Objective: To evaluate the effect of lactic acid bacteria and storage temperature on the microbial, chemical and mycotoxin composition of corn silage. Methods: Corn was harvested at 32.8% dry matter, and chopped to 1 to 2 cm. The chopped material was subjected to three treatments: i) control (distilled water); ii) $1{\times}10^6$ colony forming units (cfu)/g of Lactobacillus plantarum; iii) $1{\times}10^6cfu/g$ of Pediococcus pentosaceus. Treatments in triplicate were ensiled for 55 d at $20^{\circ}C$, $28^{\circ}C$, and $37^{\circ}C$ in 1-L polythene jars following packing to a density of approximately $800kg/m^3$ of fresh matter, respectively. At silo opening, microbial populations, fermentation characteristics, nutritive value and mycotoxins of corn silage were determined. Results: L. plantarum significantly increased yeast number, water soluble carbohydrates, nitrate and deoxynivalenol content, and significantly decreased the ammonia N value in corn silage compared with the control (p<0.05). P. pentosaceus significantly increased lactic acid bacteria and yeast number and content of deoxynivalenol, nivalenol, T-2 toxin and zearalenone, while decreasing mold population and content of nitrate and 3-acetyl-deoxynivalneol in corn silage when stored at $20^{\circ}C$ compared to the control (p<0.05). Storage temperature had a significant effect on deoxynivalenol, nivalenol, ochratoxin A, and zearalenone level in corn silage (p<0.05). Conclusion: Lactobacillus plantarum and Pediococcus pentosaceus did not decrease the contents of mycotoxins or nitrate in corn silage stored at three temperatures.
Hyeonwoo Kim;Jiwon Kim;Ji Won Cho;Kwang-Sung Ahn;Dong-Il Park;Sangsoo Kim
Genomics & Informatics
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v.21
no.3
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pp.40.1-40.11
/
2023
Microbial community profiling using 16S rRNA amplicon sequencing allows for taxonomic characterization of diverse microorganisms. While amplicon sequence variant (ASV) methods are increasingly favored for their fine-grained resolution of sequence variants, they often discard substantial portions of sequencing reads during quality control, particularly in datasets with large number samples. We present a streamlined pipeline that integrates FastP for read trimming, HmmUFOtu for operational taxonomic units (OTU) clustering, Vsearch for chimera checking, and Kraken2 for taxonomic assignment. To assess the pipeline's performance, we reprocessed two published stool datasets of normal Korean populations: one with 890 and the other with 1,462 independent samples. In the first dataset, HmmUFOtu retained 93.2% of over 104 million read pairs after quality trimming, discarding chimeric or unclassifiable reads, while DADA2, a commonly used ASV method, retained only 44.6% of the reads. Nonetheless, both methods yielded qualitatively similar β-diversity plots. For the second dataset, HmmUFOtu retained 89.2% of read pairs, while DADA2 retained a mere 18.4% of the reads. HmmUFOtu, being a closed-reference clustering method, facilitates merging separately processed datasets, with shared OTUs between the two datasets exhibiting a correlation coefficient of 0.92 in total abundance (log scale). While the first two dimensions of the β-diversity plot exhibited a cohesive mixture of the two datasets, the third dimension revealed the presence of a batch effect. Our comparative evaluation of ASV and OTU methods within this streamlined pipeline provides valuable insights into their performance when processing large-scale microbial 16S rRNA amplicon sequencing data. The strengths of HmmUFOtu and its potential for dataset merging are highlighted.
