Biological activity and chemical caracteristics of selected extracts of autochtonous macrofungi

Prema postavljenim ciljevima u ovoj doktorskoj disertaciji sakupljeno je i  determinisano ukupno sedam vrsta autohtonih gljiva sa područja Fruške gore, Tare i  Vršačkog brega, pet lignikolnih ‐ Bjerkandera adusta, Pleurocybella  porrigens, Stereum hirsutum, Stereum subtomentosum i Trametes versicolor, i  dve  terikolne ‐ Amanita  strobiliformis i Hydnum repandum. Utvrđena je  njihova biološka aktivnost (antiradikalska, antioksidativna, antimikrobna, anti‐acetilholinesterazna i citotokisčna) spram hemijskog sastava njihovih vodenih  (H2O), etanolnih (EtOH), metanolnih (MeOH) i polisaharidnih (PSH) ekstrakata.  Analiza hemijskog sastava odabranih vrsta uključila je određivanje hemijske  karakterizacije PSH ekstrakata ‐ FTIR analizom, određivanje fenolnog profila ‐  HPMC/MS‐MS, sadržaja organskih kiselina ‐ HPLC, sadržaja masnih kiselina ‐  GC‐FID i sadržaja biogenih elemenata ‐ AAS. Spektrofotometrijskim metodama određen je ukupan sadržaj proteina i ukupan sadržaj fenola i flavonoida.  Prema antiradikalskoj aktivnosti OH• , O2•‐, OH•, Asc•, DPPH•  i ABTS•+ izdvojili  su  se  ekstrakti lignikolnih vrsta:  MeOH ekstrakt vrste P. porrigens, H2O  ekstrakt P. porrigens, MeOH ekstrakt T. versicolor, H2O ekstrakt S. hirsutum, MeOH ekstrakt S. subtomentosum i H2O ekstrakt B. adusta, navedenim redom.  Najjaču antioksidativnu aktivnost dobijenu FRAP i polarografskom HPMC  metodom ispoljili su PSH i H2O ekstrakti terikolne vrste A. strobiliformis.  Antimikrobna aktivnost analiziranih ekstrakata određena je ispitivanjem  antibakterijskog, antifungalnog i antiviralnog potencijala, pri čemu se izdvojila  vrsta  H.  repandum ispoljavajući najbolji efekat na Gram‐pozitivne i Gram‐ negativne bakterije i na sve analizirane fitopatogene izolate (Fusarium i  Alternaria) i T. versicolor na analizirani bakteriofag. Anti‐acetilholinesterazna  aktivnost određena je testovima in solid i in liquid, a najbolji procenat  inhibicije AChE ispoljili su EtOH ekstrakti vrsta S. hirsutum, B. adusta, S.  subtomentosum i T. versicolor. Citotoksična aktivnost ekstrakata određena je  MTT testom, a prema najboljoj ispoljenoj aktivnosti izdvojili su se MeOH  ekstrakt P. porrigens i ekstrakti B. adusta, H2O i EtOH. Citotoksična aktivnost  ovih lignikolnih vrsta naročito je izražena nakon 72 h. Na osnovu dobijenih  rezultata, gde su se istakle različite vrste i njihovi različiti ekstrakti u  primenjenim testovima, jasno je da biološka aktivnost i hemijski sastav zavise  od porekla, vrste i tipa ekstrakta analiziranih gljiva. Na osnovu tipa rastvarača  odnosno ekstrakata vrsta, koje su pokazale najbolju aktivnost spram pomenutih  testova i na osnovu dobijenih korelacija kao i na osnovu detektovanih jedinjenja,  pretpostavljamo da su za ispoljene aktivnosti u najvećoj meri odgovorna fenolna  jedinjenja i polisaharidi.     Ispoljeni biopotencijal analiziranih vrsta gljiva upućuje na njihovu potencijalnu  upotrebu kao funkcionalne hrane i nutraceutika, kao i u biokontroli  fitopatogena.According  to  the  set  aims  of  the  presented  PhD  thesis,  seven  autochthonous fungal species from the region of Fruska Gora, Tara  and Vršac Mountains were collected and determined: five lignicolous ‐ Bjerkandera  adusta,  Pleurocybella  porrigens,  Stereum  hirsutum,  Stereum subtomentosum and Trametes versicolor, and two terricolous ‐ Amanita strobiliformis and Hydnum repandum. Biological activity of these  species  (antiradical,  antioxidant,  antimicrobial,  anti‐ acetylcholinesterase and cytotoxic) was determined in relation to the chemical composition of the extracts, aqueous (H2O), ethanolic (EtOH), methanolic (MeOH) and polysaccharide (PSH). Analysis of the chemical content of analyzed species included chemical characterization of PSH extracts  –  by  FTIR  analysis,  determination  of  phenolic  profile ‐ by HPMC/MS‐MS, content of organic acids ‐ by HPLC, fatty acid content ‐ by  GC‐FID  and  content  of  biogenic  elements ‐ by  AAS. Spectrophotometric methods were applied for determination of the content of total proteins, polyphenols and flavonoids. According to the antiradical activity obtained towards OH•, О2•‐, OH•, Asc•, DPPH• and ABTS•+ extracts of lignicolous species were singled out: P. porrigens  (MeOH  extract),  P.  porrigens  (H2O  extract), T. versicolor (MeOH   extract), S. hirsutum (H2O extract), S. subtomentosum (MeOH extract) and B. austa (H2O  extract),  respectively. The  highest antioxidant activity obtained by FRAP and the polarographic HPMC method was exhibited  for PSH  and  H2O extracts of the terricolous species A. strobiliformis. The intimicrobial activity of analyzed extracts was determined by examination of antibacterial, antifungal and antiviral potentials, whereby  the species  H.  repandum was separated by exhibiting the best effect on Gram‐positive  and Gram‐negative bacteria, and all the analyzed hytopathogenic isolates (Fusarium, Alternaria)  and T. versicolor against analyzed bacteriophage. Anti‐cetylcholinesterase activity was determined by tests in solid and in liquid, while the best  percent of AChE inhibition was showed by EtOH extracts of the species S. hirsutum, B. adusta, S. subtomentosum and T. versicolor.bThe cytotoxic activity of extracts was determined by MTT assay, and according to the best activity, the MeOH extract of P. porrigens, and H2O and EtOH extracts of B. adusta were distinguished particularly after 72 h. Based on the results obtained, favoring different species and their different extracts in the applied tests, it is clear that the biological activity and chemical composition depend on the origin, species and type of extract of the analyzed fungi. Based on the type of solvent or extract of the species that showed the best activity in relation to the above tests and on the basis of the obtained correlations as well as on  the basis of the detected compounds, we assume  that  the  phenol compounds  and  polysaccharides  are responsible for the activities performed. Demonstrated bio‐potential of analyzed fungal species indicates their  potential use as functional foods and nutraceutics, as well as in the biocontrol of phytopathogens

