Shigella Dysenteriae Research Articles

Shigellosis and Development of Multiple Antimicrobial Resistance Mechanisms of Shigella spp.

Shigellosis is a serious public health issue. Millions of people suffer from this deadly food and water borne disease each year. The main manifestations of affected persons are bloody diarrhea with excessive dehydration. The causative agent of this disease is the bacteria Shigella spp. which has four serogroups. Though Shigella flexneri and Shigella dysenteriae are the dominant serogroups in developing countries, reports of other serogroups, namely Shigella boydii and Shigella sonnei, in the food contaminations are available. There are seasonal variations of Shigella infection throughout the world. In Asian subcontinent, monsoon and post monsoon times are the ideal for infection. The transmission of the bacteria in human is usually caused by feco-oral route or by contaminated food and water. There are several groups of antibiotics like foscomycin, macrolide, amiglycoside, tetracycline etc. which were used before. But they are now become useless as Shigella spp. is getting resistant against those drugs. The quinolone groups of antibiotics like ciprofloxacin, ofloxacin, norflxacin, ceftriaxone etc. are the important drugs for the cure of the disease shigellosis but prevalence of drug resistant strains of Shigella spp. against those drugs are a great concern nowadays. The occurrence of plasmid mediated quinolone resistance genes (PMQR), efflux pump proteins and effective mutations at drug binding region of gyrA etc. are the major mechanisms for the development of drug resistance.

Antibiotic Susceptibility Pattern of Bacteria Isolated from Birds Droppings in Aliero, Kebbi State, Nigeria

Antibiotic resistant bacteria in bird droppings are an emerging problem of public health concern. This study investigated the antibiotic resistance pattern of some bacteria isolated from different poultry droppings. Twenty grams (20g) of each sample of bird droppings were obtained from chickens, guinea fowls and turkeys. Bacteria were isolated using cultural techniques. Antibiotic susceptibility testing was carried out using disk diffusion methods against the bacterial isolates. The following Gram positive and negative bacteria were isolated; Escherichia coli (26.7%), followed by Enterobacter aerogenes, Klebsiella pneumoniae, Shigella dysenteriae and Staphylococcus aureus (13.3%) respectively, Salmonella spp., Pseudomonas aeruginosa, and S. epidermidis accounted for 6.7% respectively. The Gram negative bacteria showed 100% susceptibility to Streptomycin, Gentamycin and Ciprofloxacin, followed by Chloramphenicol (83.3%), Cotrimoxazole and Perfloxacin (66.7%), Ofloxacin (50%) and the least was Amoxicillin (33.3%). Moreover, the Gram negative bacteria showed highest resistance against Augmentin (66.7%), followed by Ofloxacin (50%), Cotrimoxazole, and Perfloxacin (33.3%), and then least resistance to Chloramphenicol (16.7%).Similarly, the Gram positive bacteria showed 100% susceptibility to Streptomycin, Gentamycin, Ciprofloxacin, Cefuroxime and Perfloxacin. However, the highest percentage resistance was observed against Cotrimoxazole (50%). In conclusion, this study isolated some potentially resistant bacteria from the various samples of bird droppings. Contamination by these bird droppings may endanger the health of animals, humans and the environment as a whole. This study suggests that poultry farmers be educated about the dangers of giving medications to their birds and the significance of safe bird droppings disposal.

Microwave assisted extraction, phytochemical and antimicrobial evaluation of ethyl acetate extracts of stem bark of Ficus exasperata (Vahl)

Stem bark of Ficus exasperata was extracted using ethyl acetate by microwave-assisted solvent extraction. Phytochemical screening and antimicrobial evaluation were carried out on the extract. Phytochemical screening revealed the presence of cardiac glycosides, tannins, flavonoids, triterpenes and steroids. Antimicrobial evaluation revealed that the extract is active against Methicillin – resistant Staphylococcus aureus (MRSA), Staphylococcus aureus, Streptococcus pneumoniae, Salmonella typhi, Proteus mirabilis, Shigella dysenteriae, Candida stellatoidea and Candida tropicalis. The zones of inhibition (mm) for the test organisms were (24 - 31) mm. Minimum inhibition concentrations (mg/L) of extract against MRSA, Staphylococcus aureus, Streptococcus pneumonia, Salmonella typhi, Proteus mirabilis, Shigella dysenteriae, Candida stellatoidea and Candida tropicalis were 2.5, 1.25,1.25, 2.5, 1.25, 1.25, 1.25 and 2.5 respectively. Minimum bactericidal concentrations (mg/L) of extract against Methicillin – resistant Staphylococcus aureus (MRSA), Staphylococcus aureus, Streptococcus pneumonia, Salmonella typhi, Proteus mirabilis Shigella dysenteriae Candida stellatoidea and Candida tropicalis were 5, 2.5, 2.5, 5, 5, 2.5,2.5 and 5 respectively. The extract showed high inhibition against MRSA and Candida tropicalis.

