Publication Date: 2022
Frontiers in Nutrition (2296861X)9
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]. Copyright © 2022 Nili, Bouzari, Attaran, Ghalegolab, Rabani and Mahmoudian.
Publication Date: 2022
Scientific Reports (20452322)12(1)
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. © 2022, The Author(s).
Publication Date: 2021
Journal of Virological Methods (01660934)293
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. © 2021 Elsevier B.V.
Publication Date: 2021
Letters in Applied Microbiology (02668254)72(3)pp. 231-237
Shigella spp. are water-borne pathogens responsible for mild to severe cases bacilli dysentery all around the world known as Shigellosis. The progressively increasing of antibiotic resistance among Shigella calls for developing and establishing novel alternative therapeutic methods. The present study aimed to evaluate a novel phage cocktail of lytic phages against extended spectrum beta lactamase isolates of Shigella species in an aquatic environment. The phage cocktail containing six novel Shigella specific phages showed a broad host spectrum. The cocktail was very stable in aquatic environment. The cocktail resulted in about 99% decrease in the bacterial counts in the contaminated water by several species and strains of Shigella such as Shigella sonnei, Shigella flexneri and Shigella dysenteriae. Achieving such a high efficiency in this in-vitro study demonstrates a high potential for in-vivo and in-situ application of this phage cocktail as a bio-controlling agent against Shigella spp. contamination and infections. © 2020 The Society for Applied Microbiology
Publication Date: 2021
LWT (10961127)144
Shigella, the globally distributed causative agent of bacillary dysentery, is an important microbial contamination in raw and ready-to-eat foods. The recent commercial use of phage cocktails in the food industry is evidence that the method is a highly effective solution in controlling the pathogenic bacteria, especially the antibiotic-resistant strains. This research addresses the effects of a novel phage cocktail made of six different lytic bacteriophages to prevent drug resistant isolates of Shigella species in a wide variety of food products with a different matrix structure, viscosity, carbon content, and texture properties including cooked chicken, cherry tomato, yoghurt, cucumber and Chinese cabbage. After seven days of treatment with the phage cocktail, more than 99% of the bacterial count decreased compared to the control sample (P < 0.0001). The cocktail stability in dry conditions showed that the bacteriophages remain desirably active during food products' average shelf life. The prepared phage cocktail showed to be a very effective product for bio-controlling Shigella due to its high overlap of the host spectrums, the potential for use in a variety of foods, and stability in dry conditions. © 2021
Publication Date: 2021
FEMS Microbiology Letters (03781097)368(7)
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 β-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 β-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. © 2021 The Author(s) 2021. Published by Oxford University Press on behalf of FEMS.
Publication Date: 2021
Virus Research (01681702)298
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. © 2021 Elsevier B.V.
Publication Date: 2020
Microbial Drug Resistance (10766294)26(7)pp. 831-841
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 β-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. © Copyright 2020, Mary Ann Liebert, Inc., publishers 2020.
Publication Date: 2020
Scientific Reports (20452322)10(1)
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. © 2020, The Author(s).
Publication Date: 2019
Journal of Global Antimicrobial Resistance (22137165)19pp. 122-128
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 β-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 β-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. © 2019
Publication Date: 2019
Genomics (08887543)111(6)pp. 1283-1291
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 54,836 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. © 2018 Elsevier Inc.
Publication Date: 2019
Folia Microbiologica (00155632)64pp. 283-294
Antibiotic resistance is increasing among Staphylococcus saprophyticus strains isolated from urinary tract infection. This neces-sitates alternative therapies. For this, a lytic phage (vB_SsapS-104) against S. saprophyticus, which formed round and clear plaques on bacterial culture plates, was isolated from hospital wastewater and characterized. Microscopy analysis showed that it had a small head (about 50 nm), tail (about 80 nm), and a collar (about 22 nm in length and 19 nm in width) indicating to be a phage within Siphoviridae family. Phage vB_SsapS-104 showed a large latency period of about 40 min, rapid adsorption rate that was significantly enhanced by MgCl2 and CaCl2, and high stability to a wide range of temperatures and pH values. Restriction analyses demonstrated that phage consists of a double-stranded DNAwith an approximate genome size of 40 Kb. BLAST results did not show high similarity (megablast) with other previously identified phages. But, in Blastn, similarity with Staphylococcus phages was observed. Phage vB_SsapS-104 represented high anti-bacterial activity against S. saprophyticus isolates in vitro as it was able to lyse 8 of the 9 clinical isolates (%88.8) obtained from a hospital in Gorgan, Iran. It was a S. saprophyticus-specific phage because no lytic activity was observed on some other pathogenic bacteria tested. Therefore, phage vB_SsapS-104 can be considered as a specific virulent phage against of S. saprophyitcus isolated from urinary tract infection. This study provided the partial genomic characterization of S. saprophyticus phage and its application against urinary tract infection associated with S. saprophyticus. This phage also can be considered as a good candidate for a therapeutic alternative in the future. © Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i. 2018.
Publication Date: 2019
Microbial Pathogenesis (08824010)131pp. 175-180
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 62 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. © 2019
Publication Date: 2019
Archives of Virology (03048608)164(8)pp. 2015-2022
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. © 2019, Springer-Verlag GmbH Austria, part of Springer Nature.
Publication Date: 2019
International Journal of Food Microbiology (01681605)305
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 β-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. © 2019 Elsevier B.V.
Publication Date: 2018
Journal of Food Science and Technology (00221155)55(2)pp. 550-559
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. © 2017, Association of Food Scientists & Technologists (India).
