Dasharath Shinde
@ssbs.edu.in
PhD student and Teaching Assistant
Symbiosis School of Biological Sciences
RESEARCH INTERESTS
Lactic Acid Bcateria, Probiotics, Food-Biotechnology.
Scopus Publications
Scholar Citations
Scholar h-index
Scholar i10-index
Scopus Publications
Sarvesh V Surve, Dasharath B Shinde, Joyleen M Fernandes, Sharoni Sharma, Monty Vijayvargiya, Komal Kadam, and Ram Kulkarni
Oxford University Press (OUP)
Abstract Aims Laboratory domestication has been negligibly examined in lactic acid bacteria (LAB). Lactiplantibacillus plantarum is a highly studied and industrially relevant LAB. Here, we passaged L. plantarum JGR2 in a complex medium to study the effects of domestication on the phenotypic properties and the acquisition of mutations. Methods and results Lactiplantibacillus plantarum JGR2 was passaged in mMRS medium (deMan Rogossa Sharpe supplemented with 0.05% w/v L-cysteine) in three parallel populations for 70 days. One pure culture from each population was studied for various phenotypic properties and genomic alterations. Auto-aggregation of the evolved strains was significantly reduced, and lactic acid production and ethanol tolerance were increased. Other probiotic properties and antibiotic sensitivity were not altered. Conserved synonymous and non-synonymous mutations were observed in mobile element proteins (transposases), β-galactosidase, and phosphoketolases in all three isolates. The evolved strains lost all the repeat regions and some of the functions associated with them. Most of the conserved mutations were found in the genomes of other wild-type strains available in a public database, indicating the non-novel genomic impact of laboratory passaging. Conclusions Laboratory domestication can affect the phenotypic and genotypic traits of L. plantarum and similar studies are necessary for other important species of LAB.
Dasharath B. Shinde, Jayashri G. Mahore, Prabhanjan S. Giram, Shaktikumar L. Singh, Aditi Sharda, Divya Choyan, and Shubham Musale
Springer Science and Business Media LLC
Dasharath B. Shinde and Ram Kulkarni
Elsevier BV
Sarvesh Surve, Dasharath B. Shinde, and Ram Kulkarni
Springer Science and Business Media LLC
AbstractLactiplantibacillus plantarum is one of the most diverse species of lactic acid bacteria found in various habitats. The aim of this work was to perform preliminary phenotypic and genomic characterization of two novel and potentially probiotic L. plantarum strains isolated from Indian foods, viz., dhokla batter and jaggery. Both the strains were bile and acid tolerant, utilized various sugars, adhered to intestinal epithelial cells, produced exopolysaccharides and folate, were susceptible for tetracycline, erythromycin, and chloramphenicol, did not cause hemolysis, and exhibited antimicrobial and plant phenolics metabolizing activities. The genetic determinants of bile tolerance, cell-adhesion, bacteriocins production, riboflavin and folate biosynthesis, plant polyphenols utilization, and exopolysaccharide production were found in both the strains. One of the strains contained a large number of unique genes while the other had a simultaneous presence of glucansucrase and fructansucrase genes which is a rare trait in L. plantarum. Comparative genome analysis of 149 L. plantarum strains highlighted high variation in the cell-adhesion and sugar metabolism genes while the genomic regions for some other properties were relatively conserved. This work highlights the unique properties of our strains along with the probiotic and technically important genomic features of a large number of L. plantarum strains.
Dasharath B. Shinde, Santosh S. Koratkar, Vinay Rale, Shashikala NM, and Neetu Mishra
MDPI AG
(1) Background: Iron deficiency anemia is a significant nutritional problem all over the world. Salt formulations supplemented with encapsulated iron and iodine (double-fortified) were tested for their efficacy in managing iron deficiency anemia. In this study, we have checked the effect of these double-fortified salt formulations (iron and iodine) on hemoglobin (Hb) levels in anemic Wistar male rats. (2) Methods: The study was divided into two phases, viz., the development of anemia in the first phase and then the random division of anemic rats into five groups (Groups A to E). These rats were fed with three different salt formulations (Groups A to C); Group D was continued on a low iron diet, and Group E was on a normal pellet diet over a period of 84 days. The level of Hb was tested in each group. (3) Results: The rats in Groups A, B, C, and E recovered from anemia significantly, with higher Hb levels. On day 84, however, the Hb level in Group D continued to decrease. The bodyweight of the rats was not affected in any way. In all of the groups, histopathology examinations in various organs revealed no significant changes. (4) Conclusions: All of the three different salt formulations showed significant recovery in the anemic rats as compared to the rats fed with a normal pelleted diet.
