Sachin N. Hajare

Effectiveness of Radiation Processing in Elimination of Salmonella Typhimurium and Listeria monocytogenes from Sprouts

The effectiveness of radiation treatment in eliminating Salmonella Typhimurium and Listeria monocytogenes on laboratory inoculated ready-to-eat sprouts was studied. Decimal reduction doses (D10-values) for Salmonella Typhimurium and L. monocytogenes in dry seeds of mung (green gram), matki (dew gram), chana (chick pea), and vatana (garden pea) ranged from 0.189 to 0.303 kGy and 0.294 to 0.344 kGy, respectively. In sprouts made from these seeds, the D10-values ranged from 0.192 to 0.208 kGy for Salmonella Typhimurium and from 0.526 to 0.588 kGy for L. monocytogenes. Radiation treatment with a 2-kGy dose resulted in complete elimination of 10(4) CFU/g of Salmonella Typhimurium and 10(3) CFU/g of L. monocytogenes from all the four varieties of sprouts. No recovery of Salmonella Typhimurium and L. monocytogenes was observed in the radiation treated samples stored at 4 and 8 degrees C up to 12 days. Radiation treatment with 1 kGy and 2 kGy resulted in a reduction of aerobic plate counts and coliform counts by 2 and 4 log CFU/g, respectively; the yeast and mold counts and staphylococci counts decreased by 1 and 2 log CFU/g, respectively. However, during postirradiation storage at 4 and 8 degrees C, aerobic plate counts, coliform counts, yeast and mold counts, and staphylococci counts remained constant throughout the incubation period. This study demonstrates that a 2-kGy dose of irradiation could be an effective method of processing to ensure microbial safety of sprouts.

Radiation Processing To Ensure Safety of Minimally Processed Carrot (Daucus carota) and Cucumber (Cucumis sativus): Optimization of Dose for the Elimination of Salmonella Typhimurium and Listeria monocytogenes

Minimally processed vegetables are in demand, because they offer convenience to consumers. However, these products are often unsafe because of possible contamination with pathogens, such as Salmonella, Escherichia coli O157:H7, and Shigella species. Therefore, this study was carried out to optimize the radiation dose necessary to ensure the safety of precut carrot and cucumber. Decimal reduction doses (D-values) of Salmonella Typhimurium MTCC 98 were ca. 0.164 kGy in carrot samples and 0.178 kGy in cucumber samples. D-values of Listeria monocytogenes were determined to be 0.312 and 0.345 kGy in carrot and cucumber samples, respectively. Studies of inoculated, packaged, minimally processed carrot and cucumber samples showed that treatment with a 1-kGy dose of gamma radiation eliminated up to 4 log CFU/g of Salmonella Typhimurium and 3 log CFU/g of L. monocytogenes. However, treatment with a 2-kGy dose was necessary to eliminate these pathogens by 5 log CFU/g. Storage studies showed that both Salmonella Typhimurium and L. monocytogenes were able to grow at 10 degrees C in inoculated control samples. Neither of these pathogens could be recovered from radiation-processed samples after storage for up to 8 days.

Microbiological evaluation of sprouts marketed in mumbai, india, and its suburbs

A study was undertaken to assess the microbiological quality of sprouts marketed in Mumbai and its suburbs. A total of 124 sprout samples of four different legumes--mung (Phaseolus aureus), matki (Phaseolus aconitifolius), chana (Cicer arietinum), and vatana (Pisum sativum)--were analyzed over a period of 12 months for aerobic plate counts, coliforms, yeast and mold counts, staphylococci, Salmonella, Listeria monocytogenes, Escherichia coli, E. coli O157:H7, and coagulase-positive Staphylococcus aureus. Aerobic plate counts ranged from 7.6 to 8.9 log CFU/g, coliform counts ranged from 5.4 to 7.9 log CFU/g, yeast and mold counts ranged from 3.6 to 7.3 log CFU/g, and staphylococci counts ranged from 3.3 to 6.6 log CFU/ g. Nonpathogenic E. coli was detected in 13% of the mung, 26% of the matki, 40% of the chana, and 19% of the vatana samples. Salmonella Typhimurium was detected in 21% of the mung, 40% of the matki, and 4% of the chana samples. Salmonella Dublin was detected in 2% of the mung samples, and Salmonella Washington was detected in 4% of the matki samples. L. monocytogenes and E. coli O157:H7 were not detected in any of the samples examined. Coagulase-positive S. aureus was detected in 4% of the mung, 11% of the matki, and 4% of the chana samples. The results indicated that the marketed sprouts were of poor microbiological quality; therefore, further processing, such as radiation processing, is needed to ensure their safety.

