• Journal of Dairy Science and Biotechnology
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• v.33 no.2
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• pp.119-127
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• 2015

To date, detection of microbial populations in dairy products has been performed using culture media, which is a time-consuming and laborious method. The recently developed chromogenic media could be more rapid and specific than classical culture media. However, the newly developed molecular-based technology can detect microbial populations with greater rapidity and sensitivity than the classical method involving culture media and chromogenic media. This molecular-based technology could provide various options for monitoring the characterization of different states of bacteria and cells. Thus, it could help upgrade the processing system of the dairy industry so as to maintain the safety and quality of dairy foods. Among the various newly developed molecular-based technologies, flow cytometry can potentially be used for monitoring microbiological populations in the dairy industry if official international standards are available for this purpose. When omics technology would have biomarker identification, it could be regarded as the rapid and sensitive analytical methods. Methods based on PCR, which has become a basic technique in microbiological research, can be developed and validated as alternative methods for quantification of dairy microorganisms. This review discusses methods for monitoring microbiological populations in dairy foods and the limitations of these studies, as well as the need for further research on such methods in the dairy industry.

• Journal of Wetlands Research
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• v.26 no.2
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• pp.168-181
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• 2024

Constructed wetlands (CWs) deliver a range of ecosystem services, including the removal of contaminants, sequestration and storage of carbon, and enhancement of biodiversity. These services are facilitated through hydrological and ecological processes such as infiltration, adsorption, water retention, and evapotranspiration by plants and microorganisms. This study investigated the correlations between microbial populations, soil physicochemical properties, and treatment efficiency in a horizontal subsurface flow constructed wetland (HSSF CW) treating runoff from roads and parking lots. The methods employed included storm event monitoring, water quality analysis, soil sampling, soil quality parameter analysis, and microbial analysis. The facility achieved its highest pollutant removal efficiencies during the warm season (>15℃), with rates ranging from 33% to 74% for TSS, COD, TN, TP, and specific heavy metals including Fe, Zn, and Cd. Meanwhile, the highest removal efficiency was 35% for TOC during the cold season (≤15℃). These high removal rates can be attributed to sedimentation, adsorption, precipitation, plant uptake, and microbial transformations within the CW. Soil analysis revealed that the soil from HSSF CW had a soil organic carbon content 3.3 times higher than that of soil collected from a nearby landscape. Stoichiometric ratios of carbon (C), nitrogen (N), and phosphorus (P) in the inflow and outflow were recorded as C:N:P of 120:1.5:1 and 135.2:0.4:1, respectively, indicating an extremely low proportion of N and P compared to C, which may challenge microbial remediation efficiency. Additionally, microbial analyses indicated that the warm season was more conducive to microorganism growth, with higher abundance, richness, diversity, homogeneity, and evenness of the microbial community, as manifested in the biodiversity indices, compared to the cold season. Pollutants in stormwater runoff entering the HSSF CW fostered microbial growth, particularly for dominant phyla such as Proteobacteria, Actinobacteria, Acidobacteria, and Bacteroidetes, which have shown moderate to strong correlations with specific soil properties and changes in influent-effluent concentrations of water quality parameters.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.10
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• pp.1400-1410
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• 2004

Eight crossbred (75% Holstein Friesian) cows in mid-lactation were randomly assigned to a switchback design with a 2x2 factorial arrangement to evaluate two nonstructural carbohydrate (NSC) sources (corn meal and cassava chips) with different rumen degradability and used at two levels of NSC (55 vs. 75%) with protein source (supplied by urea in the concentrate mix). The treatments were 1) Low degradable low level of corn (55%) 2) Low degradable high level of corn (75%) 3) High degradable low level of cassava (55%) and 4) High degradable high level of cassava (75%). The cows were offered the treatment concentrate at a ratio to milk yield at 1:2. Urea-treated rice straw was offered ad libitum as the roughage and supplement with 1 kg/hd/d cassava hay. The results revealed that total DM intake, BW and digestion coefficients of DM were not affected by either level or source of energy. Rumen fermentation parameters; NH3-N, blood urea nitrogen and milk urea nitrogen were unaffected by source of energy, but were dramatically increased by level of NSC. Rumen microorganism populations were not affected (p>0.05) by source of energy, but fungal zoospores were greater for cassava-based concentrate than corn-based concentrate. Milk production and milk composition were not affected significantly by diets containing either source or level of NSC, however concentrate than corn-based concentrate averaging (4.4 and 4.2, respectively). Likewise, income over feed, as estimated from 3.5% FCM, was higher on cassava-based concentrate than corn-based concentrate averaging (54.0 and 51.4 US$/mo, respectively). These results indicate that feeding diets containing either cassava-based diets and/or a higher of oncentrates up to 75% of DM with NPN (supplied by urea up to 4.5% of DM) can be used in dairy rations without altering rumen ecology or animal performance compared with corn-based concentrate.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.9
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• pp.1332-1339
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• 2017

