getpdf  NLM-PubMed-Logo  https://doi.org/10.17113/ftb.57.01.19.5816 

Innovative Nonthermal Technologies: Chlorophyllin and Visible Light Significantly Reduce Microbial Load on Basil


Egle Paskeviciuteorcid tiny, Bernadeta Zudyteorcid tiny and Zivile Luksiene*orcid tiny


Vilnius University, Institute of Photonics and Nanotechnology, Sauletekio 10, 10223 Vilnius, Lithuania



Article history:
Received: 23 April 2018
Accepted: 21 December 2018
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Key words:
photosensitization, antimicrobial technology, fresh produce, Ocimum basilicum




Summary:
Due to the high amount of biologically active compounds, basil is one of the most popular herbs. However, several outbreaks have been reported in the world due to the consumption of basil contaminated with different food pathogens. The aim of this study is to apply nonthermal and ecologically friendly approach based on photosensitization for microbial control of basil which was naturally contaminated with mesophils and inoculated with thermoresistant food pathogen Listeria monocytogenes 56Ly. The obtained data indicate that soaking the basil in 1.5·10-4 M chlorophyllin (Chl) for 15 min and illumination with light for 15 min at 405 nm significantly reduced total aerobic microorganisms on basil by 1.3 log CFU/g, and thermoresistant L. monocytogenes 56Ly from 6.1 log CFU/g in control to 4.5 log CFU/g in the treated samples. It is important to note that this treatment had no impact on enzymatic activity of polyphenol oxidase and pectinesterase. Results obtained in this study support the idea that photosensitization technique with its high selectivity, antimicrobial efficiency and nonthermal nature can serve in the future for the development of safe nonthermal and environmentally friendly preservation technology for different fruits and vegetables.




*Corresponding author: email3 Zivile.Luksiene@tmi.vu.lt

getpdf  NLM-PubMed-Logo  https://doi.org/10.17113/ftb.57.01.19.5879  

In Vitro Assessment of the Antioxidant Properties of Aqueous Byproduct Extracts of Vitis vinifera


Roberto Puglisi1
*orcid tiny, Alex Severgnini1orcid tiny, Aldo Tava2orcid tiny and Marina Montedoro1orcid tiny


1Istituto Sperimentale Italiano Lazzaro Spallanzani, Loc. La Quercia, 26027 Rivolta d’Adda (CR), Italy
2CREA-ZA Centro di Ricerca Zootecnia e Acquacoltura, Viale Piacenza 29, 26900 Lodi, Italy



Article history:
Received: 1 June 2018
Accepted: 31 January 2019
cc



Key words:
non-cytotoxic effect, extraction at low temperature, human dermal fibroblasts, oxidative status, mitochondrial membrane




Summary:
Aqueous extracts were obtained at low temperature with the Naviglio technology from grapevine stalks (Merlot), marc (Merlot and Cabernet Sauvignon) and leaves (Merlot) as typical byproducts of winemaking industry, and their properties were evaluated cytofluorometrically on human dermal fibroblasts. Leaf extracts had the greatest total phenolic ((47.6±3.5) mg/g) and proanthocyanidin ((24.2±0.1) mg/g) contents compared to the others. The preliminary colorimetric MTT (3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide) assay individuated two consecutive non-toxic volume fractions of each extract (from 0.8 to 12.8 %) that were adopted for three cytofluorometric tests. The first cell membrane test did not evidence any harmful effects against plasma membranes at the two non-toxic volume fractions. The second mitochondrial membrane test showed a decreased (p<0.01) percentage of cells ((15.7±8.3) vs (32.5±1.3) %) with active polarized mitochondrial membranes at the higher non-cytotoxic volume fractions of extracts from Cabernet Sauvignon marc in response to 4.5 mM H2O2, and from Merlot stalks (p<0.05) at 1.5 mM H2O2 ((49.3±6.1) vs (64.6±2.4) %) and without H2O2 ((89.7±2.4) vs (96.9±1.8) %), compared to the controls submitted to the same H2O2 concentration. Conversely, mitochondrial activity of leaf extracts significantly (p<0.05) increased ((96.3±1.8) and (96.4±1.4) %) after treatment with 0.5 mM H2O2 at both non-cytotoxic volume fractions compared to control ((88.2±1.1) %). Finally, as evidenced by the third oxidative status test, stalk extracts did not evidence relevant effects on the cellular oxidative state, while the extracts of marc and leaves demonstrated significantly medium (p<0.05) to highly (p<0.001) positive effects following exposure to H2O2 ranging from 0.5 to 4.5 mM, compared to controls.




*Corresponding author:  tel3  +39036378883
                                           fax2  +390363371021
                                           email3  roberto.puglisi@istitutospallanzani.it

getpdf  NLM-PubMed-Logo  https://doi.org/10.17113/ftb.57.01.19.5865  

Development of a Transgenic Flammulina velutipes Oral Vaccine for Hepatitis B


Li-Hsin Huang1orcid tiny, Hao-Yeh Lin2orcid tiny, Ying-Tzu Lyu1orcid tiny, Chiau-Ling Gung1orcid tiny and Ching-Tsan Huang2*orcid tiny


1MycoMagic Biotechnology Co. Ltd., 8F-1, 12, Lane 270, Sec. 3, Beishen Road, New Taipei City, Taiwan
2Department of Biochemical Science and Technology, National Taiwan University, 1, Sec. 4, Roosevelt Road, Taipei, Taiwan



Article history:
Received: 24 May 2018
Accepted: 31 January 2019
cc



Key words:
Agrobacterium-mediated transformation, Flammulina velutipes, hepatitis B, mating, oral vaccine