Biological activity and chemical caracteristics of selected extracts of autochtonous macrofungi

Prema postavljenim ciljevima u ovoj doktorskoj disertaciji sakupljeno je i  determinisano ukupno sedam vrsta autohtonih gljiva sa područja Fruške gore, Tare i  Vršačkog brega, pet lignikolnih ‐ Bjerkandera adusta, Pleurocybella  porrigens, Stereum hirsutum, Stereum subtomentosum i Trametes versicolor, i  dve  terikolne ‐ Amanita  strobiliformis i Hydnum repandum. Utvrđena je  njihova biološka aktivnost (antiradikalska, antioksidativna, antimikrobna, anti‐acetilholinesterazna i citotokisčna) spram hemijskog sastava njihovih vodenih  (H2O), etanolnih (EtOH), metanolnih (MeOH) i polisaharidnih (PSH) ekstrakata.  Analiza hemijskog sastava odabranih vrsta uključila je određivanje hemijske  karakterizacije PSH ekstrakata ‐ FTIR analizom, određivanje fenolnog profila ‐  HPMC/MS‐MS, sadržaja organskih kiselina ‐ HPLC, sadržaja masnih kiselina ‐  GC‐FID i sadržaja biogenih elemenata ‐ AAS. Spektrofotometrijskim metodama određen je ukupan sadržaj proteina i ukupan sadržaj fenola i flavonoida.  Prema antiradikalskoj aktivnosti OH• , O2•‐, OH•, Asc•, DPPH•  i ABTS•+ izdvojili  su  se  ekstrakti lignikolnih vrsta:  MeOH ekstrakt vrste P. porrigens, H2O  ekstrakt P. porrigens, MeOH ekstrakt T. versicolor, H2O ekstrakt S. hirsutum, MeOH ekstrakt S. subtomentosum i H2O ekstrakt B. adusta, navedenim redom.  Najjaču antioksidativnu aktivnost dobijenu FRAP i polarografskom HPMC  metodom ispoljili su PSH i H2O ekstrakti terikolne vrste A. strobiliformis.  Antimikrobna aktivnost analiziranih ekstrakata određena je ispitivanjem  antibakterijskog, antifungalnog i antiviralnog potencijala, pri čemu se izdvojila  vrsta  H.  repandum ispoljavajući najbolji efekat na Gram‐pozitivne i Gram‐ negativne bakterije i na sve analizirane fitopatogene izolate (Fusarium i  Alternaria) i T. versicolor na analizirani bakteriofag. Anti‐acetilholinesterazna  aktivnost određena je testovima in solid i in liquid, a najbolji procenat  inhibicije AChE ispoljili su EtOH ekstrakti vrsta S. hirsutum, B. adusta, S.  subtomentosum i T. versicolor. Citotoksična aktivnost ekstrakata određena je  MTT testom, a prema najboljoj ispoljenoj aktivnosti izdvojili su se MeOH  ekstrakt P. porrigens i ekstrakti B. adusta, H2O i EtOH. Citotoksična aktivnost  ovih lignikolnih vrsta naročito je izražena nakon 72 h. Na osnovu dobijenih  rezultata, gde su se istakle različite vrste i njihovi različiti ekstrakti u  primenjenim testovima, jasno je da biološka aktivnost i hemijski sastav zavise  od porekla, vrste i tipa ekstrakta analiziranih gljiva. Na osnovu tipa rastvarača  odnosno ekstrakata vrsta, koje su pokazale najbolju aktivnost spram pomenutih  testova i na osnovu dobijenih korelacija kao i na osnovu detektovanih jedinjenja,  pretpostavljamo da su za ispoljene aktivnosti u najvećoj meri odgovorna fenolna  jedinjenja i polisaharidi.     