It is very important to differentiate of DNA derived from live or dead bacteria within mixed microbial communities in aquatic environments. Ethidium monoazide (EMA) is a DNA intercalating agent and the treatment of EMA with strong visible light cleaves the genomic DNA of bacteria. In dead bacterial cells, EMA intercalates into the genomic DNA, induces the cleavage of DNA, and inhibits the PCR amplification. In this study, we developed the EMA-PCR and EMA real-time PCR to detect the DNA derived from viable Escherichia coli (E.coli) in mixed cultures of live and dead E.coli. The treatment of EMA, $50{\mu}g/mL$, and 650 W visible halogen light exposure for 2 minutes cleaved the genomic DNA derived from heat killed E.coli but did not those of live E.coli. EMA-PCR could detect the DNA from live E.coli in mixed culture samples of live and dead E.coli at various ratio and there was no DNA amplification in only dead E.coli cultures. Similar results were observed in EMA real-time PCR. Further studies are needed to develop various EMA-PCR methods to detect viable waterborne pathogens such as Helicobacter pylori, Giardia lamblia, and so on.
Objective: Light is a significant component of housing environment in commercial poultry industry. This study was conducted to investigate whether Pekin ducks perform better under monochromatic lights than under white light with respect to their growth performance, carcass quality, eyeball development, oxidation resistance, and cecal bacterial communities. Methods: A total of 320 one-day-old male Pekin ducklings were randomly distributed into five rooms with different light treatments, white, red, yellow, green, and blue light. Each room consisted of 4 replicated pens with 16 ducklings per pen. Results: Blue light significantly decreased fat deposition by decreasing abdominal fat. Long wavelength light, such as red, green, and yellow light, considerably increased the back-to-front eyeball diameter and the red light potentially enlarged the side-to-side eyeball diameter. Besides, the blue light had adverse effects on the oxidation resistance status in terms of increasing the product malonaldehyde of lipid oxidation and decreasing the plasma concentration of total superoxide dismutase. The phyla of Firmicutes had the greatest abundance in the green and blue treatments, while Bacteroidetes in blue treatment was the least. The genus of Faecalibacterium was significantly lower under the red light. Conclusion: The high risk of cecal health status and decreased anti-oxidation activity were observed under blue light. Red, yellow, and green light might increase the risk of oversized eyeball and cecal illness. Therefore, monochromatic lights compared to white light did not show advantages on the performance of housing ducks, it turns out that the white light is the best light condition for grow-out ducks.
Soil microorganism activity in an agricultural field is affected by various factors including climate conditions, soil chemical properties, and crop cultivation. In this study, we elucidate the correlation between microorganism activity and agricultural environment factors using the dehydrogenase activity (DHA) value, which is one of the indicators of soil microbial activity. As a result, the various factors noted above were related to the DHA value. Annual rainfall, soil Mg2+, bacterial and fungal diversities, types of crops, developmental stages, seasons, and cultivation status were highly correlated with the DHA value. Furthermore, next-generation sequencing (NGS) analysis was used to identify that the type of crop affected soil microbial compositions of both bacteria and fungi. Soil used for soybean cultivation showed the highest relative abundance for Verrucomicrobia, Planctomycetes, and Acidobacteria but Actinobacteria and Firmicutes had the lowest relative abundance. In the case of soil used for potato cultivation, Actinobacteria had the highest relative abundance but Proteobacteria had the lowest relative abundance. Armatimonadetes showed the highest relative abundance in soil used for cabbage cultivation. Among the fungal communities, Mortierellomycota had the highest relative abundance for soybean cultivation but the lowest relative abundance for cabbage cultivation; further, Rozellomycota, Chytridiomycota, and Cercozoa had the highest relative abundance for cabbage cultivation. Basidiomycota had the highest relative abundance for potato cultivation but the lowest relative abundance for soybean cultivation.
