Search results for: lactic acid bacteria; multidrug-resistant pathogens

Authors: Neha Bhadauria, Indu Gaur, Shilpi Shilpi, Susmita Goswami, Prabir K. Paul

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The aerial surface of plants colonized by Human Enteric Pathogens ()has been implicated in outbreaks of enteric diseases in humans. Practice of organic farming primarily using animal dung as manure and sewage water for irrigation are the most significant source of enteric pathogens on the surface of leaves, fruits and vegetables. The present work aims to have an insight into the molecular mechanism of interaction of Human Enteric Pathogens or their metabolites with cell wall receptors in plants. Tomato plants grown under aseptic conditions at 12 hours L/D photoperiod, 25±1°C and 75% RH were inoculated individually with S. fonticola and K. pneumonia. The leaves from treated plants were sampled after 24 and 48 hours of incubation. The cell wall and cytoplasmic proteins were extracted and isocratically separated on 1D SDS-PAGE. The sampled leaves were also subjected to formaldehyde treatment prior to isolation of cytoplasmic proteins to study protein-protein interactions induced by Human Enteric Pathogens. Protein bands extracted from the gel were subjected to MALDI-TOF-TOF MS analysis. The foremost interaction of Human Enteric Pathogens on the plant surface was found to be cell wall bound receptors which possibly set ups a wave a critical protein-protein interaction in cytoplasm. The study revealed the expression and suppression of specific cytoplasmic and cell wall-bound proteins, some of them being important components of signaling pathways. The results also demonstrated HEP induced rearrangement of signaling pathways which possibly are crucial for adaptation of these pathogens to plant surface. At the end of the study, it can be concluded that controlling the over-expression or suppression of these specific proteins rearrange the signaling pathway thus reduces the outbreaks of food-borne illness.

Keywords: cytoplasmic protein, cell wall-bound protein, Human Enteric Pathogen (HEP), protein-protein interaction

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Authors: Hye Rim Kang, So Yae Koh, Ji-Yeon Ryu, Chang Kyu Lee, Ji Hee Lim, Hyeon A. Kim, Geun Hyung Im, Somi Kim Cho

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In this study, antioxidant properties and compositional analysis of fermented chocolate were examined. Chocolate was fermented with Lactobacillus plantarum CK10. As fermentation time went by, pH was decreased (5.26±0.02 to 3.98±0.06) while titratable acidity was increased (5.36±0.19 to 13.31±0.34). The total polyphenol contents were maintained through the fermentation. The contents of total polyphenol were slightly increased at 8 hr (6.34±0.12 mg GAE (Gallic acid equivalent)/g), and it reached to comparable levels of the control at 24 hr (control, 5.47±0.36 mg GAE/g); 24 hr, 5.19±0.23 mg GAE/g). Similarly, the total flavonoid contents were not significantly changed during fermentation. The pronounced radical scavenging activities of chocolate, against DPPH-, ABTS-, and Alkyl radical, were observed. The levels of antioxidant activities were not dramatically altered in the course of fermentation. By gas chromatography-mass spectrometry analysis, the increase in lactic acid was measured and four major compounds, HMF, xanthosine, caffeine, and theobromine, were identified. The relative peak area of caffeine and theobromine was considerably changed during fermentation. However, no significant difference in the levels of caffeine and theobromine were observed by high-performance liquid chromatography analysis.

Keywords: antioxidant, chocolate, compositional analysis, fermentation, Lactobaillus plantarum

Procedia PDF Downloads 287

Authors: Liang Zhou, Xiaojing Zhu, Yin Lu

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Danshen can induce the angiogenesis in advanced ischemic heart disease while inhibiting the angiogenesis in cancer. Additionally, Danshen mainly contains two groups of ingredients: the hydrophilic phenolic acids (danshensu, caffeic acid and salvianolic acid B), and the lipophilic tanshinones (dihydrotanshinone I, tanshinone II A, and cryptotanshinone). The lipophilic tanshinones reduced the VEGF- and bFGF-induced proliferation of HUVECs in dose-dependent manner, but cannot perform in others. Conversely, caffeic acid and salvianolic acid B had the opposite effect. Danshensu inhibited the VEGF- and bFGF-induced migration of HUVECs, and others were not. Most of them interrupted the forming capillary-like structures of HUVECs, except the danshensu and caffeic acid. Oppositely, caffeic acid enhanced the ability of forming capillary-like structures of HUVECs. Ultimately, the lipophilic tanshinones, danshensu and salvianolic acid B inhibited the angiogenesis, whereas the caffeic acid induced the angiogenesis. These data provide useful information for the classification of ingredients of Danshen for angiogenesis.

Keywords: angiogenesis, Danshen, HUVECs, ingredients

Procedia PDF Downloads 396

Authors: Anoff Anim, Lila A. M. Mahmoud, Maria Katsikogianni, Sanjit Nayak

Abstract:

Sustained and controlled delivery of antimicrobial drugs have been largely studied recently using metal organic frameworks (MOFs)and different polymers. However, much attention has not been given to combining both MOFs and biodegradable polymers, which would be a good strategy in providing a sustained gradual release of the drugs. Herein, we report a comparative study of the sustained and controlled release of widely used antibacterial drugs, cephalexin and metronidazole, from zinc-based MOF-5 incorporated in biodegradable polycaprolactone (PCL) and poly-lactic glycolic acid (PLGA) membranes. Cephalexin and metronidazole were separately incorporated in MOF-5 post-synthetically, followed by their integration into biodegradable PLGA and PCL membranes. The pristine MOF-5 and the loaded MOFs were thoroughly characterized by FT-IR, SEM, TGA and PXRD. Drug release studies were carried out to assess the release rate of the drugs in PBS and distilled water for up to 48 hours using UV-Vis Spectroscopy. Four bacterial strains from both the Gram-positive and Gram-negative types, Staphylococus aureus, Staphylococuss epidermidis, Escherichia coli, Acinetobacter baumanii, were tested against the pristine MOF, pure drugs, loaded MOFs and the drug-loaded MOF-polymer composites. Metronidazole-loaded MOF-5 composite of PLGA (PLGA-Met@MOF-5) was found to show highest efficiency to inhibit the growth of S. epidermidis compared to the other bacteria strains while maintaining a sustained minimum inhibitory concentration (MIC). This study demonstrates that the combination of biodegradable MOF-polymer composites can provide an efficient platform for sustained and controlled release of antimicrobial drugs and can be a potential strategy to integrate them in biomedical devices.