This study was conducted to investigate the effects of Momordica charantia saponin (MCS) on ruminal fermentation of maize stover and abundance of selected microbial populations in vitro. Five levels of MCS supplements (0, 0.01, 0.06, 0.30, 0.60 mg/mL) were tested. The pH, $NH_3-N$, and volatile fatty acid were measured at 6, 24, 48 h of in vitro mixed incubation fluids, whilst the selected microbial populations were determined at 6 and 24 h. The high dose of MCS increased the initial fractional rate of degradation at t-value = 0 ($FRD_0$) and the fractional rate of gas production (k), but decreased the theoretical maximum of gas production ($V_F$) and the half-life ($t_{0.5}$) compared with the control. The $NH_3-N$ concentration reached the lowest concentration with 0.01 mg MCS/mL at 6 h. The MSC inclusion increased (p<0.001) the molar proportion of butyrate, isovalerate at 24 h and 48 h, and the molar proportion of acetate at 24 h, but then decreased (p<0.05) them at 48 h. The molar proportion of valerate was increased (p<0.05) at 24 h. The acetate to propionate ratio (A/P; linear, p<0.01) was increased at 24 h, but reached the least value at the level of 0.30 mg/mL MCS. The MCS inclusion decreased (p<0.05) the molar proportion of propionate at 24 h and then increased it at 48 h. The concentration of total volatile fatty acid was decreased (p<0.001) at 24 h, but reached the greatest concentration at the level of 0.01 mg/mL and the least concentration at the level of 0.60 mg/mL. The relative abundance of Ruminococcus albus was increased at 6 h and 24 h, and the relative abundance of Fibrobacter succinogenes was the lowest (p<0.05) at 0.60 mg/mL at 6 h and 24 h. The relative abundance of Butyrivibrio fibrisolvens and fungus reached the greatest value (p<0.05) at low doses of MCS inclusion and the least value (p<0.05) at 0.60 mg/mL at 24 h. The present results demonstrates that a high level of MCS quickly inhibits in vitro fermentation of maize stover, while MCS at low doses has the ability to modulate the ruminal fermentation pattern by regulating the number of functional rumen microbes including cellulolytic bacteria and fungi populations, and may have potential as a feed additive applied in the diets of ruminants.
Ganjang-gejang (soy sauce-marinated crab) is a ready-to-eat (RTE) seafood and is also one of the most popular traditional dishes in Korea. It is generally prepared by washing raw blue crabs and then preserving them in soy sauce. Since this process does not involve cooking or any treatment with heat, it is difficult to control the microbiological quality of the final product. Thus, the objectives of this study were to compare the efficacies of various sanitizers in eliminating microorganisms on raw blue crab during the washing step and to evaluate the effectiveness of chitosan on the inhibition of microbial growth in the ganjang-gejang during storage. The raw blue crabs were submerged in chlorinated water (50 mg/L), peracetic acid (40 mg/L), acetic acid (5%) and lactic acid (5%) for 10 min at $25^{\circ}C$, respectively. The blue crabs treated with 5% acetic acid were marinated with soy sauce containing 0.5 and 1% of soluble chitosan, followed by storing them at 4 and $12^{\circ}C$ for up to 30 days. Results show that 5% acetic acid reduced the microbial populations on the blue crabs by 1.5 log CFU/g, which was significantly higher than those of other treatments. Based on these results, 5% acetic acid was selected for the washing step. The microbial populations of all ganjang-gejang samples significantly increased to about 8.0 CFU/g at $12^{\circ}C$ for 7 days. At $4^{\circ}C$, the microbial populations of the products containing 1% chitosan increased by about 2.9 CFU/g for 20 days, which were significantly lower than those (4.2-4.5 log CFU/g) of the products without and with 0.5% chitosan. Thus, these results suggest that 5% acetic acid washing of raw blue crabs and the addition of 1% chitosan in ganjang-gejang could improve the microbiological quality of the final products under refrigerated condition.
To use as a fundamental data for the sustainable agriculture, which is nowadays a major trend to keep the productivity and conserve the environment, 487 paddy soil samples were collected from 21 regions of the Gyeongnam Province and analyzed the chemical characteristics and microbial population of the soil. The microbial population densities were bacteria $298{\times}10^5$($4{\sim}3000{\times}10^5$ range), fungi $63{\times}10^3$($2{\sim}441{\times}10^3$ range), actinomycetes $19{\times}10^5$($0.2{\sim}1250{\times}10^5$ range), Bacillus sp. $53{\times}10^4$($4{\sim}890{\times}10^4$ range) and Pseudomonas sp. $198{\times}10^4CFU\;g^{-1}$($4{\sim}1724{\times}10^4CFU\;g^{-1}$ range), respectively. The microbial populations of the soil were in general higher in southern area than in the northern area of the Gyeongnam Province. The average ratio of bacteria/fungi population was 473. As soil clay content increased, the populations of aerobic bacteria, actinomycetes and Pseudomonas sp. were remarkably decreased. The ratio of aerobic bacteria and fungi was 1554 in sandy loam and clay loam 1144, while Bacillus sp./fungi ratio was 11 in clay loam and 10 in loam. On the topographical differences, aerobic bacteria and Bacillus sp./fungi ratio were the higher in coastal plains than any other areas. The microbial population densities from different soil types were generally lower in ill-drained paddy field than those of other paddy field. The content of $P_2O_5$, K, Ca, $NO_3-N$ and EC in soil were positively correlated to the population densities of aerobic bacteria, actinomycetes, fungi, Bacillus sp. and Pseudomonas sp.. The soil organic matter and Mg content were also positively correlated to the population densities of aerobic bacteria, actinomycetes, fungi and Bacillus sp.
