• Title/Summary/Keyword: Natural antimicrobial

Search Result 925, Processing Time 0.028 seconds

Cosmetic Efficacy of Supercritical Cannabis sativa Seed Extracts and Enhancement of Skin Permeation (초임계 대마종자 추출물의 화장품 효능과 경피흡수증진 효과)

  • Lee, Kwang Won;Park, Shinsung;Park, Su In;Shin, Moon Sam
    • The Journal of the Convergence on Culture Technology
    • /
    • v.7 no.4
    • /
    • pp.683-691
    • /
    • 2021
  • The purpose of this study is to measure the yield and to evaluate the physiological activity of Cannabis sativa seed(Hemp seed) extracts extracted using a density fluctuation supercritical carbon dioxide for each temperature condition-30℃(HSSE30), 45℃(HSSE45), 60℃(HSSE60), and to enable dissolution of the poorly water-soluble extracts by liposome formulation and to enhance the skin permeability. As a result of the yield measurement, HSSE60 showed the highest yield, and in the antioxidant activities, HSSE45 had the highest total polyphenol content, and showed the highest DPPH, ABTS+ radical scavenging activities at the highest concentration of the extracts. As a result of the antimicrobial susceptibility testing, a clear zone appeared only in the Propionibateium acnes strain. It was confirmed that particle size was reduced and the absolute value of the zeta potential increased in the case of the formulation in which the extracts were in liposomes than in the formulation in which the extracts were dissolved in deionized water, and the skin permeability was improved. Based on these experimental results, we confirmed the possibility of using the hemp seed supercritical carbon dioxide extracts, a poorly water-soluble extract, can be applied as a functional natural material for cosmetics.

Antibacterial and Antiviral Activities of Microwave-assisted Thuja orientalis Extracts (마이크로웨이브를 이용한 측백나무 추출물의 항균 및 항바이러스 특성)

  • Sangwon Ko;Jae-Young Lee;Seong-Hyeon Kim;Young-Chul Lee
    • Applied Chemistry for Engineering
    • /
    • v.34 no.2
    • /
    • pp.192-198
    • /
    • 2023
  • In this study, the components of microwave-assisted extracts obtained from Thuja orientalis leaves were analyzed, and the cytotoxicity, antibacterial and antiviral activities were evaluated. The predominant components from microwave-assisted extraction were catechin, leucopelargonidin, arecatannin, quinolone, and kaempferol derivatives, which are classified in the flavonoid and tannin groups. We observed that the 0.11 mg/mL of extract concentration did not show cytotoxicity in HaCaT cells. The antibacterial activities were tested according to the guidelines of methods for determining the bactericidal activity of antimicrobial agents. The extracts showed 99.9% antibacterial efficiency against gram-positive S. aureus, while the antibacterial effect on gram-negative E. coli was insignificant. When the extract concentration and contact time with bacteria were increased, 99.9% antibacterial efficiency was observed for E. coli as well as S. aureus. Following the standard to assess the activity of microbicides against viruses in suspension (ASTM-E1052-20), the antiviral efficiency was more than 99.99% for influenza A (H1N1) and SARS-CoV-2. These results suggest its potential use in antiviral disinfectants, surface coatings, personal protective equipment, and textiles.

Inhibitory Effects of Stewartia koreana Extracts on Pseudomonas aeruginosa Biofilm Formation (노각나무 추출물이 Pseudomonas aeruginosa의 바이오필름 형성에 미치는 영향)

  • Sang Gyun Lee;Hye Soo Kim;Soo Jeong Cho
    • Journal of Life Science
    • /
    • v.33 no.11
    • /
    • pp.936-943
    • /
    • 2023
  • This study was conducted to investigate the potential of Stewartia koreana as anti-microbial materials. The branches, stems and leaves of S. koreana were extracted into 70% ethanol and their antibacterial activity against P. aeruginosa was confirmed. The leaf, branch and stems extracts (1 mg/disc) showed the antibacterial activity against P. aeruginosa and leaf extracts showed higher antibacterial activities than those from branch extracts. The MIC against P. aeruginosa was 0.8 mg/ml and showed bacteriostatic action. The inhibitory effects of extract on biofilm formation and gene expression related to biofilm formation of P. aeruginosa was determined by biofilm biomass staining, SEM and qRT-PCR analysis. The biofilm biomass and cell growth of P. aeruginosa in the cultures treated with 0.2~2.0 mg/ml of S. koreana leaf extracts were significantly decreased in a concentration-dependent manner. We observed that the extract had an inhibitory effect on the formation of P. aeruginosa biofilms at concentrations of 0.8 mg/ml by SEM. qRT-PCR analysis showed that the lasI and rh1I gene expression associated to quorum sensing (QS) in the cultures treated with 0.2~2.0 mg/ml of S. koreana leaf extracts were suppressed in a concentration-dependent manner. Based on the above results, it can be concluded that S. koreana leaf extracts can be used as anti-microbial material derived from natural materials, as demonstrated by the antibacterial action and inhibition of biofilm formation of P. aeruginosa by QS inhibition.