Antimicrobial activity of Carum copticum and Satureja khuzestanica essential oils and acetic acid in vapor phase at different relative humidities and temperatures in peanuts

The impact of relative humidity (RH) and temperature on the effectiveness of acetic acid (AC) and essential oils of Carum copticum and Satureja khuzestanica in the vapor phase against pathogens was investigated. Inoculated peanuts were exposed to antimicrobial vapors at three humidity levels (63 ± 1, 75 ± 0, and 84 ± 1%) and three temperatures (25, 37, and 50°C) for 0, 1, and 2 hr. Results showed that increasing the RH, temperature, and fumigation time significantly affected the inhibition rate of the mentioned vapors. The highest inhibitory effect was observed under vaporizing condition including 84 ± 1% RH and 50°C, which reduced the initial population of Bacillus cereus, Staphylococcus aureus, Shigella dysenteriae, and Salmonella Typhi (8.1–8.3 log CFU/g) to 1.1, 0.5, 2.7, and 1.0 log CFU/g after 2 hr exposure to AC vapors, respectively. The results provide a promising approach to increase the inhibition rate of antimicrobial compounds in the vapor phase. Novelty impact statement Increasing the RH significantly enhanced the inhibition rate. The microbial inactivation level is enhanced with the increase of temperature. Vapors of AC exhibited higher antimicrobial activity than the two studied essential oils.

Phytochemical Analysis, Antimicrobial and Antioxidant Properties of Thymus zygis L. and Thymus willdenowii Boiss. Essential Oils.

Essential oils (EOs) are chemical products produced by odoriferous glands from a variety of plants. These essential oils have many health benefits: antiseptic, anti-inflammatory and antimicrobial activities. So due to these medicinal properties, the present study was designed to analyze essential oils of Thymus zygis L. and Thymus willdenowii Boiss. for their chemical composition and biological activities. These two thyme species were collected from the region of Ifrane, Middle Atlas of Morocco. The EO was obtained by hydrodistillation, and the yields were 5.25% for T. zygis and 3.00% for T. willdenowii. The chemical composition of the EOs was analyzed by gas chromatography coupled with mass spectrometry (GC-MS), and the results showed that T. zygis EO is dominated by carvacrol (52.5%), o-cymene (23.14%), and thymol (9.68%), while the EO of T. willdenowii contains germacrene D (16.51%), carvacrol (16.19%), and geranyl acetate (8.35%) as major compounds. The antioxidant activity assessed by Diphenylpicrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays revealed that both EOs have excellent antioxidant activities; by DPPH it resulted in IC50 = 6.13 ± 0.11 for T. zygis and 6.78 ± 0.3 µg/mL for T. willdenowii, while the one by FRAP yielded EC50 = 2.46 ± 0.01 (T. zygis) and 5.17 ± 0.2 (T. willdenowii) µg/mL. The antimicrobial activity of the two essential oils was evaluated against six bacterial strains and five fungal strains by the disk diffusion method to determine the Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and Minimum Fungicidal Concentration (MFC). The EOs revealed variable antimicrobial activities against the different tested microbial strains and showed strong antimicrobial activities, even against strains known as multi-resistant to antibiotics (Acinetobacter baumannii) at low concentrations (2 µL/mL). T. zygis EO showed the most powerful activity against all the studied bacteria, while that of T. willdenowii recorded moderate activity when tested against Shigella dysenteriae and Salmonella Typhi. With inhibition diameters that vary between 75 mm and 84 mm for concentrations of 2 µL/mL up to 12 µL/mL, S. aureus was shown to be the most sensitive to T. zygis EO. For the antifungal activity test, T. zygis EO showed the best inhibition diameters compared to T. willdenowii EO. These results showed that T. zygis EO has more powerful antioxidant and antimicrobial activities than T. willdenowii EO, therefore, we deduce that thyme EOs are excellent antioxidants, they have strong antimicrobial properties, and may in the future represent new sources of natural antiseptics that can be used in pharmaceutical and food industry.

