM ACUTE GASTROENTERITIS CASES IN COASTAL KARNATAKA, SOUTHERN INDIA

  Objective: Non-typhoidal salmonellosis is one of the leading zoonosis in the world caused by non-typhoidal Salmonella (NTS). Invasive infections with NTS serovars occurs due to the presence of virulence genes like invA along with the immunosuppressive conditions of the patient. The study was conducted to isolate and identify the NTS serovars and their antimicrobial resistance profile from patients with diarrhea and also to detect the virulence marker – invA gene among these NTS serovars.Methods: A prospective cross-sectional study was conducted from January 2015 to December 2016 at the Enteric Diseases Division, Kasturba Medical College, Manipal. 1218 fecal specimens were collected from patients with diarrhea and before antibiotic treatment. NTS serovars were identified, serotyped and then screened for the presence of invA virulence gene.Results: A total of 33 (2.7%) NTS was isolated. Salmonella typhimurium (33.34%) was predominant followed by Salmonella oslo (30.3%). Out of 33 NTS, invA was positive for 28 isolates (84.8%) of which 25 (89.3%) patients were febrile which was statistically significant (p=0.000).Conclusion: Non-typhoidal salmonellosis is an emerging global infection among immunocompromised patients. Our study showed an association between the invA gene and febrile illness among the patients suffering. Thus, this study highlights the importance of invA as a significant marker for bloodstream invasion

BERBERINE HYDROCHLORIDE COULD PROVE TO BE A PROMISING BULLET AGAINST CLOSTRIDIUM DIFFICILE INFECTION: A PRELIMINARY STUDY FROM SOUTH INDIA

Objective: Recurrent Clostridium difficile infection (CDI) and the emergence of strains with reduced susceptibility to metronidazole and vancomycinwarrants alternative therapy. Hence, we tested the potential efficacy of the natural compound berberine hydrochloride (BBRHCl) against toxigenicC. difficile.Methods: Three representative polymerase chain reaction confirmed, toxin-positive strains were included in the study. Pulsed-field gel electrophoresis(PFGE) profile and antibiogram of the strains were analyzed along with 10 other toxin positive isolates. Efficacy of BBRHCl against toxigenic C. difficilewas determined using agar diffusion by punch well method.Results: PFGE grouped the test strains into three clusters with unique susceptibility pattern toward standard antibiotics. BBRHCl was efficaciousagainst the test strains at a concentration ranging between 6.25 μg/ml and 10 mg/ml. BBRHCl's breakpoint point inhibitory zone diameter wasequivalent (p<0.001) to the epidemiological cutoff values for teicoplanin, vancomycin and 2% black seed oil. Although the predicted concentration ofBBRHCl for breakpoint zone diameter equivalent to European Committee on Antimicrobial Susceptibility Testing's epidemiological cutoff value formetronidazole was observed to fall outside the tested concentration range; it was still within the safe dosage for humans.Conclusion: The present study is promising in considering BBRHCl as a potent substitute or adjunct not only for metronidazole, vancomycin andteicoplanin but also for natural compounds like 2% black seed oil for managing resistant cases of CDI. Owing to BBRHCl's direct antibacterial and antiinflammatoryaction, further investigations will aid in the proper characterization of the therapeutic effects of similar plant compounds, to developsafe and effective drugs against the epidemiological outbreak of CDI

Non-typhoidal salmonellosis: Detection of genes responsible for virulence: A hospital-based study from Manipal, India.

