FAKTORI KOJI UTIČU NA MIKROORGANIZME

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1 FAKTORI KOJI UTIČU NA MIKROORGANIZME

2 INTRINZIČ FAKTORI EKSTRINZIČ FAKTORI IMPLICIT FAKTORI INTRINZIČ FAKTORI: nutrijenti katabolizam laktoze i proteolitička aktivnost ph i puferski kapacitet redoks potencijal aktivnost vode aw antimikrobni sastojci i strukture EKSTRINZIČ FAKTORI: temperatura relativna vlažnost IMPLICIT FAKTORI: inicijalni broj razvojne faze i ritam reprodukcije međusobni odnosi interakcije

3 ph ph = log [H + ] PODELA ACIDOFILI, NEUTROFILI I ALKALOFILI

4 TOLER ANCIJ A NA PH ODRE ĐENIH BAKTE RIJA TOLERANCIJA NA ph ODREĐENIH BAKTERIJA Organizam Min. ph Optim. ph Max. ph Lactobacillus acidophilus Escherichia coli Proteus vulgaris Enterobacter aerogenes Clostridium sporogenes Pseudomonas aeruginosa

5 REDOKS POTENCIJAL Eh VREDNOST Redoks potencijal (Eh) predstavlja meru koja pokazuje stepen oksidacije odnosno redukcije u namirnicama Eh predstavlja tendenciju supstrata da prima elektrone (redukcija) odnosno da otpušta elektrone (oksidacija) Redukciono sredstvo otpušta elektrone Oksidaciono sredstvo prima elektrone

6 Prenos elektrona sa jednog jedinjenja na drugo razlika u električnom potencijalu može da se meri i izražava se pomoću Eh vrednosti (mv) Supstrat u kome dominiraju oksidacioni procesi pozitivne vrednosti Eh Supstrat u kome dominiraju redukcioni procesi negativne vrednosti Eh

7 Redoks potencijal zavisi od: ph Hemijskog sastava namirnice Prisustva i međusobnog odnosa reduktivnih (SH grupe proteina, sumporvodonik, askorbinska kiselina, šećeri) i oksidativnih materija (nitrati i nitriti) Parcijalnog pritiska kiseonika Aktivnosti mikroorganizama Dodatak nitrata ili nitrita povećava se Eh Askorbinska kiselina smanjuje se Eh Zagrevanje mesa smanjuje se Eh (oslobađanje H 2 S) Razmnožavanje mikroorganizama smanjuje se Eh (troši se kiseonik)

8 Obligatni AEROBI rastu samo pri POZITIVNIM Eh vrednostima (prouzrokovači površinskog kvara namirnica Pseudomonadaceae, Kvasci, Plesni) Obligatni ANAEROBI rastu samo pri NEGATIVNIM Eh vrednostima (Clostridium) Fakultativni anaerobi rastu i pri pozitivnim i negativnim vrednostima Eh (Enterobacteriaceae, Staphylococcaceae) Mikroaerofili rastu u uslovima smanjenog sadržaja O 2 i povećanog sadržaja CO 2 (Lactobacillus)

9 AKTIVNOST VODE - aw Voda je neophodna za metabolizam i rast mikroorganizama s obzirom da čini 80 do 90 % njihove mase. AKTIVNOST VODE predstavlja odnos između ravnotežnog pritiska vodene pare u namirnici (p) i pritiska zasićene pare (p s ) pri istoj temperaturi: a w = p / p s ili a w = relativna vlažnost vazduha (%) / 100 a w vrednost se izražava brojevima od 0 (apsolutno suva matrija) do 1 (destilovana voda).

10 AKTIVNOST VODE - aw ak vnost vode sadržaj vode!!! Pojedine namirnice mogu imati isti sadržaj vode ali različite vrednosti aktivnosti vode Voda koja nije vezana za molekule namirnice dostupna je mikroorganizmima Aktivnost vode pokazuje u kojoj meri je voda strukturno ili hemijski vezana u namirnicama, odnosno u kojoj meri je dostupna mikroorganizmima

11 AKTIVNOST VODE - aw Aktivnost vode zavisi od: Sadržaja vode Vrste i količine u vodi rastvorljivih materija (soli, šećeri, neproteinska azotna jedinjenja) Načina vezivanja vode u namirnici (proteini, ugljeni hidrati, dispergovanje vode u emulzijama..) Snižavanje aktivnosti vode postiže se: Sušenjem ili smrzavanjem opada sadržaj vode, a raste omotski pritisak usled povećanja koncentracije rastvorljivih materija Soljenjem i dodavanjem drugih materija rastvorljivih u vodi povećava se osmotski pritisak