Ispoljeni biopotencijal analiziranih vrsta gljiva upućuje na njihovu potencijalnu  upotrebu kao funkcionalne hrane i nutraceutika, kao i u biokontroli  fitopatogena.According  to  the  set  aims  of  the  presented  PhD  thesis,  seven  autochthonous fungal species from the region of Fruska Gora, Tara  and Vršac Mountains were collected and determined: five lignicolous ‐ Bjerkandera  adusta,  Pleurocybella  porrigens,  Stereum  hirsutum,  Stereum subtomentosum and Trametes versicolor, and two terricolous ‐ Amanita strobiliformis and Hydnum repandum. Biological activity of these  species  (antiradical,  antioxidant,  antimicrobial,  anti‐ acetylcholinesterase and cytotoxic) was determined in relation to the chemical composition of the extracts, aqueous (H2O), ethanolic (EtOH), methanolic (MeOH) and polysaccharide (PSH). Analysis of the chemical content of analyzed species included chemical characterization of PSH extracts  –  by  FTIR  analysis,  determination  of  phenolic  profile ‐ by HPMC/MS‐MS, content of organic acids ‐ by HPLC, fatty acid content ‐ by  GC‐FID  and  content  of  biogenic  elements ‐ by  AAS. Spectrophotometric methods were applied for determination of the content of total proteins, polyphenols and flavonoids. According to the antiradical activity obtained towards OH•, О2•‐, OH•, Asc•, DPPH• and ABTS•+ extracts of lignicolous species were singled out: P. porrigens  (MeOH  extract),  P.  porrigens  (H2O  extract), T. versicolor (MeOH   extract), S. hirsutum (H2O extract), S. subtomentosum (MeOH extract) and B. austa (H2O  extract),  respectively. The  highest antioxidant activity obtained by FRAP and the polarographic HPMC method was exhibited  for PSH  and  H2O extracts of the terricolous species A. strobiliformis. The intimicrobial activity of analyzed extracts was determined by examination of antibacterial, antifungal and antiviral potentials, whereby  the species  H.  repandum was separated by exhibiting the best effect on Gram‐positive  and Gram‐negative bacteria, and all the analyzed hytopathogenic isolates (Fusarium, Alternaria)  and T. versicolor against analyzed bacteriophage. Anti‐cetylcholinesterase activity was determined by tests in solid and in liquid, while the best  percent of AChE inhibition was showed by EtOH extracts of the species S. hirsutum, B. adusta, S. subtomentosum and T. versicolor.bThe cytotoxic activity of extracts was determined by MTT assay, and according to the best activity, the MeOH extract of P. porrigens, and H2O and EtOH extracts of B. adusta were distinguished particularly after 72 h. Based on the results obtained, favoring different species and their different extracts in the applied tests, it is clear that the biological activity and chemical composition depend on the origin, species and type of extract of the analyzed fungi. Based on the type of solvent or extract of the species that showed the best activity in relation to the above tests and on the basis of the obtained correlations as well as on  the basis of the detected compounds, we assume  that  the  phenol compounds  and  polysaccharides  are responsible for the activities performed. Demonstrated bio‐potential of analyzed fungal species indicates their  potential use as functional foods and nutraceutics, as well as in the biocontrol of phytopathogens

