An intimate link between antimicrobial peptide sequence diversity and binding to essential components of bacterial membranes

Antimicrobial peptides and proteins (AMPs) are widespread in the living kingdom. They are key effectors of defense reactions and mediators of competitions between organisms. They are often cationic and amphiphilic, which favors their interactions with the anionic membranes of microorganisms. Several AMP families do not directly alter membrane integrity but rather target conserved components of the bacterial membranes in a process that provides them with potent and specific antimicrobial activities. Thus, lipopolysaccharides (LPS), lipoteichoic acids (LTA) or the peptidoglycan precursor Lipid II are targeted by a broad series of AMPs. Studying the functional diversity of immune effectors tells us about the essential residues involved in AMP mechanism of action. Marine invertebrates have been found to produce a remarkable diversity of AMPs. Molluscan defensins and crustacean anti-LPS factors (ALF) are diverse in terms of amino acid sequence and show contrasted phenotypes in terms of antimicrobial activity. Their activity is directed essentially against Gram-positive or Gram-negative bacteria due their specific interactions with Lipid II or Lipid A, respectively. Through those interesting examples, we discuss here how sequence diversity generated throughout evolution informs us on residues required for essential molecular interaction at the bacterial membranes and subsequent antibacterial activity. Through the analysis of molecular variants having lost antibacterial activity or shaped novel functions, we also discuss the molecular bases of functional divergence in AMPs.

Isolamento de uma quitinase extraída do cefalotórax do camarão marinho Litopenaeus schmitti (BURKENROAD, 1936) : avaliação de suas atividades antimicrobiana e larvicida

Chitinases are enzymes involved in degradation of chitin and are present in a range of organisms, including those that do not contain chitin, such as bacteria, viruses, plants and animals, and play important physiological and ecological roles. Chitin is hydrolyzed by a chitinolytic system classified as: endo-chitinases, exo-chitinases and N-acetyl-b-D-glucosaminidases. In this study a Litochitinase1 extracted from the cephalotorax of the shrimp Litopenaeus Schmitt was purified 987.32 times using ionexchange chromatography DEAE-Biogel and molecular exclusion Sephacryl S-200. These enzyme presented a molecular mass of about 28.5 kDa. The results, after kinetic assay with the Litochitinase1 using as substrate p-nitrophenyl-N-acetyl-b-Dglucosaminideo, showed apparent Km of 0.51 mM, optimal activity at pH ranging from 5.0 to 6.0, optimum temperature at 55°C and stability when pre-incubated at temperatures of 25, 37, 45, 50 and 55°C. The enzyme showed a range of stability at pH 4.0 to 5.5. HgCl2 inhibited Litochitinase1 while MgCl2 enhances its activity. Antimicrobial tests showed that Litochitinase1 present activity against gram-negative bacterium Escherichia coli in the 800 μg/mL concentration. The larvicidal activity against Aedes aegypti was investigated using crude extracts, F-III (50-80%) and Litochitinase1 at 24 and 48 hours. The results showed larvicidal activity in all these samples with EC50 values of 6.59 mg/mL for crude extract, 5.36 mg/mL for F-III and 0.71 mg/mL for Litochitinase1 at 24 hours and 3.22 and 0.49 mg/mL for the F-III and Litochitinase1 at 48 hours, respectively. Other experiments confirmed the presence of chitin in the midgut of Aedes aegypti larvae, which may be suffering the action of Litochitinase1 killing the larvae, but also the absence of contaminating proteins as serine proteinase inhibitors and lectins in the crude extract, F-III and Litochitinase1, indicating that the death of the larvae is by action of the Litochitinase1. We also observed that the enzymes extracted from intestinal homogenate of the larvae no have activity on Litochitinase1. These results indicate that the enzyme can be used as an alternative to control of infections caused by Escherichia coli and reducing the infestation of the mosquito vector of dengue.