Developments of Functional Sausage using Plant Extracts from Pine Needle and Green Tea (솔잎 및 녹차 추출물을 이용한 기능성 소시지 개발)

  • 김수민;조영석;성삼경;이일구;이신호;김대곤
    • Food Science of Animal Resources
    • /
    • v.22 no.1
    • /
    • pp.20-29
    • /
    • 2002
  • The functional sausage added to effective extracts are prepared to carried out to investigate functional and storage characteristics. This products were stored at different temperature. The changes of pH were tended to be a little ranged from pH 6.07 to pH 6.35 in control. At the same time, the pH changes treated with plant extracts showed the same tendency as control. The treatments using natural extracts revealed a little low TBARS value during storage at 10$^{\circ}C$. The nitrite scavenging ability of extracts from pine needle were higher than those of green tea extracts, irrespective of storage temperature. The VBN content was tended to be increased as storage time goes by, irrespective of storage temperature. The treatments using plant extracts revealed a little low VBN content, compared to control during storage. The changes of total bacteria were more increased to 2.2${\times}$10$^1$∼3.2 ${\times}$ 10$\^$6/ CFU/g during storage at 30$^{\circ}C$ than 2.2${\times}$10$^1$∼3.3${\times}$10$^2$CFU/g in case of storage at 10$^{\circ}C$. The treatments using plant extracts revealed an antimicrobial activity until storage at 3 days, compared to control. The lightness of sausage color were a little more decreased gradually during storage at 30$^{\circ}C$ than those of storage at 10$^{\circ}C$. Overall, the lightness of sausage color treated with pine needle extracts were a more bright than those of control. However, the redness of sausage color treated with pine needle and green tea showed the most lowest red color, compared to control. Sensory test suggested that the changes of sausage color, flavor, texture and taste were tended to be decreased gradually. In conclusion, pine needle extract was the most effective natural resources on the basis of the functional and physico-chemical properties of sausage of sausage.

Chemical Characteristics, Antimicrobial Activity and Dyeability of Gyeongsanbansi (Persimmon kaki) Unripe Juice Extraction and Fermented Liquor (경산반시의 미숙감 착즙액과 발효액의 화학적 특성, 항균성 및 염색성)

  • Heo, Buk-Gu;Park, Yun-Jum;Kim, Tae-Choon;Kim, Hyun-Ju;Park, Su-Min;Jang, Hong-Gi;Kim, Kyung-Su;Lee, Kyung-Dong;Yun, Jae-Gill
    • Korean Journal of Plant Resources
    • /
    • v.22 no.5
    • /
    • pp.438-445
    • /
    • 2009
  • This study was conducted to examine the chemical characteristics, antibacterial activity and dyeability of several juices made from Persimmon cv. 'Gyeongsanbanshi'. Young fruits of persimmon were harvested at 1st August, 2006. Fruit juice was extracted soon after harvesting, some of them were stored at $4{\sim}6^{\circ}C$ for 6 months or fermented at room temperature for 6 months. $L^*$ values of vinegar was 43.5 higher than those of fresh juice and fermented liquor. Inorganic matter contents in fresh juice, fermented liquor and vinegar were higher in the order of K ($1696{\sim}2880\;mg$/100 g), Ca ($166.7{\sim}417.9\;mg$/100 g), Mg ($203.9{\sim}214.4\;mg$/100 g), P ($37.9{\sim}109.8\;mg$/100 g), Na ($13.2{\sim}23.3\;mg$/100 g) and Fe ($8.4{\sim}14.2\;mg$/100 g). Cotton fabrics dyed with the fermented liquor and vinegar had the largest inhibitory zone against the gram-positive microorganisms with range of $16.0{\sim}35.0\;mm$. Cotton fabrics dyed with the fresh juice showed $9.0{\sim}9.5\;mm$ inhibitory zone against the gram-negative microorganisms, $15.0{\sim}21.0\;mm$ with the juice stored at $4{\sim}6^{\circ}C$ for 6 months, $22.0{\sim}23.0\;mm$ with the fermented liquor and $9.0{\sim}35.0\;mm$ with vinegar. The hue of cotton fabrics dyed with the fermented liquor had a YR levels, and antibacterial activity of them were 78.5%.

