Search results for: bacterial isolation

Authors: Manal Suleiman, George Abu-Aqil, Uraib Sharaha, Klaris Riesenberg, Itshak Lapidot, Ahmad Salman, Mahmoud Huleihel

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Klebsiella pneumoniae is one of the most aggressive multidrug-resistant bacteria associated with human infections resulting in high mortality and morbidity. Thus, for an effective treatment, it is important to diagnose both the species of infecting bacteria and their susceptibility to antibiotics. Current used methods for diagnosing the bacterial susceptibility to antibiotics are time-consuming (about 24h following the first culture). Thus, there is a clear need for rapid methods to determine the bacterial susceptibility to antibiotics. Infrared spectroscopy is a well-known method that is known as sensitive and simple which is able to detect minor biomolecular changes in biological samples associated with developing abnormalities. The main goal of this study is to evaluate the potential of infrared spectroscopy in tandem with Random Forest and XGBoost machine learning algorithms to diagnose the susceptibility of Klebsiella pneumoniae to antibiotics within approximately 20 minutes following the first culture. In this study, 1190 Klebsiella pneumoniae isolates were obtained from different patients with urinary tract infections. The isolates were measured by the infrared spectrometer, and the spectra were analyzed by machine learning algorithms Random Forest and XGBoost to determine their susceptibility regarding nine specific antibiotics. Our results confirm that it was possible to classify the isolates into sensitive and resistant to specific antibiotics with a success rate range of 80%-85% for the different tested antibiotics. These results prove the promising potential of infrared spectroscopy as a powerful diagnostic method for determining the Klebsiella pneumoniae susceptibility to antibiotics.

Keywords: urinary tract infection (UTI), Klebsiella pneumoniae, bacterial susceptibility, infrared spectroscopy, machine learning

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Authors: Fitri Ayu, Nadia, Tanti, Putri, Fatkhan, Pasid Harlisa, Suparmi

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Acne is a skin abnormal conditions experienced by many teens, this is caused by various factors such as the climate is hot, humid and excessive sun exposure can aggravate acne because it will lead to excess oil production. Flavonoids form complex compounds against extracellular proteins that disrupt the integrity of bacterial cell membrane in a way denature bacterial cell proteins and bacterial cell membrane damage. This study aimed to test the antibacterial activity of corn silk extract with a concentration of 10 %, 20 %, 30 %, 40 %, 50 %, 60 %, 70 %, 80 %, 90 % and 100 % in vitro by measuring the inhibition of the growth of bacteria Propionibacterium acne, Staphylococcus aureus and Staphylococcus epidermis then compared with the standard antibiotic clindamycin. Extracts tested by Disk Diffusion Method, in which the blank disc soaked with their respective corn silk extract concentration for 15-30 minutes and then the medium of bacteria that have been planted with Propionibacterium acne, Staphylococcus aureus and Staphylococcus epidermis in the given disk that already contains extracts with various concentration. Incubated for 24 hours and then measured the growth inhibition zone Propionibacterium acne, Staphylococcus aureus and Staphylococcus epidermidis. Corn silk contains flavonoids, is shown by the test of flavonoids in corn silk extract by using a tube heating and without heating. Flavonoid in corn silk potentially as anti acne by inhibiting the growth of bacteria that cause acne. Corn silk extract concentration which has the highest antibacterial activity is then performed in a cream formulation and evaluation test of physical and chemical properties of the resulting cream preparation.

Keywords: antibacterial, flavonoid, corn silk, acne

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Authors: R. Gounina-Allouane, N. Ouali, F. Z. Berrabah, A. Bentaleb

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For a long time, the Gram-positive spore-forming bacteria Bacillus thuringiensis (Bt) has been widely used in biological control against devastating and disease vectors insects. This is due to the insecticidal activity of its crystalline parasporal inclusion (crystals) predominantly comprised of one or more proteins (Cry and Cyt proteins) also called δ-endotoxins, produced during sporulation. The shape and composition of Bt crystals vary among strains and crystalline proteins are extremely varied (more than 475 cry gene were discovered). The insecticidal activity of Bt crystals is very well studied, thus their insecticidal mode of action is well established, however, their antimicrobial effect is largely unknown. The lack of data on the antimicrobial effect of crystalline proteins of Bt and the need for searching new antimicrobial molecules encouraged us to carried out this study. The antibacterial effect of δ-endotoxines produced by two Bt stains; a strain isolated from soil at northern of Algeria (Bt 7.2.B), and a strain isolated from a bioinsecticide (Bacillus thuringiensis var aizawai), activated by proteolysis, was assayed on clinical bacterial strains and ATCC collection ones respectively. Gram positive and negative clinical bacterial strains (Escherichia coli, Klebsiella pneumonaie, Pseudomonas aeruginosa, Staphylococcus aureus) were sensitive to activated Bt 72B endotoxins. Similarly, bacterial strains from ATCC collection (Escherichia coli ATCC 25922, Pseudomonas aerugenosa ATCC 27853, Staphylococcus aureus ATCC 25923) were sensitive to activated B. thuringiensis var aizawai δ-endotoxines. The activated δ-endotoxins were separated by SDS-PAGE.

Keywords: Bacillus thuringiensis, crystals, cry proteins, δ-endotoxins, antibacterial activity

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Authors: M. Knezevic, V. Marecic, M. Ozanic, I. Kelava, M. Mihelcic, M. Santic

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Francisella tularensis is a highly infectious, gram-negative intracellular bacterium and the causative agent of tularemia. The bacterium has been isolated from more than 250 wild species, including protozoa cells. Since Francisella is very virulent and persists in the environment for years, the aim of this study was to investigate whether Acanthamoeba castellanii-grown F. novicida exhibits an alteration in the resistance to disinfectants. It has been shown by other intracellular pathogens, including Legionella pneumophila that bacteria grown in amoeba exhibit more resistance to disinfectants. However, there is no data showing Francisella viability behaviour after intracellular life cycle in A. castellani. In this study, the bacterial suspensions of A. castellanii-grown or in vitro-grown Francisella were treated with three different disinfectants, and the bacterial viability after disinfection treatment was determined by a colony-forming unit (CFU) counting method, transmission electron microscopy (TEM), fluorescence microscopy as well as the leakage of intracellular fluid. Our results have shown that didecyldimethylammonium chloride (DDAC) combined with isopropyl alcohol was the most effective in bacterial killing; all in vitro-grown and A. castellanii-grown F. novicida were killed after only 10s. Surprisingly, in comparison to in vitro-grown bacteria, A. castellanii-grown F. novicida was more sensitive to decontamination by the benzalkonium chloride combined with DDAC and formic acid and the polyhexamethylene biguanide (PHMB). We can conclude that the tested disinfectants exhibit antimicrobial activity by causing a loss of structural organization and integrity of the Francisella cell wall and membrane and the subsequent leakage of the intracellular contents. Finally, the results of this study clearly demonstrate that Francisella grown in A. castellanii had become more susceptible to many disinfectants.

