海南粗榧内生真菌对罗非鱼无乳链球菌抑菌活性研究

利用活菌菌饼法和滤纸片法研究筛选64株海南粗榧内生真菌对罗非鱼源无乳链球菌的抑菌活性,并对活性菌株的代谢产物化学成分进行初步测试。菌饼试验结果显示,11株海南粗榧内生真菌对无乳链球菌具有明显抑制作用,占总菌株数的17.2％,抑菌圈在Φ(8.7±0.3～16.2±0.2)mm间。滤纸片试验结果显示,12株具有明显抑制作用,占总菌数的18.8％,抑菌圈在Φ（8.7±0.3～28.2±0.4）mm间。2种方法的最高活性菌株不一致,但最高活性菌株（B30、F52）和同时显示较高活性的菌株（B14、B30、B32、F7、F52）被测出都含有甾体及三萜类化合物。根据结果推测,对无乳链球菌有显著抑制作用的菌株可能含有甾体及三萜类物质。结果为深入研究内生真菌对罗非鱼无乳链球菌的抑制奠定基础,也为生物防治罗非鱼病害提供有益借鉴。     

马铃薯脱毒试管苗快繁中污染抑菌剂的筛选

在马铃薯脱毒试管苗工厂化快繁过程中,常因各种原因出现细菌污染,严重影响试管苗的生长,给生产带来损失。为了解决这一问题,本试验分别加入卡那霉素、青霉素、链霉素及青霉素和链霉素配合使用的抗生素,研究抑菌剂及抑菌剂浓度在马铃薯快繁过程中的抑制细菌的效果。结果表明:在培养基中加入卡那霉素、青霉素G钠、青霉素和链霉素的混合物都能抑制细菌的污染,但青霉素各处理不仅能抑制细菌污染,而且对试管苗的生长有明显的促进作用,其中青霉素60 mg.L-1的效果最好,同时抑菌剂的抑菌作用具有时效性。     

促生细菌Bacillus sp. QN2MO-1的鉴定及对番茄生长的影响

为分离和鉴定具有促生作用的内生菌,开发潜在功能的微生物菌株,本研究以诺尼（Morinda citrifolia L.）为研究对象,从植株不同组织中共分离得到621株内生细菌,经复筛得到具有促生和解磷作用的菌株,命名为QN₂MO-1。基于菌株的生理生化分析和16S rDNA的序列比对,该内生细菌与Bacillus siamensis KCTC 13613^T（AJVF01000043）具有99.04%的同源性。药敏性分析显示,该菌对青霉素、硫酸链霉素和氨苄西林具有明显的抗性。QN₂MO-1可促进番茄种子的萌发;与对照相比,5%菌体发酵液处理能显著提高植株的生长。     

Effects of different concentrations of bovine follicular fluid and estrous cow serum on development of murine 2-cell embryos

Murine 2-cells embryos were isolated from murine oviducts at laboratory and transferred into Ham's F-10 medium containing 0.1 mg mL(-1) streptomycin and 100 IU mL(-1) penicillin G and supplemented with 3 mg mL(-1) bovine serum albumin (BSA) or different concentrations of bovine follicular fluid (bFF) and estrous cow serum (ECS). Significantly higher (p<0.05) > or =4-cell embryos were developed when embryos were cultured 20% bFF (84.33%) comparing to 10 and 15% bFF (48.33 and 69.33%) as well as 3 mg mL(-1) BSA (65.66%). Morula rates were also lower in 10% bFF (22.33%) comparing to the other groups and were similar in 15 and 20% bFF (62.66 and 72.33% morula rates) as well as BSA containing media (55.33%). The highest (p<0.05) blastocyst rates were obtained in medium containing 20% bFF (64.33%) and the lowest belonged to 10% bFF (15%) comparing to 15% bFF (33.66%) or 3 mg mL(-1) BSA. When embryos were cultured in ECS, no significant different was observed in different culture media (76.66, 72.33, 82.5 and 65.66% > or =4-cell embryos in 10, 15 and 20% bFF and 3 mg mL(-1) BSA, respectively). Morula and blastocyst rates were also similar in all groups (32.33, 41.66 and 66.25 and 55.33% morula rates and 15.33, 27, 44.50 and 29.66% blastocyst rates for 10, 15 and 20% bFF and 3 mg mL(-1) BSA, respectively). The results of the present study demonstrated that 20% bFF could be substituted for BSA when in vitro culture of murine embryos is carried.     