Dasharath B. Shinde, Ranjitsinh Pawar, Jyotsna Vitore, Deepak Kulkarni, Shubham Musale, and Prabhanjan Giram
Wiley
Dasharath B. Shinde, Surbhi Singhvi, Santosh S. Koratkar, and Sunil D. Saroj
Veterinary World
Background and Aim: Cattle are the main reservoir of Escherichia coli O157:H7 and other verotoxigenic E. coli (VTEC); therefore, there is an increased risk of infection to humans by either direct or indirect mode of transmissions. However, the prevalence of E. coli O157:H7 in the healthy cattle population of India is yet to be ascertained. This study aimed to screen the dairy cattle in and around Pune, Maharashtra, India, for verotoxin-producing E. coli O157:H7. Materials and Methods: A total of 257 rectal swabs were collected from 15 different organized and unorganized dairy farms of Pune during the period, January-March 2015. The screening involved enrichment in EC broth followed by differential identification on MacConkey sorbitol agar. The presumptive positive isolates were further confirmed by multiplex polymerase chain reaction (PCR) using primers specific to rfbE (O157), fliC (H7), VT1 (MK1), and VT2 (MK2). Vero-toxicity and antibiotic sensitivity were examined in PCR confirmed isolates. Results: Out of the 257 samples analyzed, 1.9% (2/105) were positive for O157:H7 and 39% (41/105) were positive for VTEC. Two PCR confirmed positive O157:H7 strains and two randomly selected PCR-positive VT strains exhibited in vitro cytopathic effect on Vero cells on day-7 post-inoculation. Antibiotic sensitivity profiling of O157:H7 strains exhibited resistance against penicillin G, kanamycin, ampicillin, tetracycline, gentamycin, cefotaxime, streptomycin, and piperacillin. Conclusion: These findings reveal the presence of pathogenic E. coli O157:H7 in the healthy cattle of Pune; in a situation, wherein regular surveillance for O157:H7 is not a norm. Therefore, the findings presented herein warrant routine surveillance and public awareness to prevent the transfer of such pathogens and manage health risks to the public.
Dasharath B. Shinde, Santosh S. Koratkar, Neeti Sharma, and Ajinkya A. Shitole
Innovare Academic Sciences Pvt Ltd
<p><strong>Objective: </strong>To evaluate the <em>in vitro </em>antioxidant activity of liquorice (<em>Glycyrrhiza glabra) </em>against H<sub>2</sub>O<sub>2</sub> induced oxidative stress in HepG2 cell line.</p><p><strong>Methods: </strong>Antioxidant activity of methanolic extracts of <em>Glycyrrhiza glabra</em> was investigated by measuring total phenolic content using folin-ciocalteu reagent (FCR), free radical scavenging activity by DPPH and ferric reducing antioxidant power (FRAP). The presence of phenolic compounds and flavonoids in the extract was confirmed by Liquid Chromatography-Mass Spectrometry (LC-MS) analysis. Furthermore, the protective effect of methanolic extract of <em>Glycyrrhiza glabra</em> against oxidative stress induced by H<sub>2</sub>O<sub>2 </sub>in HepG2 cells was investigated by MTT assay. HepG2 cells were exposed with five different treatments viz. liquorice, H<sub>2</sub>O<sub>2</sub>, ascorbic acid, H<sub>2</sub>O<sub>2</sub>+liquorice and H<sub>2</sub>O<sub>2</sub>+ascorbic acid, to explore the effect of the extract on malondialdehyde (MDA) production, catalase activity, and glutathione reductase levels.<strong></strong></p><p><strong>Results: </strong>The total phenolic content estimated in <em>Glycyrrhiza glabra </em>extract was found to be 241.47 µg per 1000 µg/ml of methanolic extract. It was found that as the concentration of the extract was increased both the free radical scavenging activity and ferric ion reducing power was also found to increase. LC-MS analysis confirmed the presence of eight different phenolic compounds in the methanolic extract which are possibly contributing to the antioxidant activity exhibited by the extract. It was also observed that liquorice treated HepG2 cells showed lower MDA and higher glutathione and catalase levels as compared to only H<sub>2</sub>O<sub>2 </sub>treated HepG2 cells where increased MDA production, decreased glutathione reductase and catalase production was observed.</p><p><strong>Conclusion: </strong>Our results thus conclude that, the methanolic extract of <em>Glycyrrhiza glabra </em>can be used as natural supplements in various disease conditions where oxidative stress has been reported. <strong></strong></p><p> </p>