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The aim of this study was to screen and select potential probiotics that are able to reduce uremic toxin levels, mainly indoxyl sulphate (IS), in vitro and in vivo. Three strains, Lactobacillus plantarum subsp. plantarum BCRC12251, Lb. paracasei subsp. paracasei BCRC12188 and Streptococcus salivarius subsp. thermophilus BCRC13869, were selected to have the ability to reduce indoxyl sulphate in vitro. Combination of strains demonstrated a better IS removing ability than individual strains. In vivo, we observed that oral administration of Pm1 in cisplatin-induced acute kidney injury model significantly suppressed the accumulation of IS in the serum. Our findings suggest that treatment with the strain combination Pm1 might be a useful approach to decreasing IS accumulation in the serum and kidney. Administration of Pm1 is also likely to be beneficial to chemotherapy patients who are being treated with cisplatin.

A potential probiotic strain, Lactobacillus kefiranofaciens M1, was previously isolated from kefir grains, which are used to manufacture the traditional fermented drink kefir. The aim of this study was to investigate the effects of Lb. kefiranofaciens M1 on enterohemorrhagic Escherichia coli (EHEC) infection, using mice and intestinal cell models. BALB/c mice were daily administrated with either phosphate buffered saline or Lb. kefiranofaciens M1 at 2×10 8 cfu/mouse per day intragastrically for 7 d. Intragastric challenges with EHEC (2×10 9 cfu/mouse) were conducted on d 0, 4, and 7 after treatment. Administration of Lb. kefiranofaciens M1 was able to prevent EHEC infection-induced symptoms, intestinal damage, renal damage, bacterial translocation, and Shiga toxin penetration. Furthermore, the mucosal EHEC-specific IgA responses were increased after Lb. kefiranofaciens M1 administration in the EHEC-infected mouse system. Additionally, in vitro, Lb. kefiranofaciens M1 was shown to have a protective effect on Caco-2 intestinal epithelial cells and Caco-2 intestinal epithelial cell monolayers; the bacteria limited EHEC-induced cell death and reduced the loss of epithelial integrity. These findings support the potential of Lb. kefiranofaciens M1 treatment as an approach to preventing EHEC infection and its effects.

Lactobacillus kefiranofaciens M1 is a probiotic strain isolated from Taiwanese kefir grains. The present study evaluated the effects of heat, cold, acid and bile salt adaptations on the stress tolerance of L. kefiranofaciens M1. The regulation of protein expression of L. kefiranofaciens M1 under these adaptation conditions was also investigated. The results showed that adaptation of L. kefiranofaciens M1 to heat, cold, acid and bile salts induced homologous tolerance and cross-protection against heterologous challenge. The extent of induced tolerance varied depending on the type and condition of stress. Proteomic analysis revealed that 27 proteins exhibited differences in expression between non-adapted and stress-adapted L. kefiranofaciens M1 cells. Among these proteins, three proteins involved in carbohydrate metabolism (triosephosphate isomerase, enolase and NAD-dependent glycerol-3-phosphate dehydrogenase), two proteins involved in pH homeostasis (ATP synthase subunits AtpA and AtpB), two stress response proteins (chaperones DnaK and GroEL) and one translation-related protein (30S ribosomal protein S2) were up-regulated by three of the four adaptation treatments examined. The increased synthesis of these stress proteins might play a critical protective role in the cellular defense against heat, cold, acid and bile salt stresses.

The purpose of this study was to understand the significance of each microorganism in grain formation by evaluating their microbial aggregation and cell surface properties during co-aggregation of LAB and yeasts together with an investigation of biofilm formation. Non-grain forming strains from viili were also evaluated as a comparison. Results indicated that the kefir grain strains, Lactobacillus kefiranofaciens and Saccharomyces turicensis possess strong auto-aggregation ability and that Lactobacillus kefiri shows significant biofilm formation properties. Significant co-aggregation was noted when S. turicensis and kefir LAB strains (Lb. kefiranofaciens and Lb. kefiri) were co-cultured. Most of the tested LAB strains are hydrophilic and had a negative charge on their cell surface. Only the kefir LAB strains, Lb. kefiranofaciens HL1 and Lb. kefiri HL2, possessed very high hydrophobicity and had a positive cell surface charge at pH 4.2. In contrast, the LAB and yeasts in viili did not show any significant self-aggregation or biofilm formation. Based on the above results, we propose that grain formation begins with the self-aggregation of Lb. kefiranofaciens and S. turicensis to form small granules. At this point, the biofilm producer, Lb. kefiri, then begins to attach to the surface of granules and co-aggregates with other organisms and components in the milk to form the grains. On sub-culturing, more organisms attach to the grains resulting in grain growth. When investigated by scanning electron microscopy, it was found that short-chain lactobacilli such as Lb. kefiri occupy the surface, while long-chain lactobacilli such as Lb. kefiranofaciens have aggregated towards the center of the kefir grains. These findings agree with the above hypothesis on the formation of grains. Taken together, this study demonstrates the importance of cell surface properties together with fermentation conditions to the formation of grains in kefir.

In this research we undertake a study of image compression based on the discrete cosine transform(DCT) and discrete wavelet transform(DWT). Then a hybrid color image compression algorithm based on DCT and DWT is proposed. This algorithm is implemented through transform the color image using DWT in the YCbCr space first, and then DCT in the low frequency, adopt huffman coding, RLE and arithmetic coding in the encoded mode. In experiments, the results outperform the only DCT and the only DWT typically higher in peak signal-to-noise ratio and have better visual quality.

Vibrio parahaemolyticus strain 690 was isolated from gastroenteritis patients. Its thermal and ethanol stress responses have been reported in our previous studies. In this study, we further investigated the effects of various acid adaptation conditions including pH (5.0-6.0) and time (30-90 min) on the acid tolerance in different growth phases of V. parahaemolyticus 690. Additionally, the adaptive acid tolerance among different V. parahaemolyticus strains was compared. Results indicated that the acid tolerance of V. parahaemolyticus 690 was significantly increased after acid adaptation at pH 5.5 and 6.0 for 30-90 min. Among the various acid adaptation conditions examined, V. parahaemolyticus 690 acid-adapted at pH 5.5 for 90 min exhibited the highest acid tolerance. The acid adaptation also influenced the acid tolerance of V. parahaemolyticus 690 in different growth phases with late-exponential phase demonstrating the greatest acid tolerance response (ATR) than other phases. Additionally, the results also showed that the induction of adaptive ATR varied with different strains of V. parahaemolyticus. An increase in acid tolerance of V. parahaemolyticus was observed after prior acid adaptation in five strains (556, 690, BCRC 13023, BCRC 13025, and BCRC 12864), but not in strains 405 and BCRC 12863.

Lactobacillus kefiranofaciens M1, isolated from and identified in Taiwanese milk kefir grain, has demonstrated immune-modulating activity. In the present study, we further investigated the effects of Lb. kefiranofaciens M1 on intestinal epithelial cells in vitro and on dextran sodium sulfate (DSS)-induced colitis in vivo. The possible mechanisms regarding the cytokine products and intestinal epithelial barrier restoration as well as the putative receptor for the protective effects of Lb. kefiranofaciens M1 were investigated. In vitro results indicated that Lb. kefiranofaciens M1 could strengthen the epithelial barrier function in vitro by increasing the transepithelial electrical resistance (TEER) and significantly upregulated the level of the chemokine CCL-20 at both the apical and basolateral sites. The in vivo effects of Lb. kefiranofaciens M1 on the regulation of intestinal physiology indicate that this strain could ameliorate DSS-induced colitis with a significant attenuation of the bleeding score and colon length shortening. Production of proinflammatory cytokines was decreased and that of the antiinflammatory cytokine IL-10 was increased in the DSS-treated mice given Lb. kefiranofaciens M1. The putative receptor for the protective effects of Lb. kefiranofaciens M1 was toll-like receptor 2 (TLR2), which was involved in probiotic-induced cytokine production in vitro and in attenuation of the bleeding score and colon length shortening in vivo. In this study, the kefir lactobacillus Lb. kefiranofaciens M1 clearly demonstrated an anticolitis effect. Based on these results, Lb. kefiranofaciens M1 has the potential to be applied in fermented dairy products as an alternative therapy for intestinal disorders.

