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Tarhana is produced at batch systems in which the microbiota has changed accordingly to the microbial load from ingredients. In order to stabilize the microbiota, the effects of backslopping carried out under different temperature regimes (25 and 30 degrees C), pH (3.70 and 4.00) and inoculation rates (5, 10 and 15%) on lactic acid bacteria (LAB) diversity were determined in tarhana dough. LAB and Total Aerobic Mesophilic Bacteria (TAMB) numbers increased in all tarhana dough samples subjected to backslopping. Temperature and pH significantly affected the microbiological diversity of tarhana whereas the different inoculation rates did not. Tarhana dough showed complex tarhana microbiota following backslopping at pH 4.00 independently on the temperature applied. When backslopping was carried out at pH 3.70 and 25 degrees C, tarhana microbiota stabilized and became steady after several cycles. The LAB species found in all dough samples after the final backslopping were Lactobacillus plantarum, Lactobacillus alimentarius and Lactobacillus brevis which were able to carry out the fermentation in all conditions tested. In order to obtain a stable presence of LAB populations at industrial level for tarhana production, this work showed that backslopping is recommended at pH 3.70 and 25 degrees C with any inoculation ratios. C1 [Ozel, Burcu; Erten, Huseyin] Cukurova Univ, Fac Agr, Dept Food Engn, TR-01330 Adana, Turkey. [Ozel, Burcu] Univ Pamukkale, Cal Vocat High Sch, Dept Food Proc, TR-20700 Denizli, Turkey. [Simsek, Omer] Univ Pamukkale, Fac Engn, Dept Food Engn, TR-20160 Denizli, Turkey. [Settanni, Luca] Univ Palermo, Dipartimento Sci Agr Alimentari & Forestali, Viale Sci 4, I-90128 Palermo, Italy.

The ability ofLactobacillus plantarumto produce exopolysaccharides (EPS) of various structures and properties is effective in showing both starter and probiotic culture qualification. In this study, the potential health promoting functions of the ropy EPS produced byLactobacillus plantarumstrains isolated from tarhana were tested. A stimulation of the pro-inflammatory IL-12 and TNF-alpha cytokines was observed in the presence of the ropy EPS suggesting an in vitro immune modulation. Similarly, the tested EPS demonstrated promoted the growth of the probiotic strains in fermentation medium. A medium level of radical scavenging activities of ropy EPS was observed whereas the superoxide and hydroxyl scavenging activities were more effective. The ropy EPS also showed alpha-glucosidase inhibition and cholesterol removal characteristics depending on their concentration. These findings revealed the potential health-promoting functions of ropy EPS fromL. plantarumstrains and EPS fromL. plantarumPFC311 and PFC310 strains demonstrated multiple health-improving effects that can be further evaluated in food and other industries. C1 [Yilmaz, Tulin; Simsek, Omer] Univ Pamukkale, Dept Food Engn, TR-20070 Denizli, Turkey.

Tarhana is a traditional cereal product fermented by lactic acid bacteria (LAB) and yeast strains that has gained special interest recently as an infant nutrition. Tarhana contains wheat flour, yogurt, and various vegetables that might create a microbiological toxicological risk, especially forBacillus cereusandStaphylococcus aureus. In this study, characterization of the metabolites responsible for antibacterial activity ofPediococcus acidilacticiPFC69 andLactococcus lactisPFC77 strains obtained from tarhana was performed, and antibacterial effects were detected againstB. cereusATCC 11778 andS. aureusATCC 29213 during the fermentation. A total of 12,800 AU/mL antibacterial activity was observed for the supernatants of the PFC69 and PFC77 strains that were found to be stable at high temperature and in low pH conditions and sensitive to proteases, suggesting the antimicrobial metabolite is a bacteriocin. These bacteriocins were further purified and their molecular sizes were determined as 4.5 and 3.5 kDa, respectively. Importantly, inoculation of PFC69 and PFC77 to tarhana dough significantly decreasedB. cereusATCC 11778 andS. aureusATCC 29213 amounts from the fifth day of fermentation compared to the control dough samples.P. acidilacticiPFC69 andL. lactisPFC77 strains were concluded as bioprotective cultures for tarhana and these strains were offered for other cereal-based fermentations. C1 [Kaya, Halil Ibrahim] Univ Bayburt, Dept Food Engn, TR-69000 Bayburt, Turkey. [Simsek, Omer] Univ Pamukkale, Dept Food Engn, TR-20017 Denizli, Turkey.