Predictors of Vancomycin-Resistant Enterococcus spp. Intestinal Carriage among High-Risk Patients in University Hospitals in Serbia

The predictors of intestinal carriage of vancomycin-resistant Enterococcus spp. (VRE) among high-risk patients in the counties of the Southeast Europe Region are insufficiently investigated, yet they could be of key importance in infection control. The aim of the study was to identify risk factors associated with fecal VRE colonization among high-risk inpatients in university hospitals in Serbia. The study comprised 268 inpatients from three university hospitals. Data on patient demographics and clinical characteristics, length of hospital stay, therapy, and procedures were obtained from medical records. Chi-squared tests and univariate and multivariate logistic regressions were performed. Compared to the hemodialysis departments, stay in the geriatric departments, ICUs, and haemato-oncology departments increased the risk for VRE colonization 7.6, 5.4, and 5.5 times, respectively. Compared to inpatients who were hospitalized 48 h before stool sampling for VRE isolation, inpatients hospitalized 3–7, 8–15, and longer than 16 days before sampling had 5.0-, 4.7-, and 6.6-fold higher risk for VRE colonization, respectively. The use of cephalosporins and fluoroquinolones increased the risk for VRE colonization by 2.2 and 1.9 times, respectively. The age ≥ 65 years increased the risk for VRE colonization 2.3 times. In comparison to the University Clinical Centre of Serbia, the hospital stays at Zemun and Zvezdara University Medical Centres were identified as a protector factors. The obtained results could be valuable in predicting the fecal VRE colonization status at patient admission and consequent implementation of infection control measures targeting at-risk inpatients where VRE screening is not routinely performed

loop-mediated isothermal amplification (lamP) as a point-of-care SarS-covdetection method