Gunun, P.;Wanapat, M.;Gunun, N.;Cherdthong, A.;Sirilaophaisan, S.;Kaewwongsa, W.
Asian-Australasian Journal of Animal Sciences
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v.29
no.8
/
pp.1111-1119
/
2016
Mao seed is a by-product of the wine and juice industry, which could be used in animal nutrition. The current study was designed to determine the effect of supplementation of mao (Antidesma thwaitesianum Muell. Arg.) seed meal (MOSM) containing condensed tannins (CT) on rumen fermentation, nitrogen (N) utilization and microbial protein synthesis in goats. Four crossbred (Thai Native${\times}$Anglo Nubian) goats with initial body weight (BW) $20{\pm}2kg$ were randomly assigned to a $4{\times}4$ Latin square design. The four dietary treatments were MOSM supplementation at 0%, 0.8%, 1.6%, and 2.4% of total dry matter (DM) intake, respectively. During the experimental periods, all goats were fed a diet containing roughage to concentrate ratio of 60:40 at 3.0% BW/d and pangola grass hay was used as a roughage source. Results showed that supplementation with MOSM did not affect feed intake, nutrient intakes and apparent nutrient digestibility (p>0.05). In addition, ruminal pH and ammonia nitrogen ($NH_3$-N) were not influenced by MOSM supplementation, whilst blood urea nitrogen was decreased quadraticly (p<0.05) in goats supplemented with MOSM at 2.4% of total DM intake. Propionate was increased linearly with MOSM supplementation, whereas acetate and butyrate were remained the same. Moreover, estimated ruminal methane ($CH_4$) was decreased linearly (p<0.05) when goats were fed with MOSM at 1.6% and 2.4% of total DM intake. Numbers of bacteria and protozoa were similar among treatments (p>0.05). There were linear decreases in urinary N (p<0.01) and total N excretion (p<0.01) by MOSM supplementation. Furthermore, N retention was increased linearly (p<0.05) when goats were fed with MOSM supplementation at 1.6% and 2.4% of total DM intake. Microbial protein synthesis were not significantly different among treatments (p>0.05). From the current study, it can be concluded that supplementation of MOSM at 1.6% to 2.4% of total DM intake can be used to modify ruminal fermentation, especially propionate and N utilization in goats, without affecting the nutrient digestibility, microbial populations and microbial protein synthesis.
Kim, Ji-Sun;Jung, Jee-Yun;Cho, Seung-Kee;Kim, Ji-Eun;Kim, Tae-Jip;Kim, Beom-Soo;Han, Nam-Soo
Korean Journal of Food Science and Technology
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v.42
no.3
/
pp.281-286
/
2010
The objective of this study was to analyze the microbial populations in the raw ingredients of kimchi and their changes during an automated commercial manufacturing process. High population numbers of total aerobic bacteria, lactic acid bacteria, Leuconostoc sp., and yeast were detected in garlic, ginger, red pepper powder and this result revealed that these ingredients were the major source of microbials in kimchi. Additionally, during the salting process of Chinese cabbage, rapid microbial growth was observed and the consecutive washing process was determined to be ineffective, lowering the microbial count by only one log reduction. Yeast was also detected in various ingredients. These results strongly suggest that, in order to lower the microbial population numbers in kimchi, the side-ingredients and salting process should be subjected to the appropriate sanitization or sterilization processes at the HACCP level. Beside, treatment of salted Chinese cabbage with sodium hypochlorite solutions after the salting step is recommended. To inhibit yeast growth, appropriate chemical treatment and approval of additive uses to control microbials should be considered. These experimental results and suggestions will be used to improve the kimchi manufacturing process in factories.