Keywords: antimicrobial resistance, biodegradable polymers, cephalexin, drug release metronidazole, MOF-5, PCL, PLGA

Procedia PDF Downloads 139

Authors: Nelson R. Osungbemiro, Rafiu O. Sanni, Rotimi F. Olaniyan, Abayomi O. Olajuyigbe

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Bacteria flora of Clarias gariepinus collected from two natural habitats namely Owena River (freshwater) and Igbokoda lagoon (brackish water) were examined using standard microbiological procedures. Thirteen bacterial species were identified. The result indicated that from the identified bacteria isolated, Vibrio sp, Proteus sp. Shigella sp. and E. coli were present in both habitats (fresh and brackish waters). Others were habitat-selective such as Salmonella sp., Pseudomonas sp, Enterococcus sp, Staphylococcus sp. that were found only in freshwater habitat. While Branhamella sp, Streptococcus sp. and Micrococcus sp. were found in brackish water habitat. Bacteria load from Owena river (freshwater) was found to be the highest load recorded at 6.21 x 104cfu. T-test analysis also revealed that there was a marked significant difference between bacterial load in guts of sampled Clarias from fresh water and brackish water habitats.

Keywords: bacteria flora, gut, Clarias gariepinus, Owena river

Procedia PDF Downloads 460

Authors: Nazedah Ain Ibrahim, Mohamed Mansor Manan

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Treatment of suspected early onset neonatal sepsis (EONS) in Neonatal Intensive Care Unit (NICU) is essential. However, information regarding EONS pathogens may vary between regions. Global perspectives showed Group B Streptococcal (GBS) as the most common causative pathogens, but the widespread use of intrapartum antibiotics has changed the pathogens pattern towards gram negative microorganisms, especially E. coli. Objective of this study is to describe the pathogens isolated, to assess current treatment and risk of EONS. Records of 899 neonates born in three General Hospitals between 2009 until 2012 were retrospectively reviewed. The inclusion criteria were neonates with blood culture taken prior to empiric antibiotics administration and within 72 hours of life. Of the study group, a total of 734 (82%) cases had documented blood culture that met the inclusion criteria. Proven EONS (as confirmed by positive blood culture) was found in 22 (3%) neonates. The majority was isolated with gram positive organisms, 17 (2.3%). In addition, other common gram positive organism isolated were Coagulase negative staphylococci (7) followed by Bacillus sp. (5) and Streptococcus pneumonia (2), and only one case isolated with GBS, Streptococcus spp. and Enterococcus sp. Meanwhile, only five cases of gram negative organisms [Stenotropomonas (xantho) maltophi (1), Haemophilus influenza (1), Spingomonas paucimobilis (1), Enterobacter gergoviae (1) and E. coli (1)] were isolated. A total of 286 (39%) cases were exposed to intrapartum antibiotics and of those, 157 (21.4%) were administered prior to delivery. All grams positive and most gram negative organisms showed sensitivity to the tested antibiotics. Only two rare gram negative organisms showed total resistant. Male, surfactant, caesarean delivery and prolonged rapture of membrane >18hours were a possible risk of proven EONS. Although proven EONS remains uncommon in Malaysia, nonetheless, the effect of intrapartum antibiotics still required continuous surveillance. However, by analyzing isolated pathogens it can be used as treatment guidance in managing suspected EONS.

Keywords: early onset neonatal sepsis, neonates, pathogens, gram positive, gram negative

Procedia PDF Downloads 315

Authors: Anthony Ayodeji Adegoke, Olayinka Ayobami Aiyegoro, Thor Axel Stenstrom

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A study to investigate the species diversities, antibiogram and antibiotic resistance genes in Staphylococcus species from raw meat and dairy products collected from an abattoir and a farm shop of a research institute in Irene, South Africa over a six-month period was conducted. Polymerase Chain Reaction was used to speciate the bacteria and to detect the presence and otherwise of resistance genes. Antibiotic susceptibility testing was performed by disk diffusion method on Mueller-Hinton agar according to the Clinical Laboratory Standards Institute standards. A total of twenty-six (26) antibiotics were used to determine the antibiotic susceptibility. S. xylosus was the predominant isolate with 30% total occurrence, followed by S. epidermis, S. aureus, S. saprophyticus and S. haemolyticus with 25%, 15%, 15%, and 10% abundance respectively. The isolates were resistant to ceftezidime, gentamycin, nalidixic acid, nortrafuration, ampicillin, penicillin, oxytetracycline, tetracycline, doxycycline, clindamycin and lincomycin. mecA genes was detected among the methicillin resistant Staphylococcus species (MRSS) but no vancomycin resistance genes (van A and van B) were detected in these isolates. The presence of MRSS and multidrug resistant Staphylococcus species in meat affirms the need to avoid consumption of partially cooked meat currently rampant in South Africa, to avoid the spread of difficult to control pathogens in epidemiological proportion.