Antioxidant Activity of Extracts from Smilax china Root (청미래덩굴(Smilax china) 뿌리 추출물의 항산화활성 효과)

  • Song, Hee-Sun;Park, Yeon-Hee;Jung, Sae-Heung;Kim, Dong-Pil;Jung, Yong-Hee;Lee, Mi-Kyung;Moon, Ki-Young
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.35 no.9
    • /
    • pp.1133-1138
    • /
    • 2006
  • Smilax china root has been used as traditional medicinal remedy in China and Korea and reported to have various biological activities such as anti-inflammatory, antimutagenic and antimicrobial activities. In this study, the possibility of development as natural antioxidants of Smilax china root extracts was investigated. For the evaluation of antioxidant activity, aqueous- and 25% EtOH extract from Smilax china root were prepared and six different evaluation assay methods, i.e., measurement of total phenolics, radical scavenging effects on 1,1-diphenyl-2-picrylhydrazyl (DPPH), nitric oxide (NO) and nitrite $(NO_2)$, reducing power, and inhibitory effect on tyrosinase activity, were used. The total phenolics content of two extracts was high as the level of 36 mg of gallic acid equivalent per 1 g of dried sample tested. The radical scavenging activities of ethanol extract toward DPPH and NO were better than those of aqueous extract (p<0.05). The $NO_2$ scavenging activity of both extracts showed the highest value at pH 1.2 (98%). Especially, the $NO_2$ scavenging activities of EtOH extract were significantly stronger than those of aqueous one at pH 4.2 (51%) and pH 6.0 (32%), respectively. In the reducing power test, both extracts revealed higher ferric ion reducing activity than known antioxidant, vitamin C at the level of $0.05\sim0.1mg/mL$ (p<0.01). The 1 mL of aqueous and 25% EtOH extract showed effective inhibition activity on tyrosinase activity as 45% and 53%, respectively. Therefore, these results suggest that two extracts from Smilax china root may serve as useful natural antioxidants.

Studies on Biological Activity of Woad Extractives (XV) - Antimicrobial and antioxidative activities of extracts from diverse families - (수목 추출물의 생리활성에 관한 연구(XV) - 과별(科別)에 따른 항균 및 항산화 활성 -)

  • Lee, Sung-Suk;Lee, Hak-Ju;Choi, Don-Ha
    • Journal of the Korean Wood Science and Technology
    • /
    • v.32 no.4
    • /
    • pp.8-17
    • /
    • 2004
  • Antifungal, antibacterial, and antioxidative activities of ethanol extracts from 65 families 263 species were investigated to select tree species for the utilization of natural fungicide or preservative resources. The antifungal activities of extracts from wood, leaf and bark were measured as hyphal growth inhibition rate using four plant pathogenic and five wood rotting fungi. High inhibitory effect on the fungi growth was found in five species of Pinaceae (Pinus koraiensis, P. rigida, P. densiflora, P. banksiana. Cedrus deodara), three species of Cupressaceae (Juniperus rigida, J. chinensis, Chamaecyparis obtusa) and three species of Leguminosae (Albizzia julibrisssin, Sophora japonica, Maackia amurensis), respectively. Antibacterial activities of ethanol extracts were determined by means of disc-agar plate diffusion method using three gram-positive and five gram-negative bacteria. The ethanol extracts, which showed prominent effect on the suppression of bacteria growth, were six species of Betulaceae (Carpinus tschonoskii, C. coreana, C. laxiflora, Alnus hirsuta, A. firma, Betula schmidtii), five species of Fagaceae (Castanopsis cuspidata var. sieboldii, Quercus serrata, Q. mongolica, Q aliena, C crenata), four species of Euphorbiaceae (Aleurites fordii, Sapium sebiferum, S japonicum Mallotus japonicus) and three species of Elaeagnaceae (Elaeagnus umbellata, Elaeagnus glanbra, Elaeagnus macrophylla). According to these results, the extracts from Zelkova serrata, Pinus densiflora, Maackia amurensis, Chamaecyparis obtusa and Juniperus chinensis could be available for natural fungicide or food preservatives, because ethanol extracts from these species indicated excellent antifungal and antibacterial activities. In order to test antioxidative activities of ethanol extracts, free radical scavenging method was adopted with 1,1-diphenyl-2-picrylhydrohydrazyl (DPPH). Free radical scavenging activity was proved very high in the extracts of eight species of Rosaceae (Eriobotrya japonica, Prunus takesimensis, P yedoensis, P padus, P armeniaca var. ansu, Chaenomeles sinensis, Stephanandra incisa, Rosa multiflora) and five species of Ericaceae (Rhododenron mucronulatum, R. scblippenbacbii, R. yedoense var. poukhanense, Vaccinium bracteatum, V oldbami), resvectively. It turned out from this study that only six species among 48 species of Rosaceae showed less than 80% free radical scavenging activity. As a consequences, it could be deduced that the components effective on antioxidative activity commonly exist in Rosaceae plant family.

