RESUMEN
Proteus mirabilis is one of the most common causes of complicated urinary tract infections (UTI), especially in catheter-associated UTIs. The increased resistance to antibiotics, among P. mirabilis isolates has led us to search for alternative antibacterial agents. In this study, genome of a lytic Proteus phage VB_PmiS-Isfahan, isolated from wastewater, and active against planktonic and biofilms of P. mirabilis, isolated from UTI, was analyzed. Accordingly, the genome was sequenced and its similarity to other phages was assessed by the Mauve and EasyFig softwares. "One Click" was used for phylogenetic tree construction. The complete genome of VB_PmiS-Isfahan was 54,836â¯bp, dsDNA with a G+C content of 36.09%. Nighty-one open reading frames (ORFs) was deduced, among them, 23 were considered as functional genes, based on the homology to the previously characterized proteins. The BLASTn of VB_PmiS-Isfahan showed low similarity to complete genome of Salmonella phages VB_SenS_Sasha, 9NA, and VB_SenS-Sergei. A comparison of Nucleic acid and amino acid sequence, and phylogenetic analyses indicated that the phage is novel, significantly differs, and is distant from other genera, within Siphoviridae family. No virulence-associated and antibiotic resistance genes were detected. Thus, VB_PmiS-Isfahan phage is suggested as a potential novel candidate for the treatment of diseases, caused by P. mirabilis.
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Genoma Viral , Siphoviridae/genética , Genómica , Filogenia , Proteus mirabilis/virología , Siphoviridae/clasificación , Proteínas Virales/genéticaRESUMEN
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.
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Reacción en Cadena de la Polimerasa/métodos , Shigella dysenteriae/virología , Siphoviridae/clasificación , Siphoviridae/genética , Siphoviridae/aislamiento & purificación , ADN Viral/genética , Tamaño del Genoma , Genoma Viral , Especificidad del Huésped , Concentración de Iones de Hidrógeno , Terapia de Fagos , Shigella dysenteriae/patogenicidad , Siphoviridae/crecimiento & desarrollo , Temperatura , TermotoleranciaRESUMEN
Bacteria of the genus Raoultella are known to inhabit aquatic environments and can be found in medical samples. The pathogenicity of Raoultella ornithinolytica isolates in human has become increasingly important, and several cases of infections have been reported recently. However, there are no reports of isolation of bacteriophages infecting this bacterium. In this study, two novel phages (ISF3 and ISF6) of a methylotrophic Raoultella strain were isolated from sewage. To characterize the isolated phages, morphological features, protein profiles, restriction digestion patterns, and partial genome sequences were studied. Despite morphological differences, electron microscopy revealed that both phages had an icosahedral capsid connected to a contractile tail, suggesting that ISF3 and ISF6 both belong to the family Myoviridae. Partial nucleotide sequences of the ISF3 genome showed 99% to 100% identity to DNA of Klebsiella pneumonia phages KP15, KP27 and BMBT1; however, the restriction digestion profiles of ISF3 genome digested by EcoRI and EcoRV differed from those of Klebsiella phages KP15 and KP27. A partial sequence alignment showed that ISF6 can be classified as a member of a new viral genus due to its significant differences from other previously identified phages. To the best of our knowledge, this is the first report of the isolation and characterization of the specific Raoultella phages that have potential to be used as new pharmaceuticals against R. ornithinolytica.
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Bacteriófagos/genética , Enterobacteriaceae/virología , Secuencia de Bases , Cápside/virología , ADN Viral/genética , Humanos , Klebsiella pneumoniae/virología , Myoviridae/genéticaRESUMEN
Shigellosis (bacillary dysentery) is an acute enteric infection caused by members of Shigella genus. It causes annual deaths of approximately five million children in developing countries. Among Shigella spp., S. flexneri causes more serious forms of dysentery than other Shigella species. Due to the appearance of multidrug-resistant strains of Shigella spp., it is necessary to find alternative antimicrobial agents. The aims of this study were the isolation of a novel species-specific phage against S. flexneri and to evaluate its potential and efficacy for biocontrolling of S. flexneri in foods. Shigella flexneri PTCC 1234 was used as the host strain for bacteriophage isolation from waste water. A lytic phage of the Siphoviridae family was isolated and designated as vB_SflS-ISF001. The phage activity remained at high levels after 1 h of incubation at - 20 to 50 °C and was fairly stable for 1 h at pH values ranging from 7 to 9. The latent period and burst size were approximately 20 min and 53 ± 4 phages per host cell, respectively. Raw and cooked chicken breast were inoculated with a predetermined amount of S. flexneri and subjected to biocontrol test. The results showed that using vB_SflS-ISF001 phage led to more than two logs reduction in the count of viable S. flexneri. It was demonstrated that using vB_SflS-ISF001 phage is of high potential for developing an alternative strategy against S. flexneri contamination in foodstuffs.