Characterization and statistical optimization of γ-PGA produced by Bacillus megaterium UP47 isolated from Pentaclethra macrophylla.

Gamma-polyglutamic acid (γ-PGA) is a microbially produced non-toxic peptide biopolymer which is gaining grounds in many biotechnological fields and has a wide range of applications. In this study, the characteristics of γ-PGA produced by Bacillus megaterium isolated from an oil seed were determined, while the nutritional requirements of the bacterium were optimized using a predictive 15 factor-16 run Plackett-Burman experimental design. The main effect of each factor, the interaction and quadratic effects of the factors on optimized production were determined from Box-Benkhen model using Dell Statistica v. 12 and 13 software. Bacillus megaterium UP47 produced the highest γ-PGA (16.33 g/L) out of 56 spore-forming Bacillus strains isolated from soil, water and fermented food samples. Hydrolysates of the produced γ-PGA had a retention factor which corresponded to the L-glutamic acid standard (retention factor (rf) 0.35), while high-definition fourier transform infrared (FT-IR) spectroscopic imaging showed characteristic peaks representative of the active bonds present in γ-PGA. The γ-PGA at a concentration as low as 50 mg/100 mL exerted antimicrobial inhibitions against test pathogens. A 2.00 w/v γ-PGA solution had 11 mm and 13 mm inhibition zones against Staphylococcus aureus and Shigella dysenteriae, respectively. A second order polynomial equation for prediction of γ-PGA was derived as: γPGA yield = 3316.061 - 449.708A + 9.036A2 - 139.813B + 3.095B2 - 7.699C - 0.164C2 + 13.116AB - 0.087AB2 - 0.248A2B + 3.781AC - 0.076A2C - 0.394BC. It showed an increase in γ-PGA yield with increasing L-glutamic acid and biotin, but a decrease with yeast extract. Bacillus megaterium UP47 had a maximum γ-PGA yield of 54 g/L and 62 g/L, respectively, from the Plackett-Burman and Box-Benkhen design, thereby resulting in an appreciable increase in polymer yield after the optimization process with a 95% confidence level.

Evaluation of Antioxidant and Antibacterial Activities of White Mulberry (Morus alba L.) Fruit Extracts.

The fruit of mulberry trees (Morus sp.), mulberries, are traditionally utilised as a nutritional food and provide health benefits as well as skin nourishment in Thailand. White mulberries (Morus alba L.) from Chiang Mai and Mae Hong Son provinces were evaluated for their antioxidant and antibacterial activities. The antioxidant activities as well as the total phenolic, flavonoid and anthocyanin content of the aqueous and ethanolic extracts were determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azinobis-(3-ethylbenzothiazolin-6-sulfonic acid) (ABTS) and ferric reducing antioxidant power (FRAP) assays. The aqueous extracts of mulberries exhibited the highest antioxidant activity, which was associated with a higher phenolic and anthocyanin content. In testing the potent antibacterial activity against Escherichia coli, Salmonella Typhi, Shigella dysenteriae, Staphylococcus aureus and Vibrio cholerae, the mulberry extracts proved to be quite efficient, especially following water extraction. Time-kill and antibacterial adhesion assays further indicated that aqueous mulberry extracts could inhibit bacterial growth and prevent adhesions of pathogenic enteric bacteria on intestinal epithelial cells. It thus appears that mulberries can potentially be consumed as a good source of antioxidants, containing antimicrobial properties against some pathogenic bacteria which cause gastrointestinal tract infections.

PROBIOTIC CHARACTERIZATION OF BACILLUS SUBTILIS STRAIN ISOLATED FROM INFANT FECAL MATTER REVEALED BY 16S rRNA GENE AND PHYLOGENETIC ANALYSIS

