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Field Studios of In-situ Aerobic Cometabolism of Chlorinated Aliphatic Hydrocarbons

  • Semprini, Lewts
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.04a
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    • pp.3-4
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    • 2004
  • Results will be presented from two field studies that evaluated the in-situ treatment of chlorinated aliphatic hydrocarbons (CAHs) using aerobic cometabolism. In the first study, a cometabolic air sparging (CAS) demonstration was conducted at McClellan Air Force Base (AFB), California, to treat chlorinated aliphatic hydrocarbons (CAHs) in groundwater using propane as the cometabolic substrate. A propane-biostimulated zone was sparged with a propane/air mixture and a control zone was sparged with air alone. Propane-utilizers were effectively stimulated in the saturated zone with repeated intermediate sparging of propane and air. Propane delivery, however, was not uniform, with propane mainly observed in down-gradient observation wells. Trichloroethene (TCE), cis-1, 2-dichloroethene (c-DCE), and dissolved oxygen (DO) concentration levels decreased in proportion with propane usage, with c-DCE decreasing more rapidly than TCE. The more rapid removal of c-DCE indicated biotransformation and not just physical removal by stripping. Propane utilization rates and rates of CAH removal slowed after three to four months of repeated propane additions, which coincided with tile depletion of nitrogen (as nitrate). Ammonia was then added to the propane/air mixture as a nitrogen source. After a six-month period between propane additions, rapid propane-utilization was observed. Nitrate was present due to groundwater flow into the treatment zone and/or by the oxidation of tile previously injected ammonia. In the propane-stimulated zone, c-DCE concentrations decreased below tile detection limit (1 $\mu$g/L), and TCE concentrations ranged from less than 5 $\mu$g/L to 30 $\mu$g/L, representing removals of 90 to 97%. In the air sparged control zone, TCE was removed at only two monitoring locations nearest the sparge-well, to concentrations of 15 $\mu$g/L and 60 $\mu$g/L. The responses indicate that stripping as well as biological treatment were responsible for the removal of contaminants in the biostimulated zone, with biostimulation enhancing removals to lower contaminant levels. As part of that study bacterial population shifts that occurred in the groundwater during CAS and air sparging control were evaluated by length heterogeneity polymerase chain reaction (LH-PCR) fragment analysis. The results showed that an organism(5) that had a fragment size of 385 base pairs (385 bp) was positively correlated with propane removal rates. The 385 bp fragment consisted of up to 83% of the total fragments in the analysis when propane removal rates peaked. A 16S rRNA clone library made from the bacteria sampled in propane sparged groundwater included clones of a TM7 division bacterium that had a 385bp LH-PCR fragment; no other bacterial species with this fragment size were detected. Both propane removal rates and the 385bp LH-PCR fragment decreased as nitrate levels in the groundwater decreased. In the second study the potential for bioaugmentation of a butane culture was evaluated in a series of field tests conducted at the Moffett Field Air Station in California. A butane-utilizing mixed culture that was effective in transforming 1, 1-dichloroethene (1, 1-DCE), 1, 1, 1-trichloroethane (1, 1, 1-TCA), and 1, 1-dichloroethane (1, 1-DCA) was added to the saturated zone at the test site. This mixture of contaminants was evaluated since they are often present as together as the result of 1, 1, 1-TCA contamination and the abiotic and biotic transformation of 1, 1, 1-TCA to 1, 1-DCE and 1, 1-DCA. Model simulations were performed prior to the initiation of the field study. The simulations were performed with a transport code that included processes for in-situ cometabolism, including microbial growth and decay, substrate and oxygen utilization, and the cometabolism of dual contaminants (1, 1-DCE and 1, 1, 1-TCA). Based on the results of detailed kinetic studies with the culture, cometabolic transformation kinetics were incorporated that butane mixed-inhibition on 1, 1-DCE and 1, 1, 1-TCA transformation, and competitive inhibition of 1, 1-DCE and 1, 1, 1-TCA on butane utilization. A transformation capacity term was also included in the model formation that results in cell loss due to contaminant transformation. Parameters for the model simulations were determined independently in kinetic studies with the butane-utilizing culture and through batch microcosm tests with groundwater and aquifer solids from the field test zone with the butane-utilizing culture added. In microcosm tests, the model simulated well the repetitive utilization of butane and cometabolism of 1.1, 1-TCA and 1, 1-DCE, as well as the transformation of 1, 1-DCE as it was repeatedly transformed at increased aqueous concentrations. Model simulations were then performed under the transport conditions of the field test to explore the effects of the bioaugmentation dose and the response of the system to tile biostimulation with alternating pulses of dissolved butane and oxygen in the presence of 1, 1-DCE (50 $\mu$g/L) and 1, 1, 1-TCA (250 $\mu$g/L). A uniform aquifer bioaugmentation dose of 0.5 mg/L of cells resulted in complete utilization of the butane 2-meters downgradient of the injection well within 200-hrs of bioaugmentation and butane addition. 1, 1-DCE was much more rapidly transformed than 1, 1, 1-TCA, and efficient 1, 1, 1-TCA removal occurred only after 1, 1-DCE and butane were decreased in concentration. The simulations demonstrated the strong inhibition of both 1, 1-DCE and butane on 1, 1, 1-TCA transformation, and the more rapid 1, 1-DCE transformation kinetics. Results of tile field demonstration indicated that bioaugmentation was successfully implemented; however it was difficult to maintain effective treatment for long periods of time (50 days or more). The demonstration showed that the bioaugmented experimental leg effectively transformed 1, 1-DCE and 1, 1-DCA, and was somewhat effective in transforming 1, 1, 1-TCA. The indigenous experimental leg treated in the same way as the bioaugmented leg was much less effective in treating the contaminant mixture. The best operating performance was achieved in the bioaugmented leg with about over 90%, 80%, 60 % removal for 1, 1-DCE, 1, 1-DCA, and 1, 1, 1-TCA, respectively. Molecular methods were used to track and enumerate the bioaugmented culture in the test zone. Real Time PCR analysis was used to on enumerate the bioaugmented culture. The results show higher numbers of the bioaugmented microorganisms were present in the treatment zone groundwater when the contaminants were being effective transformed. A decrease in these numbers was associated with a reduction in treatment performance. The results of the field tests indicated that although bioaugmentation can be successfully implemented, competition for the growth substrate (butane) by the indigenous microorganisms likely lead to the decrease in long-term performance.

