Antibacterial activities of essential oils and extracts of Turkish Achillea,Satureja and Thymus species against plant pathogenic bacteria

BACKGROUND: The aims of this study were to examine the chemical composition of the essential oils and hexane extracts of the aerial parts of Satureja spicigera (C. Koch) Boiss., Thymus fallax Fisch. & CA Mey, Achillea biebersteinii Afan, and Achillea millefolium L. by GC and GC–MS, and to test antibacterial efficacy of essential oils and n‐hexane, chloroform, acetone and methanol extracts as an antibacterial and seed disinfectant against 25 agricultural plant pathogens. RESULTS: Thymol, carvacrol, p‐cymene, thymol methyl ether and γ‐terpinene were the main constituents of S. spicigera and T. fallax oils and hexane extracts. The main components of the oil of Achillea millefolium were 1,8‐cineole, δ‐cadinol and caryophyllene oxide, whereas the hexane extract of this species contained mainly n‐hexacosane, n‐tricosane and n‐heneicosane. The oils and hexane extracts of S. spicigera and T. fallax exhibited potent antibacterial activity over a broad spectrum against 25 phytopathogenic bacterial strains. Carvacrol and thymol, the major constituents of S. spicigera and T. fallax oils, also showed potent antibacterial effect against the bacteria tested. The oils of Achillea species showed weak antibacterial activity. Our results also revealed that the essential oil of S. spicigera, thymol and carvacrol could be used as potential disinfection agents against seed‐borne bacteria. CONCLUSION: Our results demonstrate that S. spicigera, T. fallax oils, carvacrol and thymol could become potentials for controlling certain important agricultural plant pathogenic bacteria and seed disinfectant. Copyright © 2009 Society of Chemical Industry     

乌贼墨抑制海洋鱼体腐败菌活性成分的分离

对乌贼墨中抑制海洋鱼体腐败菌活性成分进行了分离,并初步探讨了抑菌成分的理化特性。结果表明:乌贼墨粗提物经DEAE-纤维素和Sepharose 4B分离后获得具有较高活性的抑菌活性物质,分子量约为83589u,对四种海洋鱼体腐败菌（气单胞菌、黄杆菌、芽孢杆菌、肠杆菌）的平均抑菌率为62.36%。经紫外扫描（UV）和红外光谱（IR）分析,抑菌物质的主要成分为多糖。      

Antioxidant and inhibitory activities of thioflavanones against nitric oxide production

The antioxidant and suppressive activities of thioflavanone derivatives against production of nitric oxide were investigated. Thioflavanone derivatives were prepared with methoxy, methyl, nitro, hydroxyl, chloro, and fluoro substituents at different positions. 3′-Hydroxythioflavanone with a hydroxy group at the 3′-position of thioflavanone had a 15–43× higher antioxidant activity than unmodified thioflavanone. Among thioflavanones, 3′-nitrothioflavanone and 3′-hydroxythioflavanone showed increased inhibitory activities against nitric oxide (NO) production, compared to unmodified thioflavanone in a lipopolysaccharide stimulated culture of macrophages cells. 3′-Hydroxy-thioflavanone with a functional group at the 3′-position is a potent antioxidant with an increased inhibitory activity against NO production, compared to unmodified thioflavanone compounds.     

Growth-inhibiting activity of active component isolated from Terminalia chebula fruits against intestinal bacteria

