Enterokoki: yin - yang mikrobi

This review deals with the duality of enterococci, which can be illustrated by their yin - yang behaviour. The rough nature of this specific group of lactic acid bacteria promotes their dissemination in various environments where they significantly influence the outcome of a certain process. In the technological meaning, enterococci are leading microbes in fermentation processes of traditional foods, where their detrimental spoilage activities are equally significant. As therapeutics, enterococci manifest the probiotic properties through their positive effects on maintaining of the normal intestinal microflora, on stimulation of the immune system, on improved nutritional value of food and with the production of antimicrobial compounds (bacteriocins). At the same time, enterococci present an emerging pool of opportunistic pathogens for humans as they cause disease, possess agents for antibiotic resistance and their transfer mechanisms, and are frequently armed with potential virulence factors. Despite the yin - yang characteristics of enterococci, the long history of safe use of certain strains of enterococci in food/feed, and reliable identification and classification of enterococci with phenotypic methods supported with modern genetic tools, enables selection of promising enterococci, which could be safely used as starter cultures or food/feed additives.Ovaj revijalni prikaz opisuje dualizam enterokoka kojega možemo ilustrirati njihovim yin-yang ponašanjem. Otporna narav ove veoma specifične skupine bakterija mliječne kiseline omogućava njihovu proširenost u različitim sredinama gdje signifikantno utječu na ishod pojedinih procesa. Tako su u tehnološkom smislu enterokoki vodeći mikrobi u fermentacijskim procesima a istovremeno su i dobro poznati uzročnici kvarenja hrane. Kao terapeutici, enterokoki manifestiraju svoja probiotička svojstva pozitivnim djelovanjem na održavanje normalne intestinalne mikroflore simulirajući imunološki sustav, te poboljšavaju nutritivnu vrijednost hrane proizvodnjom antimikrobnih supstanci (bakteriocina). Pored toga enterokoki predstavljaju rastući rezervoar oportunističkih patogena za ljude jer izazivaju bolesti, sadrže činitelje za antibiotičnu rezistenciju i mehanizme njihova transfera, te često nose i potencijalne virulentne faktore. Ali usprkos yin-yang naravi enterokoka, duga povijest bezopasne upotrebe pojedinih enterokoka u hrani i krmi, te pouzdana identifikacija i klasifikacija enterokoka fenotipskim metodama poduprtim modernim genetskim tehnikama, omogućava selekciju obećavajućih enterokoka, koje bi mogli, bez posebnog rizika po zdravlje čovjeka, upotrijebiti kao starterske kulture ili dodatke hrani i krmi

Utjecaj antibiotika na bakterije mliječne kiseline probavnog sustava

Lactic acid bacteria (LAB) are common inhabitants of the gastrointestinal (GI) tract and have important role in maintaining the equilibrium of GI flora, which can be influenced by various factors like diets, antimicrobials and stress. Minimal inhibitory concentrations (MIC) and minimal bactericidal concentrations (MBC) of 6 antibiotics, commonly used in human medicine for 8 selected lactobacilli strains were determined by macrodilution and microdilution methods in liquid media and by diffusion method on agar plates. The effects of Penicillin G and Ampicillin on intestinal LAB were tested in vivo on mice as well. Lactobacilli were sensitive to Penicillin G, (penicillines and their derivatives) and Erythromycin (macrolides) by in vitro testing. Clyndamycin (pyranosid) showed moderate inhibitory effect. All lactobacilli strains were resistant to Kanamycin and Neomycin (aminoglycosides), while L. salivarius IM 124 has shown extra resistance to Erythromycin and Clyndamycin. The influence of orally administered Ampicillin showed no significant influence on LAB count in mice faeces. The effect of Penicillin G on mice LAB total count was significant, while no effect of orally administered lactobacilli was determined.Bakterije mliječne kiseline (BMK) su uobičajeni sudionici crijevne mikroflore ljudi i životinja. One imaju važnu ulogu u održavanju ravnoteže gastrointestinalne (GI) flore, na koju mogu utjecati različiti faktori poput prehrane (hrane) antimikrobnih supstancija i stresa. Pomoću makrodilucijske i mikrodilucijske metode u tekućoj podlozi te metode difuzije na agar pločama, određene su minimalne inhibicijske koncentracije (MIC) i minimalne baktericidne koncentracije (MBC) šest (6) antibiotika koji se svakodnevno upotrebljavaju u humanoj medicini za 8 sojeva BMK. Učinak penicilina G i ampicilina na BMK probavnog sustava testiran je također in vivo na laboratorijskim miševima. Laktobacili su se pokazali osjetljivim na penicilin G, na ampicilin (derivati penicilina) i na eritromicin (makrozidni antibiotik) prilikom testiranja in vitro. Klindamicin (piranozidni antibiotik) je pokazao umjereni inhibicijski učinak. Svi sojevi BMK pokazali su se otpornim na kanamicin i neomicin (aminoglikozidni antibiotici). Od svih testiranih BMK posebno se otpornim na eritromicin i klindamicin pokazao L. salivarius IM 124. Učinak oralno primjenjenog ampicilina, na broj BMK u stolici miševa nije statistički dokazan. Utjecaj penicilina G na broj BMK kod miševa pokazao se signifikantnim, ali to nije bio slučaj s oralno primijenjenim BMK