Publication Date: 2018
Journal of Molecular Microbiology and Biotechnology (14641801)28(1)pp. 37-46
Proteus mirabilis is one of the most common causes of urinary tract infection (UTI), particularly in patients undergoing long-term catheterization. Phage vB-PmiS-TH was isolated from wastewater with high lytic activity against P. mirabilis (TH) isolated from UTI. The phage had rapid adsorption, a large burst size (∼260 PFU per infected cell), and high stability at a wide range of temperatures and pH values. As analyzed by transmission electron microscopy, phage vB-PmiS-TH had an icosahedral head of ∼87 × 62 nm with a noncontractile tail about 137 nm in length and 11 nm in width. It belongs to the family Siphoviridae. Combination of the phage vB-PmiS-TH with ampicillin had a higher removal activity against planktonic cells of P. mirabilis (TH) than the phage or the antibiotic alone. Combination of the phage at a multiplicity of infection of 100 with a high dose of ampicillin (246 μg/mL) showed the highest biofilm removal activity after 24 h. This study demonstrates that using a combination of phage and antibiotic could be significantly more effective against planktonic and biofilm forms of P. mirabilis (TH). © 2018 S. Karger AG, Basel.
Publication Date: 2018
Journal of Medical Microbiology (00222615)67(3)pp. 376-386
Purpose. Shigellosis is one of the most important food-borne and water-borne diseases worldwide. Although antibiotics are considered as efficient agents for shigellosis treatment, improper use of these has led to the emergence of antibioticresistant Shigella spp. Therefore, finding a new strategy as alternative treatment seems necessary. Methodology. Different samples from a wastewater treatment plant were used to isolate Shigella spp. specific phages. Physiological properties were determined, and genomic analysis was also carried out. Results. A virulent Siphoviridae bacteriophage, vB_SsoS-ISF002, was isolated from urban wastewater in Iran and showed infectivity to different isolates of both Shigella sonnei and Shigella flexneri. vB_SsoS-ISF002 was stable at different pH values and temperatures. It had a short latent period (15 min), a large burst size (76±9 p.f.u. cell–1) and appropriate lytic activity especially at high MOI. Its genome (dsDNA) was 50 564 bp with 45.53% GC content and 76 predicted open reading frames. According to comparative genomic analysis and phylogenic tree construction, vB_SsoS-ISF002 was considered as a member of the T1virus genus. Conclusion. These results indicated that vB_SsoS-ISF002 is a novel virulent T1virus phage and may have potential as an alternative treatment for shigellosis. © 2018 The Authors.
Publication Date: 2017
Iranian Red Crescent Medical Journal (20741804)19(2)
Background: Approximately 80% of nosocomial infections are caused by strains of Klebsiella pneumoniae. Resistance to β-lactam antibiotics is a result of expression of extended-spectrum β-lactamase (ESBL) genes. Recently, phage therapy has gained increasing attention due to its many advantages over chemotherapy. Objectives: The aim of this study was to isolate ESBL-positive Klebsiella pneumoniae strains from different types of wounds, and a lytic bacteriophage against them. Methods: During a two-year period from January 2013 to February 2015, in a cross-sectional study, 41 K. pneumoniae strains were isolated from 193 categories of infected wounds at three hospitals in Isfahan, Iran. Phenotypic and genotypic methods were used to detect the ESBL-positive strains. A lytic phage against K. pneumoniae was isolated, and its host range, morphology, thermal and pH stability, saline stress, and estimated genome size were determined. Results: Of the 41 K. pneumoniae isolates, 18 were ESBL-producing and 36 carried antibiotic-resistance genes. A total of 36 out of 41 isolated samples carried one or more resistance genes. The results showed that the differences between phenotypic and genotypic identification methods were significant (P = 0.0001). The SHV, CTX-M, and TEM genes were detected in 29, 10, and 9 isolates of the tested bacteria, respectively. No bacteria contained both the SHV and the CTX-M genes. The frequency of the SHV gene was significantly higher than that of the other genes (P = 0.0001). The phage’s morphology features placed it in the Myoviridae family. Only 38 out of 41 clinical isolates were susceptible to the phage. Phage titers were completely preserved after one hour of incubation at 30°C and 40°C, and they were stable at different pH values. The phage’s survival decreased when the salt concentration was increased. Conclusions: The high rate of isolation of antibiotic-resistant strains of K. pneumoniae was consistent with other studies. As the phage was virulent and specific for K. pneumoniae, and was stable and active at different pH values, salt concentrations, and temperatures, its application in phage therapy of infected wounds is suggested. © 2016, Iranian Red Crescent Medical Journal.
Publication Date: 2017
Journal of Medical Virology (01466615)89(2)pp. 337-344
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. © 2016 Wiley Periodicals, Inc.