Exopolysaccharide producing starter cultures enable manufacturing "clean labeled" foods with improved textural and nutritional properties. The structural and technological analyses were performed on the ropy exopolysaccharides of six Lactobacillus plantarum. The incubation temperature, time and pH affected the exopolysaccharide production and high exopolysaccharide was produced in the presence of sucrose and maltose. The viscosity of exopolysaccharide was high at acidic conditions except PFC311E that showed viscous at neutral pH. Lactobacillus plantarum strains produced between 120 and 400 mg/L exopolysaccharide in which the highest was observed at L. plantarum PFC311. Exoploysaccharides were degraded over 300 degrees C except PFC311E that degraded at 295.7 degrees C. The NMR analyses revealed that the exopolysaccharides were synthesized by alpha 1-6, alpha 1-3 and alpha 1-4 bonds with glucose, galactose and fructose moieties. In conclusion, L. plantarum PFC311 produced ropy exopolysaccharide with different structural, rheological and thermal properties and reveals potential to be used in food industry. C1 [Zehir Senturk, Duygu; Simsek, Omer] Univ Pamukkale, Dept Food Engn, TR-20160 Denizli, Turkey. [Dertli, Enes] Univ Bayburt, Dept Food Engn, TR-69000 Bayburt, Turkey. [Erten, Huseyin] Univ Cukurova, Dept Food Engn, TR-01330 Adana, Turkey.

Tarhana is a traditional food produced by fermentation of a dough prepared by mixing wheat flour, yoghurt, tomato, paprika, onion and mint. The fermentation of mixture of natural products gives the tarhana an aromatized desired food characteristic. Therefore, we aimed to determine the effects of fermentation time at home and commercial scale on the aromatic volatile profile of tarhana. In this respect, nine different tarhana dough samples were prepared at home and commercial scale and analysed for volatile and organic acid composition during fermentation which were then evaluated by principal component analysis. In all dough samples, the lactic, succinic and acetic acids were the most produced organic acids while the amounts of ketones decreased, and the alcohols, in particular the esters, increased within fermentation. Particularly, ethyl-lactate and ethyl-acetate increased significantly as well as ethyl-caprylate, ethyl-capronate and ethyl-hexanote responsible for fruity and flower flavour were also accumulated. Tarhana doughs were clustered in two main groups after five days of fermentation with principal component analysis and these clusters remained stable until the end of fermentation. As a conclusion, fermentation time determines the desirable tarhana by ensuring the formation of aromatic volatiles particularly esters. It is necessary to ferment tarhana for at least five days in order to obtain typical aromatic properties. C1 [Ozdemir, Nilguen; Temiz, Hasan; Con, Ahmet Hilmi] Ondokuz Mayis Univ, Dept Food Engn, Fac Engn, Samsun, Turkey. [Simsek, Omer] Pamukkale Univ, Dept Food Engn, Fac Engn, TR-20160 Denizli, Turkey.

A wide number of Lactic Acid Bacteria (LAB) species produce alpha-glucans with their ability to synthesize glucansucrases (GS) which use sucrose as substrate for the glucan production. Recently another group of enzymes in LAB gained special interest for their ability to produce alpha-glucans targeting the substrates containing alpha 1-4-linkages and synthesizing new (alpha 1-6) or (alpha 1-3)-linkages as alpha-glucanotransferases. In this study, a putative 4,6-a-glucanotransferase (GTFB) from sourdough isolate Lactobacillus reuteri E81 was identified and expressed in Escherichia coli. The biochemical characterization of the GTFB-E81 confirmed its function as it cleaved the alpha 1-4-linkages in different substrates and produced new gluco-oligomers/polymers containing al 6 linkages together with the alpha 1-4-linkages detected by NMR analysis. GTFB-E81 produced maltooligosaccharides targeting maltose and maltoheptaose as substrates with up to DP 8 detected by TLC and ESIMS/MS analysis. The functional roles of these malto-oligosaccharides were determined by testing their immune-modulatory functions in HT29 cells and they triggered the production of anti-inflammatory 1L-4 and pro-inflammatory IL-12 cytokines. (C) 2018 Elsevier B.V. All rights reserved. C1 [Ispirli, Humeyra; Sagdic, Osman] Yildiz Tech Univ, Chem & Met Engn Fac, Dept Food Engn, TR-34000 Istanbul, Turkey. [Simsek, Omer] Pamukkale Univ, Dept Food Engn, Fac Engn, TR-20000 Denizli, Turkey. [Skory, Christopher] ARS, Renewable Prod Technol Res Unit, Natl Ctr Agr Utilizat Res, USDA, 1815 North Univ St, Peoria, IL 61604 USA. [Dertli, Enes] Bayburt Univ, Dept Food Engn, Fac Engn, TR-69000 Bayburt, Turkey.