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Effects of Replacing Lucerne (Medicago sativa L.) Hay with Fresh Citrus Pulp on Ruminal Fermentation and Ewe Performance

  • Sparkes, J.L.;Chaves, A.V.;Fung, Y.T.E.;van Ekris, I.;Bush, R.D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.2
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    • pp.197-204
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    • 2010
  • Two studies were conducted to determine the effects of replacing 30% (% in diet DM) of lucerne (Medicago sativa L.) hay with citrus pulp in Merino ewe diets: i) an in vitro study which measured ruminal fermentation; and ii) an in vivo study in which twelve Merino ewes pre- and post-lambing were fed experimental diets in a cross-over design over 120 days to evaluate effects on ewe performance (i.e. DM intake, average daily gain (ADG) and wool growth). In both the in vitro and in vivo studies, the control treatment consisted of lucerne (91.3% in diet DM), lupins (8.3% in diet DM) and phosphate (0.42% in diet DM), while the citrus pulp treatment consisted of lucerne (57.7% in diet DM), lupins (9.5% in diet DM), phosphate (0.48% in diet DM) and fresh citrus pulp (32.3% in diet DM). Data were analysed using the mixed model procedure of SAS. In the in vitro study, gas production, total volatile fatty acid (VFA) yield, proportion of propionic acid to total VFA and in vitro dry matter digestibility (IVDMD) were higher (p<0.02) in the citrus pulp treatment compared to the control treatment. In contrast, in vitro ammonia production, pH and the acetate to propionate ratio were lower (p<0.03) for the citrus pulp treatment compared to the control treatment. In the in vivo study, DM intake of ewes fed the citrus pulp diet was lower than their control ewe counterparts throughout both the pre- and post-lambing periods (928.9 vs. 1,115.0 g/d pre-; 1,285.0 vs. 1,620.3 g/d post-lambing, p<0.01), however ADG was similar (p = 0.12). Wool growth parameters and lamb performance did not differ (p>0.32) between treatments. In summary, the in vitro study demonstrated that the replacement of 30% of a lucerne diet with fresh citrus pulp improved total VFA yield, increased total gas production and improved IVDMD, while decreasing the production of ammonia, acetic acid and rumen pH. In addition, the in vivo study demonstrated that the replacement of 30% of a lucerne diet with fresh citrus pulp pre- and post-lambing decreased intake but did not affect ewe performance in terms of ADG and wool growth. These findings, of course, would be of significant interest to sheep producers endeavouring to control cost of feed ingredients whilst maintaining productivity.