Keywords: Acanthamoeba, disinfectant, Francisella, sensitivity

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Authors: Aditi Taunk, George Iskander, Kitty Ka Kit Ho, Mark Willcox, Naresh Kumar

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Bacterial infections on biomaterial implants and medical devices accounts for 60-70% of all hospital acquired infections (HAIs). Treatment or removal of these infected devices results in high patient mortality and morbidity along with increased hospital expenses. In addition, with no effective strategies currently available and rapid development of antibacterial resistance has made device-related infections extremely difficult to treat. Therefore, in this project we have developed biomaterial surfaces using antibacterial compounds that inhibit biofilm formation by interfering with the bacterial communication mechanism known as quorum sensing (QS). This study focuses on covalent attachment of potent quorum sensing (QS) inhibiting compounds, halogenated furanones (FUs) and dihydropyrrol-2-ones (DHPs), onto glass surfaces. The FUs were attached by photoactivating the azide groups on the surface, and the acid functionalized DHPs were immobilized on amine surface via EDC/NHS coupling. The modified surfaces were tested in vitro against pathogenic organisms such as Staphylococcus aureus and Pseudomonas aeruginosa using confocal laser scanning microscopy (CLSM). Successful attachment of compounds on the substrates was confirmed by X-ray photoelectron spectroscopy (XPS) and contact angle measurements. The antibacterial efficacy was assessed, and significant reduction in bacterial adhesion and biofilm formation was observed on the FU and DHP coated surfaces. The activity of the coating was dependent upon the type of substituent present on the phenyl group of the DHP compound. For example, the ortho-fluorophenyl DHP (DHP-2) exhibited 79% reduction in bacterial adhesion against S. aureus and para-fluorophenyl DHP (DHP-3) exhibited 70% reduction against P. aeruginosa. The results were found to be comparable to DHP coated surfaces prepared in earlier study via Michael addition reaction. FUs and DHPs were able to retain their in vitro antibacterial efficacy after covalent attachment via azide chemistry. This approach is a promising strategy to develop efficient antibacterial biomaterials to reduce device related infections.

Keywords: antibacterial biomaterials, biomedical device-related infections, quorum sensing, surface functionalization

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Authors: Ali Bahkali, Rayan Yousef Booq, Mohammad Khiyami

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Soil samples were collected from five different regions in the Kingdom of Saudi Arabia. Microbiological methods included dilution methods and pour plates to isolate and purify bacteria soil. The ability of isolates to develop biopolymer was investigated on petri dishes containing elements and substance concentrations stimulating developing biopolymer. Fluorescent stains, Nile red and Nile blue were used to stain the bacterial cells developing biopolymers. In addition, Sudan black was used to detect biopolymers in bacterial cells. The isolates which developed biopolymers were identified based on their gene sequence of 1 6sRNA and their ability to grow and synthesize PHAs on mineral medium supplemented with 1% dates molasses as the only carbon source under nitrogen limitation. During the study 293 bacterial isolates were isolated and detected. Through the initial survey on the petri dishes, 84 isolates showed the ability to develop biopolymers. These bacterial colonies developed a pink color due to accumulation of the biopolymers in the cells. Twenty-three isolates were able to grow on dates molasses, three strains of which showed the ability to accumulate biopolymers. These strains included Bacillus sp., Ralstonia sp. and Microbacterium sp. They were detected by Nile blue A stain with fluorescence microscopy (OLYMPUS IX 51). Among the isolated strains Ralstonia sp. was selected after its ability to grow on molasses dates in the presence of a limited nitrogen source was detected. The optimum conditions for formation of biopolymers by isolated strains were investigated. Conditions studied included, best incubation duration (2 days), temperature (30°C) and pH (7-8). The maximum PHB production was raised by 1% (v1v) when using concentrations of dates molasses 1, 2, 3, 4 and 5% in MSM. The best inoculated with 1% old inoculum (1= OD). The ideal extraction method of PHA and PHB proved to be 0.4% sodium hypochlorite solution, producing a quantity of polymer 98.79% of the cell's dry weight. The maximum PHB production was 1.79 g/L recorded by Ralstonia sp. after 48 h, while it was 1.40 g/L produced by R.eutropha ATCC 17697 after 48 h.

Keywords: bacteria forming polyhydroxyalkanoate, detection, molecular, Saudi Arabia

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Authors: Mozadded Hossen

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This study investigates the varied effects of internet addiction on college students, specifically examining the behavioral and academic outcomes. The widespread use of the Internet in academic settings has substantially impacted students' mental well-being and academic achievements. The study investigates the correlation between excessive internet usage and addiction, which manifests through symptoms including social isolation, anxiety, despair, and sleep disruptions. Additionally, the study examines the relationship between internet addiction and academic results, finding that kids with more severe addiction levels generally have lower academic performance, experience diminished focus, and show reduced involvement in academic tasks. The study intends to analyze the many consequences of internet addiction to gain insights into its ramifications. It also urges educational institutions to develop techniques that can reduce the negative impact of internet addiction and encourage healthier internet use among students. The results emphasize the necessity of implementing comprehensive measures to tackle the behavioral and academic difficulties caused by internet addiction among college students.

Keywords: internet addiction, behavioral consequences, college students, social isolation

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Authors: Sakshi Bhalla

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On the one hand, given their seemingly simplistic, near universal usage and understanding, emoji are discarded as a potential step back in the evolution of communication. On the other, their effectiveness, pervasiveness, and adaptability across and within contexts are undeniable. In this study, the responses of 40 people (categorized by age) were recorded based on a uniform two-part questionnaire where they were required to a) identify the meaning of 15 emoji when placed in isolation, and b) interpret the meaning of the same 15 emoji when placed in a context-defining posting on Twitter. Their responses were studied on the basis of deviation from their responses that identified the emoji in isolation, as well as the originally intended meaning ascribed to the emoji. Based on an analysis of these results, it was discovered that each of the five age categories uses, understands and perceives emoji differently, which could be attributed to the degree of exposure they have undergone. For example, in the case of the youngest category (aged < 20), it was observed that they were the least accurate at correctly identifying emoji in isolation (~55%). Further, their proclivity to change their response with respect to the context was also the least (~31%). However, an analysis of each of their individual responses showed that these first-borns of social media seem to have reached a point where emojis no longer inspire their most literal meanings to them. The meaning and implication of these emoji have evolved to imply their context-derived meanings, even when placed in isolation. These trends carry forward meaningfully for the other four groups as well. In the case of the oldest category (aged > 35), however, the trends indicated inaccuracy and therefore, a higher incidence of a proclivity to change their responses. When studied in a continuum, the responses indicate that slowly and steadily, emoji are evolving from pictograms to ideograms. That is to suggest that they do not just indicate a one-to-one relation between a singular form and singular meaning. In fact, they communicate increasingly complicated ideas. This is much like the evolution of ancient hieroglyphics on papyrus reed or cuneiform on Sumerian clay tablets, which evolved from simple pictograms to progressively more complex ideograms. This evolution within communication is parallel to and contingent on the simultaneous evolution of communication. What’s astounding is the capacity of humans to leverage different platforms to facilitate such changes. Twiterese, as it is now called, is one of the instances where language is adapting to the demands of the digital world. That it does not have a spoken component, an ostensible grammar, and lacks standardization of use and meaning, as some might suggest, may seem like impediments in qualifying it as the 'language' of the digital world. However, that kind of a declarative remains a function of time, and time alone.