Objective: This study investigated the effects of Bacillus subtilis CSL2 (B. subtilis CSL2) administration before Salmonella challenge on the fecal microbiota and microbial functionality of Hy-line Brown (HLB) laying hens. Methods: Fecal samples were collected from control (CON), Salmonella-infected (SAL) and Salmonella-infected, probiotic-treated (PRO) groups before and after Salmonella challenge for microbiome analysis using 16S rRNA gene pyrosequencing. Results: Infection with Salmonella led to decreased microbial diversity in hen feces; diversity was recovered with Bacillus administration. In addition, Salmonella infection triggered significant alterations in the composition of the fecal microbiota. The abundance of the phylum Firmicutes decreased while that of Proteobacteria, which includes a wide variety of pathogens, increased significantly. Bacillus administration resulted in normal levels of abundance of Firmicutes and Proteobacteria. Analysis of bacterial genera showed that Salmonella challenge decreased the population of Lactobacillus, the most abundant genus, and increased populations of Pseudomonas and Flavobacterium genera by a factor of 3 to 5. On the other hand, Bacillus administration caused the abundance of the Lactobacillus genus to recover to control levels and decreased the population of Pseudomonas significantly. Further analysis of operational taxonomic units revealed a high abundance of genes associated with two-component systems and secretion systems in the SAL group, whereas the PRO group had more genes associated with ribosomes. Conclusion: The results of this study indicate that B. subtilis CSL2 administration can modulate the microbiota in HLB laying hens, potentially acting as a probiotic to protect against Salmonella Gallinarum infection.

• The Plant Pathology Journal
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• v.25 no.1
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• pp.62-69
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• 2009

In this study, we characterized the bacterial strain KJ2C12 in relation with its biocontrol activity against Phytophthora capsici on pepper, and identified this strain using morphological, physiological, biochemical, fatty acid methyl ester, and 16S rRNA gene sequence analyses. Strain KJ2C12 significantly (P=0.05) reduced both final disease severity and areas under the disease progress curves of 5-week-old pepper plants inoculated with P. capsici compared to buffer-treated controls. As for the production of antibiotics, biofilms, biosurfactant, extracellular enzyme, HCN, and swarming activity, strain KJ2C12 produced an extracellular enzyme with protease activity, but no other productions or swarming activity. However, Escherichia coli produced weak biofilm only. Strain KJ2C12 could colonize pepper roots more effectively in a gnotobiotic system using sterile quartz sand compared to E. coli over 4 weeks after treatments. However, no bacterial populations were detected in 10 mM $MgSO_4$ buffer-treated controls. Strain KJ2C12 produced significantly higher microbial activity than the $MgSO_4$-treated control or E. coli over 4 weeks after treatments. Bacterial strain KJ2C12 was identified as Bacillus luciferensis based on morphological, physiological, and biochemical characteristics as well as FAME and 16S rRNA gene sequence analyses. In addition, these results suggested that B. luciferensis strain KJ2C12 could reduce Phytophthora blight of pepper by protecting infection courts through enhanced effective root colonization with protease production and an increase of soil microbial activity.

• Journal of Microbiology
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• v.42 no.3
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• pp.216-222
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• 2004

In a previous paper, the ogdH gene that encodes 2-oxoglutarat dehydrogenase was isolated from Salmonella typhimurium. The catalytic N-terminal region in the enzyme was found to be very specific for the Salmonella species. Therefore, the aim of the present study was to detect S. typhimurium in food sources using primers designed for OGDH-l and OGDH-2 which were based on the salmonella-specific region of the ogdH gene. A simple polymerase chain reaction (PCR) detection method was developed to detect low numbers of S. typhimurium in a chicken meat microbial consortium. Using the ogdH-specific primers under stringent amplification conditions and for gene probe analysis, fewer than 100 colony-forming units (CFUs) were detectable when pure cultures were employed. When the PCR assay was run on S. typhimurium-contaminated meat contents, only the positive meat samples containing as few as 200 CFUs reacted to the assay. The method employed for sample processing is simple and it was determined to provide a sensitive means of detecting trace amounts of S. typhimurium-specific sequences in the presence of mixed meat microbial populations. When compared with six representative intestinal gram-negative bacterial strains in foods, including Vibrio parahaemolyticus, V. vulnificus, Enterobacter cloacae, E. coli O157:H7, Pseudomonas aeruginosa, and Proteus sp., S. typhimurium had a unique and distinct PCR product (796 bp). In conclusion, the two OGDH primers were found to be rapid and sensitive detectors of Salmonella spp for the PCR method.