Kompetitivna prednost bakteriocinogenih sojeva konzorcija bakterija mliječne kiseline izoliranih iz sira proizvedenog od sirovog mlijeka

The presence of gene determinants for different bacteriocins has been already demonstrated in traditional Slovenian types of raw milk cheeses ‘Tolminc’ and ‘Kraški’. These genes were present also in the cultivable microbiota. In this research the aim was to establish how the presence of gene determinants for bacteriocins in microbial consortia is reflected in its antimicrobial activity. In addition, one of the goals was to determine whether the strains that carry gene determinants for bacteriocins have any competitive growth advantage in microbial population. Microbial consortium of ‘Tolminc’ cheese was propagated in milk and examined at the end of propagation its antimicrobial activity and the presence of gene determinants for bacteriocins. Comparison of the results obtained before and after propagation leaded to the conclusion that most of the strains possessing gene determinants for bacteriocins were unable to persist during propagation. The strains which did persist during propagation carried gene determinants for enterocins P, L50B and cytolysin. Antimicrobial activity of consortium before and after propagation was not substantially different and cannot be attributed to any of detected bacteriocins.Prisutnost genskih determinanti za stvaranje različitih vrsta bakteriocina već je ranijim istraživanjima utvrđena u slovenskim ovčjim sirevima “Tolminac” i “Kraški sir” koji se proizvode na tradicionalan način. Isti geni za izoliranu mikrobnu populaciju (konzorcij) također su potvrđeni i u kultiviranim uvjetima. U ovom radu pokušalo se utvrditi kako se prisutnost tih genskih determinanti u mikrobnom konzorciju odražava na njegovu antimikrobnu aktivnost. Osim toga, utvrđivao se i utjecaj bakterijskih sojeva koji nose gene za stvaranje bakteriocina na kompetitivnost rasta u mješovitoj populaciji. Mikrobni konzorcij izoliran iz sira “Tolminac” propagiran je u mlijeku 10 dana, nakon čega je utvrđena antimikrobna aktivnost kulture i prisutnost genskih determinanti za stvaranje bakteriocina. Usporedbom rezultata za sposobnost stvaranja bakteriocina prije i nakon precjepljivanja u mlijeku, utvrđen je gubitak te sposobnosti za većinu izoliranih sojeva. Sojevi koji su izdržali ponovljena precjepljivanja u mlijeku nosili su genske determinante za bakteriocine: enterocin P, enterocin L50B i citolizin. Antimikrobna aktivnost konzorcija prije i nakon precjepljivanja nije se značajno razlikovala te se ne može pripisati niti jednom od u ovom pokusu potvrđenih bakteriocina

In-situ inhibicija Staphylococcus aureus bakterijama mliječne kiseline iz dva tradicionalna slovenska sira proizvedena od sirovog mlijeka