Salmonella enterica serovar Typhimurium isolate HJL777 is a virulent bacterial strain in pigs. The high rate of salmonella infection are at high risk of non-typhoidal salmonella gastroenteritis development. Salmonellosis is most common in young pigs. We investigated changes in gut microbiota and biological function in piglets infected with salmonella via analysis of rectal fecal metagenome and intestinal transcriptome using 16S rRNA and RNA sequencing. We identified a decrease in Bacteroides and increase in harmful bacteria such as Spirochaetes and Proteobacteria by microbial community analysis. We predicted that reduction of Bacteroides by salmonella infection causes proliferation of salmonella and harmful bacteria that can cause an intestinal inflammatory response. Functional profiling of microbial communities in piglets with salmonella infection showed increasing lipid metabolism associated with proliferation of harmful bacteria and inflammatory responses. Transcriptome analysis identified 31 differentially expressed genes. Using gene ontology and Innate Immune Database analysis, we identified that BGN, DCN, ZFPM2 and BPI genes were involved in extracellular and immune mechanisms, specifically salmonella adhesion to host cells and inflammatory responses during infection. We confirmed alterations in gut microbiota and biological function during salmonella infection in piglets. Our findings will help prevent disease and improve productivity in the swine industry.
Objective: This study aimed to assess the impact of a hydroethanolic extract of walnut green husks (WGH) on rumen fermentation and the diversity of bacteria, methanogenic archaea, and fungi in sheep fed a high-concentrate diet. Methods: Five healthy small-tailed Han ewes with permanent rumen fistula were selected and housed in individual pens. This study adopted a self-controlled and crossover design with a control period and an experimental period. During the control period, the animals were fed a basal diet (with a ratio of concentrate to roughage of 65:35), while during the treatment period, the animals were fed the basal diet supplemented with 0.5% hydroethanolic extract of WGH. Fermentation parameters, digestive enzyme activities, and microbial diversity in rumen fluid were analyzed. Results: Supplementation of hydroethanolic extract of WGH had no significant effect on feed intake, concentrations of total volatile fatty acids, isovalerate, ammonia nitrogen, and microbial protein (p>0.05). However, the ruminal pH, concentrations of acetate, butyrate and isobutyrate, the ratio of acetate to propionate, protozoa count, and the activities of filter paper cellulase and cellobiase were significantly increased (p<0.05), while concentrations of propionate and valerate were significantly decreased (p<0.05). Moreover, 16S rRNA gene sequencing revealed that the relative abundance of rumen bacteria Christensenellaceae R7 group, Saccharofermentans, and Ruminococcaceae NK4A214 group were significantly increased, while Ruminococcus gauvreauii group, Prevotella 7 were significantly decreased (p<0.05). The relative abundance of the fungus Pseudomonas significantly increased, while Basidiomycota, Fusarium, and Alternaria significantly decreased (p<0.05). However, there was no significant change in the community structure of methanogenic archaea. Conclusion: Supplementation of hydroethanolic extract of WGH to a high-concentrate diet improved the ruminal fermentation, altered the structure of ruminal bacterial and fungal communities, and exhibited beneficial effects in alleviating subacute rumen acidosis of sheep.