Two mouse models were used to assay the antiallergic effects of the velvet antler (VA) of Formosan sambar deer (Cervus unicolor swinhoei) in this study. The results using the ovalbumin- (OVA-) sensitized mouse model showed that the levels of total IgE and OVA-specific IgE were reduced after VA powder was administrated for 4 weeks. In addition, theex vivoresults indicated that the secretion of T helper cell 1 (Th1), regulatory T (Treg), and Th17 cytokines by splenocytes was significantly increased (P<0.05) when VA powder was administered to the mice. Furthermore, OVA-allergic asthma mice that have been orally administrated with VA powder showed a strong inhibition of Th2 cytokine and proinflammatory cytokine production in bronchoalveolar fluid compared to control mice. An increase in the regulatory T-cell population of splenocytes in the allergic asthma mice after oral administration of VA was also observed. All the features of the asthmatic phenotype, including airway inflammation and the development of airway hyperresponsiveness, were reduced by treatment with VA. These findings support the hypothesis that oral feeding of VA may be an effective way of alleviating asthmatic symptoms in humans.

Kefir, a beverage produced by the action of lactic acid bacteria (LAB), yeasts and acetic acid bacteria on milk, has a long tradition of offering health benefits. The objective of this research was to optimize the best formula for producing the kefir candy with maximum viabilities of LAB and yeasts using the response surface modeling and the sequential quadratic programming (SQP) method. In this study, milk was mixed with 5% kefir grains and incubated at 20C for 24 h. The samples were blended with lyoprotectants (galactose, skim milk powder [SMP] and sucrose), freeze-dried and then mixed with sweeteners and compressed to form candies. The ratio of the lyoprotectants was determined using response surface modeling for constructing a response surface model and then using an SQP method to optimize the model. Optimization results indicated that kefir containing galactose, sucrose and SMP at 4.53, 7.0 and 5.03% (w/v), respectively, produced a chewable tablet with the highest viability of the microorganisms investigated. A relatively higher survival of microorganisms could be achieved by placing the kefir candy product in a glass bottle with deoxidant and desiccant at 4C. PRACTICAL APPLICATIONS A number of novel fermented dairy products have been developed and marketed under the concept of probiotic products, but few of these products were associated with confectionary goods. The kefir candy created in the present study with high viable cell counts (106–107 cfu/g) after extended storage (up to 2 months) provides a flavorful option to offer the health benefits of probiotics. This study not only provides an opportunity to resolve the difficulty of kefir commercialization due to its post-acidity and gas production but also increases the variety of dairy products in the market.

The aim of this study was to display a rumen bacterial β-glucanase on the cell surface of a probiotic Lactobacillus reuteri strain. The β-glucan degrading ability and the adhesion capability of the genetically modified strain were evaluated. The β-glucanase (Glu) from Fibrobacter succinogenes was fused to the C-terminus of collagen-binding protein (Cnb) from L. reuteri and then expressed by L. reuteri Pg4 as a recombinant Cnb-Glu-His(6) fusion protein. Confocal immunofluorescence microscopy and flow cytometric analysis of the transformed strain L. reuteri pNZ-cnb/glu demonstrated that Cnb-Glu-His(6) fusion protein was displayed on its cell surface. In addition, L. reuteri pNZ-cnb/glu acquired the capacity to break down barley β-glucan and showed higher adhesion capability, in comparison with the parental strain L. reuteri Pg4. To the best of the authors' knowledge, this is the first report of successful display of fibrolytic enzymes on the cell surface of intestinal lactobacilli.

The purpose of this study was to identify species of lactic acid bacteria in Taiwanese ropy fermented milk and to study their microbial dynamics during the fermentation process through conventional microbiological cultivation and PCR-denaturing gradient gel electrophoresis. Identification results indicated that Lactococcus lactis ssp. cremoris and Leuconostoc mesenteroides ssp. mesenteroides were the major lactic acid bacteria in Taiwanese ropy fermented milk. Interestingly, 3 groups were identified as Lc. lactis ssp. cremoris using 16S rDNA sequencing, but they showed different denaturing gradient gel electrophoresis patterns and assimilation of carbohydrates. In addition, the microbial dynamics study in different fermentation stages demonstrated that Lc. lactis ssp. cremoris was the most dominant bacterial species in the samples, followed by Leu. mesenteroides ssp. mesenteroides with no differences among the fermentation stages. Finally, the microbial distribution profiles showed that the microbial ecology was different in bovine, caprine, and reconstituted milk, which might further affect the characteristics of the product.

Two types of extracellular proteases with molecular mass of 50.0 and 44.8 kDa were found in H10 enzymes partially purified from Bacillus cereus H10. Further identification using liquid chromatography-tandem mass spectrometry, the enzyme with 50.0 kDa was identified as being similar to leucine dehydrogenase; while the enzyme with 44.8 kDa might be a novel keratinolytic enzyme with little similarity to other proteins. To maximize the keratinolytic and proteolytic abilities in the H10 enzymes, a combination of response surface methodology and sequential quadratic programming technique was used to study the hydrolytic pH and temperature. Results showed that the H10 enzymes could produce optimal proteolytic and keratinolytic activities at a hydrolysis temperature of 59°C at pH 7.57. Testing the protease activity on various protein substrates and temperatures indicated that the H10 enzymes showed high thermal stability and were very effective in porcine hair.

Isolation and identification of the autochthonous starter from a naturally fermented meat allows control of the fermentation process and promises microbiological safety for this specialty. Thus the purpose of this study was to identify the lactic acid bacteria and coagulase-negative cocci present in a traditional Taiwanese naturally fermented ham (TNFH) and to study the microbial dynamics at different ripening stages; the approach was a combination of conventional microbiological cultivation, polymerase chain reaction-denaturing gradient gel electrophoresis and DNA sequencing. In total, twelve different strains of lactic acid bacteria and three Staphylococcus strains were identified in the TNFH samples, whereas only 5 dominant strains were observed in the TNFH samples when the DGGE as a culture-independent method was applied. The bacterial ecology on the surface of the samples was mainly characterized by the stable presence of Lactobacillus sakei and Staphylococcus saprophyticus; nonetheless Leuconostoc mesenteroides and Carnobacterium divergens were the most abundant bacteria found in the final product. These results are also agreed with the findings of the culture-independent method. In addition, Microbacterium spp., Carnobacterium spp., Enterobacter spp., Brochothrix spp., Enterococcus spp., and Bacillus spp. were also present at the beginning of the ripening, but few bacteria were found at the center of the TNFH samples during the early ripening stages. However, after 30 days of ripening, the microbial ecology at the center of the TNFH samples paralleled that of the surface. Finally, as far as we have been able to determine, our report is the first to investigate the microbiological dynamics in fermented meat products using combination of cultivation, the Harrison disc method, DGGE and DNA sequencing as the culture-dependent method. Our report is also the first to show the presence of Staphylococcus arlettae in a fermented sausage and ham product.