Objective: The rationale of our study was to isolate and identify the putative probiotic strain from infant fecal matter exhibiting a broad range of antimicrobial activity and to analyze the effect of different culturing conditions on its probiotic properties and the production of antimicrobial metabolites. Methods: In the present study, bacterial strains were screened for probiotic properties and antimicrobial activity from infant fecal matter (6 months–2 years). The effect of varying culture conditions such as tolerance to acid, bile salt, phenol, NaCl, pH, incubation period, and temperature along with autoaggregation assay, hydrophobicity, and hemolysis was studied. The characterization of the potent strain was studied by morphological, biochemical, and 16S rRNA gene sequencing along the phylogenetic affiliation of the strain was studied. Results: Two putative probiotic bacteria (DAM and IFM) were isolated, identified, characterized, and predicted at pH 2.0, 3.0, and 4.0, the isolate IFM had 50%, 60%, and 70% survivability, while isolate DAM had 55%, 63%, and 75% survivability, respectively. At a bile salt concentration of 0.5%, both isolates had a 75% survival rate. The isolates exhibited a high percentage of hydrophobicity and autoaggregation. The isolates also had non-hemolytic activity and were susceptible to many clinical tested antibiotics (tetracycline, erythromycin, ampicillin, gentamycin, penicillin, etc.). The isolate showed antimicrobial activity against enteric pathogens such as Staphylococcus aureus, Escherichia coli, and Shigella dysenteriae. The accession number of Bacillus subtilis MT279753 and MK453362 was submitted to NCBI. Conclusion: The result revealed that isolates have potent probiotic properties and possess a direct influence on the production of antimicrobial metabolites. These parameters can be modified for the improvement of the potentiality of the isolates.

Evaluation of Antibacterial Synergism of Methanolic Extract of Dracocephalum kotschyi and Trachyspermum ammi.

BackgroundChemical preservatives are now used in various foods to increase shelf life and maintain quality instead of its natural extracts with anti-bacterial properties from plants can be used. Hence this research was planned to evaluate and study the synergistic antibacterial effect of the methanolic extracts of Dracocephalum kotschyi (D. kotschyi) and Trachyspermum ammi (T. ammi) against standard pathogenic bacteria like: Pseudomonas aeruginosa (P. aeruginosa), Shigella dysenteriae (S. dysenteriae), Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus).MethodsThe methanolic extract of D. kotschyi and T. ammi was prepared by the Soxhlet method. The minimum inhibitory concentration (MIC) of this methanolic extracts were determined by the microdilution method. Thus, by determining the amount of fractional inhibitory concentration index (FICI), the interaction between the methanolic extracts of D. kotschyi and T. ammi on the pathogenic bacteria was determined.ResultsIn this study, the MIC of the extracts of D. kotschyi and T. ammi on the pathogen; S. aureus was equal to 6.25 mg/mL and 12.5 mg/mL for S. dysenteriae, E. coli and P. aeruginosa. Hence, the combination of methanolic extracts of these plants shows a synergistic antibacterial effect (FICI < 0.5), on all tested pathogenic microorganisms was proved.ConclusionDue to the antimicrobial synergistic effect and cost-effective production process of methanolic extracts of D. kotschyi and T. ammi, they are used as natural preservatives and flavouring agents to preserve foods.

Bacteriostatic and bactericidal activities of Acalypha indica L. plant leave extracts on bacteria shigella disenteriae type 1 in vitro

This study aimed to analyze the bacteriostatic and bactericidal activity of Acalypha indica L. extract against Shigella dysenteriae type 1 bacteria in vitro. This research is a laboratory experimental study consisting of six concentration treatments, namely: (K = 0/ml, P1 = 100/ml, P2 = 200/ml, P3 = 400/ml, P4 = 800/ml, P5 = 1600 mg/ml ), and five replications each. The results showed that the herbal extract of Acalypha indica L. was bacteriostatic (inhibiting) at a concentration of 400 mg/mm, and bactericidal (killing) the bacteria Shigella dysenteriae type 1, which was tested in vitro at a concentration of at least 800 mg/ml. It is necessary to study the effect of Acalypha indica L. herbal extract from Timor in the serum of Rattus norwegicus against bacteria that cause bloody diarrhea (EHEC O157:H7, and Shigella dysenteriae type 1), MRSA bacteria, and other pathogenic bacteria.