Resposta de Chromobacterium Violaceum cultivada em alta concentração de ferro

The Chromobacterium violaceum is a β-proteobacterium Gram-negative widely found in tropical and subtropical regions, whose genome was sequenced in 2003 showing great metabolic versatility and biotechnological and pharmaceutical potential. Given the large number of ORFs related to iron metabolism described in the genome of C. violaceum, the importance of this metal for various biological processes and due to lack of data about the consequences of excess of iron in free-living organisms, it is important to study the response mechanism of this bacterium in a culture filled with iron. Previous work showed that C. violaceum is resistant to high concentrations of this metal, but has not yet been described the mechanism which is used to this survival. Thus, to elucidate the response of C. violaceum cultured in high concentrations of iron and expecting to obtain candidate genes for use in bioremediation processes, this study used a shotgun proteomics approach and systems biology to assess the response of C. violaceum grown in the presence and absence of 9 mM of iron. The analysis identified 531 proteins, being 71 exclusively expressed by the bacteria grown in the presence of the metal and 100 just in the control condition. The increase in expression of proteins related to the TCA cycle possibly represents a metabolic reprogramming of the bacteria caused by high concentration of iron in the medium. Moreover, we observed an increase in the activity assay of superoxide dismutase and catalase as well as in Total Antioxidant Activity assay, suggesting that the metal is inducing oxidative stress in C. violaceum that increases the levels of violacein and antioxidant enzymes to better adapt to the emerging conditions. Are also part of the adaptive response changes in expression of proteins related to transport, including iron, as well as an increased expression of proteins related to chemotaxis response, which would lead the bacteria to change the direction of its movement away from the metal. Systems Biology results, also suggest a metabolic reprogramming with mechanisms coordinated by bottleneck proteins involved in transcription (GreA), energy metabolism (Rpe and TpiA) and methylation (AhcY)

Chromatographic techniques to obtain the biomolecules profile of wild Suillus granulates

In recent years the interest in naturally occurring compounds has been increasing worldwide. Indeed, many of the bioactive compounds currently used as medicines have been synthesized based on the structure of natural compounds [1]. In order to obtain bioactive fractions and subsequently isolated compounds derived from natural matrices, several procedures have been carried out. One of these is to separate and assess the concentration of the active compound(s) present in the samples, a step in which the chromatographic techniques stand out [2]. In the present work the mushroom Sui/Ius granulatus (L.) Roussel was chemically characterized by chromatographic techniques coupled to different detectors, in order to evaluate the presence of nutritional and/or bioactive molecules. Some hydrophilic compounds, namely free sugars, were identified by high performance liquid chromatography coupled to a refraction index detector (HPLC-RI), and organic and phenolic acids were assessed by HPLC coupled to a photodiode array detector (HPLC-PDA). Regarding lipophilic compounds, fatty acids weredetermined by gas chromatography with a flame ionization detector (GC-FID) and tocopherols by HPLC-fluorescence detection. Mannitol and trehalose were the main free sugars detected. Different organic acids were also identified (i.e. oxalic, quinic and fumaric acids), as well as phenolic acids (i.e. gallic and p-hydroxybenzoic acids) and the related compound cinnamic acid. Mono- and polyunsaturated fatty acids were the prevailing fatty acids and a-, ~- and ~-tocopherol were the isoforms of vitamin E detected in the samples. Since this species proved to be a source of biologically active compounds, the antioxidant and antimicrobial properties were evaluated. The antioxidant activity was measured through the reducing power, free radical's scavenging activity and lipid peroxidation inhibition of its methanolic extract, and the antimicrobial activity was also tested in Gram positive and Gram negative bacteria and iri different fungi. S. granulatus presented antioxidant properties in all the performed assays, and proved to inhibit the growth of different bacterial and fungal strains. This study is a first step for classifying S. granulatus as a functional food, highlighting the potential of mushrooms as a source of nutraceutical and biologically active compounds.

Identificación bacteriana en pacientes con colecistitis aguda del servicio de cirugía Hospital Vicente Corral Moscoso, Cuenca 2001

Se realizó un estudio de tipo prospectivo descriptivo, con el objeto de identificar la presencia o no de gérmenes en la bilis de pacientes que presentaron colicistitis aguda, que acudieron al servicio de emergencia y consulta externa de Hospital Vicente Corral Moscoso. Durante el período de Enero a Mayo del 2001. Determinándose que esta patología al ser de inicio súbito, fue tratada en su mayoría antes de las 72 horas, siendo más frecuente en el sexo femenino, y con mayor incidencia entre la tercera y quinta década de la vida, de quienes se obtuvo bilis mediante punción de la vesícula biliar durante la colecistectomía. Luego se procedió a realizar Tinción de Gram y cultivos, los gérmenes encontrados en la tinción en fresco, fueron en su gran mayoría Gram negativos. Siendo además la E. Coli el gérmen más frecuente en medio de cultivo

Proteômica diferencial da Chromobacterium violaceum em resposta ao estresse oxidativo induzido por peróxido de hidrogênio