BACKGROUND: Xylan is the second most abundant renewable polysaccharide in nature and also represents an important industrial substrate. The complete degradation of xylan requires the combination of several types of xylanolytic enzymes, including endo-β-1,4-xylanases, β-xylosidases, and acetylxylan esterases. As a biocatalyst, xylanolytic enzymes with good thermal stability are of great interest, therefore, a thermo-tolerant acetylxylan esterase, AxeS20E, was investigated. RESULTS: The cDNA encoding the carbohydrate esterase (CE) domain of AxeS20E from Neocallimastix patriciarum was expressed in Escherichia coli as a recombinant His6 fusion protein. The recombinant AxeS20E protein was obtained after purification by immobilized metal ion-affinity chromatography. Response surface modeling (RSM) combined with central composite design (CCD) and regression analysis were then employed for the planned statistical optimization of the acetylxylan esterase activities of AxeS20E. The optimal conditions for the highest activity of AxeS20E were observed at 54.6 °C and pH 7.8. Furthermore, AxeS20E retained more than 85% of its initial activity after 120 min of heating at 80 °C. CONCLUSIONS: The results suggested that RSM combined with CCD and regression analysis were effective in determining optimized temperature and pH conditions for the enzyme activity of AxeS20E. The results also proved AxeS20E was thermo-tolerant and might be a good candidate for various biotechnological applications. Copyright © 2009 Society of Chemical Industry

Summary The objective of this study was to investigate the in vitro immunomodulatory effect of milk whey through enzymatic hydrolysis, microbial fermentation and two-stage hydrolytic reactions (enzymatic and microbial reactions) by analysing cytokine profiles. Results indicated that the milk whey sample fermented by Lactobacillus kefiranofaciens M1 and two-stage hydrolytic reactions (hydrolysed by Alcalase and then fermented by L. kefiranofaciens M1) could significantly induce the production of tumour necrosis factor (TNF)-α and IL-12 compared with the milk whey control and enzymatic treatments. Further characterisation of the immunomodulatory factor by membrane filtration and mutanolysin hydrolysation, the stimulatory activity for IL-12 and TNF-α production was found to be reduced and to be correlated positively with the cell wall components in L. kefiranofaciens M1. In addition, Th2-polarised splenocytes revealed that L. kefiranofaciens M1 had both IL-12 inducing and IL-4 repressing activities. These results suggested that L. kefiranofaciens M1 could direct the Th1/Th2 balance toward Th1.

BACKGROUND: 1,3-1,4-β-D-glucanase (1,3-1,4-β-D-glucan 4-glucanohydrolase; EC 3.2.1.73) has been used in a range of industrial processes. As a biocatalyst, it is better to use immobilized enzymes than free enzymes, therefore, the immobilization of 1,3-1,4-β-D-glucanase was investigated. RESULTS: A 1,3-1,4-β-D-glucanase gene from Fibrobacter succinogenes was overexpressed in Escherichia coli as a recombinant protein fused to the N terminus of oleosin, a unique structural protein of seed oil bodies. With the reconstitution of the artificial oil bodies (AOBs), refolding, purification, and immobilization of active 1,3-1,4-β-D-glucanase was accomplished simultaneously. Response surface modeling (RSM), with central composite design (CCD), and regression analysis were successfully applied to determine the optimal temperature and pH conditions of the AOB-immobilized 1,3-1,4-β-D-glucanase. The optimal conditions for the highest immobilized 1,3-1,4-β-D-glucanase activity (7.1 IU mg−1 of total protein) were observed at 39 °C and pH 8.8. Furthermore, AOB-immobilized 1,3-1,4-β-D-glucanase retained more than 70% of its initial activity after 120 min at 39 °C, and it was easily and simply recovered from the surface of the solution by brief centrifugation; it could be reused eight times while retaining more than 80% of its activity. CONCLUSIONS: These results indicate that the AOB-based system is a comparatively simple and effective method for simultaneous refolding, purification, and immobilization of 1,3-1,4-β-D-glucanase. Copyright © 2009 Society of Chemical Industry

A thermally stable and alkalophilic xylanase, XynCDBFV, from Neocallimastix patriciarum was overexpressed in Escherichia coli as a recombinant protein fused to the N-terminus of oleosin, a unique structural protein of seed oil bodies. As a result of the reconstitution of the artificial oil bodies (AOBs), the immobilization of active xylanase was accomplished. Response surface methodology (RSM) was employed for the optimization of the immobilized xylanase activity. The central composite design (CCD) and regression analysis methods were effective for determination of optimized temperature and pH conditions for the AOB-immobilized XynCDBFV. The optimal condition for the highest immobilized xylanase activity (3.93 IU/mg of total protein) was observed at 59 °C and pH 6.0. Further, AOB-immobilized XynCDBFV retained 50% of its maximal activity after 120 min at 60 °C, and it could be easily and simply recovered from the surface of the solution by brief centrifugation, and could be reused eight times while retaining more than 60% of its activity. These results proved it is a simple and effective method for direct immobilization of xylanases.

Mackerel nuggets used in this study were coated with HPMC prior to battering or coated with batter containing HPMC in order to improve crust crispness through the formation of a thermal gel barrier preventing the diffusion of water from meat into the crust during microwave reheating. A physical properties assessment and sensory analysis showed that S- and SH-pattern products exhibited a firm and brittle texture, while W- and WC-pattern batters exhibited crispy crust texture. Water molecule diffusion from fish meat into the crust was inhibited during microwave reheating in HPMC-coated products. Nevertheless, a slight toughness was observed in HPMC-coated products still hot following microwave reheating. This may relate to the thermal gelation HPMC film adhering to the crust. Nonetheless, HPMC's ability to serve as a barrier to water diffusion effectively improved the crispness of HPMC-coated mackerel nuggets that had been reheated in a microwave in comparison with non-HPMC-coated nuggets.

The objective of the present study was to investigate yeast communities in kefir grains and viili starters in Taiwan through conventional microbiological cultivation and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The DNA sequencing was used as a validity technique to ensure that all isolates within each group belonged to just one species, and to confirm the identified results of PCR-DGGE. Results indicated that a combination of conventional microbiological cultivation with PCR-DGGE and sequencing could successfully identify 4 yeast species from both types of cultures in Taiwan. Kluyveromyces marxianus, Saccharomyces turicensis, and Pichia fermentans were found in Taiwanese kefir grains with a distribution of 76, 22, and 2%, respectively, whereas Klu. marxianus, Saccharomyces unisporus and P. fermentans were identified in viili starters corresponding to 58, 11, and 31% of the total cell counts, respectively. Furthermore, the culture-independent method was applied to identify the yeast species using DGGE. Only 2 yeast species, Klu. marxianus and S. turicensis, were found in kefir grains and 2, Klu. marxianus and P. fermentans, in viili starters. These results suggest that in samples containing multiple species, PCR-DGGE may fail to detect some species. Sequences of yeast isolates reported in this study have been deposited in the GenBank database under accession nos. DQ139802, AF398485, DQ377652, and AY007920.

The objective of this research was to create a new probiotic candy with good flavor and healthy benefits by using the response surface method and a sequential quadratic programming technique. The endpoint was to increase the varieties of dairy products and enhance their market values. In this study, milk was mixed with yogurt cultures (Lactobacillus bulgaricus, Streptococcus thermophilus) and probiotics (L. paracasei, Bifidobacterium longum) and incubated at 37°C for 20 h. The samples were blended with lyoprotectants (galactose, skim milk powder and sucrose), freeze dried and then mixed with sweeteners (lactose and xylitol) to improve the texture for forming tablets. The processing conditions were optimized in two steps: the first step constructed a surface model using response surface methodology; the second step optimized the model with a sequential quadratic programming procedure. Results indicated that skim milk inoculated with L. delbrueckii subsp. Bulgaricus, S. thermophilus, L. paracasei subsp. paracasei and B. longum and blended with 6.9% of galactose, 7.0% of sucrose and 8.0% of skim milk powder would produce a new probiotic candy with the highest viability of probiotics and good flavor. A relatively higher survival of probiotics can be achieved by placing the probiotic candy product in a glass bottle with deoxidant and desiccant at 4°C. These probiotic counts remained at 10 6 -10 8 CFU/g after being stored for two months.