Pravovremeno testiranje većeg broja ljudi na SARS-CoV-2 virus je povezano sa nižim mortalitetom od COVID-19 oboljenja. Međutim, većina zemalja nema mogućnosti za takvo masivno testiranje putem metode “PCR u realnom vremenu”, zbog visoke cene neophodne opreme i potrebe za stručnim osobljem. Zbog toga se razvijaju brze i ekonomičnije metode, koje se najčešće zasnivaju na izotermalnim metodama amplifikacije DNK. Za ove metode nije potreban ciklični termostat, zbog čega su primenljivije za terensku upotrebu. Fokus je na izotermalnoj amplifikaciji posredovanoj petljom (lamP), zbog njene visoke specifičnosti, mogućnosti za upotrebu neprečišćenih uzoraka i jednostavnosti merenja signala. Autori predstavljaju pregled najvažnijih radova o LAMP metodi za detekciju SARS-CoV-2 virusa objavljenih tokom 2020. godine, kao i opise nekoliko komercijalnih kompleta na bazi LAMP metode za COVID-19 testiranje.Massive testing for SARS-CoV-2 virus is related to lower mortality rates of COVID-19. Most countries face challenges to perform massive testing with the currently available methods (real-time PCR), due to expensive equipment and requirement of highly skilled personnel. To overcome these challenges, faster and cost-effective alternative detection methods are being developed, primarily based on isothermal methods of nucleic acid amplification (isoNAAs). PCR depends on precision instruments, high cleanliness of operating conditions and cannot be easily used on-site, while isoNAAs, which do not require thermal cyclers, are more applicable for point-of-care (PoC) use. Loop-mediated isothermal amplification (LAMP) is one of the isoNAA most in focus for COVID-19 tests due to its high specificity and possibility to use unpurified specimens in combination with simplified detection setup. The article gives a review of the most significant publications on use of LAMP for SARS-CoV-2 detection and of the several commercial LAMP-based COVID-19 testing kits.Ministarstva prosvete, nauke i tehnološkog razvoja Republike Srbije – broj ugovora 451-03-68/2020-14/ 200358 za autore M. Dj, T.K i Lj.J i broj ugovora 451-03-9/2021- 14/200125 za Ž.D.P. Autori zahvaljuju Jeleni Ognjenov na pomoći u pripremi ilustracija za rad

Antifungalna aktivnost ekstrakata makrogljiva na fitopatogene sojeve gljiva rodova Fusarium sp. i Alternaria sp.