The growth-inhibitory activity of materials derived from the fruit of Terminalia chebula was evaluated against six intestinal bacteria by means of an impregnated paper disk agar diffusion method. The butanol fraction of T. chebula extract had profound growth-inhibitory activity at a concentration of 5 mg per disk. The biologically active component isolated from the T. chebula fruits was identified with a variety of spectroscopic analyses as ethanedioic acid. The growth responses varied in accordance with the bacterial strain, chemical, and dosage tested. In a test with concentrations of 2 and 1 mg per disk, ethanedioic acid had strong and moderate inhibitory activity against Clostridium perfringens and Escherichia coli, respectively, with no associated adverse effects on the growth of the four tested lactic acid-producing bacteria. Ellagic acid derived from T. chebula fruits exerted a potent inhibitory effect against C. perfringens and E. coli, but little or no inhibition was observed with treatments of behenic acid, P-caryophyllene, eugenol, isoquercitrin, oleic acid, ca-phellandrene, 3-sitosterol, stearic acid, a-terpinene, terpinen-4-ol, terpinolene, or triacontanoic acid. These results may be an indication of at least one of the pharmacological properties of T. chebula fruits.     

Antimicrobial activity of leptospermone and its derivatives against human intestinal bacteria

The anaerobic growth inhibiting activity of the essential oil of Leptospermum scoparium seeds against intestinal bacteria was evaluated in vitro. The essential oil at 5.0 mg/disc strongly (+++) inhibited the growth of Clostridium difficile and Clostridium perfringens, but did not inhibit the growth of Bifidobacterium breve, Bifidobacterium longum, Escherichia coli, or Lactobacillus casei. Spectroscopic analyses revealed that the active component of the essential oil was leptospermone. Leptospermone strongly inhibited the growth of C. difficile and C. perfringens at 1.0 mg/disc and moderately inhibited their growth at 0.5 mg/disc. However, leptospermone did not inhibit the growth of B. breve, B. longum, and L. casei. When compared with the leptospermone derivatives, 1,2,3-cyclohexanetrione-1,3-dioxime exerted strong inhibition against C. perfringens and moderate inhibition against C. difficile at 0.5 mg/disc, whereas the other derivatives exerted weak or no growth inhibition against all bacteria tested. Taken together, these results indicate that cyclohexanetrione is required for triketone derivatives to inhibit the growth of C. difficile and C. perfringens. These findings indicate that L. scoparium seed-derived materials and 1,2,3-cyclohexanetrione-1,3-dioxime are naturally occurring antimicrobial agents that could be useful in the development of new agents for the specific control of C. difficile and C. perfringens.     

Antimicrobial activities of Asian plant extracts against pathogenic and spoilage bacteria

Food Science and Biotechnology - This study was conducted to investigate the antimicrobial effects of 300 Asian plant extracts (PEs) against pathogenic and spoilage bacteria. The antimicrobial...     

Proteolytic profiles and angiotensin-I converting enzyme and alpha-glucosidase inhibitory activities of selected lactic acid bacteria

ABSTRACT:  This study was conducted to examine the growth, proteolytic profiles as well as angiotensin-I converting enzyme (ACE) and α-glucosidase (α-glu) inhibitory potentials of selected strains of lactic acid bacteria (LAB). Two strains each of yogurt bacteria ( Streptococcus thermophilus —1275 and 285, and Lactobacillus delbrueckii ssp. bulgaricus —1092 and 1368), and probiotics ( L. acidophilus —4461 and 33200, and L. casei —2607 and 15286, and 1 strain of Bifidobacterium longum 5022), were cultivated in reconstituted skim milk (RSM) at 37 °C and their proteolytic profiles and ACE as well as α-glu inhibitory activities were determined. Among all the strains of lactic acid bacteria studied, yogurt bacteria grew very well, with the exception of L. delbrueckii ssp. bulgaricus 1368 which showed a slower growth during the initial 3 h of incubation. The growth pattern corresponded well with the decrease in pH for the organisms. All the organisms showed an increase in proteolysis with time. The variations in proteolytic capabilities translated into corresponding variations in ACE inhibitory potential of these organisms. Bifidobacterium longum 5022 showed the highest ACE inhibitory potential followed by L. delbrueckii ssp. bulgaricus 1368, L. casei 15286, S. thermophilus 1275, and L. acidophilus 4461. Organisms with high intracellular enzymatic activities grew well. Also, aminopeptidases of strains of L. acidophilus 4461 and S. thermophilus 1275 that could better utilize proline containing substrates showed enhanced ACE inhibitory potential. Lactic acid bacteria possessed the ability to inhibit α-glu activity, which endowed them an antidiabetic property as well.     