Isolation, phenotypic characterization and comparative genomic analysis of 2019SD1, a polyvalent enterobacteria phage

Shigella has the remarkable capability to acquire antibiotic resistance rapidly thereby posing a significant public health challenge for the effective treatment of dysentery (Shigellosis). The phage therapy has been proven as an effective alternative strategy for controlling Shigella infections. In this study, we illustrate the isolation and detailed characterization of a polyvalent phage 2019SD1, which demonstrates lytic activity against Shigella dysenteriae, Escherichia coli, Vibrio cholerae, Enterococcus saccharolyticus and Enterococcus faecium. The newly isolated phage 2019SD1 shows adsorption time < 6 min, a latent period of 20 min and burst size of 151 PFU per bacterial cell. 2019SD1 exhibits considerable stability in a wide pH range and survives an hour at 50 °C. Under transmission electron microscope, 2019SD1 shows an icosahedral capsid (60 nm dia) and a 140 nm long tail. Further, detailed bioinformatic analyses of whole genome sequence data obtained through Oxford Nanopore platform revealed that 2019SD1 belongs to genus Hanrivervirus of subfamily Tempevirinae under the family Drexlerviridae. The concatenated protein phylogeny of 2019SD1 with the members of Drexlerviridae taking four genes (DNA Primase, ATP Dependent DNA Helicase, Large Terminase Protein, and Portal Protein) using the maximum parsimony method also suggested that 2019SD1 formed a distinct clade with the closest match of the taxa belonging to the genus Hanrivervirus. The genome analysis data indicate the occurrence of putative tail fiber proteins and DNA methylation mechanism. In addition, 2019SD1 has a well-established anti-host defence system as suggested through identification of putative anti-CRISPR and anti-restriction endonuclease systems thereby also indicating its biocontrol potential.

Development of a Self-Adjuvanting, Cross-Protective, Stable Intranasal Recombinant Vaccine for Shigellosis.

With the acquirement of antibiotic resistance, Shigella has resulted in multiple epidemics of shigellosis, an infectious diarrheal disease, causing thousands of deaths per year. Unfortunately, there are no licensed vaccines, primarily due to low or serotype-specific immunogenicity. Thus, conserved subunit vaccines utilizing recombinant invasion plasmid antigens (Ipa) have been explored as cross-protective vaccine candidates. However, achieving cross-protection against Shigella dysenteriae 1, which caused multiple pandemics/epidemics in the recent past, has been difficult. Therefore, a rational approach to improve cross-protection in the preparation for a possible pandemic should involve conserved proteins from S. dysenteriae 1 (Sd1). IpaC is one such conserved immunogenic protein that is less explored as an independent vaccine due to its instability/aggregation. Therefore, to improve cross-protection and potential immunogenicity and to be prepared for a future epidemic/pandemic, herein, we stabilized recombinant Sd1 IpaC, expressed without its chaperone, using a previously reported stabilizing detergent (LDAO) in a modified protocol and assessed its vaccine potential without an adjuvant. The protein assembled into heterogeneous complex spherical structures in the presence of LDAO and showed improved stability at storage temperatures of -80, -20, 4, 25, and 37 °C while providing enhanced yield and concentration. The protein could also be stably lyophilized and reconstituted, increasing the convenience of transportation and storage. Upon intranasal administration in BALB/c mice, the stabilized-IpaC-immunized groups generated significant antibody response and were not only protected against a high intraperitoneal dose of homologous S. dysenteriae 1 but also showed 100% survival against heterologous Shigella flexneri 2a without an adjuvant, while the control animals showed visible diarrhea (bloody-Sd1 challenge), lethargy, and weight loss with 0% survival. Overall, this work demonstrates that stabilized IpaC can be explored as a minimalist, self-adjuvanting, cross-protective, intranasal, single-antigen Shigella vaccine.

Evaluation of the antifungal, antibacterial, and anti-inflammatory activity of a halophyte plant Arthrocnemum macrostachyum Koch extracts.

Background: Indiscriminate use of conventional antibiotics has become a major factor responsible for the emergence and dissemination of multidrug-resistant strains of microorganisms. In this regard, we analyzed the plant extracts as an alternative solution for the inhibition of the resistant bugs.&#x0D; Methodology: This study was planned to investigate the antifungal, antibacterial and anti-inflammatory activities of hexane, ethyl acetate, chloroform, and methanol extracts of Arthrocnemum macrostachyum. Soxhlet apparatus was used to prepare different extracts of the whole plant using hexane, ethyl acetate, chloroform, and methanol. The antifungal and antibacterial activity was analyzed with the help of a good diffusion technique using different concentrations of plant extracts. The anti-inflammatory activity was also checked by observing the inhibition of albumin denaturation.&#x0D; Results: Maximum antifungal activity was observed by hexane, methanol, and chloroform extracts, and maximum antibacterial activity showed by the methanol extract. The most sensitive fungi found were Mucor, Aspergillus flavus, Saccharomyces, and Candida. Trichophyton mentagrophytes were found resistant to all the extracts. The most sensitive bacterial isolates found were Salmonella typhi, Proteus mirabilis, Pseudomonas aeruginosa, and Citrobacter freundii. The most resistant organisms were Streptococcus fecalis and Shigella dysenteriae. Methanol and chloroform extracts produced observable anti-inflammatory potential by inhibiting albumin denaturation. Statistical analysis was performed on SPSS version 23.0.&#x0D; Conclusion: The plant extracts showed significant antifungal, antibacterial and anti-inflammatory activity. These findings may support using the Arthrocnemum macrostachyum whole plant as an anti-inflammatory, antibacterial, and traditional antifungal plant. Our results demonstrate that this plant has considerable therapeutic effects on potential infectious agents and can be used to alternative conventional medicines.