Effects and side effects of penicillin injection in huanglongbing affected grapefruit trees

Trunk injection with penicillin has been tested to control citrus huanglongbing (HLB), but side effects and environmental safety must be assured before approval of penicillin injection can be considered. We investigated effects of penicillin injection on densities of Candidatus Liberibacter asiaticus (Las) in leaves, as well as culturable bacterial populations in rhizospheres and petioles of grapefruit trees in field and greenhouse experiments. Trees were injected with penicillin G, and leaf and root concentrations were assessed in bioassays with Bacillus subtilis. Las densities were determined by qPCR, and bacteria were isolated on a low carbon medium from roots plus rhizosphere and surface-sterilized petioles at various times after penicillin injection. Selected bacterial isolates were tested for penicillin resistance (20 μg/mL) and glyphosate resistance (7000 μg/mL), because glyphosate is widely used and cross-resistance against antibiotics had been documented. One month after penicillin injection half of the greenhouse trees were inoculated with Phytophthora nicotianae. Cycle threshold (Ct) values of Las in old and young leaves significantly increased 90 days after trunk injection with penicillin. Bacterial populations in petioles and root-rhizospheres initially increased after penicillin injections, probably due to nutrient release, then returned to control levels after one week. Penicillin resistance was common in isolates from penicillin-injected and control trees (30–94%). Significantly more glyphosate resistant than sensitive isolates were penicillin resistant (81% versus 52%). Phytophthora root rot was not increased after penicillin injection. Thus, side effects of penicillin injection tested here were minimal, while Las titers were reduced after three months.     

牛角瓜根的抑菌活性研究

以牛角瓜根为实验材料,采用索氏提取法,以4种不同溶剂(石油醚、氯仿、乙酸乙酯、甲醇)对牛角瓜的根进行提取,并采用纸片琼脂扩散法对粗提物的抑菌活性进行了评价,供试菌包括白色念珠菌等6种真菌和金黄色葡萄球菌等7种细菌。结果表明:石油醚粗提物对枯草芽孢杆菌、白色念珠菌、黑曲霉菌、大肠埃希氏菌都表现出良好的抑菌活性,且抑菌效果接近或优于阳性对照氟康唑,对尖孢镰刀菌和鲍曼不动杆菌也表现出良好的抑菌活性;氯仿粗提物对白色念珠菌及粪链球菌表现出良好的抑菌活性;乙酸乙酯粗提物对粪链球菌表现出最好的抑菌活性;甲醇粗提物的抑菌活性较弱。     

Characterization of beta-lactamases from urinary isolates of Escherichia coli in Tehran

BACKGROUND: Knowledge of antimicrobial resistance patterns in E. coli, the predominant pathogen associated with urinary tract infections (UTI) is important as a guide in selecting empirical antimicrobial therapy. METHODS: To describe the antimicrobial susceptibility of E. coli associated with UTI in a major university hospital in Tehran (Iran), seventy-six clinical isolates of E. coli were studied for susceptibility to beta-lactam antibiotics by the disc diffusion method and Minimal Inhibitory Concentrations determination. RESULTS: All isolates were resistant to ampicillin, amoxicillin and oxacillin. Resistance to the other tested antibiotics was shown to be 93.4% to cefradine, 76.3% to carbenicillin, 47.3% to cefazoline, 50% to cefalexin and 32.8% to cephalothin while 1.3% expressed resistance to cefoxitime, and 2.6% were resistant to ceftizoxime and ceftriaxone. Two isolates (2.4%) harbored extended spectrum beta-lactamases (ESBL) shown by the double disc diffusion method. Substrate hydrolysis by ultra violet spectroscopy showed that 87.4% harbored penicillinases, 9% produced cephlosporinases and 3.6% degraded both substrates. Clavulanic acid inhibited enzyme activity in 82.9%, of which 78.95% was penicillinases (group IIa) and 3.95% was cephalosporinases (group IIb) of the Bush classification system. The rest of the isolates (6.58 %) were placed in group IV beta-lactamases. No group III beta-lactamase was found, as EDTA inhibited none of the enzymes. DNA amplification by polymerase chain reaction using specific primers for ampC, TEM and SHV type beta-lactamases for all of the isolates showed that 47 organisms (60%) carried the TEM gene and 18 isolates (24%) harbored blaTEM and ampC genes. About 26% of the organisms harbored SHV type enzymes. CONCLUSION: These results indicate that E. coli can posses a variety of beta-lactamases that are responsible for beta-lactam resistance.     