A constructed sea stream in Yeongdo, Busan, Republic of Korea is mostly static due to the lifted stream bed and tidal characters, and receives domestic wastewater nearby, causing a consistent odor production and water quality degradation. Bioaugmentation of a microbial consortium was proposed as an effective and economical restoration technology to restore the polluted stream. The microbial consortium activated on site was augmented on a periodic basis (7~10 days) into the most polluted site (Site 2) which was chosen considering the pollution level and tidal movement. Physicochemical parameters of water qualities were monitored including pH, temperature, DO, ORP, SS, COD, T-N, and T-P. COD and microbial community analyses of the sediments were also performed. A significant reduction in SS, COD, T-N, and COD (sediment) at Site 2 occurred showing their removal rates 51%, 58% and 27% and 35%, respectively, in 13 months while T-P increased by 47%. In most of the test sites, population densities of sulfate reducing bacterial (SRB) groups (Desulfobacteraceae_uc_s, Desulfobacterales_uc_s, Desulfuromonadaceae_uc_s, Desulfuromonas_g1_uc, and Desulfobacter postgatei) and Anaerolinaeles was observed to generally decrease after the bioaugmentation while those of Gamma-proteobacteria (NOR5-6B_s and NOR5-6A_s), Bacteroidales_uc_s, and Flavobacteriales_uc_s appeared to generally increase. Aerobic microbial communities (Flavobacteriaceae_uc_s) were dominant in St. 4 that showed the highest level of DO and least level of COD. These microbial communities could be used as an indicator organism to monitor the restoration process. The alpha diversity indices (OTUs, Chao1, and Shannon) of microbial communities generally decreased after the augmentation. Fast uniFrac analysis of all the samples of different sites and dates showed that there was a similarity in the microbial community structures regardless of samples as the augmentation advanced in comparison with before- and early bioaugmentation event, indicating occurrence of changing of the indigenous microbial community structures. It was concluded that the bioaugmentation could improve the polluted water quality and simultaneously change the microbial community structures via their niche changes. This in situ remediation technology will contribute to an eco-friendly and economically cleaning up of polluted streams of brine water and freshwater.
Kim, Min Jeong;Shim, Chang Ki;Kim, Yong Ki;Hong, Sung Jun;Park, Jong Ho;Han, Eun Jung;Kim, Jin Ho;Kim, Suk Chul
The Plant Pathology Journal
/
v.31
no.3
/
pp.259-268
/
2015
This study investigated the chemical characteristics and microbial population during incubation of four kinds of aerated compost teas based on oriental medicinal herbs compost, vermicompost, rice straw compost, and mixtures of three composts (MOVR). It aimed to determine the effects of the aerated compost tea (ACT) based on MOVR on the growth promotion of red leaf lettuce, soybean and sweet corn. Findings showed that the pH level and EC of the compost tea slightly increased based on the incubation time except for rice straw compost tea. All compost teas except for oriental medicinal herbs and rice straw compost tea contained more ${NO^-}_3-N$ than ${NH^+}_4-N$. Plate counts of bacteria and fungi were significantly higher than the initial compost in ACT. Microbial communities of all ACT were predominantly bacteria. The dominant bacterial genera were analyzed as Bacillus (63.0%), Ochrobactrum (13.0%), Spingomonas (6.0%) and uncultured bacterium (4.0%) by 16S rDNA analysis. The effect of four concentrations, 0.1%, 0.2%, 0.4% and 0.8% MOVR on the growth of red leaf lettuce, soybean and sweet corn was also studied in the greenhouse. The red leaf lettuce with 0.4% MOVR had the most effective concentration on growth parameters in foliage part. However, 0.8% MOVR significantly promoted the growth of root and shoot of both soybean and sweet corn. The soybean treated with higher MOVR concentration was more effective in increasing the root nodule formation by 7.25 times than in the lower MOVR concentrations Results indicated that ACT could be used as liquid nutrient fertilizer with active microorganisms for culture of variable crops under organic farming condition.
