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The human gut bacteriome is believed to have pivotal influences on human health and disease while the particular roles associated with the gut phageome have not been fully characterized yet with few exceptions. It is argued that gut microbiota can have a potential role in autism spectrum disorders (ASD). The public microbiota database of ASD and typically developing (TD) Chinese individuals were analyzed for phage protein-coding units (pPCU) to find any link between the phageome and ASD. The gut phageome of ASD individuals showed a wider diversity and higher abundance compared to TD individuals. The ASD phageome was associated with a significant expansion of Caudoviricetes bacteriophages. Phages infecting Bacteroidaceae and prophages encoded within Faecalibacterium were more frequent in ASD than in TD individuals. The expansion and diversification of ASD phageome can influence the bacterial homeostasis by imposing pressure on the bacterial communities. In conclusion, the differences of phages community in in ASD and TD can be used as potential diagnosis biomarkers of ASD. Further investigations are needed to verify the role of gut phage communities in the pathogenesis of ASD.
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Here, we report the genome of phage SAP012, which was isolated against Salmonella enterica serovar Typhimurium. The SAP012 genome is 59,618 bp, with a G+C content of 56.2% and with no antibiotic resistance or virulence genes, and is quite similar at the nucleotide level to a number of previously sequenced Salmonella phage genomes, e.g., GenBank accession numbers KM366098.1 and KC139515.1.
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Water resources have long been of interest to humans and have become a serious issue in all aspects of human life. The disposal of hazardous pollutants in water resources is one of the biggest global concerns and poses many risks to human health and aquatic life. Therefore, the control of hazardous pollutants in water resources plays an important role, when it comes to evaluating water quality. Due to low toxicity, good electrical conductivity, facile functionalization, and easy preparation, magnetic materials have become a good alternative in recent years to control hazardous pollutants in water resources. In the present study, the idea of using magnetic sensors in controlling and monitoring of pharmaceuticals, pesticides, heavy metals, and organic pollutants have been reviewed. The water pollutants in drinking water, groundwater, surface water, and seawater have been discussed. The toxicology of water hazardous pollutants has also been reviewed. Then, the magnetic materials were discussed as sensors for controlling and monitoring pollutants. Finally, future remarks and perspectives on magnetic nanosensors for controlling hazardous pollutants in water resources and environmental applications were explained.
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Poluentes Ambientais , Água Subterrânea , Praguicidas , Poluentes Químicos da Água , Monitoramento Ambiental , Humanos , Fenômenos Magnéticos , Recursos HídricosRESUMO
In 2019, the world faced a serious health challenge, the rapid spreading of a life-threatening viral pneumonia, coronavirus disease 2019 (COVID-19) caused by a betacoronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). As of January 2022 WHO statistics shows more than 5.6 million death and about 350 million infection by SARS-CoV-2. One of the life threatening aspects of COVID-19 is secondary infections and reduced efficacy of antibiotics against them. Since the beginning of COVID-19 many researches have been done on identification, treatment, and vaccine development. Bacterial viruses (bacteriophages) could offer novel approaches to detect, treat and control COVID-19. Phage therapy and in particular using phage cocktails can be used to control or eliminate the bacterial pathogen as an alternative or complementary therapeutic agent. At the same time, phage interaction with the host immune system can regulate the inflammatory response. In addition, phage display and engineered synthetic phages can be utilized to develop new vaccines and antibodies, stimulate the immune system, and elicit a rapid and well-appropriate defense response. The emergence of SARS-CoV-2 new variants like delta and omicron has proved the urgent need for precise, efficient and novel approaches for vaccine development and virus detection techniques in which bacteriophages may be one of the plausible solutions. Therefore, phages with similar morphology and/or genetic content to that of coronaviruses can be used for ecological and epidemiological modeling of SARS-CoV-2 behavior and future generations of coronavirus, and in general new viral pathogens. This article is a comprehensive review/perspective of potential applications of bacteriophages in the fight against the present pandemic and the post-COVID era.