Antimicrobial Activity of Untenospongin B,a Metabolite from the Marine Sponge Hippospongia communis collected from the Atlantic Coast of Morocco

(−)-Untenospongin B isolated from the marine sponge Hippospongia communis has been tested for its antimicrobial activity against bacteria and human pathogenic fungi using agar disk method and was found to possess a broad and strong activity toward the test organisms. Its antifungal activity was further characterized by determination of the minimum inhibitory concentration (MIC) against five fungal species using broth microdilution method.     

Assessment of the Antimicrobial Activity of Algae Extracts on Bacteria Responsible of External Otitis

External otitis is a diffuse inflammation around the external auditory canal and auricle, which is often occurred by microbial infection. This disease is generally treated using antibiotics, but the frequent occurrence of antibiotic resistance requires the development of new antibiotic agents. In this context, unexplored bioactive natural candidates could be a chance for the production of targeted drugs provided with antimicrobial activity. In this paper, microbial pathogens were isolated from patients with external otitis using ear swabs for over one year, and the antimicrobial activity of the two methanol extracts from selected marine (Dunaliella salina) and freshwater (Pseudokirchneriella subcapitata) microalgae was tested on the isolated pathogens. Totally, 114 bacterial and 11 fungal strains were isolated, of which Staphylococcus spp. (28.8%) and Pseudomonas aeruginosa (P. aeruginosa) (24.8%) were the major pathogens. Only three Staphylococcus aureus (S. aureus) strains and 11 coagulase-negative Staphylococci showed resistance to methicillin. The two algal extracts showed interesting antimicrobial properties, which mostly inhibited the growth of isolated S. aureus, P. aeruginosa, Escherichia coli, and Klebsiella spp. with MICs range of 1.4 × 10⁹ to 2.2 × 10¹⁰ cells/mL. These results suggest that the two algae have potential as resources for the development of antimicrobial agents.     

A Marine Sulfate-Reducing Bacterium Producing Multiple Antibiotics: Biological and Chemical Investigation

A marine sulfate-reducing bacterium SRB-22 was isolated by means of the agar shake dilution method and identified as Desulfovibrio desulfuricans by morphological, physiological and biochemical characteristics and 16S rDNA analysis. In the bioassay, its extract showed broad-spectrum antimicrobial activity using the paper disc agar diffusion method. This isolate showed a different antimicrobial profile than either ampicillin or nystatin and was found to produce at least eight antimicrobial components by bioautography. Suitable fermentation conditions for production of the active constituents were determined to be 28 day cultivation at 25 °C to 30 °C with a 10% inoculation ratio. Under these conditions, the SRB-22 was fermented, extracted and chemically investigated. So far an antimicrobial compound, mono-n-butyl phthalate, and an inactive compound, thymine, have been isolated and characterized.     

云南省田菁根瘤菌及根瘤内生细菌遗传多样性研究

本研究采用BOX-PCR指纹图谱和16SrDNA全序列分析,对45株分离自田菁的根瘤菌及根瘤内生细菌进行了菌株多样性研究,BOX-PCR指纹图谱分析结果表明,在78%的相似性水平上,待测菌株分成了4个遗传群,其中群Ⅰ和群Ⅳ分别与根癌土壤杆菌(Agrobacterium tumefaciens)和大肠杆菌(Escherichia coli)聚在一起,其它群没有与参比菌株聚群。选取每群的代表菌株进行16SrDNA全序列测定。结果表明:待测菌株归于6个属,分别是贪铜菌属(Cupriavidus)、拜纳蒙纳斯属(Balneimonas)、土壤杆菌属(Agrobacterium)、克罗诺杆菌(Cronobacter)、肠杆菌属(Enterobacter)和泛菌属(Pantoea),菌株表现了丰富的遗传多样性;其中分离自田菁的根瘤菌隶属于贪铜菌属(Cupriavidus)和拜纳蒙纳斯属(Balneimonas)尚未见其它文章报道。     