International Journal of Industrial Entomology and Biomaterials
/
v.39
no.2
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pp.45-53
/
2019
The popularity of keeping stag beetles (Dorcus titanus castanicolor Motschulsky 1861, Coleoptera: Lucanidae) as pets has increased. Consistent with the rise in the number of insect farms using these beetles, the number of contaminated or diseased D. titanus castanicolor has also increased. This investigation was conducted to analyze the cause of D. titanus castanicolor disease. The contaminated larvae of D. titanus castanicolor showed Allomyrina nudivirus infection symptoms similar to those of Allomyrina nudivirus infection. However, the disease carried by of D. titanus castanicolor is not derived from the virus infecting Allomyrina, as determined by PCR. Our study revealed that the major gut microbes of infectious D. titanus castanicolor belonged to the phylum Proteobacteria, and specifically, Pseudomonas knackmussi (Symptom 1 - 39.62% to Symptom 2 - 41.50% to Symptom 3 - 76.76% as the disease progressed severely) and Citrobacter koseri (Symptom 1 - 1.48% to Symptom 2 - 6.04% to Symptom 3 - 6.16% as the disease progressed severely) were detected. Additionally, a high proportion of larvae from the uninfected group were found to harbor bacteria belonging to the phylum Firmicutes (72%). However, as the disease progressed severely in these beetles, the proportion of Firmicutes decreased (Symptom 1 - 72.03% to Symptom 2 - 44.7% to Symptom 3 - 26.3%). These findings imply that colonization by Firmicutes was inversely proportional to Proteobacteria colonization in the gut. This was found to be true for both the normal and disease conditions of D. titanus castanicolor. In this study, we examined the distribution of intestinal microbial communities in normal and contaminated larvae. We observed a correlation between these contaminated microbes and the overall health of the beetle, and our findings suggest that there may be a link between disease progression and the gut microbiome.
The diverse microbial communities that colonize distinct segments of the gastrointestinal tract are intimately related to aspects of physiology and the pathology of human health. However, most recent studies have focused on the rectal or fecal microbiota, and the microbial signature of the duodenum is poorly studied. In this study, we compared the microbiota in duodenal and rectal samples to illustrate the characteristic microbial signatures of the duodenum in healthy adults. Nine healthy volunteers donated biopsies and luminal contents from the duodenum and rectum. To determine the composition and diversity of the microbiota, 454-pyrosequencing of bacterial 16S rRNA was performed and multiple bioinformatics analyses were applied. The α-diversity and phylogenetic diversity of the microbiota in the duodenal samples were higher than those of the rectal samples. There was higher biodiversity among the microbiota isolated from rectal biopsies than feces. Proteobacteria were more highly represented in the duodenum than in the rectum, both in the biopsies and in the luminal contents from the healthy volunteers (38.7% versus 12.5%, 33.2% versus 5.0%, respectively). Acinetobacter and Prevotella were dominant in the duodenum, whereas Bacteroides and Prevotella were dominant in the rectum. Additionally, the percentage of OTUs shared in biopsy groups was far higher than in the luminal group (43.0% versus 26.8%) and a greater number of genera was shared among the biopsies than the luminal contents. Duodenal samples demonstrated greater biological diversity and possessed a unique microbial signature compared with the rectum. The mucosa-associated microbiota was more relatively conserved than luminal samples.
Journal of Korean Society of Environmental Engineers
/
v.36
no.9
/
pp.659-666
/
2014
The purpose of this study was to suggest a standard design procedure of landfarming for clean-up of oil-contaminated soils. The standard design procedure consisted of four main phases; soil characterization, determination of contaminated soil volume, determination of nutrient and microbial doses, and estimation of the total remedial period. This study selected standard design parameter values or ranges among various forms used in environmental engineer communities. Those were determination procedures for the contaminated soil volume, the initial contamination concentration and nutrient doses. The suggested standard design procedure were applied for a landfarm design for remediation of a real oil-contaminated site. Soil texture of the site was classified as sandy clay loam and sandy loam. Total nitrogen and total phosphorus were estimated to be 57.01 mg/kg and 83.40 mg/kg, respectively. Also the viable bacterial numbers was assessed to be $1.78{\times}10^4CFU/g$ dry soil. The amount of TPH contaminated soil was estimated to be $4,092m^3$. With the application of remedial factors, it was estimated that the contaminated soil could be treated through 9 batches with a duration of 315 days for a landfarming unit of $15m{\times}40m{\times}1m$. The amount of liquid microorganisms and fertilizers were recommended to be 4,025L and 4,641kg, respectively.
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