Radiation processing for elimination of Salmonella typhimurium from inoculated seeds used for sprout making in India and effect of irradiation on germination of seeds.

The effect of radiation processing on the germination of the sprout seeds mung (Phaseolus aureus), matki (Phaseolus aconitifolius), chana (Cicer arietinum), and vatana (Pisum sativum) in terms of percent germination, germination yield, sprout length, vitamin C content, and texture was investigated. Gradual decreases in the percent germination, germination yield, and sprout length with increases in radiation dose (0.5 to 2.0 kGy) were observed. Vitamin C content and texture remained unaffected for the seeds treated with doses of up to 2 kGy. To determine the efficacy of radiation treatment in elimination of foodborne pathogens, seeds inoculated with 4 log CFU/g of Salmonella Typhimurium were treated with radiation doses of 1 and 2 kGy. A reduction in counts of Salmonella Typhimurium in inoculated seeds after radiation treatment was observed. A radiation dose of 2 kGy resulted in the complete elimination of 4 log CFU/g of Salmonella Typhimurium from the inoculated seeds. However, on sprouting for 48 h, the count of Salmonella Typhimurium reached 8 log CFU/g for the control seeds and the seeds treated with a 1-kGy radiation dose. The aerobic plate counts for seeds were 2.0 to 2.6 log CFU/g, which were reduced to 0.9 to 1.2 log CFU/g on treatment with a 2-kGy radiation dose. On sprouting for 48 h, the aerobic plate count reached 8 log CFU/g for both the control and radiation-treated seeds. The study demonstrates that irradiation can control bacterial levels on seeds but not contamination introduced during posttreatment handling. Therefore, radiation processing of the final product (sprouts) is recommended, rather than of the seeds.

Induction of apoptosis in human cancer cells by a Bacillus lipopeptide bacillomycin D

A newly isolated and characterized Bacillus amyloliquefaciens strain fiply 3A has been found to produce an extracellular cyclic lipopeptide which structurally resembled bacillomycin D, earlier reported to be produced by Bacillus subtilis. The lipopeptide showed a dose dependent killing of three different human cancer cell lines viz. A549 (alveolar adenocarcinoma), A498 (renal carcinoma) and HCT-15 (colon adenocarcinoma), while not affecting the normal cell line L-132 (pulmonary epithelial cells) when analyzed using MTT assay and FACS analysis. Staining the cells with H2-DCFDA showed an increase in reactive oxygen species (ROS) formation in the lipopeptide treated cell population. Hoechst 33342 staining of nuclei further indicated apoptosis as a major mechanism of cell death in lipopeptide treated cells and the typical symptoms of apoptosis including cell shrinkage, nuclear condensation and fragmentation of nuclei were observed. Lipopeptide treatment induced extensive DNA damage in the treated cells, which was indicated by a TUNEL assay. Flow cytometric analysis exhibited lipopeptide concentration dependent apoptosis which was further confirmed during clonogenic assay of the lipopeptide treated cells.

Formulation of a nasogastric liquid feed and shelf-life extension using gamma radiation.

Nasogastric liquid feed formulation (NGLF) was developed for immunocompromised patients who are vulnerable targets of pathogenic assault. NGLF consisted of cereals, pulses, vegetables, and milk powder to provide balanced nutrients; however, the shelf life was only a few hours because this product was highly prone to microbial contamination and proliferation due to its high water content and rich nutrients. Postpreparation storage and distribution was very difficult, even at chilled temperatures. To overcome this problem, the NGLF was irradiated at various doses (2.5 to 10 kGy). Gamma irradiation at 10 kGy reduced the microbial load to nondetectable levels, and the product could be stored up to 1 month without any detectable increase in microbial load. The sensory evaluation did not indicate differences between the nonirradiated fresh, irradiated fresh, and stored samples. Nutritional quality in terms of total carbohydrates, dietary fiber, proteins, calories, vitamins A and C, and the micronutrients calcium, iron, and zinc was not affected by irradiation. NGLF also possessed antimutagenic potential against ethylmethanesulphonate-induced mutagenesis in Escherichia coli cells as evaluated by the rifampin resistance assay. This property of NGLF remained unchanged even after exposure to a 10-kGy dose of gamma radiation. Thus, irradiated NGLF seemed to be a safe and wholesome food for immunocompromised patients.