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Bacteriófagos , COVID-19 , Pneumonia Viral , COVID-19/terapia , Vacinas contra COVID-19 , Humanos , SARS-CoV-2RESUMO
The recent development in genetic research indicates that there exists intraspecific genetic variability in many organism groups. These variations, which result in a variety of genotypes and phenotypes within a population, are called polymorphism. Mutations in different ways can alter the organism's phenotype and affect its fitness, for example, by altering disease susceptibility or resistance. Therefore, the detection of point mutations in different genes of a population is of particular importance. The amplification-refractory mutation system technique is a PCR-based method to detect single nucleotide polymorphisms in the genome. The high repeatability, low cost, high accessibility, and no need for sophisticated technology are the main advantages of the ARMS-PCR technique, compared with other available methods such as PCR-RFLP. This chapter describes the design and analysis method of primers for the ARMS-PCR technique.
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Mutação , Primers do DNA/genética , Genótipo , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo ÚnicoRESUMO
In this study, Ti3C2 MXene nanofragments with a size distribution of about 20 nm were laden on the well-characterized bacteriophages via electrostatic bonding, introducing a new antibacterial agent as a modified virus vector to be used in high-risk bacterial environment. At > MIC of MXene, the MXene-functionalized bacteriophage would be much more active in attacking the bacteria because of the high specificity for host receptors' recognition and targeting ability of bacteriophage and bacterial surface negative charge when comparing to the phage alone. Also, the induced positive surface moieties drive MXene nanofragments toward the negative surface charge of bacteria. The main mechanisms are the specific targeting capacity of bacteriophages, often by lysing the host and bursting out, and the physical interaction of MXene nanofragments with the bacterial cell membrane, which may rupture the cell wall in microbial death. The results described that the Ti3C2 MXene significantly enhanced the bacteriophage adsorption rate and stability over long-standing cultivation in aquatic environments providing superior antibacterial efficacy against the bacterial cells target. The Ti3C2 MXene-laden bacteriophage demonstrated a fast, efficient attaching to bacterial host cells, high antibacterial potential, and reduced 99.99% of the artificial contamination in water samples. Interestingly, no re-growth of target bacteria was observed in the samples during the experiment period, and the count of bacteria constantly remained below the detection threshold. This research raises attention in proposing a novel antibacterial agent to be synthesized through a simple one-step technique devoid of shortcomings of post-treatments in conventional antibacterial treatments.
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Bacteriófagos , Antibacterianos/farmacologia , Bactérias , Titânio , ÁguaRESUMO
Bacteriophages (phages) are viruses of bacteria. Despite the growing progress in research on phage interactions with eukaryotic cells, our understanding of the roles of phages and their potential implications remains incomplete. The objective of this study was to investigate the effects of the Staphylococcus aureus phage vB_SauM_JS25 on murine norovirus (MNV) replication. Experiments were performed using the RAW 264.7 cell line. After phage treatment, MNV multiplication was significantly inhibited, as indicated by real-time quantitative polymerase chain reaction (RT-qPCR) analysis, western blotting, the 50% tissue culture infectious dose and immunofluorescence. Furthermore, we revealed transcriptional changes in phage/MNV co-incubated RAW 264.7 cells through RNA sequencing (RNA-seq) and bioinformatic analysis. Our subsequent analyses revealed that the innate immune response might play an important role in restriction of MNV replication, such as the cellular response to IFN-γ and response to IFN-γ. Additionally, gene expression of IL-10, Arg-1, Ccl22, GBP2, GBP3, GBP5, and GBP7 was increased significantly, which indicated a strong correlation between RT-qPCR and RNA-seq results. Furthermore, phage treatment activated guanylate binding proteins (GBPs), as revealed by RT-qPCR analysis, western blotting, and confocal microscopy. Taken together, these data suggest that the phage affects the innate response, such as the IFN-inducible GTPases and GBPs, and therefore exerts an antiviral effect in vitro. Collectively, our findings provide insights into the interactions of immune cells and phages, which establish phage-based antiviral effects.