12 AKTIVNOST VODE - aw Uticaj osmotskog pritiska na mikroorganizme Hipertonični rastvor voda izlazi iz ćelije Hipotonični rastvor voda ulazi u ćeliju Hipertonični rastvor Hipotonični rastvor Izotonični rastvor Strelice pokazuju kretanje vode putem osmoze

13 AKTIVNOST VODE - aw Značaj aktivnosti vode u mikrobiologiji namirnica Snižavanjem aktivnosti vode može se onemogućiti rast i razmnožavanje mikroorganizama aktivnost vode utiče na rast, sporulaciju, germinaciju, produkciju toksina, enzimske reakcije, denaturaciju proteina, geliranje membrane, degradaciju vitamina, oksidaciju masti itd. Merenjem akivnosti vode u namirnicama moguće je predvideti koji mikrooranizmi mogu biti potencijalni uzročnik kvara ili infekcije odn. trovanja

14 Opseg aktivnosti vode Mikroorganizmi koji su inhibirani najmanjom a w -vrednošću u ovom opsegu 1,00 0,95 Pseudomonas, Escherichia, Proteus, Shigella, Klebsiella, Bacillus, Clostridium perfrigens, neki kvasci 0,95 0,91 Salmonella, Vibrio, C. Botulinum, Serratia, Lactobacillus, Pediococcus, neke plesni i kvasci 0,91 0,87 Većina kvasaca (Candida, Torulopis, Hansenula), Micrococcus 0,87 0,80 Staphylococcus aureus, većina plesni (mikotoksinogene Penicillium-vrste), kvasci Saccharomyces spp, Debaryomyces 0,80 0,75 Većina halofilnih bakterija, mikotoksinogene Aspergillus-vrste Namirnice čija je a w -vrednost u ovom opsegu Sveža hrana, konzervirano voće, povrće, meso, riba i mleko; hrana koja sadrži prosečno 40 % sukroze ili 7 % NaCl-a Neki sirevi, salamureno meso, voćni koncentrati; hrana koja sadrži 55 % sukroze ili 12 % NaCl-a Fermentisane suve kobasice, suvi sirevi, margarin; Hrana koja sadrži 65 % sukroze ili 15 % NaCl-a Suve šunke, većina voćnih koncentrata, zaslađeno kondenzovano mleko, čokoladni sirupi, brašno, pirinač; voćni kolači Džem, marmelada, marcipan.. 0,75 0,65 Kserofilne plesni Suvo voće, lešnik, šećerna trska.. Hrana koja sadrži prosečno 10 % vode 0,65 0,60 Osmofilni kvasci (Saccharomyces rouxii) i neke plesni (Aspergillus chinulatus, Monascus bisporus) Suvo voće koje sadrži 15-20% vode, med, karamele 0,20 0,50 Nije moguće razmnožavanje mikroorganizama Suve testenine, začini, jaja u prahu, mleko u prahu, keks, suvo povrće, krekeri International Conference on Food Safety, 1998

15 ANTIMIKROBNI SASTOJCI LIZOCIM, LAKTOFERIN, LAKTOPEROKSIDAZA H 2 O 2 TIOCIJANAT SISTEM

16 TEMPERATURA

17

18 Market Risk Fiscal Year 2004 Fiscal Year 2005 Annual Report Quarter Ended 1 st Qtr. 2 nd Qtr. 3 rd Qtr. 4 th Qtr. Total Fiscal year 2003 Revenue $7,746 $8,541 $7,835 $8,065 $32,187 Gross profit 6,402 6,404 6,561 6,761 26,128 Net income 2,041 1,865 2,142 1,483 7,531 Basic earnings per share Diluted earnings per share Fiscal year 2004 Revenue $8,215 $10,153 $9,175 $9,292 $36,835 Gross profit 6,735 7,809 7,764 7,811 30,119 Net income 2,614 1,549 1,315 2,690 8,168 Basic earnings per share Diluted earnings per share Fiscal year 2005 Revenue $9,189 $10,818 $9,620 $10,161 $39,788 Gross profit 7,720 8,896 8,221 8,751 33,588 Net income 2,528 3,463 2,563 3,700 12,254 Basic earnings per share Diluted earnings per share (1) Includes charges totaling $750 million (pre tax) related to the Fabrikam settlement and $1.15 billion in impairments of investments. (2) Includes stock based compensation charges totaling $2.2 billion for the employee stock option transfer program. (3) Includes charges totaling $756 million (pre tax) related to Contoso subsidiaries and other matters.