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.952-964
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• 2005

The diesel-degrading activities of biofilms sampled from petroleum-contaminated groundwaters in urban subway drainage systems were examined in liquid cultures, and the microbial populations of the biofilms were characterized by denaturing gel gradient electrophoresis (DGGE) and 16S rDNA sequence analysis. Biofilm samples derived from two sites (19 K and 20 K) at subway Station N and Station I could degrade around $80\%$ of applied diesel within 20 and 40 days, respectively, at $15^{\circ}C$, and these results were strongly correlated with the growth patterns of the biofilms. The closest phylogenetic neighbor of a dominant component in the 19 K biofilm was Thiothrix fructosivorans strain Q ($100\%$ similarity). Four dominant strains in the 20 K biofilm were closely related to Thiothrix fructosivorans strain Q ($100\%$ similarity), Thiothrix sp. CC-5 ($100\%$ similarity), Sphaerotilus sp. IF14 ($99\%$ similarity), and Cytophaga-Flexibacter-Bacterioides (CFB) group bacterium RW262 ($98\%$ similarity). Three dominant members in the Station I biofilms were very similar to uncultured Cytophagales clone CRE-PA82 ($91\%$ similarity), Pseudomonas sp. WDL5 ($97\%$ similarity), and uncultured CFB group bacterium LCK-64 ($94\%$ similarity). The microbial components of the biofilms differed depending on the sampling site. This is the first report on the isolation of clones highly similar to Thiothrix fructosivorans and Thiothrix sp. from biofilms in petroleum-polluted groundwaters, and the first evidence that these organisms may play major roles in petroleum degradation and/or biofilm-development.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.365-365
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• 2017

Yogurt is a food produced by bacterial fermentation of milk and the bacteria used to make it are known as "yogurt cultures". Most of them belong to probiotics such as Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus bacteria. Domestic fermented milk market is increasing and about 30 companies are producing yogurt. The purpose of this study was to analyze the quality characteristics of domestic commercial semisolid type yogurt. We collected 20 types of commercial yogurt at local markets. Physicochemical properties including pH, sugar content, acidity, viscosity and microbial characteristics of lactic acid bacteria counts were measured. The yogurt showed pH 4.5, 7.4~18.1% of sugar contents, 0.6~1.3% of total acids and 282~748 cP of viscosities. In the microorganism populations, lactic acid bacteria count were 6.5~11.5 Log CFU/mL and anaerobic lactic acid bacteria count were 7.2 ~ 11.1 Log CFU/mL. The quality characteristics were different depending on the constituents of the sample and the microorganisms used. These results are related to the quality characteristics of yogurts which are useful information about identifying new trends in domestic fermented milk industry.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.6
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• pp.837-844
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• 2006

Four, lactating dairy cows were randomly assigned according to a $2{\times}2$ Factorial arrangement in a $4{\times}4$ Latin square design to study supplementation of urea level (U) at 2 and 4% and sodium dl-malate (M) at 10 and 20 g/hd/d in concentrate. The treatments were as follows U2M10, U2M20, U4M10 and U4M20, respectively. The cows were offered the treatment concentrate at a ratio to milk yield at 1:2.5 and urea-treated rice straw was fed ad libitum. The results have revealed that rumen fermentation and blood metabolites were similar for all treatments. The populations of protozoa and fungal zoospores were significantly different as affected by urea level and sodium dl-malate. In addition, the viable bacteria were similar for amylolytic and proteolytic bacteria. Cellulolytic bacteria were significantly affected by level of sodium dl-malate especially Selenomonas ruminantium and Megasphaera elsdenii while Butyrivibrio fibrisolvens was significantly affected by level of urea supplementation. In conclusion, the combined use of concentrate containing high level of cassava chip at 75% DM with urea at 4% in concentrate and sodium dl-malate at 20 g/hd/d with UTS as a roughage could improv rumen ecology and microbial protein synthesis efficiency in lactating dairy cows.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.40 no.6
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• pp.356-361
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• 2007

Recently, the development of various culture-independent identification techniques for environmental microbes has greatly enhanced our knowledge of microbial diversity. In particular, denaturing gradient gel electrophoresis (DGGE) of 16S rDNA fragments, generated using the polymerase chain reaction (PCR) is frequently used to examine the diversity of environmental bacterial populations. This method consists of direct extraction of the environmental DNA, amplification of the 200-600 bp 16S rDNA fragments with universal primers, and separation of the fragments according to their melting point on a denaturing gradient gel. In this study, we investigated the seaside microbial community in coastal areas of Busan, Korea, using culture-independent techniques. First, marine genomic DNA was extracted from seawater samples collected at Songjeong, Gwangahn, and Songdo Beaches. Then, PCR was used to amplify the bacterial 16S rDNA using universal primers, and DGGE was used to separate the amplified 500 bp 16S rDNA fragments. Finally, the tested 16S rDNA genes were further analyzed by sequencing. Based on these experiments, we found that DGGE analysis clearly showed variation among the regional groups. It can be used to monitor rapid changes in the bacterial diversity of various environments. In addition, the sequence analysis indicated the existence of many unculturable bacteria, in addition to Arcobacter, Pseudoaltermonas, and Vibrio species.