Determination of the Total Phenolic and Anthocyanin Contents and Antimicrobial Activity of Viburnum Opulus Fruit Juice

Viburnum opulus is a plant with fruits that are rich in biologically active substances, making it valuable to the food and pharmaceutical industries. Here, we present our study of the total phenolic and anthocyanin contents and antimicrobial activity in the fruit juice of six V. opulus L. accessions. The cultivar ??Krasnaya Grozd?? was notable for its exceptionally large amount of total phenolics, 1168?mg/100?g, with anthocyanins comprising 3?C5?% of the total phenolic content. The evaluation of the antimicrobial properties confirmed that the juice of V. opulus fruits strongly inhibited the growth of a wide range of human pathogenic bacteria, both Gram-negative (Salmonella typhimurium and S. agona) and Gram-positive (Staphylococcus aureus, Lysteria monocytogenes, and Enterococcus faecalis) organisms. Conversely, the yeasts Debaryomyces hansenii and Torulaspora delbrueckii showed complete resistance to the fruit juice, whereas a low sensitivity was demonstrated by Trichosporon cutaneum, Kluyveromyces marxianus var. lactis, Saccharomyces cerevisiae, S. cerevisiae 12R, and Candida parapsilosis.     

Chitosomes-In-Chitosan Hydrogel for Acute Skin Injuries: Prevention and Infection Control

Burns and other skin injuries are growing concerns as well as challenges in an era of antimicrobial resistance. Novel treatment options to improve the prevention and eradication of infectious skin biofilm-producing pathogens, while enhancing wound healing, are urgently needed for the timely treatment of infection-prone injuries. Treatment of acute skin injuries requires tailoring of formulation to assure both proper skin retention and the appropriate release of incorporated antimicrobials. The challenge remains to formulate antimicrobials with low water solubility, which often requires carriers as the primary vehicle, followed by a secondary skin-friendly vehicle. We focused on widely used chlorhexidine formulated in the chitosan-infused nanocarriers, chitosomes, incorporated into chitosan hydrogel for improved treatment of skin injuries. To prove our hypothesis, lipid nanocarriers and chitosan-comprising nanocarriers (≈250 nm) with membrane-active antimicrobial chlorhexidine were optimized and incorporated into chitosan hydrogel. The biological and antibacterial effects of both vesicles and a vesicles-in-hydrogel system were evaluated. The chitosomes-in-chitosan hydrogel formulation demonstrated promising physical properties and were proven safe. Additionally, the chitosan-based systems, both chitosomes and chitosan hydrogel, showed an improved antimicrobial effect against S. aureus and S. epidermidis compared to the formulations without chitosan. The novel formulation could serve as a foundation for infection prevention and bacterial eradication in acute wounds.     

Diversity and Antimicrobial Activity of Vietnamese Sponge-Associated Bacteria

This study aimed to assess the diversity and antimicrobial activity of cultivable bacteria associated with Vietnamese sponges. In total, 460 bacterial isolates were obtained from 18 marine sponges. Of these, 58.3% belonged to Proteobacteria, 16.5% to Actinobacteria, 18.0% to Firmicutes, and 7.2% to Bacteroidetes. At the genus level, isolated strains belonged to 55 genera, of which several genera, such as Bacillus, Pseudovibrio, Ruegeria, Vibrio, and Streptomyces, were the most predominant. Culture media influenced the cultivable bacterial composition, whereas, from different sponge species, similar cultivable bacteria were recovered. Interestingly, there was little overlap of bacterial composition associated with sponges when the taxa isolated were compared to cultivation-independent data. Subsequent antimicrobial assays showed that 90 isolated strains exhibited antimicrobial activity against at least one of seven indicator microorganisms. From the culture broth of the isolated strain with the strongest activity (Bacillus sp. M1_CRV_171), four secondary metabolites were isolated and identified, including cyclo(L-Pro-L-Tyr) (1), macrolactin A (2), macrolactin H (3), and 15,17-epoxy-16-hydroxy macrolactin A (4). Of these, compounds 2-4 exhibited antimicrobial activity against a broad spectrum of reference microorganisms.     