Publication Date: 2017
Iranian Endodontic Journal (17357497)12(2)pp. 226-230
Introduction: Pulp vitality and its continuous dentin prodution are essential for long-term success of direct pulp capping (DPC). The aim of present study was to evaluate the histopathological response of the canine pulp following DPC using either different dentin adhesive resins (DAR), calcium hydroxide (CH) or mineral trioxide aggregate (MTA). Methods and Materials: DPC was done on 72 dog’s teeth using 6 types of dental materials (n=12) (4 types of DAR, white MTA and CH). Therefore, six healthy dogs were anesthetized and 2 teeth from each dog were allocated to either type of mentioned DPC agents. The dental pulps were exposed mechanically by drilling in the center of class V cavities. The different types of capping materials included DARS (Clearfil S3 Bond, Optibond FL, Single Bond and Clearfil SE Bond), white MTA and CH. After 7, 21 and 63 days, two dogs were euthanized in each interval. Microscopic evaluations were done according to following criteria: intensity of inflammation, presence of necrosis and formation of hard tissue. The recorded data were analyzed by the Kruskal-Wallis, Friedman, Cochran’s and Fisher’s exact tests using SPSS software version 12 at significant level of 0.05. Results: No significant differences were found regarding necrosis among DPC materials (P>0.05). However, MTA caused higher amount of hard tissue formation after 63 days in comparison with 21 days. Conclusion: MTA provided the highest degree of hard tissue formation after 63 days. However, further studies should be performed for administering a definitive material. © 2017, Iranian Association of Endodontics. All rights reserved.
Publication Date: 2016
Iranian Biomedical Journal (1028852X)20(3)pp. 168-174
Background: SEN virus (SENV) is the latest virus proposed as a cause of unknown hepatitis cases. Among nine detected genotypes of the virus, genotypes D and H are more frequent in hepatitis cases of unknown origin. The aim of this study was to determine the frequency of SENV-D and SENV-H genotypes in the sera of healthy individuals and hepatitis B and C patients. Methods: Totally, 200 serum samples from healthy individuals as well as 50 hepatitis B and 50 hepatitis C patients were collected. Anti-HCV (hepatitis C virus), anti-human immunodeficiency virus, hepatitis B surface antigen and anti-HBV (hepatitis B virus) core antigen were detected, and serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured. Viral DNA was subjected to nested PCR. Fisher's exact and unpaired ANOVA tests were used for statistical analyses. Results: SENV was detected in 90%, 66%, and 46% of the healthy individuals HBV and HCV-positive individuals, respectively. The frequency of SENV and its two genotypes were significantly lower in hepatitis B and hepatitis C patients (P<0.01). Also, the frequency of SENV-H was higher than SENV-D in all studied groups. In SENV-positive HBV patients, the level of ALT and AST enzymes were significantly less than SENV-negative patients (P<0.05). It was the same for SENV-H-negative and -positive cases. Conclusions: The levels of liver enzymes were significantly lower in HBV patients co-infected with SENV compared to HBV patients (P<0.05), indicating a positive impact of the virus in liver pathology by decreasing liver damage and thus decreasing the liver enzymes. © 2016, Pasteur Institute of Iran. All rights reserved.
Publication Date: 2015
Jundishapur Journal Of Microbiology (20084161)8(6)pp. 1-5
acquired infections. Sewage acts as an environmental reservoir and may have a significant role in development and dissemination of antibiotic resistance. Objectives: This study was undertaken to determine the epidemiological relatedness between the MRSA isolated from sewage and human infections. Materials and Methods: Samples were collected from a referral hospital and also a sewage treatment plant in Tehran, Iran, during 2010. All the MRSA isolates were identified at the species level and typed using Phene plate (PhP) system and SCCmec typing. Antibiotic susceptibility tests were also performed. Results: Of the 1142 isolates, 200 MRSA strains from the sewage (n = 100) and the clinic (n = 100) were isolated. Distinct PhP types, consisting of 16 common types and 13 single types, and also 3 different staphylococcal cassette chromosome mec (SCCmec) types (III, IVa and IVc) were found amongst the MRSA isolated from the two different sources. The results of antibiotic susceptibility testing showed an increased resistance to penicillin, ciprofloxacin, erythromycin, clindamycin and tetracycline. In addition, none of the isolates showed resistance to vancomycin, quinupristin -dalfopristin and linezolid. Conclusions: The presence of common PhP types and also SCCmec type III, as an indicator for hospital strains, among the isolates, may indicate an epidemiological link between clinical and sewage MRSA isolates in Tehran. © 2015, Ahvaz Jundishapur University of Medical Sciences.
Publication Date: 2015
Iranian Journal Of Veterinary Research, Shiraz University (17281997)16(1)pp. 110-113
Although the infection of different animals and non-human primates with other members of Anelloviridae have already been reported there is no report about infection of animals with Torque teno midi virus/Small anellovirs (TTMDV/SAV). The aim of this study was to detect the virus in domestic village chickens. Blood samples were collected from 79 domestic village chickens in Isfahan. Blood samples of five adult laying hens and one cockerel were collected in three consecutive weeks (days 1, 8 and 14) as experimental chickens. Ten eggs were randomly collected from the eggs laid during days 12 to 17 and thin and thick egg whites and yolk samples were collected aseptically. After DNA extraction Nested-PCR was performed using SMAs/SMAr primers. In PCR, 431 bp and 441 bp products were detected. The detected bands were extracted and sequenced. Totally 26 out of 79 (32.9%) of the blood samples were positive for the virus. The frequency of the infection of the different parts of the eggs tested was 76%. For the first time TTMDV/SAV was detected in domestic village chickens which also vertically transmitted to eggs.