Lactic acid bacteria (LAB) in different original kefir grains were first assessed using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) by a culture-dependent way, and were further confirmed by DNA sequencing techniques. Results indicated that a combined method of cultivation with PCR-DGGE and subsequent DNA sequencing could successfully identify four LAB strains from three kefir grains from Taiwan (named Hsinchu, Mongolia and Ilan). Lactobacillus kefiri accounted, in the three kefir grains, for at least half of the isolated colonies while Lb. kefiranofaciens was the second most frequently isolated species. Leuconostoc mesenteroides was less frequently found but still in the three kefir grains conversely to Lactococcus lactis which based on culture-dependent isolation was only found in two of the kefir grains. It was interesting to find that all three kefir grains contain similar LAB species. Furthermore, the DGGE as a culture-independent method was also applied to detect the LAB strains. Results indicated that Lb. kefiranofaciens was found in all three kefir grains, whereas Lb. kefiri was only observed in Hsinchu kefir grain and Lc. lactis was found in both Mongolia and Ilan samples. Two additional strains, Pseudomonas spp. and E. coli, were also detected in kefir grains.

In this paper, a rapid and reliable gene-targeted species-specific polymerase chain reaction (PCR) technique based on a two-step process was established to identify bifidobacteria in dairy products. The first step was the PCR assay for genus Bifidobacterium with genus specific primers followed by the second step, which identified the species level with species-specific primer mixtures. Ten specific primer pairs, designed from nucleotide sequences of the 16-23S rRNA region, were developed for the Bifidobacterium species including B. angulatum, B. animalis, B. bifidum, B. breve, B. catenulatum, B. infantis, B. longum, B. minimum, B. subtile, and B. thermophilum. This technique was applied to the identification of Bifidobacterium species isolated from 6 probiotic products, and four different Bifidobacterium spp. (B. bifidum, B. longum, B. infantis, and B. breve) were identified. The findings indicated that the 16S-23S rDNA gene-targeted species-specific PCR technique is a simple and reliable method for identification of bifidobacteria in probiotic products. PCR combined with Denaturing Gradient Gel Electrophoresis (DGGE) for identification of the bifidobacteria was also evaluated and compared with the gene-targeted species-specific technique. Results indicated that for fermented milk products consistency was found for both species-specific PCR and PCR-DGGE in detecting species. However, in some lyophilized products, the bands corresponding to these species were not visualized in the DGGE profile but the specific PCR gave a positive result.

The objectives of this study were to microencapsulate both probiotics and culture filtrates by spray drying to maintain enzyme activity and probiotic viability during storage. Thus, probiotics and culture filtrates from lao-chao were microencapsulated by spray drying with various outlet air temperatures, and the milk-clotting activity, survival of probiotics and physical properties of the microcapsules were determined. The end purpose was to create easy-to-use probiotic Kou Woan Loa cultures. In the near future, manufacturing probiotic Kou Woan Loa could be carried out by simply mixing milk with 5% microcapsules and waiting for 1 h for coagulation, which would be time saving and convenient. The present study has shown that microencapsulation of Lactobacillus acidophilus BCRC 14079, Bifidobacterium longum BCRC 14605 and culture filtrates from lao-chao by spray drying could provide a good protection for both milk-clotting enzymes and probiotics. The average of microcapsules size and density was 10 μm and 1.68 g/cm 3 , respectively. An increase in the microencapsulation efficiency of microcapsules and lower water activity was found when the outlet air temperature was raised. However, the survival of L. acidophilus and B. longum was reduced as the outlet air temperature increased. The numbers of probiotics were maintained above the recommended therapeutic minimum (10 7 cfu/g) throughout storage.

Asia differs substantially among and within its regions populated by diverse ethnic groups, which maintain their own respective cultures and dietary habits. To address the diversity in their gut microbiota, we characterized the bacterial community in fecal samples obtained from 303 school-age children living in urban or rural regions in five countries spanning temperate and tropical areas of Asia. The microbiota profiled for the 303 subjects were classified into two enterotype-like clusters, each driven by Prevotella (P-type) or Bifidobacterium/Bacteroides (BB-type), respectively. Majority in China, Japan and Taiwan harbored BB-type, whereas those from Indonesia and Khon Kaen in Thailand mainly harbored P-type. The P-type microbiota was characterized by a more conserved bacterial community sharing a greater number of type-specific phylotypes. Predictive metagenomics suggests higher and lower activity of carbohydrate digestion and bile acid biosynthesis, respectively, in P-type subjects, reflecting their high intake of diets rich in resistant starch. Random-forest analysis classified their fecal species community as mirroring location of resident country, suggesting eco-geographical factors shaping gut microbiota. In particular, children living in Japan harbored a less diversified microbiota with high abundance of Bifidobacterium and less number of potentially pathogenic bacteria, which may reflect their living environment and unique diet.

The purpose of this research is to develop an optimal composition model for the encapsulating materials of probiotic microcapsules after one-week storage, with prebiotics and alginate as the encapsulating materials in given ranges of concentrations. The procedure includes three phases: building the response surface models, finding the optimal composition model with global optimization, and verifying the optimal model. The response surface method and the sequential quadratic programming (SQP) technique were employed to determine the optimal ratio and evaluate the tolerance of the microencapsulated probiotics to gastric conditions. Results indicated that the optimized microcapsules could increase the viability of probiotic bacteria under the simulated gastric fluid test (SGFT). The optimal combination of encapsulating materials for the probiotic microcapsules was found to be 3% sodium alginate blended with 1% peptides, 3% fructooligosaccharides (FOS), and 0% isomaltooligosaccharides (IMO). Verification experiments confirmed the SQP-based prediction with no apparent significant differences ( P > 0.05).

The purpose of this research was to develop an optimum composition model for a new synbiotic fermented dairy product with high probiotic cell counts, and to experimentally verify this model. The optimum composition model indicated the growth promoter ratio that could provide the highest growth rate for probiotics in this fermented product. Different levels of growth promoters were first blended with milk to improve the growth rates of probiotics, and the optimum composition model was determined. The probiotic viabilities and chemical properties were analyzed for the samples made using the optimal formula. The optimal combination of the growth promoters for the synbiotic fermented milk product was 1.12% peptides, 3% fructooligosaccharides (FOS), and 1.87% isomaltooligosaccharides (IMO). A product manufactured according to the formula of the optimum model was analyzed, showing that the model was effective in improving the viability of both Lactobacillus spp. and Bifidobacterium spp.