大黄抑制食源性致病菌的活性成分研究

研究了大黄对大肠杆菌、沙门氏菌和金黄色葡萄球菌的抑菌效果,初筛的结果显示大黄醇提物对这3种菌均有较强的抑菌作用,并发现其中抑菌作用最强的物质分布在大黄氯仿段萃取物中。利用UPLC-MS-MS分析大黄氯仿段萃取物,发现大黄氯仿段萃取物的主要成分分别是大黄素、芦荟大黄素、大黄酸、大黄素甲醚和大黄酚。在这五种化合物中,大黄酸对大肠杆菌和沙门氏菌具有最强的抑菌效果,对大肠杆菌和沙门氏菌的最小抑菌浓度分别为125μg/m L和250μg/m L,最小杀菌浓度分别为250μg/m L和500μg/m L,大黄素对于金黄色葡萄球菌具有较强的抑菌效果,最小抑菌浓度和最小杀菌浓度分别为62.5μg/m L和125μg/m L。      

Growth inhibitory effects of anthranilic acid and its derivatives against Legionella pneumophila

Legionella pneumophila is the principal etiologic agent of Legionnaires' disease. We found that the growth of L. pneumophila was markedly inhibited by its own cell lysate and the inhibitory effect was abolished by heat-treatment of the lysate. The genomic library of L. pneumophila was constructed in Escherichia coli and screened to determine the gene involved in the growth inhibition. A clone harboring the gene encoding anthranilate synthase (TrpE), which is involved in tryptophan biosynthesis, exhibited an inhibitory effect on the growth of L. pneumophila. Anthranilic acid exogenously added also exhibited antibacterial activity against L. pneumophila. A series of single-gene-knockout mutants of L. pneumophila lacking tryptophan synthesis genes were constructed and assessed for their susceptibility to anthranilic acid. Although the growth of mutants deficient in anthranilate phosphoribosyltransferase (TrpD) and N-(5'-phosphoribosyl)anthranilate isomerase (TrpF) was not affected by exogenous anthranilic acid, the indole-3-glycerophosphate synthase (TrpC) deficient mutant exhibited an increased susceptibility compared with the parent strain. These observations strongly indicate that 1-(2-carboxyphenylamino)-1'-deoxyribulose-5'-phosphate (CPADR-5'-P), which is an intermediate of tryptophan synthesis from anthranilic acid, is responsible for the growth inhibition of L. pneumophila.     

Proteolytic and angiotensin‐converting enzyme‐inhibitory activities of selected probiotic bacteria

This study was carried out to examine the proteolytic and angiotensin‐converting enzyme (ACE‐I) activities of probiotic lactic acid bacteria (LAB) as influenced by the type of media, fermentation time, strain type and media supplementation with a proteolytic enzyme (Flavourzyme^®). Lactobacillus casei (Lc210), Bifidobacterium animalis ssp12 (Bb12), Lactobacillus delbrueckii subsp. bulgaricus (Lb11842) and Lactobacillus acidophilus (La2410) were grown in 12% of reconstituted skim milk (RSM) or 4% of whey protein concentrates (WPC‐35) with or without combination (0.14%) of Flavourzyme^® for 12 h at 37 °C. All the strains were able to grow in both media depending on type of strains used and fermentation time. All the strains showed higher proteolytic activity and produced more antihypersensitive peptides when grown in RSM medium at 12 h, when compared to WPC. Combination with Flavourzyme^® also increased LAB growth and proteolytic and ACE‐I activities. Of the four strains used, Bb12 and La2410 outperformed Lc210 and Lb11842. The highest ACE‐I activity and proteolytic activity were found in B. animalis ssp12 combined with Flavourzyme^®. Flavourzyme^® led to an increase in the production of bioactive peptides with ACE‐I activity during 12 h at 37 °C.     