Keywords: Staphylococcus species, antibiotics, antibiotic resistance genes, food products, methicillin resistance, mecA gene

Procedia PDF Downloads 299

Authors: Bouaziz Amira, Zaatout Nawel

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Background: Antimicrobial resistance is an urgent global health challenge in human and veterinary medicine, where migratory birds play a major role in the dissemination of multi-drug-resistant bacteria. The aim of this study was to screen for the presence of carbapenemase-producing Gram-negative bacteria (GNB) in the little egret (Egrettagarzetta) migratory bird stools in Algeria. Materials/Methods: In January 2014, 12 feacal samples were collected in Garaet El-Tarf, Oum El-Bouaghi city, Algeria. Samples were subjected to selective isolation of carbapenem-resistant GNB. Representative colonies were identified using the VITEK system. The obtained isolates were subjected to antibiotic susceptibility testing using the disc-diffusion method as well as carbapenemase production was verified by the modified Carba NP test. Results: In total, ten E. coli were obtained and were resistant to amoxicillin/clavulanic acid (100%), ertapenem (70%), cefoxitin (60%) cefotaxime (20%), cefepime (20%), ciprofloxacin (20%) and aztreonam (10%). The phenotypic detection results revealed that six out of the obtained strains were positive for the modified Carba NP test. Conclusion: The present study suggests that the little egret (Egretta garzetta) could be considered a reservoir of carbapenem-resistant Gram-negative bacteria.

Keywords: antimicrobial resistance, E. coli, Egretta garzetta, carbapenem resistance, dissemination

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Authors: Hakimullah Hakim, Chiharu Toyofuku, Mari Ota, Mayuko Suzuki, Miyuki Komura, Masashi Yamada, Md. Shahin Alam, Natthanan Sangsriratanakul, Dany Shoham, Kazuaki Takehara

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Disinfectants and their application are essential part of infection control strategies and enhancement of biosecurity at farms, worldwide. Alkaline agents are well known for their strong and long term antimicrobial capacities and most frequently are applied at farms for control and prevention of biological hazards. However, inadequate information regarding such materials’ capacities to inactivate pathogens and their improper applications fail farmers to achieve the mentioned goal. Thus, this requires attention to further evaluate their efficacies, under different conditions and in different ways. Here in this study we evaluated bactericidal efficacies of food additive grade of calcium hydroxide (FdCa(OH)2) powder derived from natural calcium carbonates obtained from limestone (Fine Co., Ltd., Tokyo, Japan), and bioceramic powder (BCX) derived from chicken feces at pH 13 (NMG environmental development Co., Ltd., Tokyo, Japan), for their efficacies to inactivate bacteria in feces. [Materials & Methods] Chicken feces were inoculated by 100 µl Escherichia coli and Salmonella Infantis in falcon tubes, individually, then FdCa(OH)2 or BCX powders were individually added to make final concentration of 0, 5, 10, 20 and 30% (w/w) in total weight of 0.5g, followed by properly mixing and incubating at room temperature for certain periods of time, in a dark place. Afterwards, 10 ml 1M Tris-HCl (pH 7.2) was added onto them to reduce their pH, in order to stop powders’ activities and to harvest the remained viable bacteria, whereas using normal medium or dW2 to recover bacteria increases the mixture pH, and as a result bacteria would be inactivated soon; therefore, the latter practice brings about incorrect and misleading results. Samples were then inoculated on DHL agar plates in order to calculate colony forming units (CFU)/ml of viable bacteria. [Results and Discussion] FdCa(OH)2 powder at 10% and 5% required 3 hr and 6 hr exposure times, respectively, while BCX powder at 20% concentrations required 6 hr exposure time to kill the mentioned bacteria in feces down to lower than detectable level (≤ 3.6 log10 CFU/ml). This study confirmed capacities of FdCa(OH)2 and BCX powders to inactivate bacteria in feces, and both materials are environment friendly materials, with no risk to human or animal’s health. This finding helps farmers to properly apply alkaline agents in appropriate concentrations and exposure times in their farms, in order to prevent and control infectious diseases outbreaks and to enhance biosecurity. Finally, this finding may help farmers to implement better strategies for infections control in their livestock farms.

Keywords: bacterial inactivation, bioceramic, biosecurity at livestock farms, chicken feces

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Authors: M. Kustyawati, F. Pratama, D. Saputra, A. Wijaya

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Tempe composes of not only molds but also bacteria and yeasts. The structure of microorganisms needs to be in balance number in order the tempe to be an acceptable quality for an extended time. Sub supercritical carbon dioxide can be a promising preservation method for tempe as it induces microbial inactivation avoiding alterations of its quality attributes. Fresh tempe were processed using supercritical and sub supercritical CO2 for a defined holding times, then the growth ability of molds and bacteria were analyzed. The results showed that the supercritical CO2 processing for 5 minutes reduced the number of bacteria and molds to 0.30 log cycle and 1.17 log cycles, respectively. In addition, sub supercritical CO2 processing for 20 minutes had fungicidal effect against mold tempe; whereas, the sub supercritical CO2 for 10 minutes had reducing effect against bacteria tempe, and had fungistatic affect against mold tempe. It suggested that sub-supercritical CO2 processing for 10 min could be useful alternative technique for preservation of tempe.

Keywords: tempe, sub supercritical CO2, fungistatic effect, preservation

Procedia PDF Downloads 269

Authors: Ankita Singh, Chandan Gorain, Amirul I. Mallick

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Successful adherence of the mucosal epithelial cells is the key early step for Campylobacter jejuni pathogenesis (C. jejuni). A set of Surface Exposed Colonization Proteins (SECPs) are among the major factors involved in host cell adherence and invasion of C. jejuni. Among them, constitutively expressed surface-exposed lipoprotein adhesin of C. jejuni, JlpA, interacts with intestinal heat shock protein 90 (hsp90α) and contributes in disease progression by triggering pro-inflammatory response via activation of NF-κB and p38 MAP kinase pathway. Together with its ability to express in the bacterial surface, higher sequence conservation and predicted predominance of several B cells epitopes, JlpA protein reserves its potential to become an effective vaccine candidate against wide range of Campylobacter sps including C. jejuni. Given that chickens are the primary sources for C. jejuni and persistent gut colonization remain as major cause for foodborne pathogenesis to humans, present study explicitly used chickens as model to test the immune-protective efficacy of JlpA protein. Taking into account that gastrointestinal tract is the focal site for C. jejuni colonization, to extrapolate the benefit of mucosal (intragastric) delivery of JlpA protein, a food grade Nisin inducible Lactic acid producing bacteria, Lactococcus lactis (L. lactis) was engineered to express recombinant JlpA protein (rJlpA) in the surface of the bacteria. Following evaluation of optimal surface expression and functionality of recombinant JlpA protein expressed by recombinant L. lactis (rL. lactis), the immune-protective role of intragastric administration of live rL. lactis was assessed in commercial broiler chickens. In addition to the significant elevation of antigen specific mucosal immune responses in the intestine of chickens that received three doses of rL. lactis, marked upregulation of Toll-like receptor 2 (TLR2) gene expression in association with mixed pro-inflammatory responses (both Th1 and Th17 type) was observed. Furthermore, intragastric delivery of rJlpA expressed by rL. lactis, but not the injectable form, resulted in a significant reduction in C. jejuni colonization in chickens suggesting that mucosal delivery of live rL. lactis expressing JlpA serves as a promising vaccine platform to induce strong immune-protective responses against C. jejuni in chickens.