Background: Non-typhoidal Salmonella (NTS) is one of the four critical worldwide reasons for diarrhoeal infections and causes a noted zoonotic infection termed non-typhoidal salmonellosis. Non-typhoidal salmonellosis generally causes self-limited gastroenteritis, whereas in immunocompromised conditions can result in invasive infections. Aim and Objectives: To detect the likely NTS serovars causing non-typhoidal Salmonellosis and their virulence genes. Material and Methods: This is a prospective cross-sectional research work carried out at the Enteric Diseases Division, Central Research Lab, Kasturba Medical College, Manipal from January 2016 - June 2018. Stool samples were collected from patients with diarrhoea admitted to the tertiary care centre in Udupi district, India. Stool samples were processed according to the World Health Organisation laboratory protocol. Non-typhoidal Salmonella isolated were subjected to PCR for the detection of virulence genes. Results: Of the 1599 diarrheal samples processed, 55 NTS were isolated with a prevalence of 3.43 %. invA gene was existent in 83.6% of the isolates, spvC gene in 61.8%, stn in 100% sopB in 96.4% and sefA in 5.45%. Conclusion: The presence of virulence genes among NTS increases the complications of non-typhoidal salmonellosis. Routine antimicrobial susceptibility tests for salmonellae should be carried out and then reported to prevent the associated morbidity and mortality

The Helicobacter pylori Genome Project : insights into H. pylori population structure from analysis of a worldwide collection of complete genomes

Helicobacter pylori, a dominant member of the gastric microbiota, shares co-evolutionary history with humans. This has led to the development of genetically distinct H. pylori subpopulations associated with the geographic origin of the host and with differential gastric disease risk. Here, we provide insights into H. pylori population structure as a part of the Helicobacter pylori Genome Project (HpGP), a multi-disciplinary initiative aimed at elucidating H. pylori pathogenesis and identifying new therapeutic targets. We collected 1011 well-characterized clinical strains from 50 countries and generated high-quality genome sequences. We analysed core genome diversity and population structure of the HpGP dataset and 255 worldwide reference genomes to outline the ancestral contribution to Eurasian, African, and American populations. We found evidence of substantial contribution of population hpNorthAsia and subpopulation hspUral in Northern European H. pylori. The genomes of H. pylori isolated from northern and southern Indigenous Americans differed in that bacteria isolated in northern Indigenous communities were more similar to North Asian H. pylori while the southern had higher relatedness to hpEastAsia. Notably, we also found a highly clonal yet geographically dispersed North American subpopulation, which is negative for the cag pathogenicity island, and present in 7% of sequenced US genomes. We expect the HpGP dataset and the corresponding strains to become a major asset for H. pylori genomics

Identification of enteroaggregative Escherichia coli in infants with acute diarrhea based on biofilm production in Manipal, south India

Background: Enteroaggregative Escherichia coli (EAEC) is an emerging enteric pathogen that causes persistent diarrhea among infants, both in developing and industrialized countries. The EAEC strains adhere to epithelial cell surface, to the glass substratum and to each other in a distinctive stacked brick-formation. Thus, gold standard for identification of EAEC remains the HEp-2 cell adherence test, which is time consuming and requires specialized facilities. Aim: To evaluate the usefulness of quantitative biofilm assay to screen for EAEC from children with acute diarrhea. Materials and Methods: A total of 100 E. coli strains were collected from acute diarrheal cases from December 2005 to November 2006. The strains were screened for biofilm production using microtiter plate method. The biofilm in the microtiter plate was visualized after staining with crystal violet and was quantified using enzyme immunosorbent assay plate reader. The Aggregative plasmid and Heat stable toxin genes were evaluated by a multiplex polymerase chain reaction. The strains were identified as EAEC with an optical density at 570 nm (OD 570 ) > 0.2. Results: Of the total 100 Escherichia coli strains, 28 were positive by Polymerase Chain Reaction for two genes, AggR and EAST. Of the 28 PCR-positive strains screened for biofilm, 25 (89.2%) showed positive results by microtiter plate method. Conclusion: The quantitative biofilm assay using microtiter plate is convenient and economical and can be used as a screening method to screen E. coli isolates from acute diarrheal cases. The best use of this test is to screen large number of isolates quickly, and if positive this can be confirmed by multiplex PCR for AggR and EAST genes. This assay may contribute to demonstrating the true incidence of EAEC with and without AggR among clinically isolated E. coli strains, which can cause acute diarrhea

Biochemical Identification of Enteroaggregative … Biochemical Identification of Enteroaggregative Escherichia Coli among Infants with Acute Diarrhea from Manipal, India