During the last decades, intensive application of synthetic fungicides in the agricultural crop protection practice caused growing concern for the existence of toxic chemical residues in food as well as in the whole environment. Instead of using synthetic fungicides, it is suggested that crop protection be carried out by using preparations based on compounds of natural origin (secondary metabolites of plants or microorganisms, including macrofungi from Basidiomycota) as biological control agents. The potential of macrofungal species as biocontrol agents was analyzed in this investigation of eight autochthonous species from different locations in Serbia. Both the terricolous species: Coprinus comatus, Coprinellus truncorum, Amanita strobiliformis, Hydnum repandum and the lignicolous species: Flammulina velutipes, Stereum subtomentosum, Trametes versicolor and Bjerkandera adusta were examined, with an aim to detect some novel sources of antifungal agents. This study surveyed antifungal activity of selected macrofungal extracts (MeOH, EtOH and CHCl3) against phytopathogenic Fusarium and Alternaria strains isolated from garlic, soybean and rice: F. proliferatum, F. verticillioides, F. proliferatum, F. graminearum and A. padwickii. Microdilution method in 96 well micro-plates was applied for the estimation of antifungal effects of macrofungi extracts in the range from 24.75 to 198.00 mg/ml and determination of minimal inhibitory (MIC) and minimal fungicidal concentration (MFC). EtOH extract of mychorhizal species H. repandum showed antifungal activity against all analyzed phytopathogenic strains, with the strongest effect on Fusarium strains (MIC 24.75 mg/ml; MFC 24.75 mg/ml). Among others, MeOH extracts of S. subtomentosum and C. micaceus showed similar effects while only B. adusta showed slight effect on Fusarium strains (MIC 24.75-99.00 mg/ml; MFC 24.75-99.00 mg/ml) and none effect on A. padwickii. The obtained results indicate the possibility of using examined extracts as efficient antifungal agents and provide the basis for the new formulations of biocontrol agents against phytopathogenic fungi in the future.Tokom poslednjih decenija intezivna primena sintetičkih fungicida u zaštiti poljoprivrednih useva izaziva sve veću zabrinutost ljudi zbog prisustva toksičnih hemijskih ostataka u prehrambenim proizvodima kao i celoj okolini. Umesto korišćenja sintetičkih fungicida, preporučuje se da se zaštita useva vrši korišćenjem preparata na bazi jedinjenja prirodnog porekla (sekundarni metaboliti biljaka ili mikroorganizama uključujući i makrogljive iz razdela Basidiomycota) kao agense biološke kontrole. U ovom radu istraživan je potencijal agenasa biološke kontrole za osam autohtonih vrsta makrogljiva s različitih lokaliteta u Srbiji. Obe, terikolne vrste: Coprinus comatus, Coprinellus truncorum, Amanita strobiliformis, Hydnum repandum i lignikolne vrste: Flammulina velutipes, Stereum subtomentosum, Trametes versicolor i Bjerkandera adusta su istražene s ciljem da se otkriju neki novi izvori antifungalnih agenasa. U okviru ovog rada istražena je antifungalna aktivnost odabranih ekstrakata makrogljiva (metanolni, etanolni i hloroformski) protiv fitopatogenih sojeva Fusarium i Alternaria izolovanih s belog luka, soje i pirinča: F. proliferatum, F. verticillioides, F. proliferatum, F. graminearum i A. padwickii. Za procenu antifungalnog efekta ekstrakata makrogljiva upotrebljena je mikrodiluciona metoda mikrotitar pločama s 96 velova u opsegu koncentracija od 24,75 do 198,00 mg/ml i determinaciju minimalne inhibitorne (MIC) i minimalne fungicidne koncentracije (MFC). Etanolni ekstrakt mikorizne vrste H. repandum pokazao je atifungalnu aktivnost prema svim analiziranim fitopatogenim sojevima, sa najjačim efektom prema sojevima Fusarium (MIC 24,75 mg/ml; MFC 24,75 mg/ml). Između ostalog, sličan efekat pokazali su i metanolni ekstrakti S. subtomentosum i C. micaceus, dok je samo B. adusta imala blagi efekat na sojeve Fusarium (MIC 24,75-99,00 mg/ml; MFC 24,75-99,00 mg/ml), ali ne i na A. padwickii. Dobijeni rezultati ukazuju na mogućnost korišćenja ispitanih ekstrakata kao veoma efikasnih antifungalnih agenasa i samim tim oni predstavljaju osnovu za nove formulacije biokontrolnih agenasa protiv fitopatogenih gljiva u budućnosti

Biological activity and chemical caracteristics of selected extracts of autochtonous macrofungi