Wanapat, Metha;Anantasook, N.;Rowlinson, P.;Pilajun, R.;Gunun, P.
Asian-Australasian Journal of Animal Sciences
/
v.26
no.4
/
pp.529-536
/
2013
The objective of this study was to investigate the effect of levels of cottonseed meal with various carbohydrate sources in concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in dairy bulls. Four, 6 months old dairy bulls were randomly assigned to receive four dietary treatments according to a $2{\times}2$ factorial arrangement in a $4{\times}4$ Latin square design. Factor A was carbohydrate source; cassava chip (CC) and cassava chip+rice bran in the ratio of 3:1 (CR3:1), and factor B was cotton seed meal levels in the concentrate; 109 g CP/kg (LCM) and 328 g CP/kg (HCM) at similar overall CP levels (490 g CP/kg). Bulls received urea-lime treated rice straw ad libitum and were supplemented with 10 g of concentrate/kg BW. It was found that carbohydrate source and level of cotton seed meal did not have significant effects on ruminal pH, ammonia nitrogen concentration, microbial protein synthesis or feed intake. Animals which received CC showed significantly higher BUN concentration, ruminal propionic acid and butyric acid proportions, while dry matter, organic matter digestibility, populations of total viable bacteria and proteolytic bacteria were lower than those in the CR3:1 treatment. The concentration of total volatile fatty acids was higher in HCM than LCM treatments, while the concentration of butyric acid was higher in LCM than HCM treatments. The population of proteolytic bacteria with the LCM treatments was higher than the HCM treatments; however other bacteria groups were similar among the different levels of cotton seed meal. Bulls which received LCM had higher protein digestibility than those receiving HCM. Therefore, using high levels of cassava chip and cotton seed meal might positively impact on energy and nitrogen balance for the microbial population in the rumen of the young dairy bull.
Attempts were made to define the characteristics of microbial community as an indicator of Kimchi fermentation. Determination of communities was carried out by simple Gram-stain, followed by direct microcopic counts. In room-temperature $(15^{\circ}C)$ fermentation, microbial succession was occurred in the order of communities of Gram-positive bacteria, yeasts and Gram-negative bacteria. It was characteristic that Gram-positive bacterial community was developed during the production of lactic acid, yeasts community was developed to cause rancidity, and Gram-negative bacterial community was relevant to maceration (or softening) as well as rancidity. The fluctuation of apparent Gram-negative reaction group might be used as a criterion of death or aging of Gram-positive bacterial populations. In low-temperature fermentation $(5^{\circ}C)$, however, it was found that yeasts and Gram-negative bacterial communities did not developed but only Gram-positive bacterial community did. It follows from these results mentioned above that maturity of Kimchi depends on the development of Cram-positive bacterial community. Thus, the size and occurrence of microbial community are avaiable for an indicator of Kimchi fermentation, and also determination of community could be a useful method to predict the maturity.
Effect of chlorine dioxide ($ClO_2$) treatment on the microbial and physicochemical changes of fish paste was investigated. Fish paste samples were treated with 5, 10, and 50 ppm of $ClO_2$ solution, respectively, After $ClO_2$ treatment, fish paste samples were individually packaged and stored at 4$^{\circ}C$. The initial microbial loads of samples were 3.8 log CFU/g in total bacterial count, and 2.5 log CFU/g in yeasts and molds. Microbial growth of fish paste during storage showed that populations of total bacteria, yeast and mold were significantly reduced by $ClO_2$ treatment. In particular, the treatment of 50 ppm $ClO_2$ decreased total bacterial count the most significantly among the $ClO_2$ treated fish pastes. The pH and VBN of fish paste decreased with increasing $ClO_2$ concentration. Thiobarbituric acid reacted substance (TBARS) values of treated fish paste increased during storage, regardless of $ClO_2$ concentration. This study showed that 50 ppm chloride dioxide was the optimum dose level to extend the shelf-life of fish paste.
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