Effects of Structural Manipulation on the Bioactivity of some Coumarin-Based Products.

Coumarin (2H-1-benzopyran-2-one) is a plant-derived natural product known for its pharmacological properties, such as anti-inflammatory, anticoagulant, antibacterial, antifungal, antiviral, anticancer, antihypertensive, antitubercular, anticonvulsant, antiadipogenic, antihyperglycemic, antioxidant, and neuroprotective. Two coumarin-based products were identified in the seeds of two apple phenotypes commonly known as Granny Smith and Red Delicious. This study aimed to evaluate the chemical manipulation of these coumarin-based products to more lipophilic semisynthetic compounds and trace the role of the phenolic hydroxyl group in the bioactivity of the parent natural products. The bioactivity evaluation included studying the potentials of the natural- and semisynthetic-coumarins as antioxidant, antineoplastic, antifungal, and antibacterial agents. At the first step, the antiradical potential of these products was evaluated versus the free radicals of hydroxyl and DPPH. The second potential was investigated utilizing an MTT-based photo assay versus several cancer-line cells,including SK-OV-3, MCF-7, KYSE-30, LC540, HeLa, AR42J, AB12, and AMN3. The third and fourth potentials were recognized by conducting a disc-diffusion method against six infective bacterial strains and three fungal strains. The test bacteria were Shigella dysenteriae, Klebsiella pneumonia, Escherichia coli, Haemophilus influenzae, Salmonella typhi, and Pseudomonas aeruginosa. On the other hand, the test fungi included Aspergillus flavus, Candida albicans, and Aspergillus niger. The results arising from these biopotentials revealed that the investigated functional group exerted a positive impact on the antiradical and antineoplastic potentials of the natural derivatives; however, they had a negative consequence on their antimicrobial potentials.

Phytochemical Screening and Antibacterial Activities of Senggani (Melastoma malabathricum L.) Ethanolic Extract Leaves

The aim of the current research was to perform phytochemical screening and to know the pathogenic antibacterial activities of senggani leaves extract. Phytochemical screening was done by testing eight active compounds. Antibacterial activities testing was done by using well diffusion method at concentration level of 25%, 50%, 75%, 100%. Positive control was amoxicillin, while the negative control was aquadest. Meanwhile, pathogenic bacteria were Escherichia coli, Shigella dysenteriae, Staphylococcus aureus, and Pseudomonas aeruginosa. Research design applied was complete random design, in which the data collected were analyzed using one-way ANOVA, continued by post-hoc test through Tukey method to know which concentration provide the most significant difference. Research results showed that ethanolic extract of senggani leaves has eight active compounds, those are phenolics, alkaloids, flavonoids, tannins, triterpenoids, glycosides, steroids, and saponins. This further proved that the ethanolic extract of senggani leaves have antibacterial activity and are able to inhibit the growth of all bacteria tested. The best ability shown to inhibit E. coli bacteria was at concentration of 100%, Sh. dysenteriae and P. aeruginosa started at the concentration of 75%, and S. aureus started at the concentration of 50%. Keywords: active compounds, bacteria growth, inhibition, well diffusion

A series of nanoscaled Pt(0)‐phosphorus ylide complexes based on [60]fullerene: Synthesis, characterization, and in vitro biological assessments