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Bacteriófagos , Norovirus , Animais , Antivirais , Imunidade Inata , Camundongos , Norovirus/genética , Replicação Viral/fisiologiaRESUMO
Bacteriophages, viruses that infect and replicate within prokaryotic cells are the most abundant life forms in the environment, yet the vast majority of them have not been properly reported or even discovered. Almost all reported bacteriophages infecting the Enterobacteriaceae family, with Escherichia coli being the major subject of studies, have been isolated from wastewater, sewage, and effluent resources. In the present study, we focused on the distribution and biodiversity of Shigella phages in an aquatic ecosystem. While no Shigella bacteria was recovered from the Yangtze River, three lytic phages were isolated from this ecosystem and were subjected to biological, morphological, and genomic characteristics. Comparative genomics and phylogenetic analyses demonstrated that vB _SflM_004 isolate belongs to Myoviridae family, Felixounavirus genus of Ounavirinae subfamily, vB_SdyM_006 was classified under the same family, however, it is suggested to be in a new genus under Tevenvirinae subfamily with some other related bacteriophages. vB_SsoS_008 phage belongs to the Siphoviridae family, Tunavirus genus, Tunavirinae subfamily. The phages did not harbor any genes involved in the lysogenic cycles and showed a high temperature and pH stability. The biodiversity of the isolated phages highly suggests that continued isolation on non-model members of Enterobacteriaceae family is necessary to fully understand bacteriophage diversity in aquatic environments.
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Water pollution is one of the main challenges and water crises, which has caused the existing water resources to be unusable due to contamination. To understand the determinants of the distribution and abundance of antibiotic resistance genes (ARGs), we examined the distribution of 22 ARGs in relation to habitat type, heavy metal pollution and antibiotics concentration across six lakes and wetlands of Iran. The concentration of 13 heavy metals was determined by inductively coupled plasma atomic emission spectroscopy (ICP-AES) by Thermo Electron Corporation, and five antibiotics by online enrichment and triple-quadrupole LC-MS/MS were investigated. We further performed a global meta-analysis to evaluate the distribution of ARGs across global lakes compared with our studied lakes. While habitat type effect was negligible, we found a strong correlation between waste discharge into the lakes and the abundance of ARGs. The ARGs abundance showed stronger correlation with the concentration of heavy metals, such as Vanadium, than with that of antibiotics. Our meta-analysis also confirmed that overuse of antibiotics and discharge of heavy metals in the studied lakes. These data point to an increase in the distribution of ARGs among bacteria and their increasing resistance to various antibiotics, implying the susceptibility of aquatic environment to industrial pollution.
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Genes Bacterianos , Metais Pesados , Antibacterianos/farmacologia , Cromatografia Líquida , Resistência Microbiana a Medicamentos , Irã (Geográfico) , Metais Pesados/análise , Espectrometria de Massas em TandemRESUMO
Global spread and emergence of the extensively drug-resistant (XDR) strains of P. aeruginosa have become a concern, thus, searching for new alternative treatment approaches are required. This study was aimed to isolate and characterize a novel lytic phage against P. aeruginosa. Seventy XDR isolates of P. aeruginosa were collected from May to September 2018. Wastewater samples were used for isolation of lytic phage against XDR P. aeruginosa isolates. Host range, thermal and pH stability, adsorption rate, latent period, burst size and morphology of phage were determined following the standard protocols. Morphological characteristics of the phage revealed that it belonged to Podoviridae family and it was named vB-PaeP-007. Although the phage had a narrow host range, 47 out of 70 XDR isolates were susceptible to it. The adsorption rate, latent period and burst size of vB-PaeP-007 were approximately 89.80% in 8 min, 10 min and 93 phages per cell, respectively. Its lysis activity remained at a wide range of pH (4 up to 12) and temperature (- 20.00 up to 70.00 ËC). Regarding the physiological features and host range of the vB-PaeP-007 phage, it could be a promising candidate for phage therapy and bio-controlling of infections from XDR isolates of P. aeruginosa in human and livestock storage centers.
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Escherichia coli (E. coli) is one of the most common uropathogenic bacteria. The emergence of multi-drug resistance among these bacteria resulted in a worldwide public health problem which requires alternative treatment approaches such as phage therapy. In this study, phage VB_EcoS-Golestan, a member of Siphoviridae family, with high lytic ability against E. coli isolates, was isolated from wastewater. Its burst size was large and about 100 plaque-forming units/infected cell, rapid adsorption time, and high resistance to a broad range of pH and temperatures. Bioinformatics analysis of the genomic sequence suggests that VB_EcoS-Golestan is a new phage closely related to Escherichia phages in the Kagunavirus genus, Guernseyvirinae subfamily of Siphoviridae. The genome size was 44829 bp bp that encodes 78 putative ORFs, no tRNAs, 7 potential promoter sequences and 13 Rho-factor-independent terminators. No lysogenic mediated genes were detected in VB_EcoS-Golestan genome. Overall VB_EcoS-Golestan might be used as a potential treatment approach for controlling E. coli mediated urinary tract infection, however, further studies are essential to ensure its safety.