Bacteriocin(s) producing lactic acid bacteria naturally present in traditional cheeses represent an inexhaustive pool of microbes with safeguarding potential. Some bacteriocins produced by cheese lactic acid bacteria were already described as successful anti-staphylococcal agents. The presence of genes for bacteriocins with potential anti-staphylococcal activity was also demonstrated in two Slovenian traditional raw milk cheeses, “Tolminc” and “Kraški ovčji sir”. Same bacteriocin genes were also detected in viable lactic acid bacteria consortia’s isolated from “bacteriocin positive cheeses” on Rogosa, M17 and CATC agar media. The aim of the research was to elucidate whether or not this particular cheese consortia, in which bacteriocin genes were detected, actually exhibit anti-staphylococcal activity in milk and/or cheese. For this purpose different cheese consortia were selected in relation to versatility of detected bacteriocin genes and used to perform challenge tests against Staphylococcus aureus in milk and cheese. In milk following the time/temperature regime of traditional cheese production all cheese consortia effectively inhibited growth of Staphylococcus aureus in the range of app. 2 to 3 log. In cheese the inhibition of staphylococci was less pronounced but still evident since inhibition of app. 1.5 log was detected. Sole inhibition by lactic acid production was ruled out while linking inhibition directly to bacteriocin production would take some additional work.Bakterije mliječne kiseline (BMK), prirodno prisutne u tradicionalnim sirevima, predstavljaju neograničen izvor mikroba sa zaštitnim svojstvima. Za neke bakteriocine, koje proizvode BMK iz sira, antistafilokokna svojstva već su poznata. Prisutnost gena za bakteriocine s antistafilokoknim svojstvima potvrđena je u dva tradicionalna slovenska sira iz sirovog mlijeka; to su sir “Tolminc” i “Kraški ovčji sir”. Prisutnost istih bakteriocina potvrđena je i kod bakterija mliječne kiseline, koji su izolirali iz tih sireva na agar podlogama Rogosa, M17 i CATC. Cilj je bio provjeriti da li izolirani sojevi bakterija mliječne kiseline, koji nose gene za bakteriocine, zaista pokazuju antistafilokokna svojstva u mlijeku i/ili siru. Na temelju prisutnih gena za bakteriocine ispitana je antimikrobna aktivnost združene kulture bakterija mliječne kiseline prema bakteriji Staphylococcus aureus u mlijeku i siru. U mlijeku kojeg smo podvrgnuli vremensko-temperaturnom režimu proizvodnje tradicionalnih sireva, združena kultura bakterija mliječne kiseline inhibirala je rast bakterije Staphylococcus aureus u rasponu od 2 do 3 logaritamske jedinice. U siru je inhibicija stafilokoka bila nešto manje izražena, ali još uvijek očita sa 1,5 logaritamskih jedinica slabijim rastom S. aureus nego u kontrolnom siru. Učinak inhibicije izazvane mliječnom kiselinom bio je isključen, ali bi bilo potrebno izvesti još dodatna ispitivanja, da se utvrdi izravno inhibicijsko djelovanje bakteriocina na stafilokoke

Kompetitivna prednost bakteriocinogenih sojeva konzorcija bakterija mliječne kiseline izoliranih iz sira proizvedenog od sirovog mlijeka

The presence of gene determinants for different bacteriocins has been already demonstrated in traditional Slovenian types of raw milk cheeses ‘Tolminc’ and ‘Kraški’. These genes were present also in the cultivable microbiota. In this research the aim was to establish how the presence of gene determinants for bacteriocins in microbial consortia is reflected in its antimicrobial activity. In addition, one of the goals was to determine whether the strains that carry gene determinants for bacteriocins have any competitive growth advantage in microbial population. Microbial consortium of ‘Tolminc’ cheese was propagated in milk and examined at the end of propagation its antimicrobial activity and the presence of gene determinants for bacteriocins. Comparison of the results obtained before and after propagation leaded to the conclusion that most of the strains possessing gene determinants for bacteriocins were unable to persist during propagation. The strains which did persist during propagation carried gene determinants for enterocins P, L50B and cytolysin. Antimicrobial activity of consortium before and after propagation was not substantially different and cannot be attributed to any of detected bacteriocins.Prisutnost genskih determinanti za stvaranje različitih vrsta bakteriocina već je ranijim istraživanjima utvrđena u slovenskim ovčjim sirevima “Tolminac” i “Kraški sir” koji se proizvode na tradicionalan način. Isti geni za izoliranu mikrobnu populaciju (konzorcij) također su potvrđeni i u kultiviranim uvjetima. U ovom radu pokušalo se utvrditi kako se prisutnost tih genskih determinanti u mikrobnom konzorciju odražava na njegovu antimikrobnu aktivnost. Osim toga, utvrđivao se i utjecaj bakterijskih sojeva koji nose gene za stvaranje bakteriocina na kompetitivnost rasta u mješovitoj populaciji. Mikrobni konzorcij izoliran iz sira “Tolminac” propagiran je u mlijeku 10 dana, nakon čega je utvrđena antimikrobna aktivnost kulture i prisutnost genskih determinanti za stvaranje bakteriocina. Usporedbom rezultata za sposobnost stvaranja bakteriocina prije i nakon precjepljivanja u mlijeku, utvrđen je gubitak te sposobnosti za većinu izoliranih sojeva. Sojevi koji su izdržali ponovljena precjepljivanja u mlijeku nosili su genske determinante za bakteriocine: enterocin P, enterocin L50B i citolizin. Antimikrobna aktivnost konzorcija prije i nakon precjepljivanja nije se značajno razlikovala te se ne može pripisati niti jednom od u ovom pokusu potvrđenih bakteriocina