Prema postavljenim ciljevima u ovoj doktorskoj disertaciji sakupljeno je i  determinisano ukupno sedam vrsta autohtonih gljiva sa područja Fruške gore, Tare i  Vršačkog brega, pet lignikolnih ‐ Bjerkandera adusta, Pleurocybella  porrigens, Stereum hirsutum, Stereum subtomentosum i Trametes versicolor, i  dve  terikolne ‐ Amanita  strobiliformis i Hydnum repandum. Utvrđena je  njihova biološka aktivnost (antiradikalska, antioksidativna, antimikrobna, anti‐acetilholinesterazna i citotokisčna) spram hemijskog sastava njihovih vodenih  (H2O), etanolnih (EtOH), metanolnih (MeOH) i polisaharidnih (PSH) ekstrakata.  Analiza hemijskog sastava odabranih vrsta uključila je određivanje hemijske  karakterizacije PSH ekstrakata ‐ FTIR analizom, određivanje fenolnog profila ‐  HPMC/MS‐MS, sadržaja organskih kiselina ‐ HPLC, sadržaja masnih kiselina ‐  GC‐FID i sadržaja biogenih elemenata ‐ AAS. Spektrofotometrijskim metodama određen je ukupan sadržaj proteina i ukupan sadržaj fenola i flavonoida.  Prema antiradikalskoj aktivnosti OH• , O2•‐, OH•, Asc•, DPPH•  i ABTS•+ izdvojili  su  se  ekstrakti lignikolnih vrsta:  MeOH ekstrakt vrste P. porrigens, H2O  ekstrakt P. porrigens, MeOH ekstrakt T. versicolor, H2O ekstrakt S. hirsutum, MeOH ekstrakt S. subtomentosum i H2O ekstrakt B. adusta, navedenim redom.  Najjaču antioksidativnu aktivnost dobijenu FRAP i polarografskom HPMC  metodom ispoljili su PSH i H2O ekstrakti terikolne vrste A. strobiliformis.  Antimikrobna aktivnost analiziranih ekstrakata određena je ispitivanjem  antibakterijskog, antifungalnog i antiviralnog potencijala, pri čemu se izdvojila  vrsta  H.  repandum ispoljavajući najbolji efekat na Gram‐pozitivne i Gram‐ negativne bakterije i na sve analizirane fitopatogene izolate (Fusarium i  Alternaria) i T. versicolor na analizirani bakteriofag. Anti‐acetilholinesterazna  aktivnost određena je testovima in solid i in liquid, a najbolji procenat  inhibicije AChE ispoljili su EtOH ekstrakti vrsta S. hirsutum, B. adusta, S.  subtomentosum i T. versicolor. Citotoksična aktivnost ekstrakata određena je  MTT testom, a prema najboljoj ispoljenoj aktivnosti izdvojili su se MeOH  ekstrakt P. porrigens i ekstrakti B. adusta, H2O i EtOH. Citotoksična aktivnost  ovih lignikolnih vrsta naročito je izražena nakon 72 h. Na osnovu dobijenih  rezultata, gde su se istakle različite vrste i njihovi različiti ekstrakti u  primenjenim testovima, jasno je da biološka aktivnost i hemijski sastav zavise  od porekla, vrste i tipa ekstrakta analiziranih gljiva. Na osnovu tipa rastvarača  odnosno ekstrakata vrsta, koje su pokazale najbolju aktivnost spram pomenutih  testova i na osnovu dobijenih korelacija kao i na osnovu detektovanih jedinjenja,  pretpostavljamo da su za ispoljene aktivnosti u najvećoj meri odgovorna fenolna  jedinjenja i polisaharidi.     Ispoljeni biopotencijal analiziranih vrsta gljiva upućuje na njihovu potencijalnu  upotrebu kao funkcionalne hrane i nutraceutika, kao i u biokontroli  fitopatogena.According  to  the  set  aims  of  the  presented  PhD  thesis,  seven  autochthonous fungal species from the region of Fruska Gora, Tara  and Vršac Mountains were collected and determined: five lignicolous ‐ Bjerkandera  adusta,  Pleurocybella  porrigens,  Stereum  hirsutum,  Stereum subtomentosum and Trametes versicolor, and two terricolous ‐ Amanita strobiliformis and Hydnum repandum. Biological activity of these  species  (antiradical,  antioxidant,  antimicrobial,  anti‐ acetylcholinesterase and cytotoxic) was determined in relation to the chemical composition of the extracts, aqueous (H2O), ethanolic (EtOH), methanolic (MeOH) and polysaccharide (PSH). Analysis of the chemical content of analyzed species included chemical characterization of PSH extracts  –  by  FTIR  analysis,  determination  of  phenolic  profile ‐ by HPMC/MS‐MS, content of organic acids ‐ by HPLC, fatty acid content ‐ by  GC‐FID  and  content  of  biogenic  elements ‐ by  AAS. Spectrophotometric methods were applied for determination of the content of total proteins, polyphenols and flavonoids. According to the antiradical activity obtained towards OH•, О2•‐, OH•, Asc•, DPPH• and ABTS•+ extracts of lignicolous species were singled out: P. porrigens  (MeOH  extract),  P.  porrigens  (H2O  extract), T. versicolor (MeOH   extract), S. hirsutum (H2O extract), S. subtomentosum (MeOH extract) and B. austa (H2O  extract),  respectively. The  highest antioxidant activity obtained by FRAP and the polarographic HPMC method was exhibited  for PSH  and  H2O extracts of the terricolous species A. strobiliformis. The intimicrobial activity of analyzed extracts was determined by examination of antibacterial, antifungal and antiviral potentials, whereby  the species  H.  repandum was separated by exhibiting the best effect on Gram‐positive  and Gram‐negative bacteria, and all the analyzed hytopathogenic isolates (Fusarium, Alternaria)  and T. versicolor against analyzed bacteriophage. Anti‐cetylcholinesterase activity was determined by tests in solid and in liquid, while the best  percent of AChE inhibition was showed by EtOH extracts of the species S. hirsutum, B. adusta, S. subtomentosum and T. versicolor.bThe cytotoxic activity of extracts was determined by MTT assay, and according to the best activity, the MeOH extract of P. porrigens, and H2O and EtOH extracts of B. adusta were distinguished particularly after 72 h. Based on the results obtained, favoring different species and their different extracts in the applied tests, it is clear that the biological activity and chemical composition depend on the origin, species and type of extract of the analyzed fungi. Based on the type of solvent or extract of the species that showed the best activity in relation to the above tests and on the basis of the obtained correlations as well as on  the basis of the detected compounds, we assume  that  the  phenol compounds  and  polysaccharides  are responsible for the activities performed. Demonstrated bio‐potential of analyzed fungal species indicates their  potential use as functional foods and nutraceutics, as well as in the biocontrol of phytopathogens