Keywords: communication, emoji, language, Twitter

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Authors: Yaling Gou, Sucai Yang, Pengwei Qiao

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Integrated chemical-biological treatment is an attractive alternative to remove polycyclic aromatic hydrocarbons (PAHs) from contaminated soil; wherein indigenous bacteria is the key factor for the biodegradation of residual PAHs concentrations after the application of chemical oxidation. However, the systematical study on the impact of persulfate (PS) oxidation on indigenous bacteria as well as PAHs removal is still scarce. In this study, the influences of different PS dosages (1%, 3%, 6%, and 10% [w/w]), as well as various activation methods (native iron, H2O2, alkaline, ferrous iron, and heat) on PAHs removal and indigenous bacteria in highly contaminated aged soil were investigated. Apparent degradation of PAHs in the soil treated with PS oxidation was observed, and the removal efficiency of total PAHs in the soil ranged from 38.28% to 79.97%. The removal efficiency of total PAHs in the soil increased with increasing consumption of PS. However, the bacterial abundance in soil was negatively affected following oxidation for all of the treatments added with PS, with bacterial abundance in the soil decreased by 0.89~2.88 orders of magnitude compared to the untreated soil. Moreover, the number of total bacteria in the soil decreased as PS consumption increased. Different PS activation methods and PS dosages exhibited different influences on the bacterial community composition. Bacteria capable of degrading PAHs under anoxic conditions were composed predominantly by Proteobacteria and Firmicutes. The total amount of Proteobacteria and Firmicutes also decreased with increasing consumption of PS. The results of this study provide important insight into the design of PAHs contaminated soil remediation projects.

Keywords: activation method, chemical oxidation, indigenous bacteria, polycyclic aromatic hydrocarbon

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Authors: Abderrezzak khatib

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Biogenic amines are low molecular weight nitrogenous compounds that have the potential to accumulate in food, posing a significant risk to food safety and human health. In this study, we investigated the inhibitory activity of three strains of lactic acid bacteria (LAB), against the growth and production of biogenic amines by both foodborne pathogens and food spoilage bacteria. The foodborne pathogens studied included Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella Paratyphi, while the food spoilage bacteria comprised Enterobacter cloacae and Proteus mirabilis. The methodology involved bacterial growth determination in petri dishes, bacterial culture extraction and derivatization, and biogenic amine analysis using HPLC. Our findings revealed that the inhibitory effects of LAB on these pathogens varied, with all three LAB strains demonstrating a remarkable reduction in the total bacterial count when combined with most pathogens, compared to the individual cultures of the pathogens. Furthermore, the presence of LAB in co-cultures with the pathogens resulted in a significant decrease in the production of tyramine and other biogenic amines by the pathogens themselves. These results suggest that LAB strains hold considerable promise in preventing the accumulation of biogenic amines in food products, thereby enhancing food safety. This study provides insights into the potential utilization of LAB in the context of preserving and ensuring the safety of food products. It highlights the significance of conducting additional research endeavors to elucidate the underlying mechanisms involved and to identify the precise bioactive compounds that are responsible for the observed inhibitory effects.

Keywords: food safety, lactic acid bacteria, foodborne pathogens, food spoilage bacteria, biogenic amines, tyrosine

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Authors: Rawia M. Khalil, Ghada Abd-Elbary, Mona Basha, Ghada E. A. Awad, Hadeer A. Elhashemy

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Background: Bacterial infections of the eye are the clinical conditions responsible for ocular morbidity and blindness. Conjunctivitis is an inflammation of the conjunctiva, due to Staphylococcus aureus. Lomefloxacin HCl (LXN) is a third generation flouroquinolone antibiotic with a broad spectrum against wide range of bacteria and very effective against Staph infections especially in conjunctiva (conjunctivitis). The present study aims to develop and evaluate novel ocular proniosomal gels of Lomefloxacin Hcl (LXN); in order to improve its ocular bioavailability for the management of bacterial conjunctivitis. Materials and methods: Proniosomes were prepared by coacervation phase separation method using different types of nonionic surfactants (Span 60,40,20,Tween 20,40,60,80,Brij 35,98,72) solely and as mixtures with Span® 60. The formed gels were characterized for entrapment efficiency, vesicle size and in vitro drug release. The optimum proniosomal gel; P-LXN 7 were characterized for pH measurement, transmission electron microscopy (TEM) and differential scanning calorimetry (DSC) as well as Stability study and microbiological evaluation .The results revealed that only Span 60 was able to form stable LXN proniosomal gel when used individually while the other nonionic surfactants formed gels only in combination with Span 60 at different ratios. The optimum proniosomal gel; P-LXN 7 (Span60:Tween60, 9:1) appeared as spherical shaped vesicles having high entrapment efficiency (>80 %), appropriate vesicle size (187 nm) as well as controlled drug release over 12h. DSC confirmed the amorphous nature and the uniformity of LXN inclusion within the vesicles. Physical stability study did not show any significant changes in appearance or entrapment efficiency or vesicle size after storage for 3 months at 4°C. Ocular irritancy test revealed that P-LXN 7 was safe, well tolerable and suitable for ocular delivery. In vivo antibacterial activity of P-LXN 7 evaluated using the susceptibility test and topical therapy of induced ocular conjunctivitis confirmed the enhanced antibacterial therapeutic efficacy of the LXN-proniosomal gel compared to the commercially available LXN eye drops; Orchacin®. Conclusions: Our results suggest that proniosomal gels could provide a promising carrier of LXN for efficient ocular treatment of bacterial conjunctivitis.

Keywords: bacterial conjunctivitis, lomefloxacin HCl, ocular drug delivery, proniosomes

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Authors: Nesrine Lenchi, Salima Kebbouche-Gana, Laddada Belaid, Mohamed Lamine Khelfaoui, Mohamed Lamine Gana

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Saline lakes are extreme hypersaline environments that are considered five to ten times saltier than seawater (150 – 300 g L-1 salt concentration). Hypersaline regions differ from each other in terms of salt concentration, chemical composition and geographical location, which determine the nature of inhabitant microorganisms. In order to explore the diversity of moderate and extreme halophiles Bacteria in Chott Tinsilt (East of Algeria), an isolation program was performed. In the first time, water samples were collected from the saltern during pre-salt harvesting phase. Salinity, pH and temperature of the sampling site were determined in situ. Chemical analysis of water sample indicated that Na +and Cl- were the most abundant ions. Isolates were obtained by plating out the samples in complex and synthetic media. In this study, seven halophiles cultures of Bacteria were isolated. Isolates were studied for Gram’s reaction, cell morphology and pigmentation. Enzymatic assays (oxidase, catalase, nitrate reductase and urease), and optimization of growth conditions were done. The results indicated that the salinity optima varied from 50 to 250 g L-1, whereas the optimum of temperature range from 25°C to 35°C. Molecular identification of the isolates was performed by sequencing the 16S rRNA gene. The results showed that these cultured isolates included members belonging to the Halomonas, Staphylococcus, Salinivibrio, Idiomarina, Halobacillus Thalassobacillus and Planococcus genera and what may represent a new bacterial genus.