Bacteriocins are the most important natural preservatives against pathogens, but their limited spectrum could not fully ensure food safety. In this study, bacteriocin producers showing specific-antimicrobial activity to foodborne pathogens were isolated and the success of the cocktail of pathogen-specific bacteriocins (PSBs) in milk were demonstrated. PSB producers were isolated from 250 different foods by a new approach that screening their antimicrobial activity against a mixture of five different strains of each Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus. Lactobacillus plantarum PFC339, Enterococcus faecalis PFC340, Lactobacillus delbrueckii subsp. lactis PFC341 were identified that inhibited the growth of pathogen strains respectively, but not lactic acid bacterial strains. The molecular sizes of PSBs were 1219.021, 3346.803, 4853.768 Da respectively. They inhibited by disrupting the permeability or creating pores on the cell walls. While 100 AU/mL decreased the amount of pathogens in milk which they are specifically active, each PSBs did not show any inhibitory effect on other two pathogens. However, PSBs cocktail was able to stop the growth of all inoculated pathogens. In conclusion, three bacteriocin producers specifically inhibiting B. cereus, L. monocytogenes, S. aureus were isolated and the growth of pathogens in food systems were prevented by use of these PSBs as cocktails for the first time. © 2019 Elsevier Ltd

Bacteriocins are the most important natural preservatives against pathogens, but their limited spectrum could not fully ensure food safety. In this study, bacteriocin producers showing specific-antimicrobial activity to foodbome pathogens were isolated and the success of the cocktail of pathogen-specific bacteriocins (PSBs) in milk were demonstrated. PSB producers were isolated from 250 different foods by a new approach that screening their antimicrobial activity against a mixture of five different strains of each Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus. Lactobacillus plantarum PFC339, Enterococcus faecalis PFC340, Lactobacillus delbrueckii subsp. lactis PFC341 were identified that inhibited the growth of pathogen strains respectively, but not lactic acid bacterial strains. The molecular sizes of PSBs were 1219.021, 3346.803, 4853.768 Da respectively. They inhibited by disrupting the permeability or creating pores on the cell walls. While 100 AU/mL decreased the amount of pathogens in milk which they are specifically active, each PSBs did not show any inhibitory effect on other two pathogens. However, PSBs cocktail was able to stop the growth of all inoculated pathogens. In conclusion, three bacteriocin producers specifically inhibiting B. terms, L. monocytogenes, S. aureus were isolated and the growth of pathogens in food systems were prevented by use of these PSBs as cocktails for the first time. C1 [Kaya, Halil Ibrahim; Simsek, Omer] Univ Pamukkale, Dept Food Engn, TR-20017 Denizli, Turkey.

The antioxidant and antimicrobial potential of artichoke extract (AE) in raw beef patties (RBPs) was evaluated during the storage. The RBPs were prepared with the addition of 500 and 1000 ppm AE. Also, E.wherichia coli ATCC25922 and Listeria monocytogenes ATCC19118 were inoculated to each RBP to follow the antimicrobial activity. An evaluation of the instrumental colour, pH, total phenolic content (TPC), 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, TBARS and microbiological properties was carried out during storage. The evaluation revealed that the a* value decreased significantly, so that the reduction level of the AE500 and AE1000 samples during storage was 35, 57, and 56%, respectively, in the control. AE addition slightly decreased the pH of RBPs, which varied between 5.72 and 5.96. The TPC and DPPH values of samples with added 500 and 1000 ppm AE were 2 to 3-fold and 3 to 7-fold higher than in the control, respectively. Also, the TBARS values were determined as 43 and 54% lower than in the control at the end of storage when 500 and 1000 ppm AE were used. The AE in RBP inhibited the viability of total aerobic psychrophilic bacteria, coliform bacteria and yeast-mould in a concentration-dependent manner. AE prevented the growth of E. coli ATCC25922 and L. monocytogenes ATCC19118 inoculated to the RBPs. In conclusion, 1000 ppm AE was sufficient for antioxidant and antimicrobial activity in RBP. To our knowledge, this is the first study that presents the antioxidant and antimicrobial activity of AE used in a food model system.