19 1 st Year Sales Figures Market Distribution Financial Summary Our perennial 3 rd Quarter boost was larger than expected contributing to an exceptionally strong year. Distribution of sales across the geographic markets looks fairly steady. Our new product line, released this year, is early in its adoption phase. We expect to see substantial revenue contribution from these products over the next two years.

20 RELATIVNA VLAŽNOST DEFINICIJA... Relativna vlažnost je termin kojim opisujemo količinu vodene pare u mešavini vazduha i vode FORMULA RH = 100 x P Po a w Relativna vlažnost je mera kojom se ukazuje na količinu vodene pare u vazduhu (pri određenoj temperaturi) [P] u odnosu na maksimalnu količinu vodene pare koju vazduh može da primi pri toj temperaturi [Po] i izražava se u procentima.

21 RELATIVNA VLAŽNOST SREDINE KAO FAKTORA ZNAČAJ RH uslovi skladištenja 1 RH Primer 1. a w RH 2 Niska a w vrednost proizvoda + visok % RH (uslovi skladištenja) = adsorpcija vode namirnicom Primer 2. Visoka a w vrednost proizvoda + nizak % RH (uslovi skladištenja)= desorpcija vode/isparavanje iz namirnice Do postizanja RAVNOTEŽE A W = RV

22 SREDINE KAO FAKTORA ZNAČAJ RH RELATIVNA VLAŽNOST TAČKA ROSE je ona temperatura pri kojoj vazduh prilikom hlađenja postaje zasićen i u tom trenutku počinje izdvajanje magle ili rose na površinama (namirnica!!!) Tačka rose zavisi od tri vrednosti: Stimuliše razvoj mikroorganizama T vazduha RH BAKTERIJE KVARA Zahtevaju visoku a w Pseudomonas Acinetobacter Moraxella T proizvoda Mogu se razmnožavati i pri nižim vrednostima a w bacili, kvasci, plesni

23 UTICAJ KORISNE MIKROFLORE MEHANIZMI ANTAGONIZMA BMK Antagonizam inhibicija drugih (MO kvara ili patogenih vrsta) organizama putem kompeticije za hranjive materije, ili produkcijom metabolita sa antimikrobnim delovanjem Mikrobna interferencija efikasni nespecifični mehanizam kontrole uobičajen za sve populacije i sredine (niše) uključujući i namirnice; inhibicija rasta određenih MO od strane drugih članova habitata i to na osnovu kompeticije za hranjive materije, ostvarivanja nepovoljne sredine te kompeticije za adheziona mesta

24 ANTIMIKROBNO DELOVANJE BMK Produkt Glavni target organizmi Organske kiseline Mlečna kiselina Sirćetna kiselina Truležne i Gram negativne bakterije, pojedine plesni Truležne bakterije, klostridije, pojedini kvasci i plesni Hidrogen peroksid Enzimi Laktoperoksidaza sistem sa H 2 O 2 Lizozim (tehnologija rekombinantne DNK) Metaboliti male molekulske mase Reuterin (3-OH-propionaldehid) Diacetil Masne kiseline Patogeni i MO kvara posebno u namirnicama bogatim na sadržaju belančevina Patogeni i MO kvara (mleko i proizvodi od mleka) Nepoželjni Gram pozitivni MO Široki spektar bakterija, kvasci i plesni Gram negativne bakterije Različite bakterije Bakteriocini nizin Pojedine BMK i Gram pozitivne bakterije, posebno sporogeni MO

25 BAKTERIOCINI Heterogena grupa antimikrobnih proteina i peptida koji deluju antagonistički primarno prema blisko srodnim vrstama MO Klasifikacija (Klaenhammer, 1988) Klasa I (lantibiotici; mali, termostabilni, sadrže neuobičajene AK) Klasa II (mali termostabilni peptidi): IIapediocinu slični bakteriocini sa antilisterija efektom; IIbdvokomponentni bakteriocini; IIc ostali bakteriocini Klasa III veliki (>30kDa) termolabilni proteini Klasa IV kompleksni bakteriocini koji sadrže ugljenohidratnu i/ili lipidnu komponentu

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