Keywords: bacteria, Chott, halophilic, 16S rRNA

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Authors: Danyi Wang, Andrew Schriefer, Dennis O'Rourke, Brajendra Kumar, Yang Liu, Fei Zhong, Juergen Scheuenpflug, Zheng Feng

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Purpose: It has been recognized that the microbiome plays critical roles in disease pathogenesis, including cancer, autoimmune disease, and multiple sclerosis. To develop a clinical-grade assay for exploring microbiome-derived clinical biomarkers across disease areas, a two-phase approach is implemented. 1) Identification of the optimal sample preparation reagents using pre-mixed bacteria and healthy donor stool samples coupled with proprietary Sigma-Aldrich® bioinformatics solution. 2) Exploratory analysis of patient samples for enabling precision medicine. Study Procedure: In phase 1 study, we first compared the 16S sequencing results of two ATCC® microbiome standards (MSA 2002 and MSA 2003) across five different extraction kits (Kit A, B, C, D & E). Both microbiome standards samples were extracted in triplicate across all extraction kits. Following isolation, DNA quantity was determined by Qubit assay. DNA quality was assessed to determine purity and to confirm extracted DNA is of high molecular weight. Bacterial 16S ribosomal ribonucleic acid (rRNA) amplicons were generated via amplification of the V3/V4 hypervariable region of the 16S rRNA. Sequencing was performed using a 2x300 bp paired-end configuration on the Illumina MiSeq. Fastq files were analyzed using the Sigma-Aldrich® Microbiome Platform. The Microbiome Platform is a cloud-based service that offers best-in-class 16S-seq and WGS analysis pipelines and databases. The Platform and its methods have been extensively benchmarked using microbiome standards generated internally by MilliporeSigma and other external providers. Data Summary: The DNA yield using the extraction kit D and E is below the limit of detection (100 pg/µl) of Qubit assay as both extraction kits are intended for samples with low bacterial counts. The pre-mixed bacterial pellets at high concentrations with an input of 2 x106 cells for MSA-2002 and 1 x106 cells from MSA-2003 were not compatible with the kits. Among the remaining 3 extraction kits, kit A produced the greatest yield whereas kit B provided the least yield (Kit-A/MSA-2002: 174.25 ± 34.98; Kit-A/MSA-2003: 179.89 ± 30.18; Kit-B/MSA-2002: 27.86 ± 9.35; Kit-B/MSA-2003: 23.14 ± 6.39; Kit-C/MSA-2002: 55.19 ± 10.18; Kit-C/MSA-2003: 35.80 ± 11.41 (Mean ± SD)). Also, kit A produced the greatest yield, whereas kit B provided the least yield. The PCoA 3D visualization of the Weighted Unifrac beta diversity shows that kits A and C cluster closely together while kit B appears as an outlier. The kit A sequencing samples cluster more closely together than both the other kits. The taxonomic profiles of kit B have lower recall when compared to the known mixture profiles indicating that kit B was inefficient at detecting some of the bacteria. Conclusion: Our data demonstrated that the DNA extraction method impacts DNA concentration, purity, and microbial communities detected by next-generation sequencing analysis. Further microbiome analysis performance comparison of using healthy stool samples is underway; also, colorectal cancer patients' samples will be acquired for further explore the clinical utilities. Collectively, our comprehensive qualification approach, including the evaluation of optimal DNA extraction conditions, the inclusion of positive controls, and the implementation of a robust qualified bioinformatics pipeline, assures accurate characterization of the microbiota in a complex matrix for deciphering the deep biology and enabling precision medicine.

Keywords: 16S rRNA sequencing, analytical validation, bioinformatics pipeline, metagenomics

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Authors: Priyanka Gupta, Nilanjana Bairagi, Deepti Gupta

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New pathogenic strains of microbes are continually emerging and resistance of bacteria to antibiotics is growing. Hospitals in India have a high burden of infections in their intensive care units and general wards. Rising incidence of hospital infections is a matter of great concern in India. This growth is often attributed to the absence of effective infection control strategies in healthcare facilities. Government, therefore, is looking for cost effective strategies that are effective against HAIs. One possible method is by application of an antimicrobial finish on the uniform. But there are limited studies to show the effect of antimicrobial activity of antimicrobial finish treated nurses’ uniforms in a real hospital set up. This paper proposes a prospective non-destructive sampling technique, based on the use of a detachable fabric patch, to assess the effectiveness of silver based antimicrobial agent across five wards in a tertiary care government hospital in Delhi, India. Fabrics like polyester and polyester cotton blend fabric which are more prevalent for making coats were selected for the study. Polyester and polyester cotton blend fabric was treated with silver based antimicrobial (AM) finish. At the beginning of shift, a composite patch of untreated and treated fabric respectively was stitched on the abdominal region on the left and right side of the washed white coat of participating nurse. At the end of the shift, the patch was removed and taken for bacterial sampling on Brain Heart Infusion (BHI) plates. Microbial contamination on polyester and blend fabrics after 6 hours shift was compared in Brain Heart Infusion broth (BHI). All patches treated with silver based antimicrobial agent showed decreased bacterial counts. Percent reduction in the bacterial colonies after the antimicrobial treatment in both fabrics was 81.0 %. Antimicrobial finish was equally effective in reducing microbial adhesion on both fabric types. White coats of nurses become progressively contaminated during clinical care. Type of fabric used to make the coat can affect the extent of contamination which is higher on polyester cotton blend as compared to 100% polyester. The study highlights the importance of silver based antimicrobial finish in the area of uniform hygiene. Bacterial load can be reduced by using antimicrobial finish on hospital uniforms. Hospital staff uniforms endowed with antimicrobial properties may be of great help in reducing the occurrence and spread of infections.