AbstractHerein, A type of Pt(0) complexes based on fullerene C60 including phosphorus ylides as monodentate ligands, [(η2‐C60)Pt(Y1–4)2] (Y1 = PPh3CHC(O)C6H4‐p‐NO2, Y2 = PPh3CHC(O)C6H4‐p‐CH3, Y3 = (p‐tolyl)3P‐CHC(O)C6H4‐p‐NO2, Y4 = (p‐tolyl)3P‐CHC(O)C6H4‐p‐CH3), were synthesized via adding of related monodentate phosphorus ylide to toluene solution of C60 and Pt (dba)2 (dba = dibenzylideneacetone). The structures were identified using IR, NMR (1H, 13C, and 31P), UV–visible, ESI‐MS, and EDX spectroscopies, thermal curve TGA, and electron microscopic images (scanning electron microscopy, SEM, and transmission electron microscopy, TEM). The molecular structure of complexes clearly shows that two ylidic ligands in a cis configuration and two hexagons of C60 in the junction[6,6] coordinate to Pt(0) metal center. These complexes were treated as biological agents in antibacterial, antioxidant, and anticancer assessments. Three Gram‐negative (Escherichia coli, Salmonella typhimurium, and Shigella dysenteriae) and three Gram‐positive bacteria (Bacillus subtilis, Bacillus cereus, and Staphylococcus aureus) were used for antibacterial investigation. The activities of complexes against cancer cells were also tested using AGS (gastric carcinoma), A549 (non‐small lung carcinoma), and MCF‐7 (breast carcinoma) cell lines. All biological studies on synthesized nanocomplexes showed promising effects in comparison with standards.

Identification of Shiga-Toxin-Producing Shigella Infections in Travel and Non-Travel Related Cases in Alberta, Canada.

It has long been accepted that Shiga toxin (Stx) only exists in Shigella dysenteriae serotype 1. However, in recent decades, the presence of Shiga toxin genes (stx) in other Shigella spp. have been reported. We screened 366 Shigella flexneri strains from Alberta, Canada (2003 to 2016) for stx and 26 positive strains were identified. These isolates are highly related with the majority originating from the Dominican Republic and three isolates with Haiti origin. Both phylogenetic and spanning tree analysis of the 26 Alberta and 29 stx positive S. flexneri originating from the U.S., France, Canada (Quebec) and Haiti suggests that there are geographic specific distribution patterns (Haiti and Dominican Republic clades). This study provides the first comprehensive whole genome based phylogenetic analysis of stx positive S. flexneri strains as well as their global transmission, which signify the public health risks of global spreading of these strains.

Investigation of Azithromycin Minimum Inhibitory Concentration Values and Carbapenem Resistance in Salmonella and Shigella Clinical Isolates

Increasing resistance to first-line antibiotics used in the treatment of infections caused by Salmonella and Shigella species is emerging. Azithromycin presents a good alternative treatment option for Salmonella and Shigella infections. However, there are limited data regarding the susceptibility of azithromycin in Turkey. In this study, we aimed to evaluate the susceptibility of Salmonella and Shigella species to azithromycin, to determine and compare the minimum inhibitory concentration (MIC) values and disk diffusion zone diameters. In addition, susceptibility to meropenem and first-line antibiotic options in isolates was also investigated. A total of 170 Salmonella, 76 Shigella clinical isolates collected between 2014 and 2018 in our hospital were tested for their susceptibility to azithromycin, meropenem, ampicillin, pefloxacin, trimetoprim-sulfamethoxazole, ceftazidime, and cefotaxime. Isolates were identified by matrix-assisted laser desorption ionization time of flight mass spectrometry. The isolates were confirmed and serotyped by the reference laboratory using the conventional slide agglutination method. Susceptibility of the isolates to azithromycin and other antibiotics was evaluated by Kirby-Bauer disk diffusion method. MIC values of azithromycin were determined by the reference broth microdilution method. Combined disk diffusion test was used for the detection of extended spectrum beta-lactamase (ESBL) production. Polymerase chain reaction was performed for macrolide and carbapenem resistance genes and the detected resistance genes were confirmed by sequencing. Of the 76 Shigella isolates tested, 64 (84.2%) were identified as Shigella sonnei, 10 (13.2%) as Shigella flexneri, one (1.3%) as Shigella boydii, and one (1.3%) as Shigella dysenteriae. Among the 170 Salmonella isolates, 131 (77%) were identified as Salmonella enteritidis, 11(6.5%) as Salmonella Typhimurium, 8 (4.7%) as Salmonella Kentucky, 5 (2.9%) as Salmonella Paratyphi B, 4 (2.4%) as Salmonella Infantis, 3 (1.8%) as Salmonella Cholerasuis, and 8 (4.7%) as other serovars (Salmonella Agona, Salmonella Dabou, Salmonella Gallinarum, Salmonella Hadar, Salmonella Muenchen, Salmonella Newport, Salmonella Paratyphi C, Salmonella Senftenberg), respectively. ESBL production was determined as 7.9% (6/76) in Shigella isolates and 2.9% (5/170) in Salmonella isolates. A carbapenem resistant S.Senftenberg isolate positive for the blaOXA-48 resistance gene was detected in our study. Meropenem MIC value of the isolate was detected as > 32 µg/ml with gradient diffusion test. Among all isolates, only one S.boydii isolate was detected as resistant to azithromycin with a MIC value of 128 µg/ml. The isolate was positive for the existence of mphA gene by PCR. In the disk diffusion test, azithromycin inhibition zone diameters were ≥ 12 mm in all of the tested isolates, except for the azithromycin-resistant isolate, and the azithromycin MICs were determined as ≤ 16 µg/ ml by broth microdilution. Increasing resistance to commonly used antibiotics in Salmonella and Shigella species is emerging. The detection of a carbapenem-resistant Salmonella isolate in our study indicates that the spread of carbapenem resistance to other Enterobacterales species may cause global problems. Antimicrobial susceptibility testing of azithromycin for Salmonella and Shigella species has been difficult to establish due to the lack of approval in vitro breakpoints for all species. Consequently, our data shows that azithromycin exhibits as a good alternative therapeutic choice for the treatment of gastrointestinal diseases caused by Salmonella and Shigella species. Further studies are needed to provide appropriate in vitro breakpoints supported by clinical data.