Molecular identification of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus in nasal swabs samples from horses suffering respiratory infections in Iran

The objective of this study was to evaluate the existence of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus as probable agents associated with naturally occurring infection of the equine upper respiratory disease in Mashhad area. Nasal swabs samples from thirty horses with upper respiratory tract infections were collected. The bacteria isolated and identified were Streptococcus equi subsp. equi (1 isolate), Streptococcus equi subsp. zooepidemicus (25 isolates), Pasteurella sp. (11 isolates), Staphylococcus sp. (17 isolates), Bacillus sp. (4 isolates), Pseudomonas sp. (4 isolates), Proteus sp. (1 isolate), Neisseria sp. (1 isolate) and E. coli (1 isolate). All 25 isolates of Streptococcus equi subsp. zooepidemicus and the isolate of Streptococcus equi subsp. equi were characterized by biochemical tests and molecular techniques. For molecular identification of the subspecies S. equi and S. zooepidemicus two genomic region SeM and sodA were amplified. This study is the first report of molecular identification of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus in Iran.     

Antibiotic Supplements Affect Electrophysiological Properties and Excitability of Rat Hippocampal Pyramidal Neurons in Primary Culture

Introduction: Antibiotic supplements are regularly used in neuronal culture media to control contamination; however, they can interfere with the neuronal excitability and affect electrophysiological properties. Therefore, in this study, the effect of penicillin/streptomycin supplements on the spontaneous electrophysiological activity of hippocampal pyramidal neurons was examined. Methods: Electrophysiological whole-cell patch-clamp recordings from rat hippocampal pyramidal cells in primary culture were performed to investigate the effects of antibiotic supplements on the intrinsic excitability of cultured cells. Results: The present findings indicated that presence of antibiotic supplements (penicillin/streptomycin) in the culture medium altered the intrinsic electrical activity of hippocampal pyramidal neurons in primary culture. These alterations included: 1) depolarized resting membrane potential; 2) a significant enhancement in the after-hyperpolarization amplitude; 3) a significant increase in the area under the action potential and in the decay and rise time of the action potential; 4) a significant broadening of action potential and 5) a significant reduction in the firing frequency. Conclusion: These findings suggest that addition of antibiotic supplements to culture media influences the neuronal excitability and alters the electrophysiological properties of cultured neurons, possibly through changing the ionic conductance underlying neuronal excitability. Key Words: Primary cell culture, Patch-clamp techniques, Hippocampus     

Antimicrobial Activity of the Marine Alkaloids,Clathrodin and Oroidin,and Their Synthetic Analogues

Marine organisms produce secondary metabolites that may be valuable for the development of novel drug leads as such and can also provide structural scaffolds for the design and synthesis of novel bioactive compounds. The marine alkaloids, clathrodin and oroidin, which were originally isolated from sponges of the genus, Agelas, were prepared and evaluated for their antimicrobial activity against three bacterial strains (Enterococcus faecalis, Staphylococcus aureus and Escherichia coli) and one fungal strain (Candida albicans), and oroidin was found to possess promising Gram-positive antibacterial activity. Using oroidin as a scaffold, 34 new analogues were designed, prepared and screened for their antimicrobial properties. Of these compounds, 12 exhibited >80% inhibition of the growth of at least one microorganism at a concentration of 50 µM. The most active derivative was found to be 4-phenyl-2-aminoimidazole 6h, which exhibited MIC₉₀ (minimum inhibitory concentration) values of 12.5 µM against the Gram-positive bacteria and 50 µM against E. coli. The selectivity index between S. aureus and mammalian cells, which is important to consider in the evaluation of a compound’s potential as an antimicrobial lead, was found to be 2.9 for compound 6h.     

一株产铁载体橡胶树拮抗细菌的分离鉴定及耐药性分析

从橡胶树根部分离到一株拮抗细菌Czk1,该菌株在铬天青(CAS)检测平板上产生较大的桔黄色显色圈,具较强的铁载体产生能力.对其进行生理生化鉴定、16S rDNA序列分析,测定7种常见抗生素的抗性.结合该菌菌落、菌体特征,生理生化指标及16S rDNA序列分析,鉴定该内生菌为枯草芽孢杆菌(Bacillus subtilis),该菌对四环素、红霉素、氨苄青霉素、利福平、链霉素和卡那霉素高度敏感,对氯霉素表现抗性.