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
    • /
    • 2016.05a
    • /
    • pp.19-20
    • /
    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

  • PDF

Fish Safety and Antimicrobial Activity of Natural Sulfur Solution on Aquatic Microorganisms (Saprolegnia parasitica) Isolated from Misgurnus mizolepis (미꾸라지(Misgurnus mizolepis)에서 분리된 수생균 (Saprolegnia parasitica)에 대한 천연유황수의 항균 활성 및 처리에 대한 어류 안전성)

  • Yi, Seung-Won;Lee, Seung-Hyeop;Lee, Sang-Jong;Kim, Mi-Hee;Lee, Hye-Hyun;Chu, Saet-Byul;Kim, Kyung-Hee;Lee, Hee Jung
    • Korean Journal of Environmental Biology
    • /
    • v.35 no.2
    • /
    • pp.116-122
    • /
    • 2017
  • Basic dyes such as malachite green and methylene blue have been used as disinfectants to control water fungal infections since the 1930s. However, after succeeding reports of carcinogenicity and bioaccumulation of the dye, their use was forbidden in lieu of public health. This study undertook to evaluate the therapeutic effect of sulfur solution processed by effective microorganisms (EM-PSS) against Saprolegnia parasitica infection, and its safety in fish. In vitro antifungal evaluation of EM-PSS inhibited the growth of S. parasitica mycelia at concentrations of 50 ppm or higher. The acute toxicity test of EM-PSS to the mud fish (Misgurnus mizolepis) measured a no effect concentration (NOEC) at 100 ppm, the lowest effect concentration (LOEC) at 125 ppm, and the half-lethal concentration ($LC_{50}$) at 125 ppm in juvenile and 250 ppm in the immature stage. In addition, the ecotoxicity test of EM-PSS using Daphnia magna inhibited swimming of D. magna at concentrations of 100 ppm or less. Lastly, the EM-PSS prevented infection of S. parasitica to mud fish, at concentrations of 50 ppm. Furthermore, at 100 ppm concentration, the EM-PSS showed no acute toxicity on mud fish, nor any eco-toxic effects on D. magnano. Therefore, we conclude that carcinogenic disinfectants such as malachite green and methylene blue could be replaced by EM-PSS to remove S. parasitica in mud fish farming, and might be a potential eco-friendly disinfectant in aquaculture.

Enhancing the Effects of Zerumbone on THP-1 Cell Activation (단핵구세포주의 활성에 미치는 Zerumbone의 영향)

  • Lee, Min Ho;Kim, Sa Hyun;Ryu, Sung Ryul;Lee, Pyeongjae;Moon, Cheol
    • Korean Journal of Clinical Laboratory Science
    • /
    • v.49 no.1
    • /
    • pp.1-7
    • /
    • 2017
  • Zerumbone is a major component of the essential oil from Zingiber zerumbet Smith, which is a kind of wild ginger. In addition, various biological functions, such as liver protection, pain relief, atherosclerosis, and antimicrobial activity have been reported. It is also known to be effective in the proliferation of immune cells and the expression of cytokines. In this study, we investigated the effects of zerumbone on monocyte activation. First, it was confirmed that the proliferation of THP-1 cells was increased by zerumbone. The strongest increase in THP-1 proliferation after lipopolysaccharide treatment was observed at $5{\mu}M$ zerumbone treatment, and the increase of cell proliferation without lipopolysaccharide was the highest at $10{\mu}M$. Conversely, when treated with $50{\mu}M$ zerumbone, a rapid decrease of proliferation was observed regardless of the presence of lipopolysaccharide (LPS). The phosphorylation of signaling protein, Erk, induced by LPS was also increased by zerumbone. The strongest increase in phosphorylation was observed when treated with $50{\mu}M$ of zerumbone with reduced proliferation. The activity of transcription factor $NF-{\kappa}B$ was not significantly altered by zerumbone alone, but increased when treated with lipopolysaccharide. Furthermore, the transcription of the inflammatory cytokines $TNF-{\alpha}$ and IL-8, which are regulated by $NF-{\kappa}B$, is also increased by zerumbone. These results suggest that zerumbone can enhance the proliferation and activity of monocytes. Furthermore, it is believed that zerumbone can enhance rthe immune responses through increased monocyte activity in bacterial infections with LPS, thereby helping to treat effective bacteria.