Optimum Dietary Lipid Level and Feeding Rates of Extruded Pellets in Juvenile Flounder Paralichthys olivaceus during the Summer Season (여름철 넙치치어 배합사료의 적정지질함량 및 공급량)

  • Choi, Se-Min;Kim, Kang-Woong;Kang, Yong-Jin;Park, Hung-Sik;Bai, Sung-Chul C.
    • Journal of Aquaculture
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    • v.21 no.4
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    • pp.244-251
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    • 2008
  • We evaluated the optimum dietary lipid level and feeding rates of extruded pellets (EP) in juvenile flounder Paralichthys olivaceus during the summer season. The first experiment was conducted to determine the optimum dietary lipid level in juvenile flounder. Five isonitrogenous EP (52% crude protein) with increasing dietary lipids (6, 8, 10, 12 and 14%) were fed to satiety to triplicate groups of the juveniles ($18.4{\pm}0.11g$) twice a day for 6 weeks. Weight gain (WG) of fish fed EP with 10% lipid was significantly higher than those of fish fed EP with 6 and 14% lipid (P<0.05). Broken line model analysis suggested that the optimum dietary lipid level could be $9.08{\pm}0.37%$ for the maximum WG in juvenile flounder. The second experiment was conducted to determine the optimum feeding rate using experimental diet contained 10% lipid level that had the highest WG in first experiment. The feed intake of triplicate groups of the juveniles ($13.4{\pm}0.15g$) was restricted to four different feeding rates of 2.0, 2.5, 3.0 and 3.5% of their body weight $day^{-1}$ and to satiation using experimental diets. WG and feed efficiency (FE) of fish was affected by feeding rates. WG and specific growth rate of fish fed the diets increased with increasing feeding rate, however no significant differences (P>0.05) in WG and specific growth rate were observed between the fish fed 3.5% of their body weight $day^{-1}$ and to satiation. FE of fish fed 3.5% of their body weight $day^{-1}$ was significantly higher than those offish fed 2.0% of their body weight $day^{-1}$ and to satiation (P<0.05). Broken line model analysis suggested that the optimum dietary feeding rate could be $3.56{\pm}0.06%$ for the maximum WG in juvenile flounder. These results indicated that the optimum lipid level and feeding rate could be $9{\sim}10%$ (Energy: 4,774kcal and PIE ratio: 108mg protein/kcal in diet) and 3.5% of their body weight $day^{-1}$in juvenile flounder, respectively.

Enhancement of Scenedesmus sp. LX1 Biomass Production and Lipid Accumulation Using Iron in Artificial Wastewater and Domestic Secondary Effluent

  • Zhao, Wen-Yu;Yu, Jun-Yi;Wu, Yin-Hu;Hong, Yu;Hu, Hong-Ying
    • Microbiology and Biotechnology Letters
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    • v.42 no.2
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    • pp.131-138
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    • 2014
  • While coupling wastewater treatment with microalgal bioenergy production is very promising, new approaches are needed to enhance microalgal growth and lipid accumulation in wastewater. Therefore, this study investigated the effect of iron on the growth, nutrient removal, and lipid accumulation of Scenedesmus sp. LX1 in both artificial wastewater and domestic secondary effluents. When increasing the iron concentration from 0 to 2 mg/l in the artificial wastewater, the biomass production of Scenedesmus sp. LX1 increased from 0.17 to 0.54 g/l; the nitrogen and phosphorus removal efficiency increased from 15.7% and 80.6% to 97.0% and 99.2%, respectively; and the lipid content was enhanced 84.2%. The relationship between the carrying capacity/maximal population growth rate of Scenedesmus sp. LX1 and the initial iron concentration were also in accordance with the Monod model. Furthermore, when increasing the iron concentration to 2 mg/l in four different domestic secondary effluent samples, the lipid content and lipid production of Scenedesmus sp. LX1 was improved by 17.4-33.7% and 21.5-41.8%, respectively.

Mammary alveolar cell as in vitro evaluation system for casein gene expression involved in glucose level

