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There is increasing interest in gluten degrading enzymes for use during food and drink processing. The industrially available enzymes usually work best at low to ambient temperatures. However, food manufacturing is often conducted at higher temperatures. Therefore, thermostable gluten degrading enzymes are of great interest. We have identified a new thermostable gluten degrading proline specific prolyl endoprotease from the archaea Thermococcus kodakarensis. We then cloned and expressed it in Escherichia coli. The prolyl endoprotease was found to have a size of 70.1 kDa. The synthetic dipeptide Z-Gly-Pro-p-nitroanilide was used to characterise the prolyl endoprotease and it had maximum activity at pH 7 and 77°C. The VmaxKm and kcat values of the purified prolyl endoprotease were calculated to be 3.14 mM/s, 1.10 mM and 54 s–1 respectively. When the immunogenic gluten peptides PQPQLPYPQPQLPY (alpha-gliadin) and SQQQFPQPQQPFPQQP (gamma-hordein) were used as substrates, the prolyl endoprotease was able to degrade these. Furthermore, gluten in wort was reduced when the prolyl endoprotease was used during mashing of barley malt. The discoveries open up for new food processing possibilities and further the understanding of proline specific protease diversity.

Plant growth promoting rhizobacteria and endophytic bacteria were isolated from different varieties of turmeric (Curcuma longa L.) from South India. Totally 50 strains representing, 30 PGPR and 20 endophytic bacteria were identified based on biochemical assays and 16S rDNA sequence analysis. The isolates were screened for antagonistic activity against Pythium aphanidermatum (Edson) Fitzp., and Rhizoctonia solani Kuhn., causing rhizome rot and leaf blight diseases in turmeric, by dual culture and liquid culture assays. Results : revealed that only five isolates of PGPR and four endophytic bacteria showed more than 70% suppression of test pathogens in both assays. The SEM studies of interaction zone showed significant ultrastructural changes of the hyphae like shriveling, breakage and desication of the pathogens by PGPR B. cereus (RBac-DOB-S24) and endophyte P. aeruginosa (BacDOB-E19). Selected isolates showed multiple Plant growth promoting traits. The rhizome bacterization followed by soil application of B. cereus (RBacDOB-S24) showed lowest Percent Disease Incidence (PDI) of rhizome rot and leaf blight, 16.4% and 15.5% respectively. Similarly, P. aeruginosa (BacDOB-E19) recorded PDI of rhizome rot (17.5%) and leaf blight (17.7%). The treatment of these promising isolates exhibited significant increase in plant height and fresh rhizome yield/plant in comparison with untreated control under greenhouse condition. Thereby, these isolates can be exploited as a potential biocontrol agent for suppressing rhizome rot and leaf blight diseases in turmeric.

Globally disease outbreaks as a result of the consumption of contaminated food and feedstuffs are a regular primary problem. The foremost elements contributing to contamination are microorganisms, particularly fungi, which produce low-molecular weight secondary metabolites, with demonstrated toxic properties that are referred to as mycotoxins. Aflatoxins contaminate agricultural commodities and may cause sickness or fatality in humans and animals. Moreover, poor conditions of storage and a deficiency in regulatory measures in food quality control aggravate the main issue. For that reason, mycotoxin-related illness of nutrition represents a major health hazard for local populations. Government policies should make regulations aiming to avoid the entry of aflatoxins into food stuffs. For consumer safety, control and management strategies should be developed and implemented by regulatory authorities. There is the need for attention from farmers, scientists, government and collaborative minds throughout the country to ensure aflatoxin-free food. The present review is informative not only for health-conscious consumers, but also for relevant authorities with respect to paving the way for future research aiming to fill the existing gaps in our knowledge with regard to mycotoxins and food security. © 2016 Society of Chemical Industry.

Endophytic fungi have been isolated from the healthy turmeric (Curcuma longa L.) rhizomes from South India. Thirty-one endophytes were identified based on morphological and ITS–rDNA sequence analysis. The isolated endophytes were screened for antagonistic activity against Pythium aphanidermatum (Edson) Fitzp., and Rhizoctonia solani Kuhn., causing rhizome rot and leaf blight diseases in turmeric respectively. Results revealed that only six endophytes showed > 70% suppression of test pathogens in antagonistic dual culture assays. The endophyte T. harzianum TharDOB-31 showed significant in vitro mycelial growth inhibition of P. aphanidermatum (76.0%) and R. solani (76.9%) when tested by dual culture method. The SEM studies of interaction zone showed morphological abnormalities like parasitism, shriveling, breakage and lysis of hyphae of the pathogens by endophyte TharDOB-31. Selected endophytic isolates recorded multiple plant growth promoting traits in in vitro studies. The rhizome bacterization followed by soil application of endophyte TharDOB-31 showed lowest Percent Disease Incidence of rhizome rot and leaf blight, 13.8 and 11.6% respectively. The treatment of TharDOB-31 exhibited significant increase in plant height (85 cm) and fresh rhizome yield/plant (425 g) in comparison with untreated control under greenhouse condition. The confocal microscopy validates the colonization of the TharDOB-31 in turmeric rhizomes. The secondary metabolites in ethyl acetate extract of TharDOB-31 were found to contain higher number of antifungal compounds by high resolution liquid chromatograph mass spectrometer analysis. Thereby, endophyte T. harzianum isolate can be exploited as a potential biocontrol agent for suppressing rhizome rot and leaf blight diseases in turmeric.