The β-proteobacterium Chromobacterium violaceum is a Gram-negative, free-living, saprophytic and opportunistic pathogen that inhabits tropical and subtropical ecosystems among them, in soil and water of the Amazon. It has great biotechnological potential, and because of this potential, its genome was completely sequenced in 2003. Genome analysis showed that this bacterium has several genes with functions related to the ability to survive under different kinds of environmental stresses. In order to understand the physiological response of C. violaceum under oxidative stress, we applied the tool of shotgun proteomics. Thus, colonies of C. violaceum ATCC 12472 were grown in the presence and absence of 8 mM H2O2 for two hours, total proteins were extracted from bacteria, subjected to SDS-PAGE, stained and hydrolysed. The tryptic peptides generated were subjected to a linear-liquid chromatography (LC) followed by mass spectrometer (LTQ-XL-Orbitrap) to obtain quantitative and qualitative data. A shotgun proteomics allows to compare directly in complex samples, differential expression of proteins and found that in C. Violaceum, 131 proteins are expressed exclusively in the control condition, 177 proteins began to be expressed under oxidative stress and 1175 proteins have expression in both conditions. The results showed that, under the condition of oxidative stress, this bacterium changes its metabolism by increasing the expression of proteins capable of combating oxidative stress and decreasing the expression of proteins related processes bacterial growth and catabolism (transcription, translation, carbon metabolism and fatty acids). A tool with of proteomics as an approach of integrative biology provided an overview of the metabolic pathways involved in the response of C. violaceum to oxidative stress, as well as significantly amplified understanding physiological response to environmental stress. Biochemical and "in silico" assays with the hypothetical ORF CV_0868 found that this is part of an operon. Phylogenetic analysis of superoxide dismutase, protein belonging to the operon also showed that the gene is duplicated in genome of C. violaceum and the second copy was acquired through a horizontal transfer event. Possibly, not only the SOD gene but also all genes comprising this operon were obtained in the same manner. It was concluded that C. violaceum has complex, efficient and versatile mechanisms in oxidative stress response

Renal and antibacterial effects induced by myotoxin I and II isolated from Bothrops jararacussu venom

Bothrops jararacussu myotoxin I (BthTx-I; Lys 49) and II (BthTX-II; Asp 49) were purified by ion-exchange chromatography and reverse phase HPLC. In this work we used the isolated perfused rat kidney method to evaluate the renal effects of B. jararacussu myotoxins I (Lys49 PLA(2)) and II (Asp49 PLA(2)) and their possible blockage by indomethacin. BthTX-1 (5 mu g/ml) and BthTX-II (5 mu g/ml) increased perfusion pressure (PP; ct(120) = 110.28+/-3.70 mmHg; BthTX I = 171.28+/-6.30* mmHg; BthTX II = 175.50+/-7.20* mmHg), renal vascular resistance (RVR; ct(120) = 5.49+/-0.54 mmHg/ml.g(-1) min(-1); BthTX I = 8.62+/-0.37* mmHg/ml g(-1) min(-1); BthTX II=8.9+/-0.36* mmHg/ml g(-1) min(-1)), urinary flow (UF; ct(120)= 0.14+/-0.01 ml g(-1) min(-1); BthTX I=0.32+/-0.05* ml g(-1) min(-1); BthTX II=0.37+/-0.01* ml g(-1) min(-1)) and glomerular filtration rate (GFR; ct(120)=0.72+/-0.10 ml g(-1) min(-1); BthTX I=0.85+/-0.13* ml g(-1) min(-1); BthTX II=1.22+/-0.28* ml g(-1) min(-1)). In contrast decreased the percent of sodium tubular transport (%TNa+; ct(120)=79,76+/-0.56; BthTX I=62.23+/-4.12*; BthTX II=70.96+/-2.93*) and percent of potassium tubular transport (%TK+;ct(120)=66.80+/-3.69; BthTX I=55.76+/-5.57*; BthTX II=50.86+/-6.16*). Indomethacin antagonized the vascular, glomerular and tubular effects promoted by BthTX I and it's partially blocked the effects of BthTX II. In this work also evaluated the antibacterial effects of BthTx-I and BthTx-II against Xanthomonas axonopodis. pv. passiflorae (Gram-negative bacteria) and we observed that both PLA2 showed antibacterial activity. Also we observed that proteins Also we observed that proteins chemically modified with 4-bromophenacyl bromide (rho-BPB) decrease significantly the antibacterial effect of both PLA(2). In conclusion, BthTx I and BthTX II caused renal alteration and presented activity antimicrobial. The indomethacin was able to antagonize totally the renal effects induced by BthTx I and partially the effects promoted by BthTx II, suggesting involvement of inflammatory mediators in the renal effects caused by myotoxins. In the other hand, other effects could be independently of the enzymatic activity of the BthTX II and the C-terminal domain could be involved in both effects promoted for PLA(2). (C) 2005 Elsevier Ltd. All rights reserved.