Keywords: chickens, lipoprotein adhesion of Campylobacter jejuni, immuno-protection, Lactococcus lactis, mucosal delivery

Procedia PDF Downloads 139

Authors: Hamed Ahari, Sepideh Farokhi, Mohamad Reza Abedini

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This paper focuses on inactivation of the growth of the bacterial mixture, Salmonella enteritidis, Staphylococcus aureus, Bacillus cereus and Escherichia coli, experimentally subjected to the chicken eggshell by two types of nano particle-Ag, composite film and colloidal spray carried out at concentrations of 500, 1000 and 2000 ppm over 28 days. The GLM, Repeated Measurement-ANOVA procedure was used to analyze the effect of time and concentration of nano groups on inactivation of bacteria, simultaneously. The maximum reduction of the bacterial growth was respected to the group “spray 2000 ppm” for which the value of the bacteria reached the minimum (0.93±0.42) on day 7, calculated to be 0.0 on days14 and 28 and followed by the group “spray 1000 ppm”. It was obviously concluded that increasing the dilution of nano coating in spray and film created a significant decrease in the number of bacteria colonies on the eggshells but the effect of packaging in different concentrations of nanocomposite was not statistically significant in different days of the study.

Keywords: nano particle, composite film, eggshell, bacteria

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Authors: N. Ouslimani, T. Abadlia, M. Fadel

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This work focused on the elimination of acid textile dye (red bermacide acid dye BN-CL-200), widely used for dyeing wool and polyamide fibers, by adsorption on a natural material, wood sawdust, in the static mode by keeping under continuous stirring, a specific mass of the adsorbent, with a dye solution of known concentration. The influence of various parameters is studied like the influence of particle size, mass, pH and time. The best results were obtained with 0.4 mm grain size, mass of 3g, Temperature of 20 °C, pH 2 and Time contact of 120 min.

Keywords: acid dye, environment, wood sawdust, wastewater

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Authors: Soyeon Kang, Youn-Jee Chung, Jung Namkung

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Objective: This retrospective study investigated the association between uric acid level and bone mineral density (BMD) in the postmenopausal period. Methods: The study included 328 menopausal women (mean age, 57.3 ± 6.5 years; mean serum uric acid level, 4.6 ± 1.0 mg/dL). Patients were divided into three groups by tertile of serum uric acid level. Patients who used hormone treatment (HT), bisphosphonates, or lipid-lowering agents were included. Results: Blood urea nitrogen, serum creatinine, and serum triglyceride levels were significantly higher in the upper uric acid tertiles. No significant difference was found in the mean uric acid levels between medication users and non-users. Distinct HT regimens showed different mean serum uric acid levels. In a cross-sectional analysis, higher serum uric acid levels showed a tendency toward increased BMD in the spine and femoral neck. Longitudinal analysis of 186 women who underwent follow-up examination at a mean interval of 14.6 months revealed a trend toward a smaller reduction in femoral neck BMD in women in the upper serum uric acid tertiles. Conclusion: A positive correlation exists between serum uric acid levels and BMD in menopausal women.

Keywords: menopause, antioxidant, uric acid, bone mineral density

Procedia PDF Downloads 129

Authors: Jedli Hedi, Hedfi Hachem, Abdessalem Jbara, Slimi Khalifa

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Natural and modified clay were used as an adsorbent for CO2 capture. Sample of clay was subjected to acid treatments to improve their textural properties, namely, its surface area and pore volume. The modifications were carried out by heating the clays at 120 °C and then by acid treatment with 3M sulphuric acid solution at boiling temperature for 10 h. The CO2 adsorption capacities of the acid-treated clay were performed out in a batch reactor. It was found that the clay sample treated with 3M H2SO4 exhibited the highest Brunauer–Emmett–Teller (BET) surface area (16.29–24.68 m2/g) and pore volume (0.056–0.064 cm3/g). After the acid treatment, the CO2 adsorption capacity of clay increased. The CO2 adsorption capacity of clay increased after the acid treatment. The CO2 adsorption by clay, were characterized by SEM, FTIR, ATD-ATG and BET method. For describing the phenomenon of CO2 adsorption for these materials, the adsorption isotherms were modeled using the Freundlich and Langmuir models. CO2 adsorption isotherm was found attributable to physical adsorption.

Keywords: clay, acid treatment, CO2 capture, adsorption mechanism

Procedia PDF Downloads 211

Authors: Claire B. Salvedia, Enrico P. Supangco, Francisco B. Eligado, Renato Sa Vega, Antonio A. Rayos

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The study was conducted to evaluate the effect of probiotic feeding on weight gain, blood biochemical and hematological indices of crossbred dairy goat kids. Sixteen (16) crossbred Anglo-Nubian x Saanen dairy goat kids, 3 to 4 months old, ranging from 19 to 23kg were randomly assigned into four treatments fed with 5x109 cfu/ml probiotic supplements; Treatment 1 – control; Treatment 2 – lactic acid bacteria (L. plantarum BS and P. acidilactici 3G3); treatment 3 – S. cerevisiae 2030; Treatment 4 – multi-strain probiotics (L. plantarum BS, P. acidilactici 3G3, and S.cerevisiae 2030). Feed ration provided daily for each of the experimental animals were composed of 1kg mixed concentrate feed ((Leucaena leucocephala dried leaves and pollard), and 4 kg fresh Pennisetum purpureum and Gliciridia sepium leaves (50:50). The experimental feeding trial lasted for 9 weeks. Result revealed that treatments fed with probiotics had significantly (P≤0.05) higher weight gain compared to the control. Significant effect on plasma urea nitrogen (PUN) and triglyceride were noted during 30th and 60th day of probiotic feeding. White blood cell counts were significantly affected by probiotic feeding during the 60th day. Concentrations of glucose and cholesterol remained unchanged throughout the experimental period. The findings suggests, under the condition of the experiment, that live probiotic feeding could have a significant role in improving weight gain and metabolism of crossbred dairy goat kids.