The aim of this study was to select lactic acid bacteria (LAB) having autolytic activity on the basis of the technological aspects to accelerate the ripening of white-brined cheeses. For this purpose, mesophilic (n=526) and thermophilic (n=413) LAB isolates were collected from 34 raw milk and 16 cheese samples. 27 LAB isolates with a high autolytic activity ranging from 34 to 54 % were identified by (GTG)(5) fingerprint and 16S rDNA sequence analyses, which showed that Lactococcus lactis and Enterococcus faecium were abundant among the autolytic isolates in raw milk while E. faecium and Lactobacillus casei were in white-brined cheese. The autolysis rate of LAB strains increased by raising the temperature and pH; however, decreased somewhat at high NaCl concentrations. Additionally, the highest autolysis was observed in the presence of glucose for most of the strains. On the other hand, Lb. plantarum PFC231 was autolyzed differently, being the highest in the presence of lactose. E. durans PFC235, E. faecium PFC232 and Lb. plantarum PFC231 were autolyzed more than other LAB strains, when exposed to low pH, high NaCl concentration at 10 degrees C simultaneously. However, only Lb. plantarum PFC231, Leuconostoc mesenteroides PFC234, Lc. lactis subsp. cremoris PFC233, Pediococcus acidilactici PFC237 and Lb. rhamnosus PFC238 were able to show intracellular caseinolytic activity. In conclusion, Lb. plantarum PFC231 has the potential to be used as an adjunct starter culture in the production of white-brined cheeses because of its autolysis at low temperature, pH and high salt concentrations, as well as intracellular caseinolytic activity.

A high level of variation in microflora can be observed in profiles of lactic acid bacteria (LAB) from sourdoughs. Here, we present draft genome sequences of Lactobacillus reuteri E81, L. reuteri LR5A, L. rhamnosus LR2, L. plantarum PFC-311, and the novel Lactobacillus sp. strain PFC-70, isolated from traditional Turkish backslopped wheat sourdoughs.

The aim of this study was to select starter cultures by showing correlation between the volatiles and the starter cultures used during fermentation with the goal of producing a standard and desirable tarhana. This traditional fermented cereal product is produced with fermentation of a dough prepared by mixing wheat flour, yoghurt, some vegetables and spices. To understand the relationship between aroma development and certain starter cultures through both statistical and sensorial approaches, 11 tarhana dough samples were prepared using three lactic acid bacteria (Lactobacillus farciminis PFC83, Lactobacillus casei PFC90, Lactobacillus alimentarius PFC91) or two yeast (Pichia kudriavzevii PFC12, Candida humilis PFC138) strains, as mono- or co-culture. Ester and alcohol volatiles accumulated during fermentation in the tarhana dough samples with starter, triggered by the use of starter cultures. Additionally, it was noted that lactic, succinic and acetic acids were the most prevalent organic acids to be produced in the dough samples during fermentation, and a partial least squares regression analysis showed that L. alimentarius PFC91 and P. kudriavzevii PFC126 were correlated with the accumulation of esters and these organic acids. Moreover, a sensory analysis showed that tarhana dough samples prepared using the L. alimentarius PFC91 and P. kudriavzevii PFC126 strains obtained the most acceptable scores. In conclusion, this study showed a strong correlation between aroma development in tarhana and starter usage in which L. alimentarius PFC91 is co-cultured with P. kudriavzevii PFC126, contributing to the accumulation of the volatile aromatic compounds and organic acids that are responsible for desirable aromatic tarhana.