Publication Date: 2015
Archives of Iranian Medicine (10292977)18(10)pp. 638-642
Background: The worldwide emergence of multi-drug resistant (MDR) bacteria in recent years has caused many problems for hospitals and patients, especially intensive care unit patients. Among these clinically important MDR bacteria are Acinetobacter baumannii complex species (A. baumannii, Acinetobacter genomic species 3 and Acinetobacter genomic species 13TU) that cause a wide range of infections. Methods: The sequencing and bioinformatics analysis of a part of the Zone 1 of rpoB gene was performed for species identification of Acinetobacter isolates obtained from ICU patients with infected burns hospitalized in a hospital in Isfahan, Iran, over a 9-month period. Antibiotic sensitivity of Acinetobacter isolates was investigated using the disk diffusion method and different classes of antibiotics including amikacin, cefotaxime, ceftriaxone, ciprofloxacin, imipenem and piperacillin. Results: Acinetobacterspp. were isolated from 10 of 80 (12.5%) investigated patients. All of the 10 Ac/netobacterisolates were identified as Acinetobacter baumannii and multi-drug resistant according to antibiotic susceptibility tests. Conclusion: Of the Acinetobacter baumannii complex members, only A. baumannii species was identified among the isolates obtained from patients with infected burns in an Isfahan hospital over a 9-month period. © 2015 Academy of Medical Sciences of I.R. Iran. All rights reserved.
Publication Date: 2015
Journal of Environmental Radioactivity (18791700)144pp. 113-119
A new actinobacterial strain was isolated from Ab-e-Siah spring (dark water) taken from the Ramsar city in Iran, and subjected to several stress conditions investigation. The isolate, named MG2 strain, was Gram-positive, aerobic, diplococci or tetrad shaped, non-spore forming and non-motile. Phylogenetic analysis of the isolate using 16S rDNA sequence indicated that the organism matched best with the genus Kocuria and the highest sequence similarities (98.55%) being found with Kocuria rosea. The 16S rDNA sequence determined in this study has been deposited in the NCBI database with the accession no. JX534199, K. rosea strain MG2. The isolated strain was an alkaliphilic-mesophilic bacterium because the optimal growth was observed at pH 9.2 and temperature of 28°C under aerobic condition. MG2 was a halotolerant strain and tolerated maximally to 15% NaCl concentraion. Viability analysis by flow cytometry indicated that this strain had highly resistance to UV-C radiation and moderately resistance to desiccation after 28 days. The viability of K. rosea strains MG2 and Deinococcus radiodurans R1 were determined D87 and D98 according to D index, respectively, by a dose radiation 25J/cm (Appukuttan etal., 2006). Thus the UV resistance of strain MG2 was comparable with representative radiation resistant Deinococcus. Also MG2 was grown at 1-4% of H2O2 as an oxidant agent. This research is the first study on multiple extreme resistance of Kocuria rosea new strain (MG2) isolated in Iran. © 2015 Elsevier Ltd.
Publication Date: 2015
Brazilian Journal Of Microbiology (15178382)46(3)pp. 791-797
One of the most economically important bacterial pathogens of plants and plant products is Dickeya dadantii. This bacterium causes soft rot disease in tubers and other parts of the potato and other plants of the Solanaceae family. The application of restricted host range bacteriophages as biocontrol agents has recently gained widespread interest. This study purposed to isolate the infectious agent of the potato and evaluate its biocontrol by bacteriophages. Two phytopathogenic strains were isolated from infected potatoes, identified based on biochemical and 16S rRNA gene sequencing, and submitted to GenBank as D. dadantii strain pis3 (accession no. HQ423668) and D. dadantii strain sip4 (accession no. HQ423669). Their bacteriophages were isolated from Caspian Sea water by enriching the water filtrate with D. dadantii strains as hosts using spot or overlay methods. On the basis of morphotypes, the isolated bacteriophages were identified as members of the Myoviridae and Siphoviridae families and could inhibit the growth of antibiotic resistant D. dadantii strains in culture medium. Moreover, in Dickeya infected plants treated with bacteriophage, no disease progression was detected. No significant difference was seen between phage-treated and control plants. Thus, isolated bacteriophages can be suggested for the biocontrol of plant disease caused by Dickeya strains. © 2015, Sociedade Brasileira de Microbiologia.
Publication Date: 2015
Jundishapur Journal Of Microbiology (20083645)8(3)
Background: Halomethanes are toxic and carcinogenic chemicals, which are widely used in industry. Also they can be formed during water disinfection by chlorine. Biodegradation by methylotrophs is the most important way to remove these pollutants from the environment. Objectives: This study aimed to represent a simple and rapid method for quantitative study of halomethanes utilizing bacteria in drinking water and also a method to facilitate the biodegradation of these compounds in the environment compared to cometabolism. Materials and Methods: Enumeration of chlorinated methane utilizing bacteria in drinking water was carried out by most probable number (MPN) method in two steps. First, the presence and the number of methylotroph bacteria were confirmed on methanolcontaining medium. Then, utilization of dichloromethane was determined by measuring the released chloride after the addition of 0.04 mol/L of it to the growth medium. Also, the effect of nanosilver particles on biodegradation of multiple chlorinated methanes was studied by bacterial growth on Bushnell-Haas Broth containing chloroform (trichloromethane) that was treated with 0.2 ppm nanosilver. Results: Most probable number of methylotrophs and chlorinated methane utilizing bacteria in tested drinking water were 10 and 4 MPN Index/L, respectively. Chloroform treatment by nanosilver leads to dechlorination and the production of formaldehyde. The highest growth of bacteria and formic acid production were observed in the tubes containing 1% chloroform treated with nanosilver. Conclusions: By combining the two tests, a rapid approach to estimation of most probable number of chlorinated methane utilizing bacteria is introduced. Treatment by nanosilver particles was resulted in the easier and faster biodegradation of chloroform by bacteria. Thus, degradation of these chlorinated compounds is more efficient compared to cometabolism. © 2015, Ahvaz Jundishapur University of Medical Sciences.