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Escherichia coli/virologia , Genoma Viral , Siphoviridae/genética , Infecções Urinárias/microbiologia , DNA Viral/genética , Escherichia coli/isolamento & purificação , Humanos , Terapia por Fagos , Análise de Sequência de DNA , Siphoviridae/isolamento & purificação , Águas Residuárias/virologiaRESUMO
S. Enteritidis continues to be the most common pathogen of farm animals and a major public health burden worldwide. Using bacteriophages is a potential alternative to antibiotics against S. Enteritidis infection. In this study, the genome analysis of the lytic phage vB_SenM-PA13076 (PA13076) infecting S. Enteritidis revealed a linear, double-stranded DNA genome, which comprised of 52,474 bp and contained 69 ORFs. It belongs to the order Caudovirales; family Myoviridae, genus unclassified. The genes coded for DNA packaging, phage structural proteins, lysis components, DNA recombination, regulation, modification, and replication. No bacterial virulence or drug-resistance genes were detected. The phage PA13076 protected mice from a lethal dose of S. Enteritidis 13076Amp (5 × 108 CFU) by reducing the concentration of bacterial cells in blood, intestine, liver, spleen, and kidney. The phage PA13076 achieved at least 2.5 log reductions of S. Enteritidis cells in infected mice within 24 h (P < 0.05) when compared to the organs of control mice. The data also indicated that phage PA13076 could rapidly enter the blood and four organs of infected mice, remaining therein at concentrations of>104 PFU/g for at least 72 h. These results show that phage PA13076 has definite potential as an antibacterial therapeutic agent for attenuating S. Enteritidis infections.
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Terapia por Fagos , Infecções por Salmonella/terapia , Fagos de Salmonella , Salmonella enteritidis , Animais , Antibacterianos/isolamento & purificação , Bacteriemia/virologia , Galinhas/virologia , Fezes/virologia , Genoma Viral , Intestinos/microbiologia , Intestinos/virologia , Rim/microbiologia , Rim/virologia , Fígado/microbiologia , Fígado/virologia , Camundongos , Myoviridae/genética , Myoviridae/isolamento & purificação , Salmonelose Animal/terapia , Fagos de Salmonella/genética , Fagos de Salmonella/isolamento & purificação , Salmonella enteritidis/patogenicidade , Salmonella enteritidis/virologia , Baço/microbiologia , Baço/virologiaRESUMO
AIM: To investigate the potential effects of carboxyl and amino termini of HCV core protein on the HSCs activation. BACKGROUND: The core protein is recognized as the most important fibrosis inducer of Hepatitis C virus (HCV). While the exogenous fibrotic effect of HCV core protein has been reported earlier, the endogenous effect and the role of two termini must still be investigated. METHODS: Plasmids expressing full length, carboxyl-truncated (T1), or amino-truncated (T3) versions of the core were transfected into LX 2 cells. MTT assay was performed to evaluate the cytotoxicity of the endogenous expression of different regions of core protein on these cells. Afterwards, the total RNA was reversely transcribed and introduced into quantitative polymerase chain reaction (qPCR) to measure the expression level of collagen type I (COL1A1), α-smooth muscle actin (-SMA), tissue metalloproteinase inhibitor 1 (TIMP-1), and transforming growth factor-ß1 (TGF-ß1). In addition, TGF-ß1 as a fibrotic factor, was also assessed in the supernatant of LX-2 cells using ELISA method. RESULTS: The full and T1 versions of the core exhibited a measurable proliferative effect on LX 2 cells (P<0.05). Analysis of the gene expression was also showed that, in spite of amino-truncated version, these constructs represented a significant activation impact compared to the empty plasmid. Moreover, the result of TGF ß assay was in agreement with the results of mRNA expression analysis. CONCLUSION: The endogenous expression of the full and carboxyl-truncated versions of the core exhibited a significant activator effect on HSCs. Therefore, it can be concluded that, amino domain of HCV core protein performs a stellate cell activation role.