This study aims to determine the effects of inulin and fructo-oligosaccharide (FOS) on the probiotic properties of five Lactobacillus spp. isolated from human milk. Lactobacillus spp. were isolated and identified, and the growth characteristics, acid and bile salt tolerance, antagonistic effects, and cholesterol assimilation of Lactobacillus strains were investigated in the presence of inulin and FOS. Lactobacillus casei L1 was able to utilize inulin and FOS as carbon source as well as glucose even other strains were able to use, including Lactobacillus rhamnosus GG. This strain also showed high tolerance to acid and bile salt, even at pH 2.5 and 0.5% bile salt levels, respectively. Inulin and FOS promoted the antimicrobial activity of L. casei L1 against pathogenic bacteria. Cholesterol assimilation was higher than in the other and control probiotic strains in the presence inulin and FOS, which were measured as 14 and 25 mg/dL, respectively. In conclusion, L. casei L1 can use both inulin and FOS to maintain its viability both at digestive conditions and also the relevant prebiotics, and show broad antagonistic activity and cholesterol assimilation.

The phenol degradation capacity of Serratia marcescens NSO9-1 isolated from olive mill wastewater was evaluated and optimized in this study. Plackett-Burman design coupled with Box-Behnken methodology was used to evaluate the effects of medium components and significant parameters on phenol degradation by this relevant strain. According to Plackett-Burmanbased statistical screening, seven of the eleven components of the medium had a significant effect on the metabolism of phenol degradation. The most important factors were MgSO4, NaCl, CaCl2, and molybdenum salt, which had an effective contribution of 90.12%. Additionally, Box-Behnken methodology using a quadratic model was adopted to investigate the mutual interactions between process parameters. The analysis results indicated that interactions between pH and temperature, pH and inoculum amount, and incubation time and inoculum amount critically affected the response variable. The experimental results showed that under the determined conditions, 41.66% of the maximum removal efficiency of phenol was achieved. The optimal conditions were 8.94, 30.22 degrees C, 4.19 days, and 4.68% (v/v) for pH, temperature, incubation time, and inoculum amount, respectively. The validity and practicability of this statistical optimization strategy confirmed the relation between predicted and experimental values. Using a selective isolation method, the performance of this indigenous strain isolated from olive oil mill wastewater, which contained polyphenolic compounds, is comparable to the reported literature at higher phenol concentrations.

In this study, lactic acid bacteria diversity during the fermentation of homemade and commercially prepared Tarhana, a traditional fermented cereal food from Anatolia, was determined and compared. The isolates collected from Tarhana dough were differentiated according to their (GTG)(5) profiles and then identified using 16S rDNA and pheS gene sequences. The variation of lactic acid bacteria during fermentation was also screened using PCR-DGGE. Commercially prepared Tarhana dough was fermented with higher Lactobacillus spp. diversity than homemade Tarhana dough. Lactobacillus casei, L. alimentarius, L. fabifermentas, and L. paralimentarius were identified differently from the fermentation of commercially prepared Tarhana dough. PCR-DGGE analysis revealed that L. plantarum was the main strain for homemade Tarhana, whereas L. brevis and L. alimentarius were observed in commercially prepared Tarhana dough fermentation. In conclusion, L. plantarum, L. brevis and L. alimentarius can be useful as a potential starter culture for the industrial production of Tarhana.

The aim of this study was to evaluate the use of mono and mixed lactic acid bacteria (LAB) cultures to determine suitable LAB combinations for a type II sourdough system. In this context, previously isolated sourdough LAB strains with antimicrobial activity, which included Lactobacillus plantarum PFC22, Lactobacillus brevis PFC31, Pediococcus acidilactici PFC38, and Lactobacillus sanfranciscensis PFC80, were used as mono or mixed culture combinations in a fermentation system to produce type II sourdough, and subsequently in bread dough production. Compared to the monoculture fermentation of dough, the use of mixed cultures shortened the adaptation period by half. In addition, the use of mixed cultures ensured higher microbial viability, and enhanced the fruity flavor during bread dough production. It was determined that the combination of L. plantarum PFC22+P. acidilactici PFC38+L. sanfranciscensis PFC80 is a promising culture mixture that can be used in the production of type II sourdough systems, and that may also contribute to an increase in metabolic activity during bread production process.