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In healthy individuals John Cunningham virus is latent without any clinical signs, but in the cases of the use of immunosuppressive drugs in graft recipients, autoimmune diseases and also increasing of age, that the immune system is suppressed it may cause disease in reactivation. Progressive multifocal leukoencephalopathy (PML) is the well-known disease caused by the virus. It has also been associated with nephropathy and tumorogensis. At present, based on vp1 capsid gene 7 genotypes have been detected. Genetic variations of JC virus in different geographical areas and the presence of different subtypes is a useful tool for reconstructing of the genetic information of JC virus and understanding of its evolution. The aim of this study was to investigate different genotypes of the JC virus in the urine of 100 kidney transplant recipients, 43 rheumatoid arthritis patients, and 100 healthy individuals as control group in Isfahan. DNA was extracted by phenol-chloroform method and subjected to a nested PCR using specific primer for vp1 capsid gene designed by Oligo 7 software. Fisher's exact test was used for statistical analyses. Using MEGA 6 software the sequences were aligned using Clustal W tool and phylogenetic trees were constructed by neighbor joining method. Thirty-one positive samples were sequenced. Genotypes 1, 3, and 4 of the virus were detected for the first time in Iran. For the first time genotype 3 was reported as the dominant genotype in Iran. For the first time in the world, genotype 4 was detected in rheumatoid arthritis patients. J. Med. Virol. 89:337-344, 2017. © 2016 Wiley Periodicals, Inc.
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Genotipo , Virus JC/clasificación , Virus JC/genética , Trasplante de Riñón/efectos adversos , Infecciones por Polyomavirus/virología , Receptores de Trasplantes , Infecciones Tumorales por Virus/virología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , ADN Viral/química , ADN Viral/genética , ADN Viral/aislamiento & purificación , Femenino , Variación Genética , Humanos , Lactante , Recién Nacido , Irán , Virus JC/aislamiento & purificación , Masculino , Persona de Mediana Edad , Filogenia , Reacción en Cadena de la Polimerasa , Adulto JovenRESUMEN
Recently, the relationship between immunoreactivity to Epstein-Barr virus (EBV) and hypo-vitamin D in multiple sclerosis (MS) patients has been described. The aim of this study was to investigate whether vitamin D3 supplementation in MS patients could influence the immune response against latent EBV infection. Forty MS patients were recruited in this study. Twenty-seven patients were supplemented with 50,000 IU/week of vitamin D3 for 6 months and thirteen enrolled as controls. 25-Hydroxyvitamin D (25OHD) levels and IgG titers against EBNA1 and VCA were determined pre- and post-supplementation. All the patients were seropositive for EBV prior to vitamin D supplementation. In this cohort, 22.5% and 47.5% of the MS patients had deficient and insufficient levels of 25OHD, respectively. Our findings confirm that antibody titers against EBV in MS patients rise after the onset of the disease and indicate that vitamin D3 supplementation could limit augmentation of these titers in MS patients.