A PCR-free genosensing platform for detection of Shigella dysenteriae in human plasma samples by porous and honeycomb-like biochar decorated with ultrathin flower-like MoS2 nanosheets incorporated with Au nanoparticles

Shigella dysenteriae, a gram-negative bacterium, which results in the most infectious of bacterial shigellosis and dysenteries. In this study, an innovative gene detection platform based on label-free DNA sequences was developed to detect Shigella dysenteriae in human plasma samples. The porous and honeycomb-like structure of biochar (BC) was first synthesized through a pyrolysis process. Then, the produced biochar was effectively decorated with flower-like MoS2 nanosheets (MoS2/BC). The resulting nanocomposite was incorporated with Au nanoparticles (AuNPs) by applying chronoamperometry technique, and then the subsequent product including MoS2 nanosheets, biochar and AuNPs were immobilized on the Au electrode surface and used for modifier agent in electrochemical bio-assays. Structural and morphological study of the synthesized compounds were investigated using various characterization methods such as FE-SEM, TEM, EDS, FTIR, and XRD. Various electrochemical techniques including cyclic voltammetry (CV) and Differential pulse anodic stripping voltammetry (DPASV) have been used to investigate the applicability of the fabricated genosensing bio-assay. Under optimal conditions, LOD and LOQ were calculated 9.14 fM and 0.018 pM respectively. In addition, a linear range from 0.01 to 100 pM was obtained for single stranded-target DNA (ss-tDNA), with R2 of 0.9992. The recoveries ranged from 98.0 to 101.3%. The fabricated bio-detection assay demonstrated high selectivity for 1, 2, and 3 base mismatch sequences. In addition, a negative control of the gene detection platform which was performed to study selectivity was provided by ss-tDNA from Haemophilusinfluenzae, and Salmonella typhimurium. Moreover, it is important to mention that the organized bioassay is simply reusable and reproducible with the RSD% (relative standard deviation) ˂ 5 to next detection assays.

A New Isoflavan From the Heartwood of Dalbergia cochinchinensis

A new isoflavan, named dalbergiacochan A (1), was isolated from the heartwood of Dalbergia cochinchinensis, along with two known isoflavans, namely mucronulatol (2) and 2′-O-methylsepiol (3). Their structures were elucidated based on extensive spectroscopic analyses, including one-dimensional (1D) and two-dimensional nuclear magnetic resonance (2D NMR), and mass spectroscopy (MS) data, and the absolute stereochemistry of compound 1 was determined as R-3′,6-dihydroxy-2′,4′,8-trimethoxyisoflavan from its circular dichroism spectrum. Compound 1 was inactive against Escherichia coli, Bacillus thuringiensis, and Shigella dysenteriae.