Keywords: probiotics, weight gain, blood biochemical indices, crossbred dairy goat kids

Procedia PDF Downloads 491

Authors: Anoff Anim, Lila Mahmound, Maria Katsikogianni, Sanjit Nayak

Abstract:

Sustained and controlled delivery of antimicrobial drugs have been largely studied recently using metal organic frameworks (MOFs)and different polymers. However, much attention has not been given to combining both MOFs and biodegradable polymers which would be a good strategy in providing a sustained gradual release of the drugs. Herein, we report a comparative study of the sustained and controlled release of widely used antibacterial drugs, cephalexin and metronidazole, from zinc-based MOF-5 incorporated in biodegradable polycaprolactone (PCL) and poly-lactic glycolic acid (PLGA) membranes. Cephalexin and metronidazole were separately incorporated in MOF-5 post-synthetically, followed by their integration into biodegradable PLGA and PCL membranes. The pristine MOF-5 and the loaded MOFs were thoroughly characterized by FT-IR, SEM, TGA and PXRD. Drug release studies were carried out to assess the release rate of the drugs in PBS and distilled water for up to 48 hours using UV-Vis Spectroscopy. Four bacterial strains from both the Gram-positive and Gram-negative types, Staphylococus aureus, Staphylococuss epidermidis, Escherichia coli, Acinetobacter baumanii, were tested against the pristine MOF, pure drugs, loaded MOFs and the drug-loaded MOF-polymer composites. Metronidazole-loaded MOF-5 composite of PLGA (PLGA-Met@MOF-5) was found to show highest efficiency to inhibit the growth of S. epidermidis compared to the other bacteria strains while maintaining a sustained minimum inhibitory concentration (MIC). This study demonstrates that the combination of biodegradable MOF-polymer composites can provide an efficient platform for sustained and controlled release of antimicrobial drugs, and can be a potential strategy to integrate them in biomedical devices.

Keywords: antimicrobial resistance, biodegradable polymers, cephalexin, drug release metronidazole, MOF-5, PCL, PLGA

Procedia PDF Downloads 85

Authors: Yousef Naserzadeh, Niloufar Mahmoudi

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Oxidation resistance is one of the important parameters in maintaining the quality of olive oil during its storage. Ensuring the stability of the quality of olive oil is one of the important concerns of the producers and consumers. Prooxidants such as iron and copper accelerate the oxidation reaction, and also anti-oxidants and chelating compounds delay it. In this study, chelating effect of tea extract which contains significant amounts of tannic acid is investigated in comparison with citric acid. To do it, 0.1 ppm copper was added to these four kinds of oil, sunflower, olive, canola, and tallow, and then chelating effect of citric acid (0.01%), tannic acid (0.01%) and tea extract (0.1%) were measured by adding to this composition. To this end, the resistance time of the oils against oxidation was measured at 120 °C and an air flow of 20 liters per hour. And the value of peroxide was measured by oven test in six periods of 24 hours at 105 °C. The results showed that citric acid, tannic acid and tea extract had chelating property and increased the resistance time of the studied oils. As a result, considering chelating property and increasing resistance of oil, tannic acid showed better effect than tea extract and tea extract had better effect than citric acid.

Keywords: tannic acid, chelate, edible oils, black tea extract, TBHQ

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Authors: V. V. Lakshmi, Y. V. S. Annapurna

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Urinary tract infection (UTI) is one of the most common infectious diseases seen in the community. Susceptible individuals experience multiple episodes, and progress to acute pyelonephritis or uro-sepsis or develop asymptomatic bacteriuria (ABU). Ability to cause extraintestinal infections depends on several virulence factors required for survival at extraintestinal sites. Presence of virulence phenotypes enhances the pathogenicity of these otherwise commensal organisms and thus augments its ability to cause extraintestinal infections, the most frequent in urinary tract infections(UTI). The present study focuses on detection of the virulence characters exhibited by the uropathogenic organism and most common factors exhibited in the local pathogens. A total of 700 isolates of E.coli and Klebsiella spp were included in the study. These were isolated from patients from local hospitals reported to be suffering with UTI over a period of three years. Isolation and identification was done based on Gram character and IMVIC reactions. Antibiotic sensitivity profile was carried out by disc diffusion method and multi drug resistant strains with MAR index of 0.7 were further selected.. Virulence features examined included their ability to produce exopolysaccharides, protease- gelatinase production, hemolysin production, haemagglutination and hydrophobicity test. Exopolysaccharide production was most predominant virulence feature among the isolates when checked by congo red method. The biofilms production examined by microtitre plates using ELISA reader confirmed that this is the major factor contributing to virulencity of the pathogens followed by hemolysin production

Keywords: Escherichia coli, Klebsiella sp, Uropathogens, Virulence features.