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Anticuerpos Antivirales/sangre , Colecalciferol/administración & dosificación , Herpesvirus Humano 4/inmunología , Esclerosis Múltiple/tratamiento farmacológico , Vitamina D/análogos & derivados , Adulto , Anticuerpos Antivirales/inmunología , Antígenos Virales/inmunología , Proteínas de la Cápside/inmunología , Suplementos Dietéticos , Infecciones por Virus de Epstein-Barr/inmunología , Antígenos Nucleares del Virus de Epstein-Barr/inmunología , Femenino , Herpesvirus Humano 4/efectos de los fármacos , Humanos , Inmunoglobulina G/sangre , Masculino , Esclerosis Múltiple/inmunología , Esclerosis Múltiple/virología , Vitamina D/sangreRESUMEN
Lactococcus garvieae is an emerging pathogen responsible for lactococcosis, a serious disease in trout aquaculture. The identification of new bacteriophages against L. garvieae strains may be an effective way to fight this disease and to study the pathogen's biology. Three L. garvieae phages, termed WP-1, WWP-2 and SP-2, were isolated from different environments, and their morphological features, genome restriction profiles and structural protein patterns were studied. Random cloning of HindIII-cut fragments was performed, and the fragments were partially sequenced for each phage. Although slight differences were observed by transmission electron microscopy, all of the phages had hexagonal heads and short non-contractile tails and were classified as members of the family Podoviridae. Restriction digestion analysis of the nucleic acids of the different phages revealed that the HindIII and AseI digests produced similar DNA fragment patterns. Additionally, SDS-PAGE analysis indicated that the isolated phages have similar structural proteins. The sequence BLAST results did not show any significant similarity with other previously identified phages. To the best of our knowledge, this study provides the first molecular characterization of L. garvieae phages.
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Bacteriófagos/clasificación , Bacteriófagos/aislamiento & purificación , Lactococcus/virología , Podoviridae/clasificación , Podoviridae/aislamiento & purificación , Bacteriófagos/genética , Datos de Secuencia Molecular , Filogenia , Podoviridae/genética , Proteínas Virales/genética , Proteínas Virales/metabolismoRESUMEN
Here, we report the first genome sequence of a new virulent phage of Microbacterium oxydans, termed vB_MoxS-ISF9, which was isolated from sewage. Transmission electron microscopy showed that the isolated phage, which has a hexagonal head of about 80 nm in diameter and a long non-contractile tail of about 240 nm, belongs to the family Siphoviridae. The vB_MoxS-ISF9 DNA was completely sequenced and found to be 59,254 bp in length, with a G+C content of 62.76% and 120 putative open reading frames (ORFs). The predicted protein products of the ORFs were identified, and their sequences were analyzed. In a comparison with all available phage genomes, vB_MoxS-ISF9 did not show any significant similarity to other previously reported bacteriophages. To the beast of our knowledge, this is the first report of the isolation and complete genomic sequencing of a virulent phage against a member of the genus Microbacterium.
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Actinomycetales/virología , Bacteriófagos/genética , Bacteriófagos/aislamiento & purificación , ADN Viral/química , ADN Viral/genética , Genoma Viral , Bacteriófagos/ultraestructura , Composición de Base , Orden Génico , Microscopía Electrónica de Transmisión , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Análisis de Secuencia de ADN , Aguas del Alcantarillado/virología , Siphoviridae/genética , Siphoviridae/aislamiento & purificación , Siphoviridae/ultraestructura , Virión/ultraestructuraRESUMEN
The most frequent infections caused by Pseudomonas aeruginosa are local infections in soft tissues, including burns. Today, phage use is considered a suitable alternative to cure infections caused by multi-drug-resistant (MDR) and extensively drug-resistant (XDR) bacteria. We investigated the potential of a novel phage (vB_PaS-HSN4) belonging to Caudoviricetes class, against XDR and MDR P. aeruginosa strains in vivo and in vitro. Its biological and genetic characteristics were investigated. The phage burst size and latent were 119 and 20 min, respectively. It could tolerate a broad range of salt concentrations, pH values, and temperatures. The combination with ciprofloxacin significantly enhanced biofilm removal after 24 h. The genome was dsDNA with a size of 44,534 bp and encoded 61 ORFs with 3 tRNA and 5 promoters. No virulence factor was observed in the phage genome. In the in vivo infection model, treatment with vB_PaS-HSN4 increased Galleria mellonella larvae survival (80%, 66%, and 60%) (MOI 100) and (60%, 40%, and 26%) (MOI 1) in the pre-treatment, co-treatment, and post-treatment experiments, respectively. Based on these characteristics, it can be considered for the cure of infections of burns caused by P. aeruginosa.