Nisin is the first bacteriocin identified in Lactococcus lactis and belongs to type 1 lanthibiotic group. High nisin production in cultured media is related with the composition of fermentation medium, pH, produced nisin concentration and most importantly growth amount of cell. For industrial purpose, batch, fed-batch and continue fermentation systems were developed by regarding these factors. Maintaining efficient production of nisin having important potential at preservation of foods is important for both industrial production and using as starter culture. In this review the fermentation factors at nisin production were outlined and constructed model systems were compared.

Tarhana is a traditional cereal-based fermented food produced with a mixture of yoghurt and flour. The main microbiota in the fermentation of tarhana is yeast, together with lactic acid bacteria. In this study, the yeast microbiota of home-made tarhana (HMT) and plant-type tarhana (PTT) dough samples was evaluated and compared during fermentation. Culture-dependent LSU and ITS-5.8S rDNA sequence analysis of yeast isolates collected during the tarhana dough fermentation clarified 45 selected isolates representing different clusters. These yeast isolates displayed high homologywith species Pichia kudriavzevii (11), Candida glabrata (11), Candida humilis (10), Saccharomyces cerevisiae (7), Kluyveromyces marxianus (4), Kazachstania servazzi (1), and Kazachstania unispora (1). Additionally, both culture-dependent and PCR-Denaturated Gradient Gel Electrophoresis (PCR-DGGE) analyses showed that S. cerevisiae, P. kudriavzevii and K. marxianus were abundant in the fermentation of HMT dough samples whereas P. kudriavzevii, C. humilis, and C. glabrata dominated the PTT dough samples. It was concluded that tarhana fermentation was accomplished with the presence of a wide variety of yeast species that mainly included P. kudriavzevii in both HMT and PTT dough samples.

In this study, nisin production of Lactococcus lactis N8 was optimized by independent variables of glucose, hemin and oxygen concentrations in fed-batch fermentation in which respiration of cells was stimulated with hemin. Response surface model was able to explain the changes of the nisin production of L. lactis N8 in fed-batch fermentation system with high fidelity (R-2 98%) and insignificant lack of fit. Accordingly, the equation developed indicated the optimum parameters for glucose, hemin, and dissolved oxygen were 8 g L-1 h(-1), 3 mu g mL(-1) and 40%, respectively. While 1711 IU mL(-1) nisin was produced by L. lactis N8 in control fed-batch fermentation, 5410 IU mL(-1) nisin production was achieved within the relevant optimum parameters where the respiration of cell was stimulated with hemin. Accordingly, nisin production was enhanced 3.1 fold in fed-batch fermentation using hemin. In conclusion the nisin production of L. lactis N8 was enhanced extensively as a result of increasing the biomass by stimulating the cell respiration with adding the hemin in the fed-batch fermentation. (C) 2015 American Institute of Chemical Engineers

Clementine mandarins, is a very important fruit group due the production of high quality fruits that can be seedless as consequence to their sexual self-incompatibility. In this study, the self-compatible/incompatible status of 40 clementine mandarins provided by the Cukurova University collection was determined by histological analysis. For this purpose, self-pollinations were performed and pistil samples were taken 3, 5, 7, 9, 11, 13, 15 and 20 days after pollination. All samples were fixed in FPA70 solution and stored at 4°C. Pistil samples were stained with aniline blue and observed by fluorescence microscope following the staining and squashing. All pollen tubes of the genotypes arrested growth in the upper or middle part of style and none reached the base of the style. As a result, all the 40 clementine cultivars tested have been identified as self-incompatible.

Utilisation of antimicrobial agents at pre- or post-smoking processes was considered for the microbial quality of hot-smoked trout fillets prepared at plant scale. Potassium sorbate (PS) and sodium lactate (SL) were used separately or in combinations at 3% (w/v) in brine for pre- smoking or with spraying at post-smoking. Pre-smoking usage could reduce the total aerobic mesophilic bacteria (TAMB) values better than the post-smoking usage during storage. However, post-smoking application was more effective at preventing the moulds and yeasts (MY) growth. PS in brine at pre- smoking was most efficient, which could keep trout fillets for even four weeks at 6 +/- 1 degrees C storage within the range of consumable limits. Additionally, PS usage reduced the contaminated bacterial diversity and especially eliminated the outgrowth of Serratia liquefaciens. As a conclusion, pre-smoking application repressed the contaminated microbial growth during the storage of smoked trout fillets, which may eliminate the microbial risks.