Publication Date: 2015
Cellular Immunology (00088749)294(1)pp. 9-12
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. © 2015 Elsevier Inc.
Publication Date: 2015
Jundishapur Journal Of Microbiology (20084161)8(11)
Background: Torque Teno Midi Virus/Small Anellovirus (TTMDV/SAV) is a member of the Gammatorquevirus genus within the family Anelloviridae. It is detected in healthy, Hepatitis B Virus, Hepatitis C Virus and HIV infected individuals and also patients with acute respiratory disease in different countries, but its role in clinical diseases and its full geographical distribution is still unclear. Objectives: The current study aimed to detect the frequency of infection with TTMDV/SAV in the sera of healthy blood donors, hepatitis C infected and HIV positive individuals in Lorestan province, Iran; and also investigate the possible role of TTMDV/SAV virus in liver diseases. Materials and Methods: Fifty two, 36, 4, and 110 serum samples from HIV positive, patients with HIV/HCV and HIV/HCV/HBV co-infections, and healthy individuals were collected in Khorramabad city, respectively. Nested-polymerase chain reaction was performed using SMAs/SMAr primers to detect TTMDV/SAV DNA. Serum aminotransferases were measured. Results: In the HIV/HCV, HIV/HCV/HBV, HIV, and control cases, 29 (80.5%), 3 (75%), 43 (82.7%), and 16 (14.5%) were positive for DNA of TTMDV/SAV, respectively. In the HIV/HCV infected cases and HIV positive cases the level of Alanine aminotransferase (ALT) and Aspartate aminotransferase (AST) were not significantly different in TTMDV/SAV infected and non-infected individuals (P > 0.05). Conclusions: Although significant differences (P < 0.01) were observed in the frequency of TTMDV/SAV between healthy controls and each of the HIV positive and HIV/HCV co-infected individuals, no significant difference was observed between HIV positive and HIV/HCV co-infected cases, which may be due to HIV associated immunodeficiency. This is the first time that TTMDV/SAV is reported in HIV infected individuals worldwide. Interpretation of the high frequency of the virus (82.7%) in HIV cases needs more detailed studies. © 2015, Ahvaz Jundishapur University of Medical Sciences.
Publication Date: 2014
Gene (18790038)551(2)pp. 222-229
To date, a few numbers of bacteriophages that infect Lactococcus garvieae have been identified, but their complete genome sequences have not yet been investigated. For the first time, herein, the complete DNA sequence of a new phage of L. garvieae (phage WP-2) is reported and analyzed. The morphological characteristics indicated that the phage had a small isometric head along with a short and non-contractile tail, suggesting that WP-2 belongs to the family Podoviridae. Bioinformatic analysis revealed that phage WP-2 can be classified as a new member of Ahjdlikevirus in the Picovirinae subfamily because it had a small dsDNA of 18,899. bp with 24 open reading frames and a protein-primed DNA polymerase. The phage nucleotide sequence and predicted protein products have been identified to share very limited evidence of homology with complete genome and proteome of other phages. To our knowledge, this is the first Ahjdlikevirus bacteriophage which can infect a member of the Lactococcus genus. © 2014 Elsevier B.V.All rights reserved.
Publication Date: 2014
Archives of virology (14328798)159(9)pp. 2537-2540
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.
Publication Date: 2014
Jundishapur Journal Of Microbiology (20083645)7(5)
Background: Cervical cancer is the second most common cancer in women worldwide. Recent studies show that human papillomavirus (HPV) DNA is present in all cervical carcinomas and in some cervicitis cases, with some geographical variation in viral subtypes. Therefore determination of the presence of HPV in the general population of each region can help reveal the role of these viruses in tumors. Objectives: This study aimed to estimate the frequency of infection with HPV in cervicitis, cervical adenocarcinoma, intraepithelial neoplasia and squamus cell carcinoma samples from the Isfahan Province, Iran. Patients and Methods: One hundred and twenty two formalin fixed paraffin embedded tissue samples of crevicitis cases and different cervix tumors including cervical intraepithelial neoplasia (CIN) (I, II, III), squamus cell carcinoma (SCC) and adenocarcinoma were collected from histopathological files of Al-Zahra Hospital in Isfahan. Data about histopathological changes were collected by reexamination of the hematoxylin and eosin stained sections. DNA was extracted and subjected to Nested PCR using consensus primers, MY09/MY11 and GP5+/GP6+, designed for amplification of a conserved region of the genome coding for L1 protein. Results: In total 74.5% of the tested samples were positive for HPV. Amongst the tested tumors 8 out of 20 (40%) of CIN (I, II, III), 5 out of 21 (23.8%) of adenocarcinoma cases and 78 out of 79 chronic cervicitis cases were positive for HPV. Conclusions: The rate of different carcinomas and also the rate of HPV infection in each case were lower than other reports from different countries. This could be correlated with the social behavior of women in the area, where they mostly have only one partner throughout their life, and also the rate of smoking behavior of women in the studied population. On the other hand the rate of HPV infection in chronic cervicitis cases was much higher than cases reported by previous studies. This necessitates more attention to the role of human papillomaviruses in the their induction in the studied area. © 2014, Ahvaz Jundishapur University of Medical Sciences; Published by Kowsar Corp.