  • Heo, Young Tae;Ha, Woo Tae;Lee, Ran;Lee, Won-Young;Jeong, Ha Yeon;Hwang, Kyu Chan;Song, Hyuk
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.6
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    • pp.878-885
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    • 2017
  • Objective: Glucose is an essential fuel in the energy metabolism and synthesis pathways of all mammalian cells. In lactating animals, glucose is the major precursor for lactose and is a substrate for the synthesis of milk proteins and fat in mammary secretory (alveolar) epithelial cells. However, clear utilization of glucose in mammary cells during lactogenesis is still unknown, due to the lack of in vitro analyzing models. Therefore, the objective of this study was to test the reliability of the mammary alveolar (MAC-T) cell as an in vitro study model for glucose metabolism and lactating system. Methods: Undifferentiated MAC-T cells were cultured in three types of Dulbecco's modified Eagle's medium with varying levels of glucose (no-glucose: 0 g/L, low-glucose: 1 g/L, and high-glucose: 4.5 g/L) for 8 d, after which differentiation to casein secretion was induced. Cell proliferation and expression levels of apoptotic genes, Insulin like growth factor-1 (IGF1) receptor, oxytocin receptor, ${\alpha}S1$, ${\alpha}S2$, and ${\beta}$ casein genes were analyzed at 1, 2, 4, and 8 d after differentiation. Results: The proliferation of MAC-T cells with high-glucose treatment was seen to be significantly higher. Expression of apoptotic genes was not affected in any group. However, expression levels of the mammary development related gene (IGF1 receptor) and lactation related gene (oxytocin receptor) were significantly higher in the low-glucose group. Expressions of ${\alpha}S1-casein$, ${\alpha}S2-casein$, and ${\beta}-casein$ were also higher in the low-glucose treated group as compared to that in the no-glucose and high-glucose groups. Conclusion: The results demonstrated that although a high-glucose environment increases cell proliferation in MAC-T cells, a low-glucose treatment to MAC-T cells induces higher expression of casein genes. Our results suggest that the MAC-T cells may be used as an in vitro model to analyze mammary cell development and lactation connected with precise biological effects.

Ecological Study of Shortnecked Clam Ruditapes philippinarum from the Jindu Coast of Hansan Island, Korea (한산도 진두해역 양식 바지락의 자원생태학적 연구)

  • Cho, Sang-Man;Jeong, Woo-Geon
    • The Korean Journal of Malacology
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    • v.23 no.1
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    • pp.25-30
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    • 2007
  • In order to estimate the ecological parameters of the population, short-necked clams, Ruditapes philippinarum, were collected from Jindu coast at Hansan Island, from April to December in 2005. The growth of the clam was estimated by von Bertalanffy growth model and each parameters were obtained from a nonlinear regression as $L_{\infty}\;=\;48.8\;mm$, k = 0.28 and $t_0\;=\;-0.88\;(R^2\;=\;0.994)$. Instantaneous coefficient of total mortality (Z) and fishing mortality (F) was calculated to be 0.8614/yr. and 0.6122/yr, respectively. The age at the first capture was estimated to be 2.88 year. Using the present parameter for clam fishing, the yield-per recruit (Y/R) was estimated to be 3.44g. Although the maximum Y/R was obtained under the parameters ($t_c\;=\;3.48\;yr$. and F = 0.9295/yr), it is favorable to maintain the current fishing effort considering the annual fluctuation of the parameters.

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Growth Modeling of Perilla frutescens (L.) Britt. Using Expolinear Function in a Closed-type Plant Factory System (완전제어형 식물공장에서 선형지수함수를 이용한 들깨의 생육 모델링)

  • Seounggwan Sul;Youngtaek Baek;Young-Yeol Cho
    • Journal of Bio-Environment Control
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    • v.32 no.1
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    • pp.34-39
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    • 2023
  • Growth modeling in plant factories can not only control stable production and yield, but also control environmental conditions by considering the relationship between environmental factors and plant growth rate. In this study, using the expolinear function, we modeled perilla [Perilla frutescens (L.) Britt.] cultivated in a plant factory. Perilla growth was investigated 12 times until flower bud differentiation occurred after planting under light intensity, photoperiod, and the ratio of mixed light conditions of 130 μmol·m-2·s-1, 12/12 h, red:green:blue (7:1:2), respectively. Additionally, modeling was performed to predict dry and fresh weights using the expolinear function. Fresh and dry weights were strongly positively correlated (r = 0.996). Except for dry weight, fresh weight showed a high positive correlation with leaf area, followed by plant height, number of leaves, number of nodes, leaf length, and leaf width. When the number of days after transplanting, leaf area, and plant height were used as independent variables for growth prediction, leaf area was found to be an appropriate independent variable for growth prediction. However, additional destructive or non-destructive methods for predicting growth should be considered. In this study, we created a growth model formula to predict perilla growth in plant factories.