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The globally increasing incidence of antibiotic resistance in pathogenic microorganisms such as Shigella, a cause of human acute gastrointestinal infections, calls for developing effective alternatives. In this study, the antibiotic resistance pattern, extended-spectrum ß-lactamase (ESBL)-production, and molecular characteristics of 70 multidrug-resistant isolates belong to the two most frequent species of Shigella genus, that is, Shigella sonnei (44 isolates) and Shigella flexneri (26 isolates) were investigated. These isolates were used to evaluate both specificity and activity of Shigella-specific bacteriophages, vB_SflS-ISF001, vB_SsoS-ISF002, and a cocktail of both. Twelve out of the 21 tested resistance genes were detected in the isolates. About 59% of S. sonnei and 46% of S. flexneri isolates were identified as ESBL producers. The bacteriophages showed a high efficiency of plating (EOP ≥0.5) in about 75% of the isolates. Moreover, the growth of >85% of the isolates was inhibited by the phage cocktail of vB_SflS-ISF001 and vB_SsoS-ISF002. The phage cocktail was effective against a wide range of ESBL-positive and -negative isolates of S. sonnei and S. flexneri. Therefore, this phage cocktail has the potential to inhibit or significantly decrease the spread of drug-resistant Shigella in humans, food chains, and water/wastewater sanitation systems.
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Antibacterianos/farmacologia , Terapia por Fagos/métodos , Shigella flexneri/efeitos dos fármacos , Shigella sonnei/efeitos dos fármacos , beta-Lactamases/efeitos dos fármacos , Genes Bacterianos , Humanos , Testes de Sensibilidade Microbiana , Shigella flexneri/genética , Shigella sonnei/genética , beta-Lactamases/genéticaRESUMO
Titanium does not react well with the human tissues and due to its bio-inert nature the surface modification has yet to be well-studied. In this study, the sonoelectrochemical process has been carried out to generate TiO2 nanotube arrays on implantable Ti 6-4. All the prepared nanotubes fill with the vancomycin by immersion and electrophoresis method. Drug-releasing properties, antibacterial behavior, protein adsorption and cell attachment of drug-modified nanotubes are examined by UV-vis, flow cytometry, modified disc diffusion, BSA adsorption, and FESEM, respectively. The most uniform morphology, appropriate drug release, cell viability behavior and antibacterial properties can be achieved by samples anodized in the range of 60-75â¯V. Also improves the adsorption of BSA protein in bone healing and promotes osteoblast activity and osseointegration. Drug loading efficiency increases up to 60% via electrophoresis comparing the immersion method for anodized sample in 75â¯V. While electrophoresis does not affect the amount of vancomycin adsorption for lower voltages. Besides, the present study indicates that an anodized sample without drug loading has no antibacterial activity. Moreover, 28-days drug releasing from nanotubes is investigated by mathematical formula according to Fickian's law to find an effective dose of loaded drug.
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Portadores de Fármacos/química , Nanotecnologia , Nanotubos/química , Titânio/química , Ondas Ultrassônicas , Antibacterianos/química , Antibacterianos/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/farmacologia , Liberação Controlada de Fármacos , Eletroquímica , Humanos , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacosRESUMO
Shigellosis is one of the most important acute enteric infections caused by different species of Shigella, such as Shigella flexneri. Despite the use of antibiotic therapy to reduce disease duration, this approach is becoming less effective due to the emergence of antibiotic resistance among Shigella spp. Bacteriophages have been introduced as an alternative for controlling shigellosis. However, the bacteriophages must be without any lysogenic or virulence factors, toxin coding, or antibiotic-resistant genes. In this study, the whole genome sequence of vB-SflS-ISF001, a virulent Siphoviridae bacteriophage specific for Shigella flexneri, was obtained, and a comparative genomic analysis was carried out to identify its properties and safety. vB-SflS-ISF001 genomic DNA was measured at 50,552 bp with 78 deduced open reading frames (ORFs), with 24 ORFs (30.77%) sharing similarities with proteins from the genomes of homologous phages that had been reported earlier. Genetic analysis classifies it under the genus T1virus of the subfamily Tunavirinae . Moreover, comparative genomic analysis revealed no undesirable genes in the genome of vB-SflS-ISF001, such as antibiotic resistance, virulence, lysogeny, or toxin-coding genes. The results of this investigation indicate that vB-SflS-ISF001 is a new species, and confirm its safety for the biocontrol of S. flexneri.