A microbiology study of the surgical site infection, at a hospital in northern Portugal

Surgical site infections (SSIs) remain a major clinical problem in terms of morbidity, mortality, time spent in hospital and overall direct and indirect costs. Objectives: To measure the prevalence of the SSI, by type of surgery and microbiologically characterization, in adult patients undergoing surgery during 2015 at a public hospital in northern Portugal. Methods: A prospective study, attended by 609 adult patients, undergoing surgery. The sociodemographic and clinical data of the population, as well as the surgical procedure and microbiological study were analyzed using Microsoft Access 2013. Results: In the sample of 609 adults undergoing surgery, it was found that 62.89% of surgical wound were cleaned, 15.8% were clean-contaminated, 8.70% were contaminated and 9.36% infected. About 62.52% of the intervened patients had antibiotic prophylaxis prior to surgery. Out of all surgeries, 33.3% were laparoscopic. The percentage of SSI was 5.74%; In these positive cases, only 3.61% was identiied the responsible bacteria. The urgent surgeries have more infections when compared to the programmed ones. In colon surgery the number of infections was 60% after cholecystectomy (22.86%). In hernioplasty, infection occurred in only 2.86% of the patients. The most isolated bacteria was Escherichia coli with 59%, in which 30% are -producing-lactamases of extended spectrum, followed by Pseudomonas aeruginosa (13.6%) and Serratia marcescens (13.6%). The mortality rate was 14.8%. Pseudomonas aeruginosa was isolated in 3 of 4 patients who died. Conclusions: The most microorganisms belong to the group of Gram-negative and are usually linked to infections associated with health care.

Patogenia de piscirickettsiosis: estudio sobre la infectividad de Piscirickettsia salmonis y el efecto de sus productos extracelulares

La piscirickettsiosis es una enfermedad contagiosa sistémica de los peces teleósteos cuyo agente causal es Piscirickettsia salmonis, una bacteria gram negativa intracelular facultativa. Esta enfermedad se ha descrito esporádicamente en distintas áreas geográficas y especies de peces en el mundo, pero es endémica y particularmente severa en salmónidos criados en agua de mar en Chile. En esta tesis se investigaron algunos aspectos de la patogénesis de esta enfermedad, estudiándose la infectividad de P. salmonis, tanto in vitro como in vivo, y buscándose además evidencias de la capacidad de secretar exotoxinas por parte de esta bacteria. Los ensayos de infectividad en células CHSE-214, procedentes de embrión de salmón chinook (Oncorhynchus tshawytscha), mostraron que existe una rápida adherencia de la bacteria a la superficie de la membrana plasmática (≤ 5 min posinoculación) seguida de su incorporación al citoplasma de estas células, proceso que ocurre entre las 3 y las 6 h posinoculación. Por su parte, el estudio de infectividad in vivo, que se realizó en trucha arcoiris (O. mykiss), reveló que este proceso comprende tres etapas principales: (i) una fase de rápida adhesión a células epiteliales principalmente de piel y branquias, pero también del canal alimentario; (ii) una invasión progresiva desde los sitios de entrada hacia tejidos más profundos hasta alcanzar el torrente sanguíneo y; (iii) una rápida diseminación vía hematógena para alcanzar virtualmente todos los tejidos corporales. Finalmente, se demostró que P. salmonis puede secretar exotoxinas termolábiles que tienen un efecto citotóxico selectivo según la célula blanco expuesta y que, probablemente, son parte de los factores de virulencia involucrados en la patogénesis de la piscirickettsiosis.

Complete genome of the multidrug-resistant Acinetobacter baumannii strain KBN10P02143 isolated from Korea

Acinetobacter baumannii , a strictly aerobic, non-fermentative, Gram-negative coccobacillary rod-shaped bacterium, is an opportunistic pathogen in humans. We recently isolated a multidrug-resistant A. baumannii strain KBN10P02143 from the pus sample drawn from a surgical patient in South Korea. We report the complete genome of this strain, which consists of 4,139,396 bp (G + C content, 39.08%) with 3,868 protein-coding genes, 73 tRNAs and six rRNA operons. Identification of the genes related to multidrug resistance from this genome and the discovery of a novel conjugative plasmid will increase our understanding of the pathogenicity associated with this species.