Enhancement of Transduction Efficiency and Antitumor Effects of IL-12N220L-expressing Adenovirus by Co-delivery of DOTAP

  • Youn, Je-In;Jin, Hyun-Tak;Sung, Young-Chul
    • IMMUNE NETWORK
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    • v.7 no.4
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    • pp.179-185
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    • 2007
  • Background: Adenovirus (Ad) vectors have been widely used for many gene therapy applications because of their high transduction ability and broad tropism. However, their utility for cancer gene therapy is limited by their poor transduction into cancer cells lacking the primary receptor, coxsackievirus and adenovirus receptor (CAR). Methods: To achieve CAR-independent gene transfer via Ad, we pretreated Ad with 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) and analyzed their transduction efficiency into cancer cells in vitro and in vivo comparing with the virus alone. Results: Treatment of DOTAP significantly increased adenoviral gene transfer in tumor cells in vitro. Moreover, DOTAP at an optimum dose $(10{\mu}g/ml)$ enhanced IL-12 transgene expression by fivefold in tumor, and twofold in serum after intratumoral injection of adenovirus expressing IL-12N220L (Ad/IL-12N220L). In addition, cotreatment of DOTAP decreased tumor growth rate in the Ad/IL-12N220L-transduced tumor model, finally leading to enhanced survival rate. Conclusion: Our results strongly suggest that DOTAP could be of great utility for improving adenovirus-mediated cancer gene therapy.

Biodegradation Kinetics of Phenol and pcresol by Micrococcus sp. M1 (Micrococcus sp. M1에 의한 Phenol과 p-Creso의 생분해 Kinetics)

  • Son, Hong-Joo;Jang, Woong-Seok;Lee, Geon;Lee, Sang-Joon
    • Journal of Environmental Science International
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    • v.6 no.2
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    • pp.153-163
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    • 1997
  • In order to fad the most fitted biodegradation model, biodegradation kinetics model to the initial phenol and p-cresot concentrations were investigated and had been fitted by the linear regression. Bacteria capable of degrading p-cresol were isolated from soil by enrichment culture technique. Among them, strain Ml capable of degradillg p.rcresol has also degraded phenal and was identified as the genus Micrococcus from the results from of taxonomical studies. The optimal tonditlons for the biodegradation of phenal and p-cresol by Micrococcus sp. Ml were $NH_4NO_3$ 0.05%, pH 7.0, 3$0^{\circ}C$, respectively, and medium volume 100m1/250m1 shaking flask. iwicrococcus sp. Ml was able to grow on phenal concentration up to 14mM and p-cresol concelltration up to 0.8mM. With increasing substrate concentraction, the lag period increased, but the maximum specific growth rates decreased. The yield coefficient decreased with increasing substrate concentation. The biodegradation kinetics of phenol and p-cresol were best described by Monod with growth model for every experimented concentration. In cultivation of mixed substrate, p-cresol was degraded first and phenol was second. This result implies that p-cresol and phenol was not degraded simultaneously.

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Heat-Killed and Live Enterococcus faecalis Attenuates Enlarged Prostate in an Animal Model of Benign Prostatic Hyperplasia

  • Choi, Young-Jin;Fan, Meiqi;Tang, Yujiao;Iwasa, Masahiro;Han, Kwon-Il;Lee, Hongchan;Hwang, Ji-Young;Lee, Bokyung;Kim, Eun-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1134-1143
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    • 2021
  • In the present study, we investigated the inhibitory effect of heat-killed Enterococcus faecalis (E. faecalis) and live E. faecalis on benign prostatic hyperplasia (BPH). The BPH rat model was established by administering male rats with testosterone propionate (TP, 5 mg/kg, in corn oil) via subcutaneous injections daily for four weeks after castration. The rats were divided into five groups: Con, corn oil-injected (s.c.) + DW administration; BPH, TP (5 mg/kg, s.c.) + DW administration; BPH+K_EF, TP (5 mg/kg, s.c.) + heat-killed E. faecalis (7.5 × 1012 CFU/g, 2.21 mg/kg) administration; BPH+L_EF, TP (5 mg/kg, s.c.) + live E. faecalis (1 × 1011 CFU/g, 166 mg/kg) administration; BPH+Fi, TP (5 mg/kg, s.c.) + finasteride (1 mg/kg) administration. In both of BPH+K_EF and BPH+L_EF groups, the prostate weight decreased and histological changes due to TP treatment recovered to the level of the Con group. Both of these groups also showed regulation of androgen-signaling factors, growth factors, and apoptosis-related factors in prostate tissue. E. faecalis exhibited an inhibitory effect on benign prostatic hyperplasia, and even heat-killed E. faecalis showed similar efficacy on the live cells in the BPH rat model. As the first investigation into the effect of heat-killed and live E. faecalis on BPH, our study suggests that heat-killed E. faecalis might be a food additive candidate for use in various foods, regardless of heat processing.