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For effective use of phages as antimicrobial agents for controlling multidrug resistant S. Pullorum, it is important to understand phage biology. A lytic S. Pullorum phage was isolated and characterized from chicken feces, and its whole genome was sequenced and analyzed. A new lytic phage-vB_SPuM_SP116 (in brief SP116)- isolated and characterized using S. Pullorum SPu-116 as its host belongs to Myoviridae A1 group. Phage SP116 had a lytic effect on 27 of 37 (72.9%) different serotypes of clinical Salmonella strains. It showed a high bactericidal activity in killing all pathogens in cultures containing 5â¯×â¯105â¯cfu/mL and achieved more than 6.58 and 5.97 log unit reductions in cultures containing 5â¯×â¯106â¯cfu/mL and 5â¯×â¯107 cfu/mL, respectively. The one-step growth curve showed that the burst size was up to 118 pfu/bacterial cell. Complete genome sequence analysis revealed a linear, double-stranded DNA genome of 87,510 bp with an average G + C content of 38.84%, including 128 predicted open reading frames (ORFs) and 22 tRNA genes. SP116 was classified as a Felix O1 virus based upon the general phage characterization and the genomic information. Regarding its high efficacy in preventing especially S. Pullorum infection and its lack of any bacterial virulence, antimicrobial resistance, and lysogenesis genes, it could be a potential alternative candidate for the treatment of S. Pullorum infections.
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Especificidade de Hospedeiro , Myoviridae/genética , Myoviridae/ultraestrutura , Fagos de Salmonella/genética , Fagos de Salmonella/ultraestrutura , Salmonella enterica/virologia , Animais , Bacteriólise , Composição de Bases , Galinhas , Contagem de Colônia Microbiana , DNA Viral/química , DNA Viral/genética , Fezes/virologia , Genoma Viral , Viabilidade Microbiana , Myoviridae/isolamento & purificação , Myoviridae/fisiologia , Fases de Leitura Aberta , Terapia por Fagos , RNA de Transferência/genética , Infecções por Salmonella/terapia , Fagos de Salmonella/isolamento & purificação , Fagos de Salmonella/fisiologia , Sequenciamento Completo do GenomaRESUMO
Shigella spp. can be isolated from various food sources and is responsible for many outbreaks and sporadic cases of foodborne diseases worldwide. Although Shigella species are known as one of the major foodborne pathogens, a few studies have characterized the prevalence and molecular basis of antibiotic resistance of Shigella spp. isolated from food origins. This study investigated the prevalence of Shigella spp. in a wide range of food samples (1400 samples), and the phenotypic and genotypic basis of antimicrobial resistance of the isolates. In addition, the potential of two Shigella specific phages (vB_SflS-ISF001 and vB_SsoS-ISF002) to control the growth of the isolates in food was tested. Shigella sonnei and Shigella flexneri were detected in 11 (0.8%) and 8 (0.6%) samples, respectively. The highest prevalence of Shigella spp. was observed in vegetables. Multidrug resistance phenotypes were noticeably frequent and observed in 17 isolates (89.5%) out of 19 isolates. Moreover, 13 (68.4%), 9 (47.4%) and 17 (89.5%) isolates were positive for ß-lactamase-encoding, plasmid-mediated quinolone resistance and tetracycline resistance genes, respectively. Treatment with the phages reduced bacterial counts up to 3 and 4 log when used individually or in cocktail form, respectively. The findings of this study indicate the prevalence of Shigella spp. in food sources and also provide useful information for a better understanding of the molecular aspects of antimicrobial resistance in Shigella spp.. The results also suggest that the combination of vB_SflS-ISF001 and vB_SsoS-ISF002 phages can effectively reduce contamination of two important species of Shigella in food.