Characterization of silver nanoparticles prepared by wet chemical method and their antibacterial and cytotoxicity activities

Purpose: To investigate the efficiency of silver nanoparticles synthesized by wet chemical method, and evaluate their antibacterial and anti-cancer activities. Methods: Wet chemical method was used to synthesize silver nanoparticles (AgNPs) from silver nitrate, trisodium citrate dehydrate (C6H5O7Na3.2H2O) and sodium borohydride (NaBH4) as reducing agent. The AgNPs and the reaction process were characterized by UV–visible spectrometry, zetasizer, transmission electron microscopy (TEM) and scanning electron microscopy (SEM) equipped with energy dispersive spectroscopy (EDS). The antibacterial and cytotoxic effects of the synthesized nanoparticles were investigated by agar diffusion method and MTT assay respectively. Results: The silver nanoparticles formed were spherical in shape with mean size of 10.3 nm. The results showed good antibacterial properties, killing both Gram-positive and Gram-negative bacteria, and its aqueous suspension displayed cytotoxic activity against colon adenocarcinoma (HCT-116) cell line. Conclusion: The findings indicate that silver nanoparticles synthesized by wet chemical method demonstrate good cytotoxic activity in colon adenocarcinoma (HCT-116) cell lines and strong antibacterial activity against various strains of bacteria.

Endotoxinas bacterianas versus resposta inflamatória e alérgica

Endotoxinas bacterianas são lipopolissacáridos da membrana externa das bactérias Gram-negativas e são responsáveis pela libertação de citocinas no organismo humano. Estas citocinas podem exacerbar a sintomatologia de determinadas patologias, nomeadamente do foro pulmonar, agravando a sua situação clínica. A procura de melhor conforto habitacional, com a criação de micro ambientes "artificiais" dentro de casa, oferece bem-estar, mas por vezes arrasta consigo uma maior quantidade de poluentes aéreos indoor, nomeadamente bioaerossóis. A influência das bactérias Oram­ negativas em suspensão indoor nos níveis basais dos marcadores analíticos sanguíneos de resposta inflamatória e alérgica, é um ponto importante a ter em consideração quando se pretende elevar o grau de conforto de uma habitação sem implicações negativas para a saúde humana. Este estudo demonstra que a presença das endotoxinas bacterianas indoor, na ausência de patologia aguda, não prejudica o ser humano e a melhoria das condições habitacionais não degrada o estado fisiológico dos indivíduos. /Summary: Bacterial endotoxins are lipopolysaccharides found in the outer membrane of Gram­ negative bacteria and are responsible for the release of cytokines in the human body. These cytokines may exacerbate the symptoms of certain diseases, including pulmonary events, worsening their medical condition. The demand for better housing comfort with the creation of "artificial" micro-environments in the house, offers well-being, but often brings a greater quantity of indoor air pollutants, including bioaerosols. The influence of suspended indoor Gram-negative bacteria in basal levels of blood analytical markers of inflammatory and allergic rhinitis, is an important point to consider when attempting to raise the comfort level of a dwelling without negative implications for human health. This study proves that the presence of indoor bacterial endotoxin in the absence of acute pathology, do not affect humans and the improvement of living conditions does not degrade the physiological status of individuals.

Green synthesis of silver nanoparticles using Arbutus andrachne leaf extract and its antimicrobial activity

Purpose: To synthesize silver nanoparticles (AgNPs) of Arbutus andrachne leaf water extract (LE) and to evaluate the antimicrobial activity of both LE and AgNPs. Methods: The synthesized AgNPs were characterized using the following techniques: ultraviolet-visible spectroscopy (UV-vis), Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), thermal gravimetric analysis (TGA), X-ray diffraction (XRD) analysis, and analysis of particle size (PS) and zeta potential (ZP). The antimicrobial activities of LE and NPs were assessed by Kirby-Bauer disc diffusion (DD) and broth microdilution (MD) methods according to the recommendations of the Clinical and Laboratory Standards Institute (CLSI). LE and AgNPs were examined against fresh cultures of four Gram-positive and five Gram-negative bacteria, and three yeast strains. Results: AgNPs were successfully synthesized and characterized using Arbutus andrachne LE. The AgNPs showed moderate antibacterial activity against Staphylococcus aureus ATCC 6538p, S. epidermidis ATCC 12228, Escherichia coli ATCC 29998, Klebsiella pnemoniae ATCC 13883 and Pseudomonas aeruginosa ATCC 27853, and also antifungal activity against Candida albicans ATCC 10239 and C. krusei ATCC 6258. Conclusions: Due to the potent activity of AgNPs against Gram-positive and Gram-negative bacteria, and yeast strains, it is suggested that AgNPs are potential broad spectrum antimicrobial agents.