Keywords: antimicrobial finish, bacteria, infection control, silver, white coat

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Authors: Pimpak Phumat, Sakornrat Khongkhunthian, Thomas Rades, Anette Müllertz, Siriporn Okonogi

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Self-nanoemulsifying drug delivery systems (SNEDDS) containing 4-allylpyrocatechol (AP) extracted from Piper betle were developed to enhance water solubility of AP by using modeling and design (MODDE) program. The amount of AP in each SNEDDS formulation was determined by using high-performance liquid chromatography. The formulation consisted of 20% Miglyol®812N, 40 % Kolliphor®RH40, 30 % Maisine®35-1 and 10 % ethanol was found to be the best SNEDDS that provided the highest loading capacity of AP. (141.48±15.64 mg/g SNEDDS). The system also showed miscibility with water. The particle shape and size of the AP-SNEDDS after dispersing in water was investigated by using a transmission electron microscope and photon correlation spectrophotometer, respectively. The results showed that they were a spherical shape, having a particle size of 34.27 ± 1.14 nm with a narrow size distribution of 0.17 ± 0.04. The particles showed negative zeta potential with a value of -21.66 ± 2.09 mV. Antibacterial activity of AP-SNEDDS containing 1.5 mg/mL of AP was investigated against Streptococcus intermedius. The effect of this system on S. intermedius cells was observed by a scanning electron microscope (SEM). The results from SEM revealed that the bacterial cells were obviously destroyed. Killing kinetic study of AP-SNEDDS was carried out. It was found that the killing rate of AP-SNEDDS against S. intermedius was dose-dependent and the bacterial reduction was 79.86 ± 0.45 % within 30 min. In comparison with chlorhexidine (CHX), AP-SNEDDS showed similar antibacterial effects against S. intermedius. It is concluded that SNEDDS is a potential system for enhancing water solubility of AP. The antibacterial study reveals that AP-SNEDDS can be a promising system to treat bacterial infection caused by S. intermedius.

Keywords: SNEDDS, 4-allylpyrocathecol, solubility, antibacterial activity, Streptococcus intermedius

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Authors: Sun Jin Park, Kyung-Ja Cho

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This study focused on characteristics related to hasty behaviour. To investigate the relation between personal characteristics and hasty behaviour, 601 data were collected, 335 males and 256 females answered their own 'social avoidance and distress’, ‘anxiety’, ‘sensation seeking', 'hope', and ' hasty behaviour. And then 591 data were used for the analysis. The factor analysis resulted hasty behaviour consisted of 5 factors, time pressure, isolation, uncomfortable situation, boring condition, and expectation of reward. The result showed anxiety, sensation seeking, and hope related to hasty behaviour. Specifically, anxiety was involved in every hasty behaviour. This result means that psychological tension and worry are related to hasty behaviour in common. 'Social avoidance and distress', 'sensation seeking' and 'hope' influenced on hasty behaviour under time pressure, in isolation, in expectation of rewards respectively. This means that each factor of hasty behaviour has anxiety as its basis, expressed through a varied nature.

Keywords: hasty behaviour, social avoidance and distress, anxiety, sensation seeking, hope

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Authors: Ahmed Al-Hajjaj, Alaa A-Maatoq

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Introduction Urinary tract infections (UTIs) are most common bacterial infection in women. Recurrent urinary tract infections ( rUTI) are defined by the occurrence of at least two episodes of UTI in six months or three in 12 months. Low dose prophylactic antibiotics were used for rUTI prevention. However, this is associated with increased risk of adverse effects and bacterial resistance. Therefore, exploring non-antibiotic prophylaxis is necessary. One of these is D-Mannose which is natural monosaccharide. The proposed mechanism of action of D-Mannose is inhibition of bacterial adhesion to the urothelium. The available data about its effectiveness is conflicting. Methods the systematically searched the literature for studies using D-mannose alone in prevention of rUTI in females aging 18 years or above in comparison to antibiotics, placebo or control. Results Systematic search revealed total of five eligible studies. Three studies were randomized trials (RCTs). There was one prospective study and one retrospective cohort study. Two out of three RCTs were open label trials which concluded D-Mannose effectiveness however both have poor quality of evidence. Both prospective and retrospective studies included demonstrated effectiveness of D Mannose, however both have small sample size (total 45 and 27 patients). The highest quality and largest RCT (total 598 patients) compared D-Mannose to placebo concluded there is no difference between D-Mannose and placebo in rUTI prevention. Meta-analysis of three studies revealed no statistical difference in rUTI prevention between D Mannose and placebo/control Conclusion There is currently no solid evidence to support the effectiveness of D-Mannose in prevention of recurrent urinary tract infections.

Keywords: d-mannose, urinary tract infection, recurrent cystitis, antibiotics prophylaxis

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Authors: Karen L. Rincon-Granados, América R. Vázquez-Olmos, Adriana-Patricia Rodríguez-Hernández, Gina Prado-Prone, Margarita Rivera, Roberto Y. Sato-Berrú

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In this work, a hybrid film based on poly-3-hydroxybutyrate (P3HB) and nickel ferrite (NiFe₂O₄) nanoparticles (NPs) was obtained by a simple and reproducible methodology in order to study its antibacterial and cytotoxic properties. The motivation for this research is the current antimicrobial resistance (RAM). This is a threat to human health and development worldwide. RAM is caused by the emergence of bacterial strains resistant to traditional antibiotics that were used as treatment. Due to this, the need to investigate new alternatives for preventing and treating bacterial infections emerges. In this sense, metal oxide NPs have aroused great interest due to their unique physicochemical properties. However, their use is limited by the nanostructured nature, commonly obtained by chemical and physical synthesis methods, as powders or colloidal dispersions. Therefore, the incorporation of nanostructured materials in polymer matrices to obtain hybrid materials that allow disinfecting and preventing the spread of bacteria on various surfaces. Accordingly, this work presents the synthesis and study of the antibacterial properties of the P3HB@NiFe₂O₄ hybrid film as a potential material to inhibit bacterial growth. The NiFe₂O₄ NPs were previously synthesized by a mechanochemical method. The P3HB and P3HB@NiFe₂O₄ films were obtained by the solvent casting method. The films were characterized by X-ray diffraction (XRD), Raman scattering, and scanning electron microscopy (SEM). The XRD pattern showed that the NiFe₂O₄ NPs were incorporated into the P3HB polymer matrix and retained their nanometric sizes. By energy dispersive X-ray spectroscopy (EDS), it was observed that the NPs are homogeneously distributed in the film. The bactericidal effect of the films obtained was evaluated in vitro using the broth surface method against two opportunistic and nosocomial pathogens, Staphylococcus aureus and Pseudomonas aeruginosa. The bacterial growth results showed that the P3HB@NiFe₂O₄ hybrid film was inhibited by 97% and 96% for S. aureus and P. aeruginosa, respectively. Surprisingly, the P3HB film inhibited both bacterial strains by around 90%. The cytotoxicity of the NiFe₂O₄ NPs, P3HB@NiFe₂O₄ hybrid film, and the P3HB film was evaluated using human skin cells, keratinocytes, and fibroblasts, finding that the NPs are biocompatible. The P3HB film and hybrids are cytotoxic, which demonstrated that although P3HB is known and reported as a biocompatible polymer, under our work conditions, P3HB was cytotoxic. Its bactericidal effect could be related to this activity. Its films are bactericidal and cytotoxic to keratinocytes and fibroblasts, the first barrier of human skin. Despite this, the hybrid film of P3HB@NiFe₂O₄ presents synergy with the bactericidal effect between P3HB and NPs, increasing bacterial inhibition. In addition, NPs decrease the cytotoxicity of P3HB to keratinocytes. The methodology used in this work was successful in producing hybrid films with antibacterial activity. However, future challenges are generated to find relationships between NPs and P3HB that allow taking advantage of their bactericidal properties and do not compromise biocompatibility.