In-situ inhibicija Staphylococcus aureus bakterijama mliječne kiseline iz dva tradicionalna slovenska sira proizvedena od sirovog mlijeka

Bacteriocin(s) producing lactic acid bacteria naturally present in traditional cheeses represent an inexhaustive pool of microbes with safeguarding potential. Some bacteriocins produced by cheese lactic acid bacteria were already described as successful anti-staphylococcal agents. The presence of genes for bacteriocins with potential anti-staphylococcal activity was also demonstrated in two Slovenian traditional raw milk cheeses, “Tolminc” and “Kraški ovčji sir”. Same bacteriocin genes were also detected in viable lactic acid bacteria consortia’s isolated from “bacteriocin positive cheeses” on Rogosa, M17 and CATC agar media. The aim of the research was to elucidate whether or not this particular cheese consortia, in which bacteriocin genes were detected, actually exhibit anti-staphylococcal activity in milk and/or cheese. For this purpose different cheese consortia were selected in relation to versatility of detected bacteriocin genes and used to perform challenge tests against Staphylococcus aureus in milk and cheese. In milk following the time/temperature regime of traditional cheese production all cheese consortia effectively inhibited growth of Staphylococcus aureus in the range of app. 2 to 3 log. In cheese the inhibition of staphylococci was less pronounced but still evident since inhibition of app. 1.5 log was detected. Sole inhibition by lactic acid production was ruled out while linking inhibition directly to bacteriocin production would take some additional work.Bakterije mliječne kiseline (BMK), prirodno prisutne u tradicionalnim sirevima, predstavljaju neograničen izvor mikroba sa zaštitnim svojstvima. Za neke bakteriocine, koje proizvode BMK iz sira, antistafilokokna svojstva već su poznata. Prisutnost gena za bakteriocine s antistafilokoknim svojstvima potvrđena je u dva tradicionalna slovenska sira iz sirovog mlijeka; to su sir “Tolminc” i “Kraški ovčji sir”. Prisutnost istih bakteriocina potvrđena je i kod bakterija mliječne kiseline, koji su izolirali iz tih sireva na agar podlogama Rogosa, M17 i CATC. Cilj je bio provjeriti da li izolirani sojevi bakterija mliječne kiseline, koji nose gene za bakteriocine, zaista pokazuju antistafilokokna svojstva u mlijeku i/ili siru. Na temelju prisutnih gena za bakteriocine ispitana je antimikrobna aktivnost združene kulture bakterija mliječne kiseline prema bakteriji Staphylococcus aureus u mlijeku i siru. U mlijeku kojeg smo podvrgnuli vremensko-temperaturnom režimu proizvodnje tradicionalnih sireva, združena kultura bakterija mliječne kiseline inhibirala je rast bakterije Staphylococcus aureus u rasponu od 2 do 3 logaritamske jedinice. U siru je inhibicija stafilokoka bila nešto manje izražena, ali još uvijek očita sa 1,5 logaritamskih jedinica slabijim rastom S. aureus nego u kontrolnom siru. Učinak inhibicije izazvane mliječnom kiselinom bio je isključen, ali bi bilo potrebno izvesti još dodatna ispitivanja, da se utvrdi izravno inhibicijsko djelovanje bakteriocina na stafilokoke

Karakterizacija bakteriocinogenih sojeva bakterija mliječne kiseline iz tradicionalnog slovenskog sira ‘Tolminc’