Abstract: Background : The EAEC strains have been associated classically with persistent diarrhea which represents a disproportionate share of diarrheal mortality. EAEC strains have been shown to elicit damage to intestinal mucosa and growth retardation in infants. Detection of EAEC strains can make a significant contribution to public health in many areas. The use of biofilm assays as a screening method for EAEC from clinical isolates and multiplex PCR as confirmatory method may be useful

Distribution of Candida Species in Different Clinical Samples and Their Virulence: Biofilm Formation, Proteinase and Phospholipase Production: A Study on Hospitalized Patients in Southern India

Introduction: Candida species are normal inhabitants of the skin and mucosa. The importance of epidemiological monitoring of yeasts involved in pathogenic processes is unquestionable due to the increase of these infections over the last decade; Materials and Methods: The clinical samples from the respiratory tract (sputum, bronchial wash, tracheal secretions), saliva, blood, urine, middle ear discharge, vitreous fluid, corneal ulcer, and plastic devices (endotracheal tube, catheter tip, suction tip) were collected and cultured. The species of Candida isolated were identified. Results: A total of 111 isolates of Candida species were recovered from 250 diverse clinical sources. C. albicans (39.64%) was the most isolated species, although the Candida non albicans species with 60.36% showed the major prevalence. In blood cultures, C. krusei (38.23%) and C. albicans (20.58%) were isolated frequently. C. albicans (63.27%) was the predominant species in mucosal surface. Urinary tract infections caused by yeasts were more frequent in hospitalized patients, C. krusei (50.0%) being commonly isolated, followed by C. albicans (25.0%). Discussion: Several virulence factors like, biofilm, proteinase, phospholipase, etc. contribute to the pathogenecity. Early detection of virulence factors by Candida is useful in clinical decision making. We therefore have aimed at demonstrating the formation of biofilm using the method proposed by Branchini et al, (1994). The proteinase produced by Candida was estimated as per the method of Staib et al, (1965). Phospholipase assay was carried out as per the method of Samaranayake et al, (2005). Conclusions : The data suggests that the capacity of Candida species to produce biofilm may be a reflection of the pathogenic potential of the isolates. C. krusei and C. tropicalis showed strong slime production. The non-Candida albicans produced more proteinase than C. albicans. C. albicans produced higher levels of phospholipase than non Candida albicans in this study

Probable Mode of Action of an Indigenous Drug Formulation in Garbhini Pandu wsr Iron Deficiency Anemia

Pregnancy marks a significant milestone in a woman's life, transitioning her from a woman to a mother. While it is a joyous journey, it brings about numerous physical and physiological changes that can lead to certain health challenges. Iron deficiency anemia is a prevalent health concern during pregnancy, particularly widespread in developing nations, necessitating urgent intervention to safeguard the lives of both the mother and the child. About 4 to 16% of maternal deaths are due to anemia. In contemporary science, to prevent complications like maternal and fetal mortality and morbidity, increased risk of infection, spontaneous abortion, intrauterine growth retardation, intrauterine death, premature delivery, low birth weight, postpartum hemorrhage etc during antenatal and postnatal period, various haematinic drugs are prescribed from 3rd month onwards. In Ayurveda this condition is considered as Garbhini Pandu. The Rasa and Rakta of the mother are carried to the fetus for its proper growth and development. If not, it leads to Rasa dhatu kshaya in Garbhini which finally leads to Garbhini Pandu. Ayurveda highlights the contradiction of Shodana in pregnant ladies, hence Shamana chikitsa can be followed judiciously. Considering the necessity of utilizing a safe and beneficial drug for Garbhini Pandu during pregnancy, certain indigenous drugs such as Agasthya, Amalaki, Draksha, Musta, Pippali, Sigru, Shuddha Kasisa, and Bhavana dravyas like Dadima, Bhringaraja, and Mandukaparni have been incorporated. This article highlights about the probable mode of action of indigenous drug formulation in Garbhini Pandu