Molecular Approaches for Detection of <i>Trichoderma</i> Green Mold Disease in Edible Mushroom Production

Due to the evident aggressive nature of green mold and the consequently huge economic damage it causes for producers of edible mushrooms, there is an urgent need for prevention and infection control measures, which should be based on the early detection of various Trichoderma spp. as green mold causative agents. The most promising current diagnostic tools are based on molecular methods, although additional optimization for real-time, in-field detection is still required. In the first part of this review, we briefly discuss cultivation-based methods and continue with the secondary metabolite-based methods. Furthermore, we present an overview of the commonly used molecular methods for Trichoderma species/strain detection. Additionally, we also comment on the potential of genomic approaches for green mold detection. In the last part, we discuss fast screening molecular methods for the early detection of Trichoderma infestation with the potential for in-field, point-of-need (PON) application, focusing on isothermal amplification methods. Finally, current challenges and future perspectives in Trichoderma diagnostics are summarized in the conclusions

Using Vertebrate Stem and Progenitor Cells for Cellular Agriculture, State-of-the-Art, Challenges, and Future Perspectives

Global food systems are under significant pressure to provide enough food, particularly protein-rich foods whose demand is on the rise in times of crisis and inflation, as presently existing due to post-COVID-19 pandemic effects and ongoing conflict in Ukraine and resulting in looming food insecurity, according to FAO. Cultivated meat (CM) and cultivated seafood (CS) are protein-rich alternatives for traditional meat and fish that are obtained via cellular agriculture (CA) i.e., tissue engineering for food applications. Stem and progenitor cells are the building blocks and starting point for any CA bioprocess. This review presents CA-relevant vertebrate cell types and procedures needed for their myogenic and adipogenic differentiation since muscle and fat tissue are the primary target tissues for CM/CS production. The review also describes existing challenges, such as a need for immortalized cell lines, or physical and biochemical parameters needed for enhanced meat/fat culture efficiency and ways to address them