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Authors: Onur Yel

Abstract:

This study represents the removal of harmful dye substances from wastewaters by using waste and cheap adsorbents. Rapid population growth and industrialization occasion anthropogenic pollution which gives irreversible damage to the environment. One of the ways in which water pollution occurs is caused by the release of the dyestuffs in the textile industry. The release of dyestuffs to the environment directly damages the living creatures that have acquired water habitat. Especially, wastewater cannot be used for nutritional purposes. In addition, some adsorbents have mutagenic and/or carcinogenic effects. By blocking photosynthesis, it hinders the inhibition of photosynthetic bacteria in the water, which damages the ecological balance and also causes the formation of malodorous compounds. Moreover, the lack of oxygen can pose a serious danger to the lives of other living organisms that need oxygen. In recent years, some physical and chemical methods are preferred for the removal of dyestuffs. However, the utilization of these methods is expensive. For this reason, the availability of new and cheap adsorbents becomes the more significant issue. In this study, an investigation of various variables on the removal of Methylene Blue and Acid Red-88 dyestuffs from wastewaters by the usage of pulverized cypress cones has been carried out. Thus, various masses of absorbent (0.1-0.25-0.5-1-2-4-5 grams) are used in 50, 100, 150, 200, 300 ppm concentrations of Methylene Blue and Acid Red-88 dyestuffs’ solutions, and with a variety of the interaction time (0.25-0.5-1-2-4-5 hours). The mixtures were centrifuged and the absorbance of the filtrates was measured on a UV spectrophotometer to determine their remaining concentrations. In the study, the highest removal ratio of Acid Red-88 dyestuff was found to be 81% at 200 ppm of dyestuff with 2 grams of adsorbent at 300 minutes. For Methylene Blue experiments, the removal percentage was found as 98% where 2 grams of adsorbent is used in 200 ppm dyestuff solution at 120 minutes of interaction.

Keywords: acid red-88, biosorption, methylene blue, cypress cones, water pollution

Procedia PDF Downloads 141

Authors: V. Baweja, K. K. Gupta, V. Dubey, C. Keshavam

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Antimicrobial activity of six indigenous plants such as Tulsi Ocimum sanctum, Neem Azadirachta indica, Aloe vera, Turmeric Curcuma longa, Lantana Lantana camara, and Clove Syzygium aromaticum was assessed against the gut microbiota of the dengue fever mosquito Aedes aegypti, keeping in view that the presence of midgut bacteria may affect the ability of the vector to transmit pathogens. Eleven different types of bacterial clones were isolated from the midgut of lab-reared fourth instar larvae of Aedes aegypti and were grown on LB agar medium at an optimum temperature of 25 ºC. Identification of these bacteria was done on the basis of their colony characteristic such as colony size, shape, opacity, elevation, consistency, and growth. Light microscopic studies of the gut microbiota revealed dominance of Gram-negative cocci over gram positive cocci and bacilli and Gram-negative bacilli. Identification of species was done by chemical characterization of the colonies. Crude extracts of all test plants were screened for their antimicrobial activities against gut microbiota by disc diffusion assay. The zone of exclusion seen after 24 hr of incubation in different assays revealed the most potent antibacterial activities in neem followed by clove and turmeric. Lantana and Aloe vera were least effective.

Keywords: plant extract, aedes, dengue, antimicrobial activity

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Authors: Miaomiao Zhang, Sabrina Beckmann, Haluk Ertan, Rocky Chau, Mike Manefield

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Phenazines are a large class of nitrogen-containing aromatic heterocyclic compounds, which are almost exclusively produced by bacteria from diverse genera including Pseudomonas and Streptomyces. Phenazine-1-carboxylic acid (PCA) as one of 'core' phenazines are converted from chorismic acid before modified to other phenazine derivatives in different cells. Phenazines have attracted enormous interests because of their multiple roles on biocontrol, bacterial interaction, biofilm formation and fitness of their producers. However, in spite of ecological importance, degradation as a part of phenazines’ fate only have extremely limited attention now. Here, to isolate PCA-degrading bacteria, 200 mg L-1 PCA was supplied as sole carbon, nitrogen and energy source in minimal mineral medium. Quantitative PCR and Reverse-transcript PCR were employed to study abundance and activity of functional gene MFORT 16269 in PCA degradation, respectively. Intermediates and products of PCA degradation were identified with LC-MS/MS. After enrichment and isolation, a PCA-degrading strain was selected from soil and was designated as Rhodanobacter sp. PCA2 based on full 16S rRNA sequencing. As determined by HPLC, strain PCA2 consumed 200 mg L-1 (836 µM) PCA at a rate of 17.4 µM h-1, accompanying with significant cells yield from 1.92 × 105 to 3.11 × 106 cells per mL. Strain PCA2 was capable of degrading other phenazines as well, including phenazine (4.27 µM h-1), pyocyanin (2.72 µM h-1), neutral red (1.30 µM h-1) and 1-hydroxyphenazine (0.55 µM h-1). Moreover, during the incubation, transcript copies of MFORT 16269 gene increased significantly from 2.13 × 106 to 8.82 × 107 copies mL-1, which was 2.77 times faster than that of the corresponding gene copy number (2.20 × 106 to 3.32 × 107 copies mL-1), indicating that MFORT 16269 gene was activated and played roles on PCA degradation. As analyzed by LC-MS/MS, decarboxylation from the ring structure was determined as the first step of PCA degradation, followed by cleavage of nitrogen-containing ring by dioxygenase which catalyzed phenazine to nitrosobenzene. Subsequently, phenylhydroxylamine was detected after incubation for two days and was then transferred to aniline and catechol. Additionally, genomic and proteomic analyses were also carried out for strain PCA2. Overall, the findings presented here showed that a newly isolated strain Rhodanobacter sp. PCA2 was capable of degrading phenazines through decarboxylation and cleavage of nitrogen-containing ring, during which MFORT 16269 gene was activated and played important roles.