Publication Date: 2014
Iranian Journal Of Blood And Cancer (20084609)6(3)pp. 119-126
Background: Recently, some new viruses have been identified for their association with hepatitis which Torque Teno Mini Virus being among them. The aim of this study was to determine the frequency of Torque Teno Mini Virus in healthy individuals and hepatitis B and C patients in Isfahan, Iran. Materials and Methods: One hundred serum samples of healthy individuals from Isfahan Blood Transfusion Organization were collected. A total of 25 human serum samples from hepatitis B and 25 samples from hepatitis C infected patients were also collected from Mahdieh diagnostic laboratory in Isfahan, Iran. Viral DNA was extracted and Torque Teno Mini Virus DNA was detected using a nested PCR with primer sets designed for a conserved region of the Torque Teno Mini Virus genome. PCR and Reverse transcriptase PCR were used for detection of HBV and HCV respectively. Results: Torque Teno Mini Virus -DNA was detected in 17% of healthy individuals. It also was detected in 20% and 48% of serum samples from hepatitis B and C infected individuals, respectively. The frequency of Torque Teno Mini Virus was significantly higher in hepatitis C patients versus healthy individuals (P < 0.05). Also, the frequency of TTMV in hepatitis C patients was significantly higher than hepatitis B patients (P < 0.05). Conclusions: The difference in Torque Teno Mini Virus frequency between the hepatitis C and healthy group was significant (P< 0.05). The etiology of the higher infection rate in hepatitis C individuals needs to be determined.
Publication Date: 2014
Archives of Virology (03048608)159(11)pp. 2909-2915
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. © 2014, Springer-Verlag Wien.
Publication Date: 2014
Clinical and Experimental Neuroimmunology (17591961)5(1)pp. 77-83
Objectives A growing body of evidence shows that leptin acts as a pro-inflammatory cytokine in autoimmune disorders and is related to multiple sclerosis (MS) pathogenesis. The present study was an analysis of serum leptin levels among healthy volunteers and patients with different subtypes of MS, opticospinal MS (OSMS) and neuromyelitis optica (NMO). Methods Leptin concentrations in the sera of 121 healthy volunteers and 201 patients with different subtypes of MS, as well as in 27 NMO and 27 OSMS, were measured. Results Significant differences in leptin serum levels were observed between healthy volunteers, and MS, OSMS and NMO patients (P < 0.001). Furthermore, leptin serum concentration was in correlation with expanded disability status scale (EDSS) in primary progressive MS and secondary progressive MS groups. Interestingly, while the female-to-male ratio of leptin was approximately 2 in each group, the NMO female patients showed sevenfold higher levels of leptin than males. Conclusion The present results show that leptin concentration is important in the pathogenesis of different neuroinflammatory diseases of the central nervous system, in particular NMO. © 2014 Japanese Society for Neuroimmunology.
Publication Date: 2014
Aquaculture International (09676120)22(4)pp. 1469-1480
Lactococcosis, a significant emerging disease of fish caused by Lactococcus garvieae, has become one of the devastating problems due to its serious economic damage in aquaculture. The aim of this study was to isolate and characterize a lytic phage infecting L. garvieae as a potential bioagent for the treatment of lactococcosis. In this regard, one strain of L. garvieae was isolated from diseased rainbow trout, and then, following biochemical and molecular identifications, its specific phage, WWP-1, which was able to destroy L. garvieae cells through the lytic cycle, was isolated from a municipal wastewater sample. Transmission electron microscope revealed that the isolated phage possesses an icosahedral head and a non-contractile short tail, resembled to members of the family Podoviridae. Moreover, phage WWP-1 represented optimal antibacterial activity at temperatures ranging from 15 to 30 °C, suggesting that it could be very effective at rainbow trout rearing temperature. Restriction profile analysis revealed that NdeI can digest WWP-1 genome while EcoRI, EcoRV, and BamHI were incapable of cutting its DNA. According to the in vivo experiment result, WWP-1 could decrease mortality rate of infected rainbow trout in aquaculture. The results suggest that this naturally occurring bacteriophage could be considered as a promising agent to control the disease caused by L. garvieae strains in rainbow trout rearing. © 2014 Springer International Publishing Switzerland.
Publication Date: 2014
Journal of Pure and Applied Microbiology (09737510)8(1)pp. 797-806
Yoghurt is an important source of Lactobacilli and other lactic acid bacteria. In Iran, native yoghurts naturally contain some valuable probiotic lactobacilli. The aim of this study was to isolate and identify Lactobacilli strains with high quality probiotic potentials from all kinds of yoghurts made by traditional dairy producers of Isfahan, Iran. Lactobacillus strains were isolated from various traditional yoghurts. Then probiotic properties of the selected lactobacilli were determined. Strong acid and bile salt tolerant strains were considered as high quality probiotics, and identified at the species level by biochemical tests and were further identified according to16s rRNA specific sequences. A total of 82 Lactobacillus strains were isolated. Fourteen strains were graded as high quality probiotic lactobacilli (strong acid and bile tolerants). The majority of high quality probitics were sensitive to the most commonly used antibiotics. The phenotypic characterization of high quality probiotics resulted in identification of different lactobacillus species including 3 Lactobacillus casei, 8 Lactobacillus plantarum, and 3 Lactobacillus pentosus. The results of 16s rRNA gene sequencing assay confirmed the biochemical tests. The results indicated that, the Lactobacilli isolated from native yoghurts of Isfahan have great probiotic potentials, and could be used for production of different probiotic products.