Keywords: poly-3-hydroxybutyrate, nanoparticles, hybrid film, antibacterial

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Authors: Delsy Salinas, Andreia Garcês, Augusto Silva, Paula Brilhante Simões

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Urinary tract infection (UTI) is a common disease affecting dogs and cats in both community and hospital environment. Bacteria is the most frequent agent isolated, fewer than 1% of infections are due to parasitic, fungal, or viral agents. Common symptoms and laboratory abnormalities includeabdominal pain, pyrexia, renomegaly, and neutrophilia with left shift. A rapid and precise identification of the bacterial agent is still a challenge in veterinarian laboratories. Therefore, this cross-sectional study aims to describe bacterial colony patterns of urine samples by using chromID™ CPS® EliteAgar (BioMérieux, France) from canine and feline specimens submitted to a veterinary laboratory in Portugal (INNO Veterinary Laboratory, Braga)from January to March2022. All urine samples were cultivated in CPS Elite Agar with calibrated 1 µL inoculating loop and incubated at 37ºC for 18-24h. Color,size, and shape (regular or irregular outline)were recorded for all samples. All colonies were classified as Gram-negative or Gram-positive bacteriausing Gram stain (PREVI® Color BioMérieux, France) and determined if they were pure colonies. Identification of bacteria species was performed using GP and GN cards inVitek 2® Compact(BioMérieux, France). A total of 256/1003 submitted urine samples presented bacterial growth, from which 172 isolates were included in this study. The sample’s population included 111 dogs (n=45 males and n=66 females) and 61 cats (n=35 males and n=26 females). The most frequent isolated bacteria wasEscherichia coli (44,7%), followed by Proteus mirabilis (13,4%). All Escherichia coli isolates presented red to burgundy colonies, a colony diameter between 2 to 6 mm, and regular or irregular outlines. Similarly, 100% of Proteus mirabilis isolates were dark yellow colonies with a diffuse pigment and the same size and shape as Escherichia coli. White and pink pale colonies where Staphylococcus species exclusively and S. pseudintermedius was the most frequent (8,2 %). Cian to blue colonies were mostly Enterococcusspp. (8,2%) and Streptococcus spp. (4,6%). Beige to brown colonies were Pseudomonas aeruginosa (2,9%) and Citrobacter spp. (1,2%).Klebsiella spp.,Serratia spp. and Enterobacter spp were green colonies. All Gram-positive isolates were 1 to 2 mm diameter long and had a regular outline, meanwhile, Gram-negative rods presented variable patterns. This results showed that theprevalence of E coli and P. mirabilis as uropathogenic agents follows the same trends in Europe as previously described in other studies. Both agents presented a particular color pattern in CPS Elite Agar to identify them without needing complementary tests. No other bacteria genus could be correlated strongly to a specific color pattern, and similar results have been observed instudies using human’s samples. Chromogenic media shows a great advantage for common urine bacteria isolation than traditional COS, McConkey, and CLEDAgar mediums in a routine context, especially when mixed fermentative Gram-negative agents grow simultaneously. In addition, CPS Elite Agar is versatile for Artificial Intelligent Reading Plates Systems. Routine veterinarian laboratories could use CPS Elite Agar for a rapid screening for bacteria identification,mainlyE coli and P.mirabilis, saving 6h to 10h of automatized identification.

Keywords: cats, CPS elite agar, dogs, urine pathogens

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Authors: M. S. Raza, A. Kapil, Sonu Tyagi, H. Gautam, S. Mohapatra, R. Chaudhry, S. Sood, V. Goyal, R. Lodha, V. Sreenivas, B. K. Das

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Background: Acute bacterial meningitis remains the major cause of mortality and morbidity. More than half of the survivors develop the significant lifelong neurological abnormalities. Diagnosis of the hospital acquired acute bacterial meningitis (HAABM) is challenging as it appears either in the post operative patients or patients acquire the organisms from the hospital environment. In both the situations, pathogens are exposed to high dose of antibiotics. Chances of getting multidrug resistance organism are very high. We have performed this experiment to find out the etiological agents of HAABM and its antibiotics susceptibility pattern. Methodology: A perspective study was conducted at the Department of Microbiology, All India Institute of Medical Sciences, New Delhi. From March 2015 to April 2018 total 400 Cerebro spinal fluid samples were collected aseptically. Samples were processed for cell count, Gram staining, and culture. Culture plates were incubated at 37°C for 18-24 hours. Organism grown on blood and MacConkey agar were identified by MALDI-TOF Vitek MS (BioMerieux, France) and antibiotic susceptibility tests were performed by Kirby Bauer disc diffusion method as per CLSI 2015 guideline. Results: Of the 400 CSF samples processed, 43 (10.75%) were culture positive for different bacteria. Out of 43 isolates, the most prevalent Gram-positive organisms were S. aureus 4 (9.30%) followed by E. faecium 3 (6.97%) & CONS 2 (4.65%). Similarly, E. coli 13 (30.23%) was the commonest Gram-negative isolates followed by A. baumannii 12 (27.90%), K. pneumonia 5 (11.62%) and P. aeruginosa 4(9.30%). Most of the antibiotics tested against the Gram-negative isolates were resistance to them. Colistin was most effective followed by Meropenem and Imepenim for all Gram-negative HAABM isolates. Similarly, most of antibiotics tested were susceptible to S. aureus and CONS. However, E. faecium (100%) were only susceptible to vancomycin and teicoplanin. Conclusion: Hospital acquired acute bacterial meningitis (HAABM) is becoming the emerging challenge as most of isolates are showing resistance to commonly used antibiotics. Gram-negative organisms are emerging as the major player of HAABM. Great care needs to be taken especially in tertiary care hospitals. Similarly, antibiotic stewardship should be followed and antibiotic susceptibility test (AST) should be performed regularly to update the antibiotic patter and to prevent from the emergence of resistance. Updated information of the AST will be helpful for the better management of the meningitis patient.

Keywords: CSF, MALDI-TOF, hospital acquired acute bacterial meningitis, AST

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Authors: Sana Mohammad Jafar

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Salmonella is one of the major causes of foodborne diseases throughout the world. Shrimp are an important commodity in world fishery trade. The microbiological quality of shrimp must be evaluated for assurance of shrimp. The aim of this study was to evaluate the microbiological quality and to determine the prevalence of Salmonella in shrimp sold in Khuzestan province. In this study, a total of 245 samples of shrimp sold in Khuzestan province were tested for Salmonella prevalence and total microbial population. The mean aerobic bacterial count in 50.2% of samples was 2200, in 29.8% of samples was 13,600, in 20% of samples was 36,700, and the mean aerobic bacterial count in the total samples was 20,000. (20,000 cfu/cc). Of the total samples, 33 samples were positive for Salmonella and the prevalence of Salmonella was determined 13.4%. These results indicate the possibility that shrimp contribute to foodborne infections. The improvement of shrimp quality is an important issue, and shrimp before consuming should be washed with water containing chlorine, with the aim of increasing safety. In addition, it should be avoided to eat shrimp as raw or not cooked properly.