Antimicrobial activities of polyhexamethylene guanidine hydrochloride-based disinfectant against fungi isolated from cocoa beans and reference strains of bacteria

This study was conducted to assess the antibacterial and the antifungal activity of a polyhexamethylene guanidine hydrochloride (PHMGH)-based disinfectant and to determine if it could be used as a disinfectant for the treatment of cocoa beans. The activity of PHMGH was tested in vitro for efficacy against five reference strains of pathogenic bacteria and six strains of fungi isolated from cocoa beans. All the strains tested were sensitive to the disinfectant. The MICs reported were between 0.01 and 1.9 mg/ml and equal to the MBC or minimum fungicidal concentration (MFC) regardless of the strains of those microorganisms. The bacteria were more sensitive to PHMGH than were the fungi. Enterobacter cloacae was the most sensitive bacterium with a MIC and MBC of 0.01 mg/ml, whereas the genus Aspergillus was the least susceptible of the microorganisms tested, with a MIC and MFC from 1.0 to 1.9 mg/ml. The time required for the activity of PHMGH varies from 2 min for Enterobacter cloacae to 12 min for Aspergillus tamarii and generally increases with the MBC or the MFC. Through this in vitro study, the PHMGH has been proved to be bactericidal and fungicidal on the strains studied. Hence, it could probably serve as a fungicidal disinfectant for the treatment of cocoa beans after harvesting.     

The inhibitory effects of wine phenolics on lysozyme activity against lactic acid bacteria

The lysozyme of hen's egg white is used in winemaking to control spontaneous lactic acid bacteria (LAB). A total of eight LAB strains, isolated from grape must and wine, were used to assess the inhibitory effects of wine phenolics on lysozyme activity. The presence of phenolics, extracted from grape pomace, in growth medium reduced the mortality rate due to the lysozyme activity. This effect was especially clear in the case of strains belonging to Lactobacillus uvarum, Pediococcus parvulus and Oenococccus oeni, which are more sensitive to lysozyme than L. plantarum and L. hilgardii strains. Cell lysis assays carried out on four strains sensitive to lysozyme and Micrococcus lysodeikticus ATCC 4698, used as a reference strain, confirmed the inhibition of grape pomace phenolics on the muramidase. There was no interference from non-flavonoids, flavanols and flavonol compounds, when they were tested individually, on the lysozyme activity against the strains. Anthocyanins extracted from grape skins slightly inhibited the activity only against M. lysodeikticus. However, proanthocyanidins extracted from seed berries, strongly inhibited the lysozyme. In this extract, dimers were the predominant oligomers of flavan-3-ol. The study demonstrated that the effectiveness of lysozyme against LAB in red winemaking is related to the amount of low molecular weight proanthocyanidins that are released when the grapes are macerating.     

芝麻叶多酚的纯化及对蛋白质非酶糖基化的抑制作用

从D101、AB-8、NKA-9、S-8、ADS-5大孔吸附树脂中通过静态实验筛选出AB-8树脂,研究其对芝麻叶粗提物中多酚的静态吸附和解吸性能,并通过单因素实验确定AB-8树脂柱的最佳纯化工艺条件;然后,考察了芝麻叶多酚对体外蛋白质非酶糖基化终末产物（AGEs）生成的抑制作用。结果表明:AB-8大孔吸附树脂对芝麻叶多酚粗提物具有较好的分离纯化效果;AB-8树脂柱对芝麻叶多酚粗提物的最佳纯化条件为:上柱液体积12mL,上柱液浓度3.5mg/mL,pH5.0,洗脱体积为3BV（柱体积）,依次以30%（v/v）和50%（v/v）乙醇溶液进行梯度洗脱,多酚纯度从16.88%分别提高到28.43%和45.71%,纯化产物分别命名为SPP1和SPP2。芝麻叶多酚粗提物、SPP1、SPP2对蛋白质非酶糖基化终产物的生成具有明显的抑制作用,抑制效果均优于阳性对照品氨基胍。      