Summary:
Orally administered fungal vaccines show promise for the prevention of infectious diseases. Edible mushrooms are deemed appropriate hosts to produce oral vaccines due to their low production cost and low risk of gene contamination. However, their low expression level of antigens has limited the potential development of oral vaccines using mushrooms. The low expression level might result from impurity of the transgenic mycelia since dikaryotic mycelia are commonly used as transformation materials. In this study, stable transgenic hepatitis B virus surface antigen (HBsAg) in Flammulina velutipes transformants was obtained by Agrobacterium-mediated transformation, followed by fruiting and basidiospore mating. The formation of HBsAg was detected by western blot analysis. The expression levels of HBsAg in transgenic F. velutipes fruiting bodies were (129.3±15.1), (110.9±1.7) and (161.1±8.5) ng/g total soluble protein. However, the values may be underestimated due to incomplete protein extraction. Two of the four pigs in the experimental group produced positive anti-HBsAg-specific IgG after being fed the HBsAg transgenic F. velutipes fruiting bodies for 20 weeks, while no anti-HBsAg antibody was detected in the control group. One of the positive pigs had HBsAg titres of 5.36 and 14.9 mIU/mL in weeks 10 and 14, respectively, but expression faded thereafter. The other positive pig displayed HBsAg titres of 9.75, 17.86 and 39.87 mIU/mL in weeks 14, 18 and 20, respectively. The successful immunogenicity in pigs fed transgenic F. velutipes fruiting bodies demonstrated the potential of using the fungus as an oral vaccine.



*Corresponding author:  tel3 +886233664454
                                           fax2 +886223634796
                                            email3 cthuang@ntu.edu.tw

getpdf  NLM-PubMed-Logo  https://doi.org/10.17113/ftb.57.01.19.5909  

Proteinaceous Pancreatic Lipase Inhibitor from the Seed of Litchi chinensis


Sveeta V. Mhatre1,2orcid tiny, Amita A. Bhagit1,2orcid tiny and Raman P. Yadav2*orcid tiny


1Department of Medical Biotechnology, MGM School of Biomedical Sciences, MGM Institute of Health Sciences, Sector 1, Kamothe 410209, Navi Mumbai, Maharashtra, India
2MGMIHS OMICS Research Center, MGM Central Research Laboratory, MGM Medical College and Hospital, MGM Institute of Health Sciences, Sector 1, Kamothe 410209, Navi Mumbai, Maharashtra, India



Article history:
Received: 22 June 2018
Accepted: 11 December 2018
cc



Key words
:
fruit seed, Litchi chinensis, proteinaceous pancreatic lipase inhibitor, obesity treatment




Summary:
A study of the pancreatic lipase inhibitory activity of a protein from the seed of Litchi chinensis was carried out. Protein was isolated by 70 % ammonium sulphate precipitation followed by dialysis. Lipase inhibitory activity of the protein was evaluated using both synthetic (p-nitrophenyl palmitate) and natural (olive oil) substrates. Protein at the final concentration of 100 μg/mL was able to inhibit 68.2 % pancreatic lipase on synthetic substrate and 60.0 % on natural substrate. Proteinaceous nature of the inhibitor was determined using trypsinization assay. Pancreatic lipase inhibitory protein was sensitive to 0.05 % trypsin treatment with the loss of 61.9 % activity. IC50 of this proteinaceous pancreatic lipase inhibitor was 73.1 μg/mL using synthetic substrate. This inhibitory protein was sensitive to pH, with the highest inhibitory activity at pH=8.0 and the lowest at pH=3.0. Protein was further analyzed using 10 % non-reducing sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and, interestingly, it showed the presence of a single band of (61±2) kDa when stained with Coomassie brilliant blue. The isolated protein was finally crystallized to see its homogeneity by batch crystallization method. Crystals were well formed with distinct edges. The isolated protein showed good pancreatic lipase inhibitory activity.



*Corresponding author: tel3 +912227432890
                                                +912227437693
                                          fax2 +912227431094
                                          email3 ramanpyadav@gmail.com

getpdf  NLM-PubMed-Logo  https://doi.org/10.17113/ftb.57.01.19.5983  

Multiplex Real-Time PCR with HRM for Detection of Lactobacillus sakei and Lactobacillus curvatus in Food Samples


Konstantin A. Kurbakov1
*orcid tiny, Evgenii A. Konorov1,2,3orcid tiny, Mikhail Y. Minaev1orcid tiny and Oksana A. Kuznetsova1orcid tiny


1V.M. Gorbatov Federal Research Center for Food Systems of Russian Academy of Sciences, Talalikhina 26, 109316, Moscow, Russian Federation
2Vavilov Institute of General Genetics, Gubkina 3, 119333, Moscow, Russian Federation
3Martsinovsky Institute of Medical Parasitology, Tropical and Vector-Borne Diseases, Sechenov University, Malaya Pirogovskaya 20-1, 119435 Moscow, Russian Federation



Article history:
Received: 9 August 2018
Accepted: 17 January 2019
cc



Key words:
real-time PCR, HRM, starter cultures, fermented sausages, Lactobacillus sakeiLactobacillus curvatus




Summary:
Optimization of fermentation processes requires monitoring the species composition of starter cultures and their growth during fermentation. Most starter cultures contain closely related species. Nowadays, high-resolution melting (HRM) analysis is extensively used for multiplex identification of closely related species. In the present paper, we applied real-time polymerase chain reaction (PCR) with HRM analysis for the detection and differentiation of Lactobacillus sakei and L. curvatus. A primer pair was selected for the site of the rpoA gene of Lactobacillus spp. Eleven starter cultures and fifteen fermented sausages with a known bacterial composition were successfully tested using real-time PCR with HRM analysis with the developed primer pair.




*Corresponding author: tel3 +79629529223
                                          email3 homo_ludens@vniimp.ru

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