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Bacteriófagos/fisiologia , Queijo/microbiologia , Farmacorresistência Bacteriana , Fast Foods/microbiologia , Carne/microbiologia , Leite/microbiologia , Shigella/virologia , Verduras/microbiologia , Animais , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Bovinos , Contaminação de Alimentos/análise , Contaminação de Alimentos/estatística & dados numéricos , Humanos , Testes de Sensibilidade Microbiana , Prevalência , Shigella/efeitos dos fármacos , Shigella/genética , Shigella/isolamento & purificação , Tetraciclina/farmacologia , beta-Lactamases/genética , beta-Lactamases/metabolismoRESUMO
OBJECTIVES: Shigella spp. are an important group of waterborne pathogens worldwide. This study aimed to determine the frequency of Shigella spp. in a large collection of water samples and to uncover molecular aspects of antimicrobial resistance in the recovered isolates. METHODS: The antimicrobial resistance patterns, antimicrobial resistance genes (ARGs), including ß-lactamases (blaTEM, blaSHV, blaCTX-M, blaOXA, blaPER, blaVEB, blaGES and blaCMY), carbapenemases (blaKPC, blaNDM and blaIMP), plasmid-mediated quinolone resistance (PMQR) genes [qnrA, qnrB, qnrS and aac(6')-Ib] and tetracycline resistance genes [tet(A), tet(B), tet(C) and tet(D)], as well as class 1 and 2 integrons were analysed in Shigella spp. isolated from different water sources in Iran. RESULTS: Of 788 tested samples, Shigella sonnei and Shigella flexneri were detected in 9 (1.1%) and 6 (0.8%) samples, respectively. A multidrug-resistant (MDR) phenotype was observed in all of the isolates. Among the 15 Shigella isolates, 12 (80.0%), 5 (33.3%) and 7 (46.7%) were positive for genes encoding ß-lactam resistance, PMQR and tetracycline resistance, respectively. Class 1 integrons were more frequently detected among the isolates (8/15; 53.3%), consisting of 7 isolates (87.5%) with dfrA17-aadA5 and 1 isolate (12.5%) with sat1-aadA1 gene cassettes. The class 2 integron was detected in 3 isolates (20.0%) with the classic gene cassette array dfrA1-sat2-aadA1. CONCLUSIONS: Overall, this study showed that Shigella spp. are prevalent in water sources in Iran. Furthermore, the potential role of ARGs and integrons in the emergence of a MDR phenotype in Shigella isolates of water origin was demonstrated.
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Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Farmacorresistência Bacteriana Múltipla , Integrons , Shigella/classificação , Prevalência , Quinolonas/farmacologia , Análise de Sequência de DNA/métodos , Shigella/efeitos dos fármacos , Shigella/genética , Shigella/isolamento & purificação , Shigella flexneri/efeitos dos fármacos , Shigella flexneri/genética , Shigella flexneri/isolamento & purificação , Shigella sonnei/efeitos dos fármacos , Shigella sonnei/genética , Shigella sonnei/isolamento & purificação , Tetraciclinas/farmacologia , Água , Microbiologia da Água , beta-Lactamases/genéticaRESUMO
BACKGROUND: Shigella dysenteriae is one of the members of Shigella genus which was the main responsible of different Shigellosis outbreaks worldwide. The increasing consumption of antibiotics has led to the emergence and spreading of antibiotic-resistant strains. Therefore, finding new alternatives for infection control is essential, one of which is using bacteriophages. MATERIALS AND METHODS: Lytic bacteriophage against Shigella dysenteriae was isolated from petroleum refinery wastewater. Phage morphological and genetic characteristics were studied using TEM, and sequencing, respectively. In addition, the genome size was estimated, and phage resistance to different temperatures and pH, host range, adsorption rate, and one-step growth were investigated. RESULTS: According to the morphology and genetic results, this phage was named vB-SdyS-ISF003. Sequencing of the PCR products revealed that the vB-SdyS-ISF003 phage belongs to the species T1virus, subfamily Tunavirinae of family Siphoviridae. This was the first detected bacteriophage against S. dysenteriae, which belongs to the family Siphoviridae. In addition, its host range was limited to S. dysenteriae. The genome size was about 62â¯kb. vB-SdyS-ISF003 phage has a number of desirable characteristics including the limited host range to S. dysenteriae, very short connection time, a relatively wide range of temperature tolerance -20 to 50⯰C, pH tolerance of 7-9 without significant reduction in the phage titer. CONCLUSION: vB-SdyS-ISF003 is a novel virulent T1virus phage and has the appropriate potential for being used in bio controlling of S. dysenteriae in different condition.