Publication Date: 2014
Iranian Journal Of Blood And Cancer (20084595)7(1)pp. 25-29
Background: Approximately 600,000 deaths occur every year as a result of the acute and chronic consequences of hepatitis B virus infection. Ten different hepatitis B virus genotypes have been identified with distinct geographical distributions. Different clinical outcomes, including the rate of mutations, development of hepatocellular carcinoma, chronicity, response to treatment, transplantation rejection and occult infections, are affected by specific genotypes. The aim of the present study was to determine the frequency of genotype D of the virus in Isfahan, Iran. Patients and Methods: In this study primarily hepatitis B virus positive patients were identified by the detection of HBs antigen using ELISA test and then PCR was used as a confirmatory test. Fifty five patients that were identified as hepatitis B positive were tested for hepatitis D genotype using type - specific PCR. Results: The patients included 30 (54.5%) females and 25 (45.5%) males. In total, frequency of genotype D was 29 out of 55 cases (52.7%). Genotype D was detected in 19 (63.3%) females and 10 (40.0%) males indicating no statistically significant difference. The difference in the level of liver enzymes in patients infected with genotype D and non-genotype D hepatitis B virus were not significant. Conclusion: In the present study the frequency of genotype D among patients with hepatitis B virus infection in Isfahan, Iran, was 52.7%. No significant relation was observed between the level of liver enzymes and infection with the genotype D. © 2014, Iranian Pediatric Hematology and Oncology Society. All rights reserved.
Publication Date: 2013
Jundishapur Journal Of Microbiology (20083645)6(2)pp. 144-149
Background: Staphylococcus aureus is associated with different infections ranging from skin and soft tissue infections to endocarditis and fatal pneumonia. S. aureus is still the most common bacterial species isolated from inpatient specimens and the second most common from outpatient specimens. Today, methicillin resistant S. aureus (MRSA) isolates are present in the hospitals of most countries and are often resistant to several antibiotics. Objectives: This study was conducted from 2007 to 2011 to detect prevalence and antibiotic resistance patterns among MRSA and methicillin sensitive S. aureus (MSSA) isolated from hospitals in Tehran, Iran. Materials and Methods: Totally 726 isolates of S. aureus were collected from three referral hospitals in Tehran. All isolates were identified at the species level by standard biochemical tests. Susceptibility to eighteen antibiotics was determined by disc diffusion method. Then oxacillin and vancomycin minimum inhibitory concentration (MIC) of resistant isolates was also determined using Etest. mecA gene was detected using specific primers. Results: A total of 216 (30%) strains were found to be MRSA isolates. The highest antibiotic resistance was to penicillin, clindamycin, tobramycin and tetracycline respectively. Ninety three and 61% of MRSA and MSSA isolates were multidrug resistant (MDR) respectively. However, no strain was resistant to vancomycin, synercid, linezolid and chloramphenicol. Sixty nine percent of MRSA isolates showed high level of resistance to oxacillin (MIC ≥ 256 μg/mL). mecA gene was detected among all MRSA isolates. Conclusions: Although the frequency of MRSA isolates in the current study was low, resistance to other antibiotics was high and most of the isolates were found to be MDR. Regular surveillance of hospital-associated infections and monitoring of their antibiotic sensitivity patterns are required to reduce MRSA prevalence. High frequency of MDR isolates of S. aureus could be considered as an urgent warning for public health. © 2013, Kowsar Corp.; Published by Kowsar Corp.
Publication Date: 2013
Jundishapur Journal Of Microbiology (20083645)6(1)pp. 80-85
Background: Staphylococcus aureus is a common cause of infections among humans and animals and it is known as a community-acquired and nosocomial pathogen. Most of the isolates contain lysogenic phages which are responsible for production of various virulence factors such as enterotoxins, staphylokinase, β-lysin, lipase, exfoliative toxin A and Pantone-vlaentine leukociden (PVL). All staphylococcus isolates are classified in 6 Objectives: This study was performed to detect the presence of bacteriophage types and determine antibiotic resistance pattern of methicillin resistant S. aureus (MRSA) isolates obtained from a tertiary care hospital in Tehran, Iran from 2008 to 2010. Materials and Methods: A total of 968 S. aureus isolates were collected from a tertiary care hospital in Tehran, Iran and identified at the species level by PCR and biochemical Susceptibility to 17 antibiotics was determined. Then oxacillin and vancomycin minimum inhibitory concentration (MIC) of the resistant isolates were determined. Multiplex-PCR was used to detect 6 classes of prophages. Results: Out of the 968 isolates 247 isolates were resistant to methicillin. Highest antibiotic resistance was seen to penicillin (100%), erythromycin (89.8%), kanamycin (89.4%), ciprofloxacin (88.6%) and tobramycin (87.4) respectively. None of the MRSA isolates showed resistance to vancomycin, synercid and linezolid. MIC results indicated that 46.1 and 4.4% of isolates with high (MIC ≥ 128 μg/ml) and low level (MIC ≥ 4 μg/ml) showed resistance to oxacillin, respectively. Four different phage types and eight patterns of prophages were detected. All MRSA isolates contained at least one prophage. Totally, 2.8, 69.2 and 27.9% contained 5, 4 and 3 different prophage types, respectively. Conclusions: High prevalence of different classes of prophages indicating the potential to carry a broad spectrum of virulence factors and high oxacillin resistance were found in the MRSA isolates. Detection of SGF phage in 100% of the isolates indicates the ability of these isolates to produce virulence factors. © 2013 Ahvaz Jundishapur University of Medical Sciences; Published by Kowsar Corp.
Publication Date: 2013
Intervirology (14230100)56(4)pp. 265-270
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. Copyright © 2013 S. Karger AG, Basel.