The purpose of this research was to examine traditional Slovenian ‘Tolminc’ cheese for the presence of lactic acid bacteria that produce several bacteriocins. The presence of gene determinants for different bacteriocins in this type of cheese and in the cultivable population of ‘Tolminc’ microbiota, have already been demonstrated, as well as its antimicrobial activity. Due to the difficulties in connecting the presence of gene determinants for bacteriocins with the observed antimicrobial activity it was decided to examine in this study the same features on the level of individual bacteriocinogenic strains. Like in previous results, enterococci and their bacteriocins prevailed in cheese microbial consortia. None of isolated strains inhibited growth of Staphylococcus aureus, while the other indicator strains were inhibited in a strain specific manner. Most of isolated strains carried gene determinants for cytolysin. On the basis of gene determinants for bacteriocins, antimicrobial activity, phenotyping by PhP (PhenePlateTM) system and PCR identification, some similarities found were among Enterococcus isolates.Cilj ovog rada bio je provjeriti prisutnost sojeva bakterija mliječne kiseline koji proizvode različite bakteriocine u tradicionalnom slovenskom siru ‘Tolminc’. Prisutnost genskih determinanti za pojedine bakteriocine u ovoj vrsti sira i izoliranoj populaciji mikrobiote sira ‘Tolminc’ već je bila prikazana, a također i njihova antimikrobna aktivnost. Zbog poteškoća pri povezivanju detektiranih genskih determinanti za bakteriocine i antimikrobne aktivnosti, odlučeno je u ovom radu analizirati ista svojstva i kod pojedinačnih bakteriocinogenih sojeva. Slično prethodnim istraživanjima, enterokoki i njihovi bakteriocini najbolje su bili zastupljeni. Nijedan od izoliranih sojeva nije inhibirao bakteriju Staphylococcus aureus, dok su ostale indikatorske mikroorganizme inhibirali različito. Većina sojeva nosila je genske determinante za bakteriocin citolizin. Na temelju genskih determinanti za bakteriocine, antimikrobne aktivnosti, fenotipizacije s PhP (PhenePlateTM) sistemom i identifikacije roda i vrste sojeva, mogu se naći neke sličnosti između Enterococcus sojeva

Influence of two feed supplements on technological properties of goat’s milk

Hranidba koza na dnevnoj bazi sastoji se od ispaše, sijena i/ili suplemenata. Suplementi su neophodni u hranidbi visokoproduktivnih pasmina koza u svrhu postizanja genetskog potencijala i uzgajivači moraju odabrati režim hranidbe koji osigurava proizvodnju velikih količina mlijeka bez neželjenih promjena u tehnološkoj kvaliteti mlijeka. U ovom istraživanju određivan je utjecaj dva komercijalno dostupna suplementa na sposobnost koagulacije kozjeg mlijeka te na reološka svojstva jogurta od kozjeg mlijeka. Ukupno 61 koza pasmine slovenska alpska koza hranjena je s dva suplementa tijekom tri godine. Prvi suplement (FS1) imao je viši udjel ječma i lucerne, dok je drugi suplement (FS2) imao dodane mineralne tvari i vitamine te više udjele pšenice i suncokretove pogače. Posljedično, FS1 je imao više sirovih vlakana, što je vrlo vjerojatno rezultiralo 15 % većom čvrstoćom, konzistencijom i kohezivnosti (P<0,05) jogurta iz skupine FS1 u odnosu na jogurte iz skupine FS2. K tomu je i vrijeme koagulacije (r) bilo kraće (P<0,05) kod jogurta iz skupine FS1 u usporedbi s jogurtima iz skupine FS2. Čvrstoća gruša 30 min nakon dodatka enzima (a30) također je bila veća u skupini FS1, iako te razlike nisu bile statistički značajne. Uzimajući sve navedeno u obzir, dobiveni rezultati ukazuju kako su koze hranjenje suplementom FS1 proizvodile mlijeko boljih tehnoloških svojstava nego koze hranjene suplementom FS2, unatoč činjenici da nisu utvrđene statistički značajne razlike u kemijskom sastavu mlijeka iz obiju navedenih skupina. Ova studija pokazala je da se pažljivim odabirom suplementa u hranidbi mogu poboljšati tehnološka svojstva kozjeg mlijeka. Međutim, potrebno je provođenje daljnjih istraživanja u svrhu određivanja specifičnih mehanizama koji su doveli do prethodno navedenih razlika.Goat’s daily diet is usually based on grazing, hay and/or feed supplements. Feed supplements are crucial in the diet of high productive goats to achieve their genetic potential and breeders must choose balanced feeding regime to produce large quantities of milk without affecting the technological quality of milk. In the present study, we evaluated the influence of two commercially available feed supplements on goat milk coagulation properties and rheological properties of yoghurts. Goats of the Slovenian Alpine breed (61) were fed with two feed supplements during the 3-year experiment. Feed supplement 1 (FS1) had higher proportions of barley and alfalfa, while feed supplement 2 (FS2) had added premix of minerals and vitamins and had higher proportions of wheat and sunflower meal. Con¬sequently, FS1 had more crude fibres, which is the most probable reason for approximately 15 % higher firmness, consistency and cohesiveness (P<0.05) of yoghurts in FS1 group, compared to the FS2 group. Moreover, the rennet coagulation time (r) was shorter (P<0.05) in the FS1 group, compared to the FS2 group. Curd firmness 30 min after enzyme addition (a30) was also higher in FS1 group although the results were not statistically significant. Taking together, our results indicate that goats fed with FS1 produced milk with better technological properties compared to those fed with FS2, despite the fact that there were no significant differences in chemical composition of milk from each group. We showed that careful selection of feed supplement’s constituents could improve technological properties of goat milk. However, further studies are needed to evaluate the mechanisms of the observed differences