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Bacteriófagos , Quemaduras , Infecciones por Pseudomonas , Humanos , Pseudomonas aeruginosa , Genómica , AntibacterianosRESUMEN
BACKGROUND: Cervical cancer is one of the most common malignancies in women, and its treatment has many side effects. Therefore, in this research, the effects of the LaSota strain of oncolytic Newcastle disease virus vaccine on cervical intraepithelial neoplasia (CIN) patients were investigated. METHODS: 15 patients who met the inclusion criteria and diagnosed as CIN II and CIN III were included in the study. The vaccine was injected inside the cervix (neoplasia site) at increasing doses during 21 days, and they were evaluated for adverse events. NDV antibody titer was measured in 90 days and the levels of ki-67 and p16 proteins were studied by immunohistochemistry. Also, the levels of some important inflammatory cytokines in the serum of CIN patients were measured and finally the patients were evaluated according to the final outcomes and the reduction of tumor lesions. RESULTS: Only in the first dose of vaccine some patients showed flu-like symptoms. The accumulation of NDV antibodies started on the 7th day of the study and increased until the 90th day. Administration of LaSota vaccine had no significant effect on the expressions of Ki-67 and p16 proteins. Nevertheless, a decrease in the serum levels of Il-1ß was observed in patients after the administration of the vaccine, but the serum levels of both Il-2 and INF-γ upregulated significantly. Also, vaccine administration had no significant effect in reducing CIN grades and lesions. CONCLUSIONS: In general, we concluded that LaSota strain of NDV vaccine has no therapeutic effectiveness in CIN patients.
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Enfermedad de Newcastle , Displasia del Cuello del Útero , Vacunas Virales , Animales , Humanos , Femenino , Virus de la Enfermedad de Newcastle , Enfermedad de Newcastle/prevención & control , Antígeno Ki-67 , Estudios de Cohortes , Displasia del Cuello del Útero/metabolismo , Anticuerpos Antivirales , Inhibidor p16 de la Quinasa Dependiente de Ciclina/metabolismoRESUMEN
OBJECTIVES: Torque teno mini virus (TTMV) is classified as the Betatorquevirus genus of Anelloviridae. Little is known about the prevalence of TTMV in humans. Worldwide, cervical cancer is the second most common cancer affecting women. This study aimed to estimate the TTMV infection frequency in cervicitis cases and cervical tumors including intraepithelial neoplasia (CIN), squamous cell carcinoma (SCC) and adenocarcinoma, and the possible role of this virus in the etiology of them in an Isfahan population. METHODS: 79 cervicitis cases and 42 tumors were collected from histopathological files of Al-Zahra Hospital in Isfahan, Iran. DNA was extracted and subjected to nested polymerase chain reaction. RESULTS: Totally 62% of the tested samples were positive for TTMV. It was positive in 52.4% of adenocarcinoma, 68.4% of CIN and 100% SCC cases. In cervicitis, 48% of the cases were positive. In the phylogenetic construct two of the cervical tumor isolates and two of the cervicitis isolates were placed in the same cluster with already reported isolates from Japan (EF538880 and AB041962). Also, three of the cervical tumors isolated (JQ734980, JQ734981 and JQ734982) were placed in another cluster. CONCLUSION: The presence of the virus in cervical tissues suggests possible sexual transmission of the virus.
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Infecciones por Virus ADN/epidemiología , Torque teno virus/aislamiento & purificación , Neoplasias del Cuello Uterino/epidemiología , Cervicitis Uterina/epidemiología , Adulto , Anciano , Cuello del Útero/patología , Cuello del Útero/virología , Infecciones por Virus ADN/transmisión , Infecciones por Virus ADN/virología , ADN Viral/aislamiento & purificación , Femenino , Humanos , Irán/epidemiología , Persona de Mediana Edad , Prevalencia , Enfermedades Virales de Transmisión Sexual/epidemiología , Enfermedades Virales de Transmisión Sexual/virología , Neoplasias del Cuello Uterino/virología , Cervicitis Uterina/virología , Adulto JovenRESUMEN
Torque teno midi virus and small anellovirus (TTMDV/SAV) are members of the genus Gammatorquevirus within the family Anelloviridae. Cervical cancer is the second most prevalent cancer after breast cancer. The aim of this study was to determine the frequency of infection by these viruses in cervicitis and cervical tumors of women from Isfahan, Iran. Formalin-fixed, paraffin-embedded tissue samples from cervical cancers (n = 42) and cervicitis cases (n = 79) were subjected to nested PCR to identify TTMDV/SAV viral sequences. Of the 42 tumor cases, 22, 18 and 2 were diagnosed as adenocarcinoma, cervical intraepithelial neoplasia and squamous cell carcinoma, respectively. In total, 23 (55%) of the tumor samples were positive for TTMDV/SAV. Of the 79 cervicitis cases, 38 (48%) were also positive for TTMDV/SAV. This is the first report of TTMDV/SAV in cervicitis and cervical tumors of women.