Keywords: determination, total microbial, Salmonella, shrimp

Procedia PDF Downloads 243

Authors: Ghadah Abusalim, Suliman Alharbi, Hesham Khalil, Milton Wainwright, Mohammad A. Khiyami

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The use of implantable foreign devices in medicine has recently increased dramatically. Intravascular cannulae and catheters are used to administer fluids, medications, parenteral nutrition, and blood products in order to monitor hemodynamic status and also to provide hemodialysis. The early and late failure of inserted or implanted devices is largely the result of bacterial infection and may lead to the disruption of integration between the device and the tissues which surround it. Staphylococcus aureus and Staphylococcus epidermidis are widely considered to be the most common organisms causing device-related infection. Our study showed that S. aureus and S. epidermidis adhered to intravascular cannulae made up of PTFE, SPTFE and vialon. Adhesion of S. epidermidis and S. aureus to intravascular cannulae varied significantly depending upon the type of material used and the presence of coating materials. Both bacteria adhered less to PTFE followed by Vialon and SPTFE and the adhesion capacity of S. aureus and S. epidermidis increased over time. Coating intravascular cannulae with human serum albumin inhibited the adhesion of S. aureus and S. epidermidis to these cannulae, and pretreatment of cannulae with fibronectin inhibited the adhesion of S. epidermidis but increased the adhesion of S. aureus to all types of cannulae. Pretreatment of cannulae surface with potassium chloride or calcium chloride increased the adhesion of S. aureus and S. epidermidis to cannulae, suggesting a role for electrostatic forces in the mechanism of such adhesion. This study will hopefully clarify the mechanism of adhesion and provide possible means of preventing such adhesion either by the use of better material coatings or by interfering with the process of adhesion by targeting bacterial structures responsible for it. Currently we recommend the use of PTFE cannulae as they exhibit a lower bacterial adhesion capacity compared to the other tested cannulae.

Keywords: Staphylococcus epidermidis, Staphylococcus aureus, adhesion, cannulae, PTFE, Vialon

Procedia PDF Downloads 351

Authors: E. Sobhani-Tehrani, K. Khorasani, N. Meskin

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This paper presents a novel integrated hybrid approach for fault diagnosis (FD) of nonlinear systems. Unlike most FD techniques, the proposed solution simultaneously accomplishes fault detection, isolation, and identification (FDII) within a unified diagnostic module. At the core of this solution is a bank of adaptive neural parameter estimators (NPE) associated with a set of single-parameter fault models. The NPEs continuously estimate unknown fault parameters (FP) that are indicators of faults in the system. Two NPE structures including series-parallel and parallel are developed with their exclusive set of desirable attributes. The parallel scheme is extremely robust to measurement noise and possesses a simpler, yet more solid, fault isolation logic. On the contrary, the series-parallel scheme displays short FD delays and is robust to closed-loop system transients due to changes in control commands. Finally, a fault tolerant observer (FTO) is designed to extend the capability of the NPEs to systems with partial-state measurement.

Keywords: hybrid fault diagnosis, dynamic neural networks, nonlinear systems, fault tolerant observer

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Authors: T. Thompson, E. F. Zegeye

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Bacterial Cellulose (BC) is a structural organic compound produced in the anaerobic process. This material can be a useful eco-friendly substitute for commercial textiles that are used in industries today. BC is easily and sustainably produced and has the capabilities to be used as a replacement in textiles. However, BC is extremely fragile when it completely dries. This research was conducted to improve the mechanical properties of the BC by reinforcing with an organic polymer and exfoliated graphite (EG). The BC films were grown over a period of weeks in a green tea and kombucha solution at 30 °C, then cleaned and added to an enhancing solution. The enhancing solutions were a mixture of 2.5 wt% polymer and 2.5 wt% latex solution, a 5 wt% polymer solution, a 0.20 wt% graphite solution and were each allowed to sit in a furnace for 48 h at 50 °C. Tensile test samples were prepared and tested until fracture at a strain rate of 8 mm/min. From the research with the addition of a 5 wt% polymer solution, the flexibility of the BC has significantly improved with the maximum strain significantly larger than that of the base sample. The addition of EG has also increased the modulus of elasticity of the BC by about 25%.

Keywords: bacterial cellulose, exfoliated graphite, kombucha scoby, tensile test

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Authors: Junkal Gutierrez, Hernane S. Barud, Sidney J. L. Ribeiro, Agnieszka Tercjak

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Nowadays the interest on green nanocomposites and on the development of more environmental friendly products has been increased. Bacterial cellulose has been recently investigated as an attractive environmentally friendly material for the preparation of low-cost nanocomposites. The formation of cellulose by laboratory bacterial cultures is an interesting and attractive biomimetic access to obtain pure cellulose with excellent properties. Additionally, properties as molar mass, molar mass distribution, and the supramolecular structure could be control using different bacterial strain, culture mediums and conditions, including the incorporation of different additives. This kind of cellulose is a natural nanomaterial, and therefore, it has a high surface-to-volume ratio which is highly advantageous in composites production. Such property combined with good biocompatibility, high tensile strength, and high crystallinity makes bacterial cellulose a potential material for applications in different fields. The aim of this investigation work was the fabrication of novel hybrid inorganic-organic composites based on bacterial cellulose, cultivated in our laboratory, as a template. This kind of biohybrid nanocomposites gathers together excellent properties of bacterial cellulose with the ones displayed by typical inorganic nanoparticles like optical, magnetic and electrical properties, luminescence, ionic conductivity and selectivity, as well as chemical or biochemical activity. In addition, the functionalization of cellulose with inorganic materials opens new pathways for the fabrication of novel multifunctional hybrid materials with promising properties for a wide range of applications namely electronic paper, flexible displays, solar cells, sensors, among others. In this work, different pathways for fabrication of multifunctional biohybrid nanopapers with tunable properties based on BC modified with amphiphilic poly(ethylene oxide-b-propylene oxide-b-ethylene oxide) (EPE) block copolymer, sol-gel synthesized nanoparticles (titanium, vanadium and a mixture of both oxides) and functionalized iron oxide nanoparticles will be presented. In situ (biosynthesized) and ex situ (at post-production level) approaches were successfully used to modify BC membranes. Bacterial cellulose based biocomposites modified with different EPE block copolymer contents were developed by in situ technique. Thus, BC growth conditions were manipulated to fabricate EPE/BC nanocomposite during the biosynthesis. Additionally, hybrid inorganic/organic nanocomposites based on BC membranes and inorganic nanoparticles were designed via ex-situ method, by immersion of never-dried BC membranes into different nanoparticle solutions. On the one hand, sol-gel synthesized nanoparticles (titanium, vanadium and a mixture of both oxides) and on the other hand superparamagnetic iron oxide nanoparticles (SPION), Fe2O3-PEO solution. The morphology of designed novel bionanocomposites hybrid materials was investigated by atomic force microscopy (AFM) and scanning electron microscopy (SEM). In order to characterized obtained materials from the point of view of future applications different techniques were employed. On the one hand, optical properties were analyzed by UV-vis spectroscopy and spectrofluorimetry and on the other hand electrical properties were studied at nano and macroscale using electric force microscopy (EFM), tunneling atomic force microscopy (TUNA) and Keithley semiconductor analyzer, respectively. Magnetic properties were measured by means of magnetic force microscopy (MFM). Additionally, mechanical properties were also analyzed.