Fenotipska i genotipska karakterizacija autohtone zajednice laktobacila izolirane iz tradicionalnog istarskog ovčjeg sira

The objective of this study is to describe the diversity of indigenous cultivable community of the lactobacilli associated with the production of traditional Istrian cheese and to get a collection of well characterized strains. Raw milk and cheese samples were collected from three different farms in Istria during ripening. A total of 212 mesophilic and thermophilic Lactobacillus isolates as well as bulk colonies (consortia) were investigated using culture-dependent approach combining phenotyping and genotyping. Biochemical fingerprinting with PhenePlate-LB system preliminary grouped 212 isolates in 16 distinct PhP types. Only one representative isolate from each PhP cluster was further analyzed by genotyping for a reliable identification at the genus and species level by employing PCR techniques and sequencing of 16S rRNA genes. Sequence analysis of 16S rRNA revealed the presence of Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus casei, Lactobacillus paracasei and Lactobacillus rhamnosus. Lactobacilli were screened for possible resistance against seven selected antibiotics: ampicillin, tetracycline, penicillin, rifampin, clindamycin, erythromycin and vancomycin. Although there was no clear pattern of antimicrobial susceptibility to most tested antibiotics, all representative isolates were resistant to vancomycin. The analysis of bulk colonies by denaturing gradient gel electrophoresis (DGGE) identified Lactobacillus plantarum and Lactobacillus brevis as predominant members of Lactobacillus population. Pediococcus pentosaceus, Pediococcus acidilactici, Streptococcus sp. and Leuconostoc mesenteroides were also detected as part of the analysed consortia. The prevalence of identified species and community members of lactobacilli agrees with other studies of raw milk cheese and represents a useful base for further selection.Svrha je ovog istraživanja bila utvrditi raznolikost autohtone mikrobne zajednice laktobacila u istarskom siru te detaljno genetički definirati sojeve. Uzorkovanje mlijeka i svježeg sira provodilo se na tri obiteljska poljoprivredna gospodarstva na području Istre. Ukupno 212 izolata mezofilnih i termofilnih laktobacila, te ukupna zajednica (konzorcij) istraženi su pomoću fenotipskih i genotipskih metoda ovisno o izolaciji na selektivnim hranjivim podlogama. Primjenom biokemijskog testa PhenePlate-LB, 212 izolata preliminarno je svrstano u 16 skupina prema načinu korištenja supstrata. Po jedan reprezentativni izolat laktobacila iz svake PhP skupine genotipski je analiziran PCR analizom i sekvencioniranjem 16S rRNA gena te je određeno kojem rodu i vrsti pripadaju. Sekvencijskom su analizom određene ove vrste: Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus casei, Lactobacillus paracasei i Lactobacillus rhamnosus. Dodatno je testirana osjetljivost svakog reprezentativnog izolata na sljedeće antibiotike: ampicilin, tetraciklin, penicilin, rifampicin, klindamicin, eritromicin i vankomicin. Kod svih izolata utvrđena je rezistencija na vankomicin, iako nije jasno utvrđen mehanizam osjetljivosti na ostale testirane antibiotike. PCR DGGE analizom ukupne zajednice laktobacila (konzorcija) utvrđeno je da prevladavaju vrste Lactobacillus plantarum i Lactobacillus brevis. Također su pronađene vrste Pediococcus pentosaceus, Pediococcus acidilactici, Streptococcus sp. i Leuconostoc mesenteroides. Prisutnost određenih vrsta u kompleksnoj mikrobnoj zajednici laktobacila istarskog sira potvrđuje rezultate brojnih studija o tradicijskim sirevima, što predstavlja važan autohtoni genetički potencijal za daljnju selekciju izolata