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Torque teno virus/aislamiento & purificación , Neoplasias del Cuello Uterino/virología , Cervicitis Uterina/virología , Carcinoma de Células Escamosas/virología , Femenino , Humanos , Irán , Datos de Secuencia Molecular , Filogenia , Torque teno virus/clasificación , Torque teno virus/genética , Neoplasias del Cuello Uterino/patologíaRESUMEN
A total of 114 isolates of methicillin-resistant Staphylococcus aureus (MRSA) were collected from hospitals in Tehran, Iran. A multiplex PCR was designed to examine the presence of six different prophage classes. The results showed high diversity of bacteriophages, with four different prophage types and eight prophage patterns. An important S. aureus phage coding for several virulence factors, Φ-77-like phage, was detected in 97 % of the isolates. We found a high rate of resistance of MRSA isolates to penicillin, ciprofloxacin, tobramycin and kanamycin. This is the first study showing high prevalence and diverse bacteriophage populations in MRSA strains in Iranian hospitals.
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Antibacterianos/farmacología , Farmacorresistencia Bacteriana Múltiple , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Staphylococcus aureus Resistente a Meticilina/genética , Profagos/genética , Infecciones Estafilocócicas/microbiología , Variación Genética , Hospitales , Humanos , Irán , Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Pruebas de Sensibilidad Microbiana , Fagos de Staphylococcus/genética , Factores de Virulencia/genéticaRESUMEN
Periodontitis is a chronic inflammatory condition that can damage soft tissues and supporting teeth. Enterococcus faecalis is an opportunistic pathogen usually living in the oral cavity and plays a critical role in apical periodontitis that significantly threatens human health. The use of bacteriophages as an alternative way to eliminate bacterial infections is a promising approach. E. faecalis was isolated from the depth of dental packets of patients with periodontitis. Antimicrobial susceptibility was tested using 16 antimicrobial agents. Also, a specific virulent bacteriophage (vB_EfaS-SRH2) with an irregular pentagonal morphology of the head and a non-contractile tail belonging to the Siphoviridae, was isolated from wastewater in East of Isfahan, Iran, and its physiological and genomic specifications were investigated. The genome was double-strand DNA with 38,746 bp length and encoded 62 putative ORFs. In addition, eight Anti-CRISPERs and 30 Rho-dependent terminators were found. No tRNA was found. It had a short latent period of 15 min and a large burst size of ~ 125. No undesirable genes (antibiotic resistance, lysogenic dependence, and virulence factors) were identified in the genome. Based on physiological properties and genomic characteristics, this phage can be used as a suitable choice in phage therapy for periodontitis and root canal infection.
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Bacteriófagos , Periodontitis , Siphoviridae , Enterococcus faecalis/genética , Genoma Viral , Humanos , Periodontitis/genética , Periodontitis/terapia , Siphoviridae/genéticaRESUMEN
Many different strategies have been used to fight against the Coronavirus disease (COVID-19) pandemic as a therapeutics or prophylaxis approaches. However, not enough attention has been paid to general and specific immune factors and nutritional components found in hyper-immunized dairy products. Hyper-immune bovine colostrum (HBC) has been used against many different respiratory and gastrointestinal tracts infections during past decades. An isolated dairy farm was established, and nine mixed Holstein X Simmental dairy cattle in their 6-7 months of gestation period were chosen for hyper-immunization with inactivated Severe acute respiratory syndrome corona virus-2 (SARS-CoV-2). For this, six cows were inoculated with 2 ml of 109.4/ml (TCID50) of the virus. As a control group, three cows were inoculated with the carrier without virus. Specific IgG level against the SARS-CoV-2 was measured before and after immunization in the sera, and in the colostrum and milk following parturition in hyper-immunized cows using indirect Enzyme-linked immunosorbent assay (ELISA). Neutralizing antibodies in the serum and colostrum was measured by a quantitative ELISA. The safety of the product was determined in40 healthy volunteers aged between 18-65 years old (13 females and 27 males) in the phase 1 clinical trial (https://www.irct.ir/trial/51259). No adverse effects were observed in the experimental cows. A very high level of IgG was observed in the first colostrum that sharply decreased in the following 7 days in the milk. The titer of specific neutralizing antibody in the colostrum samples was 69 times higher than the sera. No adverse effects and clinical complications were reported by the authorized ethics committee, and an official certificate on the safety of the product was issued. Beside other strategies, this approach could be used for large-scale and low-cost production of immune components to be used as a nutritional supplement to confront current SARS-CoV-2 and future pandemics. Clinical Trial Registration: [https://www.irct.ir/trial/51259].