M.-J. CHEN, K.-N. CHEN, AND C.-W. LIN ABSTRACT: The purpose of this research was to evaluate the effects of various concentrations of glucono-- lactone (GDL) and skim milk powder, as well as the addition of prebiotics, on the rheology and probiotic viabilities of dairy tofu. Additionally, modern optimization techniques were applied to attempt to determine the optimal processing conditions and growth rate for the selected probiotics (Lactobacillus. acidophilus, L. casei, Bifidobacteria bifidum, and B. longum). There were 2 stages in this research to accomplish the goal. The 1st stage was to derive surface models using response surface methodology (RSM); the 2nd stage performed optimi- zation on the models using sequential quadratic programming (SQP) techniques. The results were demon- strated to be effective. The most favorable production conditions of dairy tofu were 1% GDL, 0% peptides, 3% isomaltooligosaccharides (IMO), and 18% milk, as confirmed by subsequent verification experiments. Analysis of the sensory evaluation results revealed no significant difference between the probiotic dairy tofu and the GDL analog in terms of texture and appearance (P > 0.05). The viable numbers of probiotics were well above the recommended limit of 10 6 CFUg for the probiotic dairy tofu throughout the tested storage period.

Food allergy is now recognized as a worldwide problem, and like other atopic disorders its incidence appears to be increasing. Kefir is reported to possess the ability to reduce intestinal permeation of food antigens; however, no experimental study has clearly evaluated the relationships between kefir consumption, allergen-specific IgE response, and intestinal microflora. The aim of this study was to evaluate the effect of oral consumption of milk kefir and soymilk kefir on in vivo IgE and IgG1 production induced by ovalbumin (OVA) in mice. The effects of kefir administration on the murine intestinal microflora were also examined. Oral administration of milk kefir and soymilk kefir for 28 days significantly increased the fecal populations of bifidobacteria and lactobacilli, while it significantly decreased those of Clostridium perfringens. Milk kefir and soymilk kefir also significantly decreased the serum OVA-specific IgE and IgG1 levels for both groups, but not those of the IgG2a analogues. Consumption of milk kefir and soymilk kefir suppressed the IgE and IgG1 responses and altered the intestinal microflora in our supplemented group, suggesting that milk kefir and soymilk kefir may be considered among the more promising food components in terms of preventing food allergy and enhancement of mucosal resistance to gastrointestinal pathogen infection. Copyright © 2006 Society of Chemical Industry

The purpose of this research was to study the effect of freeze drying on the microorganisms in kefir. Influences of lyoprotectants and rehydrated media (water at 4°C, 25°C; 10% reconstituted milk at 4°C, 25°C) on the viability of lactic acid bacteria and yeasts in freeze-dried kefir were investigated. Kefir was made from cow milk which was inoculated with 5% kefir grains, and incubated at 20°C for 20 h. Lyoprotectants (galactose, lactose, maltose, sucrose and trehalose) were added independently before dehydration of kefir by freeze drying. Results indicated significant loss in viability of microorganisms in kefir after freeze-drying. Addition of 10% galactose or 10% sucrose as lyoprotectants significantly increased the survival rates of both lactic acid bacteria and yeasts (p 0.05). (Asian-Aust. J. Anim. Sci. 2006. Vol 19, No. 1 : 126-130)

One of the prerequisites for the successful implementation of industrial-scale goat kefir production is to understand the effects of different kefir grains and culture conditions on the microbial and chemical properties of the goat kefir. Thus, the objectives of the present study were to evaluate the characteristics of kefir grains in Taiwan on the microbial and chemical properties of goat milk kefir, as well as to understand the influence of culture conditions on production of medium chain-length triglycerides (MCT). Kefir grains were collected from households in northern Taiwan. Heat-treated goat milk was inoculated with 3-5% (V/W) kefir grains incubated at 15, 17.5, 20 or 22.5°C for 20 h, and the microflora count, ethanol content, and caproic (C6), caprylic (C8), and capric acid (C10) levels measured at 4 h intervals. Our results indicate that incubation with kefir grains results in 10 6 -10 7 CFU/ml microflora count and 1.18 g/L of ethanol content at 20 h of fermentation. Incubation with 5% kefir grain at 20-22.5°C produces the highest MCT levels. (Asian-Aust. J. Anim. Sci. 2005. Vol 18, No. 5 : 711-715)

ABSTRACT ABSTRACT ABSTRACT: The purpose of this research was to improve probiotic microencapsulation using prebiotics and : The purpose of this research was to improve probiotic microencapsulation using prebiotics and : The purpose of this research was to improve probiotic microencapsulation using prebiotics and : The purpose of this research was to improve probiotic microencapsulation using prebiotics and : The purpose of this research was to improve probiotic microencapsulation using prebiotics and modern optimization techniques to determine optimal processing conditions, performance, and survival rates. modern optimization techniques to determine optimal processing conditions, performance, and survival rates. modern optimization techniques to determine optimal processing conditions, performance, and survival rates. modern optimization techniques to determine optimal processing conditions, performance, and survival rates. modern optimization techniques to determine optimal processing conditions, performance, and survival rates. Prebiotics (fructooligosaccharides or isomaltooligosaccharides), growth promoter (peptide), and sodium algi- Prebiotics (fructooligosaccharides or isomaltooligosaccharides), growth promoter (peptide), and sodium algi- Prebiotics (fructooligosaccharides or isomaltooligosaccharides), growth promoter (peptide), and sodium algi- Prebiotics (fructooligosaccharides or isomaltooligosaccharides), growth promoter (peptide), and sodium algi- Prebiotics (fructooligosaccharides or isomaltooligosaccharides), growth promoter (peptide), and sodium algi- nate were incorporated as coating materials to microencapsulate 4 probiotics ( nate were incorporated as coating materials to microencapsulate 4 probiotics ( nate were incorporated as coating materials to microencapsulate 4 probiotics ( nate were incorporated as coating materials to microencapsulate 4 probiotics ( nate were incorporated as coating materials to microencapsulate 4 probiotics (Lactobacillus acidophilus Lactobacillus acidophilus Lactobacillus acidophilus Lactobacillus acidophilus Lactobacillus acidophilus, Lacto- Lacto- Lacto- Lacto- bacillus casei bacillus casei bacillus casei bacillus casei bacillus casei, Bifidobacterium bifidum Bifidobacterium bifidum Bifidobacterium bifidum Bifidobacterium bifidum Bifidobacterium bifidum, and Bifidobacterium longum Bifidobacterium longum Bifidobacterium longum Bifidobacterium longum Bifidobacterium longum). The proportion of the prebiotics, pep- ). The proportion of the prebiotics, pep- ). The proportion of the prebiotics, pep- ). The proportion of the prebiotics, pep- ). The proportion of the prebiotics, pep- tide, and sodium alginate was optimized using response surface methodology (RSM) to 1st construct a surface tide, and sodium alginate was optimized using response surface methodology (RSM) to 1st construct a surface tide, and sodium alginate was optimized using response surface methodology (RSM) to 1st construct a surface tide, and sodium alginate was optimized using response surface methodology (RSM) to 1st construct a surface tide, and sodium alginate was optimized using response surface methodology (RSM) to 1st construct a surface model, with sequential quadratic programming (SQP) subsequently adopted to optimize the model and evaluate model, with sequential quadratic programming (SQP) subsequently adopted to optimize the model and evaluate model, with sequential quadratic programming (SQP) subsequently adopted to optimize the model and evaluate model, with sequential quadratic programming (SQP) subsequently adopted to optimize the model and evaluate model, with sequential quadratic programming (SQP) subsequently adopted to optimize the model and evaluate the survival of microencapsulated probiotics under simulated gastric fluid test. Optimization results indicated the survival of microencapsulated probiotics under simulated gastric fluid test. Optimization results indicated the survival of microencapsulated probiotics under simulated gastric fluid test. Optimization results indicated the survival of microencapsulated probiotics under simulated gastric fluid test. Optimization results indicated the survival of microencapsulated probiotics under simulated gastric fluid test. Optimization results indicated that 1% sodium alginate mixed with 1% peptide and 3% fructooligosaccharides as coating materials would that 1% sodium alginate mixed with 1% peptide and 3% fructooligosaccharides as coating materials would that 1% sodium alginate mixed with 1% peptide and 3% fructooligosaccharides as coating materials would that 1% sodium alginate mixed with 1% peptide and 3% fructooligosaccharides as coating materials would that 1% sodium alginate mixed with 1% peptide and 3% fructooligosaccharides as coating materials would produce the highest survival in terms of probiotic count. The verification experiment yielded a result close to the produce the highest survival in terms of probiotic count. The verification experiment yielded a result close to the produce the highest survival in terms of probiotic count. The verification experiment yielded a result close to the produce the highest survival in terms of probiotic count. The verification experiment yielded a result close to the produce the highest survival in terms of probiotic count. The verification experiment yielded a result close to the predicted values, with no significant difference ( predicted values, with no significant difference ( predicted values, with no significant difference ( predicted values, with no significant difference ( predicted values, with no significant difference (P > 0.05). The storage results also demonstrated that addition > 0.05). The storage results also demonstrated that addition > 0.05). The storage results also demonstrated that addition > 0.05). The storage results also demonstrated that addition > 0.05). The storage results also demonstrated that addition of prebiotics in the walls of probiotic microcapsules provided improved protection for the active organisms. of prebiotics in the walls of probiotic microcapsules provided improved protection for the active organisms. of prebiotics in the walls of probiotic microcapsules provided improved protection for the active organisms. of prebiotics in the walls of probiotic microcapsules provided improved protection for the active organisms. of prebiotics in the walls of probiotic microcapsules provided improved protection for the active organisms. These probiotic counts remained at 10 These probiotic counts remained at 10 These probiotic counts remained at 10 These probiotic counts remained at 10 These probiotic counts remained at 10 6 to 10 to 10 to 10 to 10 to 10 7 colony-forming units (CFU)/g for microcapsules stored for 1 mo colony-forming units (CFU)/g for microcapsules stored for 1 mo colony-forming units (CFU)/g for microcapsules stored for 1 mo colony-forming units (CFU)/g for microcapsules stored for 1 mo colony-forming units (CFU)/g for microcapsules stored for 1 mo and then treated in simulated gastric fluid test and bile salt test. and then treated in simulated gastric fluid test and bile salt test. and then treated in simulated gastric fluid test and bile salt test. and then treated in simulated gastric fluid test and bile salt test. and then treated in simulated gastric fluid test and bile salt test. K K K K Keywords: microencapsulation, prebiotics, probiotics, response surface methodology microencapsulation, prebiotics, probiotics, response surface methodology microencapsulation, prebiotics, probiotics, response surface methodology microencapsulation, prebiotics, probiotics, response surface methodology microencapsulation, prebiotics, probiotics, response surface methodology