Keywords: bacterial cellulose, block copolymer, advanced characterization techniques, nanoparticles

Procedia PDF Downloads 235

Authors: Muhammad Naveed, Sohail Yousaf, Zahir Ahmad Zahir, Birgit Mitter, Angela Sessitsch

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Most bacterial endophytes originate from the soil and enter plants via the roots followed by further spread through the inner tissues. The mechanisms allowing bacteria to colonize plants endophytically are still poorly understood for most bacterial and plant species. Specific bacterial functions are required for plant colonization, but also the plant itself is a determining factor as bacterial ability to establish endophytic populations is very often dependent on the plant genotype (cultivar) and inoculums density. The effect of inoculum density (107, 108, 109 CFU mL-1) of Burkholderia phytofirmans strain PsJN was evaluated on growth and endophytic colonization of different maize and potato cultivars under axenic and natural soil conditions. PsJN inoculation significantly increased maize seedling growth and tuber yield of potato at all inoculum density compared to uninoculated control. Under axenic condition, PsJN inoculation (108 CFU mL-1) significantly improved the germination, root/shoot length and biomass up to 62, 115, 98 and 135% of maize seedling compared to uninoculated control. In case of potato, PsJN inoculation (109 CFU mL-1) showed maximum response and significantly increased root/shoot biomass and tuber yield under natural soil condition. We confirmed that PsJN is able to colonize the rhizosphere, roots and shoots of maize and potato cultivars. The endophytic colonization increased linearly with increasing inoculum density (within a range of 8 x 104 – 3 x 107 CFU mL-1) and were highest for maize (Morignon) and potato (Romina) as compared to other cultivars. Efficient colonization of cv. Morignon and Romina by strain PsJN indicates the specific cultivar colonizing capacity of the bacteria. The findings of the study indicate the non-significant relationship between colonization and plant growth promotion in maize under axenic conditions. However, the inoculum level (109 CFU mL-1) that promoted colonization of rhizosphere and plant interior (endophytic) also best promoted growth and tuber yield of potato under natural soil conditions.

Keywords: crop genotype, inoculum density, Burkholderia phytofirmans PsJN, colonization, growth, potato

Procedia PDF Downloads 489

Authors: D. Zahran, M. AlKhazindar, M. Khalil, E. T. A. Sayed

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Streptomyces scabies is a pathogenic actinomycete that infects potato crop causing severe production losses. Actinophages affect the composition and diversity of the bacterial population, thereby, can be used as a biological control. Samples of actinomycetes and phages were collected from different cultivated soils including farms of Faculty of Science, Faculty of Agriculture and different locations in Giza, Egypt. Actinomycetes were identified by using biochemical, morphological tests and molecular studies using 16S rRNA sequencing. Two specific phages (E1 and E2) against Streptomyces scabies and other hosts were isolated. Phages were identified using dilution end point (DEP), longevity in vitro (LIV), thermal inactivation point (TIP), host range and electron microscopy. PhageE1 was characterized by 10-8 (DEP),180 days(LIV), 95°C(TIP), narrow host range and electron microscopy showed ahead (59.9 nm) and neck (10.4nm). On the other hand phageE2 had 10-20 (DEP),180 days(LIV), 90°C(TIP), and the size of head was (67.2 nm) and tail (114nm). Antiviral activity was also studied using different chemicals (NaCL, KCL, CaCL2, BaCL2, CoCL2, AgNO3, ALCL3and HgCL2) with different concentrations and different plant extracts with different concentrations (star anise, tea, tillia, peppermint, ginger, cumin, chamomile, turmeric cinnamon, marjoram and black cumin). Both Phage E1and phage E2 were vulnerable to (cumin, ginger, chamomile, guavas leaves and star anise) but resistant to (Tillie, marjoram, fennelflower seeds, peppermint, and cinnamon).

Keywords: potato scab, actinophages, biological control, electron microscopy, TIP, DEP, LIV, antiviral activity

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Authors: Abdela Bulbula

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Background: Marek’s disease virus is a devastating infection, causing high morbidity and mortality in chickens in Ethiopia. Methods: The current study was conducted from March to November, 2021 with the general objective of performing antemortem and postmortem, isolation, and molecular detection of Marek’s disease virus from outbreak cases in southwestern Ethiopia. Accordingly, based on outbreak information reported from the study sites namely, Bedelle, Yayo, and Bonga towns in southwestern Ethiopia, 50 sick chickens were sampled. The backyard and intensive farming systems of chickens were included in the sampling and priorities were given for chickens that showed clinical signs that are characteristics of Marek’s disease. Results: By clinical examinations, paralysis of legs and wings, gray eye, loss of weight, difficulty in breathing, and depression were recorded on all chickens sampled for this study and death of diseased chickens was observed. In addition, enlargement of the spleen and gross lesions of the liver and heart were recorded during postmortem examination. The death of infected chickens was observed in both vaccinated and non-vaccinated flocks. Out of 50 pooled feather follicle samples, Marek’s disease virus was isolated from 14/50 (28%) by cell culture method and out of six tissue samples, the virus was isolated from 5/6(83.30%). By Real time polymerization chain reaction technique, which was targeted to detect the Meq gene, Marek’s disease virus was detected from 18/50 feather follicles which accounts for 36% of sampled chickens. Conclusion: In general, the current study showed that the circulating Marek’s disease virus in southwestern Ethiopia was caused by the oncogenic Gallid herpesvirus-2 (Serotype-1). Further research on molecular characterization of revolving virus in current and other regions is recommended for effective control of the disease through vaccination.

Keywords: Ethioi, Marek's disease, isolation, molecular

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Authors: Haitham Qaralleh, Syed Z. Idid, Shahbudin Saad, Deny Susanti, Osama Althunibat

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This study was carried out to isolate the active antimicrobial compounds from Neopetrosia exigua using bio-guided assay isolation against Staphylococcus aureus. N. exigua was extracted using methanol and subjected to liquid-liquid extraction using solvents with different polarity (n-hexane, carbon tetrachloride, dichloromethane, n-butanol and water). Purification of the active components of n-butanol and dichloromethane fractions was done using Sephadex LH-20 and reverse phase chromatography. Based on the biological guided fractionation results, dichloromethane and n-butanol fractions showed the highest antimicrobial activity. Purification of the active components of n-butanol and dichloromethane fractions yielded three compounds. The structure of the isolated compounds were elucidated and found to be 5-hydroxy-1H-indole-3-carboxylic acid methyl ester, cyclo-1`-demethylcystalgerone and avarol derivative. Avarol was showed potent bactericidal effect against S. aureus. N. exigua appears to be rich source of natural antimicrobial agents. Further studies are needed to investigate the mode of action of these compounds.

Keywords: antimicrobial, avarol, Neopetrosia exigua, Staphylococcus aureus

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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

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