Background: The protein/isozyme fingerprint is useful in differentiating the species and acts as a biochemical marker for identification and systematic studies of medicinal plant species. Objective: In the present study, protein and isozyme profiles for peroxidase, esterase, acid phosphatase, polyphenol oxidase, alcohol dehydrogenase, and alkaline phosphatase of five species of Memecylon (Melastomataceae), Memecylon umbellatum, Memecylon edule, Memecylon talbotianum, Memecylon malabaricum, and Memecylon wightii were investigated. Materials and Methods: Fresh leaves were used to prepare crude enzyme extract for analyzing the five enzymes isozyme variations. Separation of isozymes was carried out using polyacrylamide gel electrophoresis (PAGE) and the banding patterns of protein were scored. Pair-wise comparisons of genotypes, based on the presence or absence of unique and shared polymorphic products, were used to regenerate similarity coefficients. The similarity coefficients were then used to construct dendrograms, using the unweighted pair group method with arithmetic averages. Results: A total of 50 bands with various Rf values and molecular weight were obtained through PAGE analysis. Among the five Memecylon species, more number of bands was produced in M. wightii and less number of bands was observed in M. edule. The results of similarity indices grouped M. malabaricum and M. wightii in one cluster with 98% similarity and M. umbellatum, M. edule, and M. talbotianum are grouped in another cluster with 79% similarity showing close genetic similarities which is in accordance with the morphological identification of Memecylon species. Conclusion: The protein/isozyme fingerprint is useful in differentiating the species and acts as a biochemical marker for identification of Memecylon species. SUMMARY Biochemical characterization of Memecylon species was evaluated by SDS-PAGE of extracted protein and isozyme profiling on native PAGE.After electrophoresis, each gel was stained with specific stains. Genetic distance relationships were evaluated based on the banding patterns of protein on isozymes.Unique banding pattern of esterase, peroxidase, acid phosphatase, alcohol dehydrogenase and polyphenol oxidase are observed in all the five species of Memecylon, which represent the fingerprint of Memecylon species.SDS-PAGE and isozyme profiling of five Memecylon species revealed that M. malabaricum and M. wightii grouped in one cluster and M. umbellatum, M. edule and M. talbotianum grouped in another cluster showing close genetic similarities which is in accordance with the morphological identification of Memecylon species.This is the first report on the comparison of protein and isozyme profile of five different Memecylon species. Abbreviations Used: SDS-PAGE: Sodium docecyl sulfate polyacrylamide gel electrophoresis; NTSYS PC2: Numerical taxonomy system, version 2.2 for Windows XP, Vista, Win7, Win 8 and Win10 including 64 bit

The genus Memecylon L. (Melastomataceae), an important source of traditional medicine is mainly distributed in the Western Ghats region of Karnataka. In Indian ethnomedical practices, this plant is reported to be effective for skin and viral diseases including herpes and chickenpox. The objective of the present study was to screen Memecylon species namely M. umbellatum, M. edule, M. talbotianum, M. malabaricum and M. wightii for the bioactive compounds with special reference to skin diseases. Leaf samples were dried and extracted using methanol. TLC analysis of methanol extracts of five Memecylon species showed single band with RF value 0.61 detected at 260 nm. This band was eluted and subjected to high performance liquid chromatography. Profiles of Memecylon species confirmed the presence of α-tocopherol on comparison with the standard. Among five Memecylon species, M. malabaricum (0.724%) and M. talbotianum (0.362%) showed highest α-tocopherol concentration. For the first time α-tocopherol was reported from Memecylon species. Further, the structure of α-tocopherol was confirmed by liquid chromatography-mass spectrometry and nuclear magnetic resonance spectroscopy analysis. Presence of α-tocopherol (vitamin E) in Memecylon species validates the traditional use of Memecylon species in the treatment of skin diseases.