Antibacterial and cytotoxic properties of isoprenoids from the red sea soft coral, Lobophytum sp

Purpose: To evaluate the antibacterial and cytotoxic activities of the secondary metabolites of Lobophytum sp. Methods: Maceration with methanol: chloroform (1:1) was applied to extract the coral material. Chromatographic and spectroscopic techniques were employed for fractionation, isolation and elucidation of pure compounds. Antibacterial activities were performed by well diffusion method against three Gram-positive and four Gram-negative bacteria. Brine shrimp lethality test was employed to predict toxicity, while antitumor activity were tested by 3-(4, 5-dimethylthiazol-2-yl)-2, 5- diphenyltetrazolium bromide (MTT) method against Ehrlich carcinoma cells. Results: Four sesquiterpenes, one cembranoid type diterpenes and two steroids were isolated. 1 exhibited significant antibacterial activity against four tested bacteria (P. aeruginosa, S. aureus, S. epidermis, and S. pneumonia) with MIC value of 15 μg/mL. Moreover, 1 showed high diameter zone of inhibition ranging from 16 - 18 mm against test bacteria. Compounds 4 and 5 displayed moderate antibacterial activity against all test bacteria with inhibition zone diameter (IZD) ranging from 11 – 15 mm and MIC values of 30 μg/mL. 2, 3, 6 and 7 exhibited weak antibacterial activity (IZD, 7 - 11 mm; MIC ≥ 30 μg/mL). In addition, only diterpene compound (4) showed high toxicity against A. Salina and antitumor activity against Erhlich carcinoma cells with the LD50 of 25 and 50 μg/mL, respectively. Conclusion: This study reveals the strong antibacterial activity of sesquiterpene alismol (1) and the potential antibacterial and antitumor activity of cembranoid type diterpene, cembrene A (4).

Synthesis and antimicrobial activity of some 2- Piperidinomethylamino-4-(7-H/substituted coumarin-3-yl)- 6-chlorosubstitutedphenyl pyrimidines

Purpose: To prepare and evaluate some 2-piperidinomethylamino-4-(7-H/substitutedcoumarin-3-yl)-6- chlorosubstitutedphenyl pyrimidines as antimicrobial agents. Methods: Some 2-piperidinomethylamino-4-(7-H/substitutedcoumarin-3-yl)-6-chlorosubstitutedphenyl pyrimidines were prepared by reacting 2-amino-4-(7-H/substitutedcoumarin-3-yl)-6- (chlorosubstitutedphenyl) pyrimidines with piperidine and formaldehyde. The chemical structures of the synthesized compounds were elucidated by Fourier transform infrared (FTIR), 1H-nuclear magnetic resonance (1H-NMR), mass spectrometry and elemental analysis. These compounds were investigated for their antimicrobial activity against ten bacteria and five fungi by serial plate dilution method using standard drugs, namely, ofloxacin and ketoconazole, respectively, and their minimum inhibitory concentrations (MICs) were also determined. Results: A total of eighteen new compounds (1a-18a) were synthesized. Compound 6a (MIC = 50 μg/mL; p < 0.05 or less) displayed the highest activity against S. aureus , E. faecalis , Staphylococcus epidermidis , B. subtilis , and B. cereus . Compound 6a further showed good activity (MIC = 25 μg/mL; p < 0.05 or less) against E. coli ; P. aeruginosa K. pneumonia , B. bronchiseptica , and P. vulgaris . Compounds 6a (MIC = 25 μg/mL; p < 0.0001) and 17a (MIC = 25 μg/mL; p < 0.0001) displayed very good activity against C. albicans , A. niger , A. flavus , M. purpureous , and P. citrinum , respectively. Analysis of structure-activity relationship revealed that the presence of bromo group at 7-postion of the coumarin moiety along with the 4-chlorophenyl group at position-6 of the pyrimidine ring is critical for antimicrobial activity against Gram-positive bacteria, Gram negative bacteria and fungi. Conclusion: The synthesized 2-piperidino derivatives are better antifungal and antibacterial agents than the earlier reported 2-morpholino derivatives, but require further investigations against other microbial strains to ascertain their broad spectrum antimicrobial activity.