抑制阪崎肠杆菌的乳酸菌筛选及其抗菌物质特性研究

目的 从酸菜和大弹涂鱼肠道中分离可产抑制阪崎肠杆菌活性的乳酸菌,并对抑菌物质的特性和抑菌机制进行研究,为其在食品保鲜中的应用奠定基础。方法 利用抑菌圈实验从分离所得的多株乳酸菌中筛选具有抑制阪崎肠杆菌活性的菌株并鉴定,排除酸及过氧化氢的影响后,分析其抑菌产物的特性。分离纯化抑菌物质,并初步探讨抑菌机制。结果 筛选得到一株抑菌效果最好的乳酸菌SA3,经鉴定为发酵黏液乳杆菌(Limosilactobacillus fermentum),其上清液对阪崎肠杆菌抑菌圈直径达到16.94 mm。上清液具有蛋白特性和热稳定性,以乙酸乙酯粗提出的抑菌物质对鼠伤寒沙门氏菌、白色念珠菌、大肠埃希氏菌等致病菌表现出较强抑菌活性。其抑菌机制为乳酸菌所产抑菌物质可破坏阪崎肠杆菌的细胞壁及内容物。结论 本研究分离到的发酵黏液乳杆菌SA3有望应用于乳制品中阪崎肠杆菌的生物防腐。     

Growth inhibition of Listeria spp. on Camembert cheese by bacteria producing inhibitory substances.

Bacterial strains exhibiting antimicrobial activity towards other bacteria are quite common in nature. During the past few years several genera have been shown to exert inhibitory action against Listeria. spp. In the present work strains of Enterococcus, Lactobacillus and Lactococcus were tested for their influence on the development of Listeria spp. on Camembert cheese. Partial or complete inhibition of growth of Listeria spp. was observed using various inhibitory bacteria. Complete inhibition occurred when the inhibitory strain was used as a starter culture and there was a low level of contamination with Listeria spp. during the first stage of ripening. Very little inhibition occurred if the inhibitory strain was added together with the starter culture.     

Potential competitive exclusion bacteria from poultry inhibitory to Campylobacter jejuni and Salmonella

The objective of this study was to isolate from chickens potential competitive exclusion bacteria (CE) that are inhibitory to Campylobacter jejuni or Salmonella, or to both, for subsequent development of a defined CE product for use in poultry. Adult chickens from family farms, commercial farms, and broiler chicken research centers were sampled to identify and select C. jejuni-free donor chickens. A challenge treatment, which included administering perorally 106 CFU C. jejuni per chicken and determining undetectable cecal shedding of campylobacters at 4 weeks, was important for identifying the best CE donor chickens. Screening of bacterial colonies obtained from nine donor chickens by using selective and nonselective media yielded 636 isolates inhibitory to six C. jejuni strains in vitro, with 194 isolates being strongly inhibitory. Of the 194 isolates, 145 were from ceca, and 117 were facultative anaerobic bacteria. One hundred forty-three isolates were inhibitory to six strains of Salmonella (including five different serotypes) in vitro. Of these, 41 were strongly inhibitory to all C. jejuni and Salmonella strains evaluated, and most were Lactobacillus salivarius. A direct overlay method, which involved directly applying soft agar on plates with discrete colonies from mucus scrapings of gastrointestinal tracts, was more effective in isolating CE than was the frequently practiced isolation method of picking and transferring discrete colonies and then overlaying them with soft agar. The best approach for obtaining bacteria highly inhibitory to Salmonella and C. jejuni from chickens was to isolate bacteria from ceca under anaerobic conditions. Free-range chickens from family farms were better donors of potential CE strongly inhibitory to both Salmonella and Campylobacter than were chickens from commercial farms and broiler chicken research centers.