This study aims to evaluate the antioxidative activities of cow-milk kefir and goat-milk kefir. Antioxidative mechanisms, including radical-scavenging effects, ferrous-ion chelating ability, reducing power and antioxidant activity, were investigated herein. Kefirs demonstrated significantly greater scavenging effects upon 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide radicals, an inhibition effect upon linoleic-acid peroxidation, and more substantial reducing power, but reduced glutathione peroxidase (GSH-Px) activity than was the case for milks. There was no significant difference between milks and kefirs as regards ferrous-ion chelating ability and superoxide dismutase (SOD) activity. These findings have demonstrated that kefirs possess antioxidant activity, thereby suggesting that kefirs are potential candidates for the role of useful natural antioxidant supplements for the human diet. (Asian-Aust. J. Anim. Sci. 2005. Vol 18, No. 4 : 567-573)

Growth promoters were added to skim milk to retain the viability of Lactobacillus acidophilus and Bifidobacterium longum to help the product meet the "therapeutic minimum" at the time of consumption. The experiments were divided into two parts. The first part of the study used chicory inulin, isomalto-oligosaccharides and sucrose to investigate the effects of sugars on the activity of L. acidophilus and B. longum. The results indicated that the addition of isomalto-oligosaccharides stimulated growth of L. acidophilus and B. longum, resulting in a higher level of the probiotics after one month storage and yielded better β-galactosidase activity during fermentation. The second part studied the effects of three growth promoters on the viability of the probiotic cultures and the response surface method was employed to find the optimal ratio for addition of the growth promoters. The optimal ratio for added calcium gluconate, sodium gluconate and N-acetylglucosamine in fermented milk drinks were established. The response surface method proved to be a very effective way of optimizing the activity of probiotic cultures when developing a new fermented milk drink. (Asian-Aust. J. Anim. Sci. 2004. Vol 17, No. 5 : 705-711)

Calcium gluconate (0.0 to 0.5%), sodium gluconate (0.0 to 1.0%), and N-acetylglucosamine (0.0 to1.0%) were added to skim milk to retain the viability of Lactobacillus acidophilus and Bifidobacterium longum .To carry out response surface modeling, the regression method was performed on experimental results to buildmathematical models. The models were then formulated as an objective function in an optimization problemthat was consequently optimized using a genetic algorithm approach to obtain the maximum viability of theprobiotics. The genetic algorithms (GAs) were examined to search for the optimal value. The results indicatedthat GAs were very effective for optimizing the activity of probiotic cultures.Keywords: probiotics, optimization, genetic algorithms, response surface modeling Introduction L ACTOBACILLUS ACIDOPHILUS AND BIFIDOBACTERIA MUST RETAIN viability and activity in food carriers to meet the suggested“therapeutic minimum” at the time of consumption (Playne 1994).Several factors have been claimed to affect the viability of probioticcultures in fermented milk products. The culture conditions, chem-ical composition of the fermentation medium (for example, carbo-hydrate source), final acidity, growth promoters and inhibitors, in-cubation temperature, and fermentation time and storagetemperature have all been identified as having effects on the man-ufacturing and storage of yogurt (Young and Nelson 1978; Hammanand Marth 1983; Kneifel and others 1993; Lankaputhra and Shah1995).In food microbiology, during the past few years, much effort hasbeen directed to developing models describing the combined ef-fects of the factors for microbe growth. Response surface method-ology (RSM) is a collection of statistical and mathematical tech-niques useful for developing, improving, and optimizing processes.The main advantage of RSM is the reduced number of experimentaltrials needed to evaluate multiple parameters and their interactions(Porretta and others 1995; Lee and others 2000). It was successfullyused for application in finding the optimum producing conditionsof the dairy product Kou Woan Lao (Weng and others 2001). The ex-perimental data were used to build mathematical models using theregression method. Once an appropriate approximating model isobtained, this model may be analyzed to determine the optimumconditions for the process.Genetic algorithms (GAs) are search procedures that imitate thenatural evolution process and can be used for the computation ofthe global maximum or minimum of a function (Mitchell 1996).Genetic algorithms differ from other search techniques in that theysearch among a population of points and use probabilistic ratherthan deterministic transition rules. As a result, genetic algorithmssearch more globally (Wang 1997). More details regarding GAs willbe included in the next section.The purpose of this research was to study the addition of calci-um gluconate (0.0 to 0.5%), sodium gluconate (0.0 to 1.0%) and N-acetylglucosamine (0.0 to 1.0%) in milk to optimize the viability ofthe probiotics by RSM and genetic algorithms. The experimentaldata were used to build mathematical models using the regressionmethod. The mathematical models were formulated as an objectivefunction (as in an optimization problem) and were optimized us-ing the genetic algorithm approach to obtain the maximum viabilityfor the probiotics. The endpoint was to provide a new optimizationmethod to improve the growth of probiotics in dairy products.