Keywords: decarboxylation, MFORT16269 gene, phenazine-1-carboxylic acid degradation, Rhodanobacter sp. PCA2

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Authors: Muhammad Saad Shoaib Khan, Rasbin Basnet, Qingyao Shu

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Phospholipids are one of the major classes of lipid comprising 10% of total grain lipid in rice. Phospholipids are the main phosphorus containing lipid in the rice endosperm, contributing to rice palatability and seed storage property. However, in the rice grain, the majority of phosphorus occur in the form of phytic acid and are highly abundant in the bran. Phytic acid, also known as hexaphosphorylated inositol (IP6), are strong chelating agents which reduces the bioavailability of essential dietary nutrients and are therefore less desirable by rice breeders. We used the CRISPR/Cas9 system to generate mutants of a phospholipase D gene (PLDα1), which is responsible for the degradation of phospholipids into phosphatidic acid (PA). In the mutants, we found a significant reduction in the concentration of phytic acid in the grain as compared to the wild-type. The biochemical analysis of the PLDα1 mutants showed that the decrease in production of phosphatidic acid is due to reduced accumulation of CDP-diacylglycerolderived phosphatidylinositol (PI), ultimately leading to lower accumulation of phytic acid in mutants. These results showed that loss of function of PLD in rice leads to lower production of phytic acid, suggesting the potential application of Ospldα1 in breeding rice with less phytic acid.

Keywords: CRISPR/Cas9, phospholipase D, phytic acid, rice

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Authors: Kumudini Apsara Munasinghe, Devin Gregory Lissau, Ryan Thomas Wade

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Fresh fruits and vegetables are rich in vitamins, minerals, and fibers and help maintain a healthy weight over high-calorie food. Eating fruits and vegetables protects us from free radicals produced by metabolic reactions and safeguards us from cardiovascular disease and cancer. However, there has been an increased concern about foodborne diseases tied to contaminated farmers’ market produce. In addition, very little information is available about the contribution of eating raw fruits and vegetables to human exposure to antibiotic-resistant bacteria. This research aims to identify bacteria isolated from farmers’ market fruits and vegetables and understand their antibiotic resistance. Vegetables and fruits were collected from farmers’ markets around Frostburg and Cumberland areas in Maryland and transported to the microbiology lab at Frostburg State University for the isolation of bacteria. Bacteria were extracted from tomatoes, cucumber, strawberry, and lettuce using Tryptic soy broth overnight at 37°C, and Tryptic Soy agar was used for the streak plate technique to isolate bacteria. Pure cultures were used to identify bacteria using biochemical reactions after conducting Gram staining technique. The research used many biochemical reactions, including Mannitol Salt agar, MacConkey agar, and Eosin Methylene blue agar, for identification. Antibiotic sensitivity was tested for many different types of antibiotics, including amoxicillin, penicillin, tetracycline, ampicillin, and erythromycin. Most prevalent bacteria in the isolates were Staphylococcus, Bacillus, Micrococcus, Enterococcus, Enterobacter, Citrobacter, and other bacteria from the family Enterobacteriaceae. The data obtained from this research will be useful to educate and train farmers and individuals involved in post-harvest processes such as transportation and selling in farmers’ markets. Further results for bacterial antibiotic resistance will be obtained, and unculturable bacteria will be identified by next-generation DNA sequencing.

Keywords: antibiotic resistance, farmers markets, fruits, bacteria, vegetables

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Authors: Bahdja Guerfi, N. Hadhoum, I. Azouz, M. Bendoumia, S. Bouafia, F. Z. Hadjadj Aoul

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Background: 'Penicillin V' is a narrow, bactericidal antibiotic of the beta-lactam family of the naturally occurring penicillin group. It is limited to infections due to the germs defined as sensitive. The objective of this work was to identify and to characterize Penicillin V acid and its related compounds by High-performance liquid chromatography (HPLC). Methods: Firstly phenoxymethylpenicillin was identified by an infrared absorption. The organoleptic characteristics, pH, and determination of water content were also studied. The dosage of Penicillin V acid active substance and the determination of its related compounds were carried on waters HPLC, equipped with a UV detector at 254 nm and Discovery HS C18 column (250 mm X 4.6 mm X 5 µm) which is maintained at room temperature. The flow rate was about 1 ml per min. A mixture of water, acetonitrile and acetic acid (65:35:01) was used as mobile phase for phenoxyacetic acid ‘impurity B' and a mixture of water, acetonitrile and acetic acid (650:150:5.75) for the assay and 4-hydroxypenicillin V 'impurity D'. Results: The identification of Penicillin V acid active substance and the evaluation of its chemical quality showed conformity with USP 35th edition. The Penicillin V acid content in the raw material is equal to 1692.22 UI/mg. The percentage content of phenoxyacetic acid and 4-hydroxypenicillin V was respectively: 0.035% and 0.323%. Conclusion: Through these results, we can conclude that the Penicillin V acid active substance tested is of good physicochemical quality.

Keywords: characterization, HPLC, Penicillin V acid, related substances

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Authors: Hyun Ji Hyun, Hyo Sun Suh, Min Kook Kim, Yong Chan Kwon, Byung-Mu Lee

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Scope: This study is focused on the effect of myristic acid on LPS-induced inflammation in RAW 264.7 macrophage cells. Methods and results: For the experiment, RAW 264.7 mouse macrophage cell line was used. Results showed that treatment with myristic acid can attenuate LPS-induced inflammation. Moreover, myristic acid significantly suppressed expression of inflammatory mediators and down-regulating UVB-induced intracellular ROS generation. Furthermore, myristic acid reduced the expression of NF-κB by inhibiting degradation of IκB-α and ERK, JNK, and p38 pathways by inhibiting phosphorylation in RAW 264.7 macrophage cells. Conclusion: Overall, these data suggest that the myristic acid could reduce LPS-induced inflammation. Acknowledgment: This research was supported by the Ministry of Trade, Industry & Energy(MOTIE), Korea Institute for Advancement of Technology(KIAT) through the Encouragement Program for The Industries of Economic Cooperation Region

Keywords: anti-inflammation, myristic acid, ROS, ultraviolet light

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Authors: Bashistha Kumar Kanth, Seung Pil Pack

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Carbonic anhydrase is ubiquitously distributed in organisms, and is fundamental to many eukaryotic biological processes such as photosynthesis, respiration, CO2 and ion transport, calcification and acid–base balance. However, CA occurs across the spectrum of prokaryotic metabolism in both the archaea and bacteria domains and many individual species contain more than one class. In this review, various roles of CA involved in cellular mechanism are presented to find out the CA functions applicable for industrial use.