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Multiple sclerosis (MS) is a disease of young adults which is characterized by autoimmune demyelination of the central nervous system. Interaction of genetics and environmental factors are required to cause MS. Among the proposed environmental factors for MS, viral infections are thought to play a role in the pathogenesis of the disease. Torque teno mini virus (TTMV), which has recently been shown to infect humans, is a member of circoviridae, and has a circular DNA with 2860 nucleotides. Since there are a few data about the pathogenicity of this virus, this study sought to investigate the presence of TTMV in sera from MS patients and healthy individuals. We studied 149 serum samples from MS patients and 150 sera of healthy individuals. Serum DNA was extracted using phenol-chloroform and was subjected to nested polymerase chain reaction. TTMV-DNA was detected in 24 (16%) sera of the healthy blood donors and in 21 (14.1%) samples of the MS patients, where the difference did not reach significance (p > .05). The result of this study could not establish an association between TTMV infection and MS.
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Infecciones por Virus ADN/etiología , Esclerosis Múltiple/etiología , Esclerosis Múltiple/virología , Torque teno virus/genética , Torque teno virus/patogenicidad , Adulto , ADN Viral/análisis , Femenino , Humanos , Masculino , Análisis de Secuencia de ADN/métodosRESUMEN
Multiple sclerosis (MS) is the most common autoimmune disease characterized by multifocal areas of inflammatory demyelination within the central nervous system. Cytomegalovirus (CMV) has a complex pathobiology and in most cases is simply asymptomatic. There is some recent controversy over the role of CMV in the pathology of MS. The aim of this study was to evaluate active CMV infection and its effect on the humoral immunity in patients with MS. Serum, plasma, peripheral blood mononuclear cells (PBMCs), saliva and urine collected from MS patients (n=78) and healthy subjects (n=123) were screened for the presence of anti-CMV antibodies and CMV-DNA by nephelometric and PCR methods. Concentrations of total antibodies in MS subtypes were measured using both nephelometric and enzyme linked fluorescent assay (ELFA) techniques. The results extend the observation of an increased frequency of CMV-DNA in patients, in contrast with controls (p<0.001). Furthermore, systemic CMV infections were found in 25.5% of patients and only 3.2% of controls (p<0.001). There was significant difference in the titers of anti-CMV IgG and total IgE in patient and controls (P<0.001). These results support the hypothesis that CMV may contribute to MS thought to establish systemic infection process and induce immune response.
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Infecciones por Citomegalovirus/inmunología , Infecciones por Citomegalovirus/virología , Inmunoglobulinas/inmunología , Esclerosis Múltiple/inmunología , Esclerosis Múltiple/virología , Adolescente , Adulto , Infecciones por Citomegalovirus/epidemiología , Femenino , Humanos , Inmunoglobulina E/inmunología , Inmunoglobulina M/inmunología , Masculino , Persona de Mediana Edad , Esclerosis Múltiple/epidemiología , Reacción en Cadena de la Polimerasa , Adulto JovenRESUMEN
Salmonellosis is an important worldwide food-borne disease. Increasing resistance to Salmonella spp. has been reported in recent years, and now the prevalence of multidrug-resistant Salmonella spp. is a worldwide problem. This necessitates alternative approaches like phage therapy. This study aimed to isolate bacteriophages specific for Salmonella enterica serovar Paratyphi B and S. enterica serovar Typhimurium isolated from different sources (chicken meat, beef and eggshells). The antibiotic resistance profiles of the bacteria were determined by phenotypic and genotypic methods. The prevalence of extended-spectrum ß-lactamase genes was examined by polymerase chain reaction. In total, 75% of the isolated Salmonella strains were resistant to tetracycline, whereas 70% of them were resistant to azithromycin. All of the isolates from beef were resistant to nalidixic acid. The most common extended-spectrum ß-lactamase genes among the isolates were blaSHV (15%) followed by blaTEM (10%) and blaCTX (5%). Two specific bacteriophages were isolated and characterized. The host range for vB_SparS-ui was Salmonella Paratyphi B, S. enterica serovar Paratyphi A and S. enterica, while that for vB_StyS-sam phage was Salmonella Typhimurium and S. enterica serovar Enteritidis. The characteristics of the isolated phages indicate that they are proper candidates to be used to control some foodstuff contaminations and also phage therapy of infected animals.