Prebiotics (peptides, N-acetyglucoamine, fructo-oligosaccharides, isomalto-oligosaccharides and galacto- oligosaccharides) were added to skim milk in order to improve the growth rate of contained Lactobacillus acidophilus, Lactobacillus casei, Bifidobacterium longum and Bifidobacterium bifidum. The purpose of this research was to study the potential synergy between probiotics and prebiotics when present in milk, and to apply modern optimization techniques to obtain optimal design and performance for the growth rate of the probiotics using a response surface-modeling technique. To carry out response surface modeling, the regression method was performed on experimental results to build mathematical models. The models were then formulated as an objective function in an optimization problem that was consequently optimized using a genetic algorithm and sequential quadratic programming approach to obtain the maximum growth rate of the probiotics. The results showed that the quadratic models appeared to have the most accurate response surface fit. Both SQP and GA were able to identify the optimal combination of prebiotics to stimulate the growth of probiotics in milk. Comparing both methods, SQP appeared to be more efficient than GA at such a task. (Asian-Aust. J. Anim. Sci. 2003. Vol 16, No. 6 : 894-902)

Response surface methodology (RSM) was applied to optimize the water-holding capacity (WHC) of fibrinogen/plasma protein gels heated at 75°C for 10 min to develop a new edible gel. The effects of varying the amount of fibrinogen, plasma protein, calcium, and pH were evaluated to optimize the WHC using RSM. Estimation of the overall effects of the 4 factors on the gels revealed that the pH and concentrations of fibrinogen and plasma protein significantly affected the WHC of the gels (p 0.05).

Lao-chao is a traditional Chinese fermented rice product with a sweet and fruity flavor, containing high levels of glucose, a little alcohol and milk-clotting characteristics. In order to optimize commercial production of lao-chao, Rhizopus javanicus and Saccharomyces cerevisiae were selected as the mold and yeast starter, respectively. A commercial mixed starter (chiu-yao) was used as control. Fermentation of the experimental combination revealed a sharp drop in pH (to 4.5) on the fourth day, remaining constant thereafter. Content of reducing sugars gradually decreased throughout the entire fermentation period. Of the free amino acids, higher quantities of alanine, leucine, proline, glutamic acid, glutamine and NH3 were noted. For sugars, glucose revealed the highest concentration, while organic acid levels, including those for oxalic, lactic, citric and pyroglutamic acid, increased throughout the fermentation period. Twenty-one compounds were identified by gas chromatography from aroma concentrates of the lao-chao culture filtrate, prepared using the headspace method. For the flavor components, higher quantities of ethanol, fusel oil and ester were determined in both culture filtrates. In regard to the evaluation of yogurt-like product, there were significant differences in alcoholic smell, texture and curd firmness. (Asian-Aust. J. Anim. Sci. 2002. Vol 15, No. 4 :602-609)

The characteristics of polysaccharides isolated from milk and soymilk kefir grains, and the composition of flavor and volatile compounds from soymilk kefic were investigated. Soymilk kefir grains revealed lower polysaccharide content than milk kefir grains, with a polysaccharide profile consisting primarily of glucose and galactose, with the former predominating. The apparent molecular weight of the polysaccharide was estimated at 1.7 × 10 4 Da. In comparison to nonfermented sbymilk, the concentrations of the key volatile compounds for soymilk kefir (such as acetaldehyde, acetone, diacetyl, and ethanol) increased, while n-hexanal concentration decreased. The acetaldehyde level for soymilk kefir was slightly higher, but levels for the other volatile compounds were lower than for milk kefir. Les polyosides et composes volatils responsables de la flaveur du kefir provenant de lait ou de lait de soja fermente sont compares. La teneur en glucides est plus faible dans les grains de kefir de lait du lait de soja (glucose et galactose). La teneur en composes volatils est superieure dans le lait de soja fermente que dans le produit non fermente.

Three strains of Vibrio parahaemolyticus (690, BCRC 13023 and BCRC 13025) implicated in food poisoning outbreaks in Taiwan were subjected to acid adaptation at pH 5.5 for 90 min. The growth behaviors of acid-adapted and non-adapted V. parahaemolyticus in the media supplemented with various nitrogen and carbon sources were investigated. The effects of acid adaptation on the thermostable direct hemolysin (TDH) secretion and fatty acid profiles of V. parahaemolyticus were also examined. Results showed that acid-adapted and non-adapted V. parahaemolyticus 690, BCRC 13023 and BCRC 13025 grew similarly in TSB-3% NaCl and basal media supplemented with various carbon and nitrogen sources during incubation period. Higher TDH secretion was noted with V. parahaemolyticus 690 among the three strains. However, acid-adapted strains produced less amounts of TDH than non-adapted strains when they were grown in TSB-3% NaCl. 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Intestinal protection is an important function of probiotics. However, there is no evidence that the potential probiotic Lactobacillus paracasei 01 is able to improve intestinal function. In the present study, the protective effect and underlying mechanisms of L. paracasei 01 on intestinal epithelial cell in vitro were investigated.A fermented milk beverage containing L. paracasei 01 at 3 × 10(8) CFU mL(-1) was made from skim milk powder, glucose, sucrose and isomaltooligosaccharide. Results indicated that L. paracasei 01 fermented milk beverage could reduce macromolecule permeability caused by dextran sodium sulfate-induced damage. Besides, 50 mL L(-1) fermented milk beverage with live L. paracasei 01 in the cell medium could increase intestinal epithelial cell Caco-2 growth, decrease lipopolysaccharide/tumor necrosis factor-α (TNF-α)/interferon-γ (IFN-γ)-induced Caco-2 cell death and chemokine CCL-20 production. The protecting mechanisms involve promoting intestinal epithelial cell growth and intestinal epithelial integrity to strengthen the intestinal barrier against chemical and inflammatory stimuli-induced damage. Inhibition of inflammatory cytokines and chemokine such as IFN-γ, TNF-α and CCL-20 also contributes to the beneficial effects of this product on epithelial function.Lactobacillus paracasei 01 fermented milk beverage may be a new functional food with intestinal protecting effects in vitro.

Lactobacilli kefiranofaciens M1 has shown novel immunomodulation and anti-allergy probiotic attributes in cell and animal models. An acute oral toxicity assessment of L. kefiranofaciens M1 was evaluated in Sprague-Dawley rats. The rats were randomly assigned to four groups (12 rats/sex/group): the low dose group was orally gavaged with L. kefiranofaciens M1 at 3.0×10(8)cfu/kg bw while the medium dose and high dose groups received 9.0×10(9)cfu/kg bw and 1.8×10(10)cfu/kg bw, respectively, for 28days. The control group received phosphate buffer saline. The body weights were measured weekly while blood samples were collected for haematology and serum biochemistry tests. Histopathology of the organs (heart, liver, kidney, adrenal glands, spleen, ovary, testis), and urinalysis were conducted on study termination. The body weight gain of the L. kefiranofaciens M1 and control groups were comparable during the administration period. Overall, L. kefiranofaciens M1 did not induce adverse effects on haematology, serum biochemistry, and urinalysis parameters. Gross and microscopic histopathology of the organs revealed no toxicity effect of L. kefiranofaciens M1. In conclusion, 1.8×10(10)cfu/kg bw of L. kefiranofaciens M1 was considered as the no-observed-adverse-effect-level (NOAEL), which was the highest dose tested in the present study.