Seed Mycoflora of Brinjal (Solanum melongena L)

Growth Stages in Pearl Millet [Pennisetum typhoides (Stapf & Hubb)] and Susceptibility to Infection of Downy Mildew Pathogen (Sclerospora graminicola (Sacc) Schroet)

Histology of Carpel Infection by Claviceps fusiformis in Pearl Millet

Background: Appendicitis is one of the commonest causes of abdominal pain requiring emergency surgery. Diagnosing acute appendicitis clinically still remains a common surgical problem as the clinical signs and symptoms of other abdominal pathologic conditions mimic the diagnosis of acute appendicitis. Delay in diagnosis and surgery for this condition may lead to various complications like perforation, abdominal abscess etc. By knowing perforation prior to the exploration, we can manage the condition very effectively in terms explaining the prognosis of disease, morbidity of surgery, wound infection, and requirement of emergent nature of surgery. Aim of the study is to determine of role of hyperbilirubinemia as a new diagnostic marker to predict gangrenous/perforated appendicitis.Methods: It’s a retrospective study was conducted in the department of surgery, MMCRI, Mysore during the period of January 2015 to December 2015 total of 100 patients with clinical as well as ultrasonographic diagnosis of acute appendicitis or appendiceal perforation were made. The serum bilirubin test was carried out in all the patients pre-operatively.Results: In the study, the total 100 patients enrolled for the study, hyperbilirubinemia (> 1.2 mg/dL) in present study was found in 47 patients with 19 acute and 28 patients perforated appendicitis of all the 100 patients while 53 patients had normal bilirubin levels (≤ 1.2 mg/dL) 51 patients acute and 2 patients perforated appendix.Conclusions: Total serum bilirubin appears to be a new promising laboratory marker for diagnosing appendicular perforation. The patients with clinical signs and symptoms of appendicitis and with hyperbilirubinemia without elevation of liver enzymes should be identified as having a higher probability of appendicular perforation, suggesting total serum bilirubin levels have a predictive potential for the diagnosis of appendicular perforation.

Background: The preterm infants admitted to a high-tech Neonatal Intensive Care Unit (NICU) are vulnerable to noise in the environment. Aim of the study: To determine the physiological responses of preterm infants exposed to noise in the NICU. Materials and Methods: Noise levels in the NICU were measured using calibrated Sound Level Meter (SLM) i.e, Hand Held Analyzer Type 2250, Bruel and Kjaer, Denmark. The physiological responses of preterm infants were recorded in the Phillips IntelliVue MP20 Junior multichannel monitors. Results: The maximum hourly equivalent sound LAeq of 62.12 + 2.24 dB A was found on the 2nd day of the measurement that was carried out for a week’s time. The preterm infants cared for in the open care system were exposed to noise levels as high as 78.89 dB A over a period of 24 hours. There was a slight increase in heart rate (HR) on the second day, third day and sixth day of the measurement. Respiratory rate (RR) and Oxygen saturation (SpO2) were within normal range. Conclusion: The study has found that preterm infants were exposed to noise above the recommended levels on a day-to-day basis.

In the recent past, consumers in India have been at a point where there is a multiplicative effect in income growth, aspirations, and changed consumption patterns across income level segments. Therefore, understanding the rural consumer behavior and the process of decision-making has acquired significant attention and there has been a rise in the importance of private corporate big giants as the rural consumers started buying/consuming everything that was meant for the urban market. Keeping these changes in mind, the present paper tried to ascertain whether - there is any significant difference of opinion between the genders regarding the decision making process; the paper intended to find the different factors' (primitive) influence on the rural consumers and the places where these primitive factors influence; and also attempted to gain knowledge regarding the exclusive consumer decision making process of the rural consumer. The study took into account the opinion of 125 respondents from two taluks of Shimoga district, Karnataka. The data were analyzed with the help of SPSS v 20 software. Mean, standard deviation, chi-square, and factor analysis techniques were used to test the hypotheses, and based on the test results, an exclusive model (HSP Model) of rural consumers' factors of influence, place of influence, and decision-making process was developed.

To develop a highly sensitive and rapid protocol for simultaneous detection and differentiation of Tobacco mosaic virus (TMV) and Tomato mosaic virus (ToMV) in pepper and tomato. In this study, we use the multiplex PCR technique to detect dual infection of these two viruses.A multiplex RT-PCR method consisting of one-tube reaction with two primer pairs targeted to replicase genes was developed to simultaneously detect TMV and ToMV in seed samples of pepper and tomato. Specific primers were designed from conserved regions of each of the virus genomes, and their specificity was confirmed by sequencing PCR products. RT-PCR detected up to 10(-6) dilution of total RNA extracted from infected leaves. Multiplex RT-PCR revealed the presence of both TMV and ToMV in three of 18 seed samples of tomato and one of 18 seed samples of pepper.The multiplex PCR assay was a cost effective, quick diagnostic technique, which was helpful in differentiating TMV and ToMV accurately.The multiplex PCR assay described in this study is a valuable tool for plant pathology and basic research studies. This method may facilitate better recognition and distinction of TMV and ToMV in both pepper and tomato.

The uninucleate ascospores of Claviceps fusiformis germinate to give primary and secondary ‘conidia’. The nucleus of most ascospores migrated into primary ‘conidia’ and then into secondary ‘conidia’ without undergoing any division. In some the ascospore nucleus divided into two and this resulted in two primary conidia. The asexual conidia of honeydew-origin germinated to give secondary, tertiary and quaternary conidia. The nucleus from the primary conidium migrated into the successive conidia without division. Inoculation studies have shown that only the last formed conidia are infective whether they are of ascospore or conidial origin.