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Fagos de Salmonella/aislamiento & purificación , Fagos de Salmonella/fisiología , Salmonella paratyphi B/virología , Salmonella typhimurium/virología , Antibacterianos/farmacología , Proteínas Bacterianas/genética , Farmacorresistencia Bacteriana/genética , Genoma Viral/genética , Especificidad del Huésped , Concentración de Iones de Hidrógeno , Fagos de Salmonella/ultraestructura , Salmonella enterica/efectos de los fármacos , Salmonella enterica/genética , Salmonella enterica/virología , Salmonella paratyphi B/efectos de los fármacos , Salmonella paratyphi B/genética , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/genética , Serogrupo , Cloruro de Sodio , Temperatura , Proteínas Virales/química , Proteínas Virales/genéticaRESUMEN
The bacteriophage (phage) DNA extraction methods for genomics analysis is a critical and time-consuming process. Hence, a rapid and cost-effective method for DNA extraction of phages is favorable for phage biologists. In the present study, a cost-effective, simple and rapid procedure for phage genome extraction in less than 10 min is introduced. Highly concentrated phage lysates were prepared using acetone precipitation followed by extraction using various methods such as commercial kits, TES lysis buffer, potassium iodide, and sodium iodide. The quality of the extracted DNA was analyzed by agarose gel electrophoresis and UV absorbance of DNA at 260 and 280 nm. Finally, the extracted DNA was subjected to restriction digestion and next-generation sequencing to approve the efficiency of the method. Based on the time, cost, and quality of obtained DNA, the acetone precipitation of phages and extraction by potassium iodide or sodium iodide method was determined to be the best method for phage DNA extraction tested in this study. Moreover, the extracted genomic DNA using this method is suitable for phage genomic analysis such as restriction enzyme studies, preparation of DNA library, and also next-generation sequencing.
Asunto(s)
Bacteriófagos , Bacteriófagos/genética , ADN , Genómica , Secuenciación de Nucleótidos de Alto RendimientoRESUMEN
Enterococcus faecalis is an environmental agent of bovine mastitis in cows and has many cytopathic effects on the urinary tract in both humans and animals. In this study, a novel lytic bacteriophage, vB_EfaS-DELF1, was isolated against 21 E. faecalis isolated from bovine mastitis, including vancomycin-resistant E. faecalis (VRE). vB_EfaS-DELF1 bacteriophage was specific for E. faecalis and showed no lytic effects against other tested Enterococcus spp., Gram-negative, or Gram-positive bacteria. Moreover, no activity was observed against yogurt starters. The phage suspension was stable in a wide range of pH, salinity, and temperature. It retained its activity in 3.5 % fat milk. vB_EfaS-DELF1 has the common phenotypic features of Siphoviridae with a double-strand DNA of 40,248 bp in length and a G + C content of 34.9 %. The genome encodes 62 putative ORFs and no tRNA. No undesirable genes such as lysogenic mediators, antibiotic resistance, or virulence factor genes were detected in the genome. The comparative genomic analysis demonstrated similarity to the other available phage genomes. The highest similarity was observed with two other phages (50 % coverage and 82.38 % identities with IME-EFm1; 35 % coverage and 86.22 % identities with IME-EFm5) that were placed in the same clade. The differences with the other aligned phages were high and were placed in distant clusters. Regarding the specificity of this new bacteriophage against all of the tested E. faecalis isolates and, in particular, against the vancomycin-resistant ones, and also the absence of antibiotic resistance or virulence genes in its genome, vB_EfaS-DELF1 is suggested as a potential candidate for biocontrol of E. faecalis infections.