Lactobacillus kefiranofaciens M1 is a novel probiotic strain that was isolated from kefir grains. Previously, we have demonstrated the immunoregulatory, anti-allergic, anti-asthmatic and anti-colitis abilities of L. kefiranofaciens M1 in a number of in-vitro and in-vivo experiments. However, whether the effects of L. kefiranofaciens M1 are elicited directly on the host or act by regulating the host's microbiota remains unknown. A number of studies have used germ-free or gnotobiotic animals to investigate the relationship between probiotics and colitis; therefore the aim of this study was to investigate the effects of L. kefiranofaciens M1 on germ-free mice. Such an approach should help in determining the direct effects of L. kefiranofaciens M1 on the host itself. Four-week-old female germ-free mice were inoculated intragastrically with 2×10(8) CFU/mouse L. kefiranofaciens M1 once or at 2-day intervals for 14 days. Bacterial colonization, the Th1/Th2 cytokine profile of the mice's splenocytes and the anti-colitis effect of L. kefiranofaciens M1 were investigated. The strongest response in terms of splenic Th1 cytokine IFN-γ and IL-12 production upon TLR activation was detected in the continuous treatment group when comparing to the single inoculation group and the germ-free control. In addition, continuous inoculation with L. kefiranofaciens M1 was found to ameliorate the symptoms of DSS-induced colitis in germ-free mice. However, L. kefiranofaciens M1 failed to colonize the host. Thus it would seem that L. kefiranofaciens M1 is likely to act directly on the host and not be involved in microbiota regulation.

Bacillus cereus strain C1L, screened from the rhizosphere of Lilium formosanum, prevents severe lily leaf blight. The purpose of this study was to formulate a new biopesticide by microencapsulating the C1L strain to enhance residual stability. In addition, since the viable cell number of Bacillus cereus C1L is an important factor affecting the biological effects of this fungicide, this study also used the response surface methodology and sequential quadratic programming methods to optimize the best formula for maximum viability of Bacillus cereus C1L microencapsulated by spray drying. Optimization results revealed that the strain could maintain 42% viability with 18.3% maltodextrin and 12.5% gum arabic as coating materials and spray drying at an outlet temperature of 73.5 oC. A comparison with chemical pesticides showed that maneb was the most efficient disease inhibitor, followed by B. cereus C1L encapsulated by spray drying. The chemical pesticide probenazole and B. cereus C1L encapsulated by extrusion revealed no significant differences in disease control.

UNLABELLED The purpose of this study was to manufacture new functional low-fat cheeses using Taiwanese ropy fermented milk (TRFM) and Lactococcus lactis subsp. cremoris strains isolated from TRFM. After 28 d of ripening and storage, the viable populations of lactic acid bacteria (LAB) in the low-fat cheeses made with L. lactis subsp. cremoris TL1 (TL1), L. lactis subsp. cremoris TL4 (TL4), and TRFM still maintained above 10(8) CFU/g. The low-fat cheeses made with TL1 and TRFM showed higher moisture contents than the cheeses made with TL4, full-fat, and low-fat cheese controls. The low-fat cheeses made with TL1 and TL4 had higher customer preferential scores similar to full-fat cheese control in the sensory evaluation. Additionally, the low-fat cheeses fermented with TL1, TL4, and TRFM for 4 h had higher 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical-scavenging and ferrous ion-chelating abilities than the cheeses fermented with the starters for 8 h, full-fat, and low-fat cheese controls. A better angiotensin-converting enzyme (ACE) inhibition activity was also observed in the low-fat cheeses made with TL1, TL4, and TRFM than that in the full-fat and low-fat cheese controls during ripening and storage period. PRACTICAL APPLICATION As health-conscious consumers continue to seek low-fat alternatives in their diets, there remain strong interests for the dairy industry to develop low-fat cheeses to meet the demands. This study clearly demonstrated that the low-fat cheeses fermented with TL1 for 4 h showed a better overall acceptability and possessed antioxidative abilities and ACE inhibitory activities than other cheeses tested in this study. By improving its flavor and investigating the possible mechanisms of its functionalities in the future, this low-fat cheese might possibly be commercialized and give a positive impact on cheese consumption in the future.

In the present study, we have investigated the importance of fermentation media on grain formation and the microbial characteristics of sugary kefir. The sugary kefir grains were fermented in brown sugar, cow's milk or goat's milk. Using culture-dependent and culture-independent methods, we identified the microorganisms present in both the grains and filtrate and then evaluated their distribution. The structure of the grains was also observed by scanning electronic microscopy (SEM). The identification results indicated that there were remarkable changes in microbial ecological profiles of the sugary kefir grains and their filtrates when brown sugar and milk were compared as fermentation media. Three lactic acid bacteria (LAB) species (Leuconostoc mesenteroides, Lactobacillus mali and Lactobacillus hordei) were found in the grains fermented using brown sugar. However, four species, named Leu. mesenteroides, Lactococcus lactis, Bifidobacterium psychraerophilum and Enterococcus faecalis, were identified in the grains fermented using either cow's or goat's milk. The size and structure of the kefir grains were also significantly influenced by the culture medium. We hypothesize that the grains originally may contain many different microorganisms and the identified changes are an adaption to each specific medium during grain formation and growth. The distribution of strains thus may vary depending on the carbon and energy sources available for grain fermentation and these microbial changes will further affect the granulation and growth of the grains. This study is important to our understanding of the mechanism of kefir grain formation and growth because it explores the relationship between fermentation media and kefir microorganisms.

In this study, we assessed the anti-asthmatic effects of heat-inactivated Lactobacillus kefiranofaciens M1 (HI-M1) and its fermented milk using different feeding procedures and at various dosage levels. The possible mechanisms whereby HI-M1 has anti-allergic asthmatic effects were also evaluated. Ovalbumin (OVA)-allergic asthma mice that have been orally administrated the HI-M1 samples showed strong inhibition of production of T helper cell (Th) 2 cytokines, pro-inflammatory cytokines, and Th17 cytokines in splenocytes and bronchoalveolar fluid compared to control mice. An increase in regulatory T cell population in splenocytes in the allergic asthma mice after oral administration of H1-M1 was also observed. In addition, all of the features of the asthmatic phenotype, including specific IgE production, airway inflammation, and development of airway hyperresponsiveness, were depressed in a dose-dependent manner by treatment. These findings support the possibility that oral feeding of H1-M1 may be an effective way of alleviating asthmatic symptoms in humans.

This study demonstrated that oral feeding of heat-inactivated Lactobacillus (L b.) kefiranofaciens M1 from kefir grains effectively inhibited immunoglobulin (Ig) E production in response to ovalbumin (OVA) in vivo. The pattern of cytokine production by splenocyte cells revealed that the levels of cytokines produced by T helper (Th) 1 cells increased, and those of cytokines produced by Th2 cells decreased in the heat-inactivated M1 feeding group. These findings indicated that Lactobacillus kefiranofaciens M1 in the kefir played an important role in antiallergic activities. By additional analysis using flow cytometry and microarray, the mechanism of suppression of IgE production by oral feeding of the heat-inactivated M1 probably occurs because of upregulation of the expression of Cd2, Stat4, and Ifnr leading to skewing the Th1/Th2 balance toward Th1 dominance, elevation of the CD4+CD25+ regulatory T (Treg) percentage, and reduction of activated CD19+ B cells. Downregulation of complement system and components was also involved in suppression of IgE production. Practical Application: Kefir has long been considered good for health. Its health benefits include immunoregulatory effects. However, there is a lack of knowledge concerning the immunoregulatory effects induced by kefir lactic acid bacteria (LAB). Our data clearly demonstrated the antiallergic activity of kefir LAB, Lactobacillus (L b.) kefiranofaciens M1. By additional analysis using flow cytometry and microarray, the possible mechanism of suppression of IgE production by oral feeding of the heat-inactivated M1 was also elucidated. Our findings indicated that Lactobacillus kefiranofaciens M1 may have a great potential for utilization in functional food products.