Isolation and identification of antibacterial compound from Indo- Himalayan Aconitum nagarum

Abstract Objective To isolate and identify the antibacterial compound from Aconitum nagarum Stapf., a little known medicinal plant in Manipur (Indo–Himalaya). Methods The enriched alkaloid extract from Aconitum nagarum root was fractionated through silica gel column chromatography and analysed for antibacterial activity against bacterial species including human pathogens, Staphylococcus aureus, Salmonella typhimurium, Escherichia coli and Bacillus subtilis . The most potent compound (F 3 C 2 ) isolated from this fraction was purified by chromatography and identified by spectral (IR and NMR) analysis. Results Among different fractions (F1-F6), ether fraction (F3) was found to have maximum antibacterial activity. Based on activity guided fractionation, the compound was identified as a diterpenoid alkaloid aconitine [(1α,3α,6α,14α,16β)–8–(acetyloxy)–20–ethyl–3,13,15–trihydroxy–1,6,16–trimethoxy–4–(methoxymethyl)aconitan–14–yl benzoate]. Conclusion The antibacterial compound showed activities against different human pathogens and has been identified as aconitine.

MYCOTOXINS | Immunological Techniques for Detection and Analysis

Mycotoxins are toxic secondary metabolites produced by certain fungi in agricultural commodities. Mycotoxins can induce both acute and chronic effects on human and animal health, and hence it is important to control mycotoxin contamination in foods and feeds. Among all the analytical techniques available for mycotoxin detection, immunoassays offer higher sensitivity and specificity. These antigen–antibody–based assays include enzyme-linked immunosorbent assays, immunoradiometric assay, and radioimmunoassay. Recently, newer techniques are introduced that include enzyme-linked immuno-magnetic electrochemical array, lateral flow devices, and surface plasmon resonance immunobiosensors. Optical-based immunological techniques involve fiber-optic immunosensors and fluorescent polarization immunoassays. Most of these assays are rapid and can perform real-time analysis of mycotoxins.

Lack of induced mutagenesis in E. coli or human lymphoblast cell line upon long-term sub-culturing in medium from irradiated meat

Abstract Purpose: Current study was aimed to enhance the confidence of consumers as well as entrepreneurs towards food irradiation program. Materials and methods: In this work, safety of high dose (25 kGy) irradiated meat samples (HDIMS) was ascertained by scoring mutation frequency through a long-term sub-culturing study in Escherichia coli MG1655 cells (ATCC 700926) up to 1500 generations (at 1%), 250 generations (at 5% and 10%) and human lymphoblast thymidine kinase heterozygote (TK6) cell line (ATCC CRL-8015) [at two gene loci, tk−/+ (thymidine kinase) and hprt+ (Hypoxanthine Phosphoribosyltransferase)] up to 156 generations using goat meat sample. Also these samples were assayed at further radiation doses of 10, 45 and 70 kGy at 2% concentration (in cell line), and 1% (in E. coli). Study was also performed with other meat samples such as chicken, fishes (pomfret and rohu) and shrimps by carrying out limited long-term sub-culturing trials in human lymphoblast cell line. Mutation analysis was also carried out using a novel DPAR (Differential loss of Plasmid Antibiotic Resistance) assay followed by sequencing of tcR (tetracycline resistance) gene of pBR322 plasmid isolated from E. coli cells grown for 1500 generations on HDIMS medium and RAPD (Random Amplified Polymorphic DNA) analysis of the genome. Results and conclusion: None of the assays exhibited any induced mutation when analyzed at regular time intervals. RAPD analysis also did not indicate any change in its nucleotide sequence, ruling out the occurrence of any silent mutation. Thus, the present findings report absence of mutagenic effect of high dose irradiated meat samples.