Keywords: carbonic anhydrase, mechanism, CO2 sequestration, respiration

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Authors: Astrid Tannert, Anuradha Ramoji, Christina Ebert, Frederike Gladigau, Lorena Tuchscherr, Jürgen Popp, Ute Neugebauer

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Staphylococcus aureus is a facultative intracellular pathogen, which by entering host cells may evade immunologic host response as well as antimicrobial treatment. In that way, S. aureus can cause persistent intracellular infections which are difficult to treat. Depending on the strain, S. aureus may persist at different intracellular locations like the phagolysosome. The first barrier invading pathogens from the blood stream that they have to cross are the endothelial cells lining the inner surface of blood and lymphatic vessels. Upon proceeding from an acute to a chronic infection, intracellular pathogens undergo certain biochemical and structural changes including a deceleration of metabolic processes to adopt for long-term intracellular survival and the development of a special phenotype designated as small colony variant. In this study, the endothelial cell line Ea.hy 926 was used as a model for acute and chronic S. aureus infection. To this end, Ea.hy 926 cells were cultured on QIAscout™ Microraft Arrays, a special graded cell culture substrate that contains around 12,000 microrafts of 200 µm edge length. After attachment to the substrate, the endothelial cells were infected with GFP-expressing S. aureus for 3 weeks. The acute infection and the development of persistent bacteria was followed by confocal laser scanning microscopy, scanning the whole Microraft Array for the presence and for detailed determination of the intracellular location of fluorescent intracellular bacteria every second day. After three weeks of infection representative microrafts containing infected cells, cells with protruded infections and cells that did never show any infection were isolated and fixed for Raman micro-spectroscopic investigation. For comparison, also microrafts with acute infection were isolated. The acquired Raman spectra are correlated with the fluorescence microscopic images to give hints about a) the molecular alterations in endothelial cells during acute and chronic infection compared to non-infected cells, and b) metabolic and structural changes within the pathogen when entering a mode of persistence within host cells. We thank Dr. Ruth Kläver from QIAGEN GmbH for her support regarding QIAscout technology. Financial support by the BMBF via the CSCC (FKZ 01EO1502) and from the DFG via the Jena Biophotonic and Imaging Laboratory (JBIL, FKZ PO 633/29-1, BA 1601/10-1) is highly acknowledged.

Keywords: correlative image analysis, intracellular infection, pathogen-host adaption, Raman micro-spectroscopy

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Authors: Kelli G. Thorup, Kristopher A. Blee

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Climate change enhances the occurrence of extreme weather: wildfires, drought, rising summer temperatures, all of which dramatically decline forest growth and increase tree mortality in the mixed-conifer forests of Sierra Nevada, California. However, microbiota living in mutualistic relations with plant rhizospheres have been found to mitigate the effects of suboptimal environmental conditions. The goal of this research is to isolate native beneficial bacteria, plant-growth promoting rhizobacteria (PGPR), that can alleviate heat stress in Pinus ponderosa seedlings. Bacteria were isolated from the rhizosphere of Pinus ponderosa juveniles located in mixed-conifer stand and further characterized for PGP potential based on their ability to produce key growth regulatory phytohormones including auxin, cytokinin, and gibberellic acid. Out of ten soil samples taken, sixteen colonies were isolated and qualitatively confirmed to produce indole-3-acetic acid (auxin) using Salkowski’s reagent. Future testing will be conducted to quantitatively assess phytohormone production in bacterial isolates. Furthermore, bioassays will be performed to determine isolates abilities to increase tolerance in heat-stressed Pinus ponderosa seedlings. Upon completion of this research, a PGPR could be utilized to support the growth and transplantation of conifer seedlings as summer temperatures continue to rise due to the effects of climate change.

Keywords: conifer, heat-stressed, phytohormones, Pinus ponderosa, plant-growth promoting rhizobacteria

Procedia PDF Downloads 118

Authors: Frans J. Smit, Wisdom A. Munzeiwa, Hermanus C. M. Vosloo, Lyn-Marie Birkholtz, Richard K. Haynes

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Malaria and antimicrobial infections remain significant global health concerns, necessitating the continuous search for novel therapeutic approaches. This abstract presents an overview of the potential use of triazenes as effective agents against malaria and various antimicrobial pathogens. Triazenes are a class of compounds characterized by a linear arrangement of three nitrogen atoms, rendering them structurally distinct from their cyclic counterparts. This study investigates the efficacy of triazenes against malaria and explores their antimicrobial activity. Preliminary results revealed significant antimalarial activity of the triazenes, as evidenced by in vitro screening against P. falciparum, the causative agent of malaria. Furthermore, the compounds exhibited broad-spectrum antimicrobial activity, indicating their potential as effective antimicrobial agents. These compounds have shown inhibitory effects on various essential enzymes and processes involved in parasite survival, replication, and transmission. The mechanism of action of triazenes against malaria involves interactions with critical molecular targets, such as enzymes involved in the parasite's metabolic pathways and proteins responsible for host cell invasion. The antimicrobial activity of the triazenes against bacteria and fungi was investigated through disc diffusion screening. The antimicrobial efficacy of triazenes has been observed against both Gram-positive and Gram-negative bacteria, as well as multidrug-resistant strains, making them potential candidates for combating drug-resistant infections. Furthermore, triazenes possess favourable physicochemical properties, such as good stability, solubility, and low toxicity, which are essential for drug development. The structural versatility of triazenes allows for the modification of their chemical composition to enhance their potency, selectivity, and pharmacokinetic properties. These modifications can be tailored to target specific pathogens, increasing the potential for personalized treatment strategies. In conclusion, this study highlights the potential of triazenes as promising candidates for the development of novel antimalarial and antimicrobial therapeutics. Further investigations are necessary to determine the structure-activity relationships and optimize the pharmacological properties of these compounds. The results warrant additional research, including MIC studies, to further explore the antimicrobial activity of the triazenes. Ultimately, these findings contribute to the development of more effective strategies for combating malaria and microbial infections.

Keywords: malaria, anti-microbials, triazene, resistance

Procedia PDF Downloads 102