Publication Date: 2012
Journal of Pure and Applied Microbiology (09737510)6(2)pp. 621-625
Enterococci are part of the normal flora of humans and animals. Recently, Enterococci have caused great concern due to developing of resistance to many antimicrobial agents. Their ubiquity in animal and human digestive tracts , medical importance, frequent multiple antibiotic resistance and their seemingly unlimited capacity for horizontal gene transfer via numerous mobile genetic elements make this bacterial group ideal for investigating the ecology of antibiotic resistance. The aim of this study was to investigate and identify the prevalence of VRE (Vancomycin Resistant Enterococcus) within isolated Enterococci taken from a number of Tehran Livestock husbandry units. Putative Enterococci (n = 242) were isolated on Membrane Filter Enterococcus Selective Agar Medium , supplemented with 2 , 4 and 8 μgr/ml vancomycin from cow samples. A total isolates passed the standard biochemistry tests for the genus and species as well as specific genus and species primers. The antibiotic susceptibility was determined by the disc diffusion method for 6 antibiotics. MIC of vancomycin was also done using broth micro-dilution assay by CLSI recommendations. Results showed that 138, 90 and 14 of the isolates were E. faecium , E. feacalis and E. gallinarum, respectively. 41, 25, 18, 10, 21 and 22 of the isolates were resistant to vancomycin , tetracycline, gentamicin , chroramphenicol, ciprofloxacin and erythromycin, respectively. An MIC test on 65% of the isolates was > 256 μgr/ml. Diversity of VRE isolates was restricted to 3 species. E. faecium had high resistance to a broad range of antibiotics. The results of this study suggest that more attention should be paid to the livestock samples as a reservoir of resistance elements. Surveillance of VRE reservoirs in animal husbandry to clarify the mechanism of transfer are urgently required.
Publication Date: 2012
Archives of Virology (03048608)157(2)pp. 291-295
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. © 2011 Springer-Verlag.
Publication Date: 2012
Archives of Virology (03048608)157(9)pp. 1807-1811
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. © 2012 Springer-Verlag.
Publication Date: 2012
Iranian Journal Of Basic Medical Sciences (20083874)15(5)pp. 1097-1101
Objective(s) In this study we investigated the expression of GABAA receptor subunits during brain development. These receptors may change in the embryonic chick forebrain. Materials and Methodes The expression levels of four types of GABAA receptor gamma subunits (γ1, γ2, γ3 and γ4) were quantified in the embryonic chick forebrain at 32 hr, 3, 7, 14, and 20 days of incubation and day one after hatching. The expression level of mRNA in the forebrain of embryonic chicken was measured using real-time RT-PCR. Results The expression level of each subunit increased gradually with development and reached a plateau on 20th day of embryonic development. A reduction was observed on day one after hatching in all gamma subunits. Conclusion This may explain the different physiological and pharmacological function of GABA receptor gamma subunits before and after hatching.
Publication Date: 2012
Jundishapur Journal Of Microbiology (20084161)5(4)pp. 550-554
Background: In recent decades bacteriophages have been used as treating agents against some pathogens. Also, bacteriophages could be considered as alternatives of antimicrobial agents in plant protection. Objectives: This study aimed to isolate a pathogen from potato (Solanum tubersom) tubers (Pseudomonas putida), and of its specific bacteriophage from soil and wastewater. Materials and Methods: Infected samples of soft rot potato tubers were collected from Flavarjan farms (Isfahan province, Iran) to isolate and identify P. putida by biochemical and molecular methods. Soil and wastewater samples were obtained locally to isolate the bacteriophage attributed to P. putida. Soil suspension was centrifuged and then filtrated by 0.2 micrometer Millipore filter. The wastewater was directly filtrated by 0.2 micrometer filter after centrifugation. After incubation of isolated bacteria together with phage contained solution, plaques were detected in nutrient agar. Subsequently, clearance of P. putida liquid culture incubated by added filtrated bacteriophage was observed. The structure and morphological characteristics of P. putida related bacteriophage was remarked by transmission electron microscopy (TEM). Results: Isolated strain sda2 was identified as Pseudomonas putida with related accession number HQ423667. The result showed that isolated bacteriophage belonged to Cystoviridae family. Conclusions: We isolated a new strain of P. putida as well as a novel bacteriophage through which potato disease caused by the bacterium could be treated. © 2012 Ahvaz Jundishapur University of Medical Sciences; Published by Kowsar Corp.
Publication Date: 2011
Journal of Isfahan Medical School (10277595)29(147)pp. 912-921
Background: SEN virus is mainly transmitted via blood and its relationship with different diseases is under investigation. The prevalence of SEN-V in healthy individuals, including blood donors, clearly differs geographically. Female genital lesions range from mild infections without symptoms such as chronic cervicitis to various cancers. This study was done to investigate the possible correlation of SEN virus infection with cervicitis and cervical tumors. Methods: In this study, 117 samples of formalin fixed paraffin embedded biopsies of uterus and cervix were subjected to nested PCR to detect D and H strains of SEN-V. After extraction from gel, two PCR products were sequenced and compared with the sequences in Gene Bank. Fisher's exact test was used for statistical analyses. Findings: High homology with D and H strains was observed. SEN-V was detected only in 4 cases of 45 cancer samples which one was positive for H, one for D and one for both strains. Out of 72 cervicitis cases 13 and 9 subjects were positive for D and H strains respectively. Compare to cervical cancers, the frequency of infection was higher in cervicitis cases (30%) (P > 0.05). Conclusion: May be there is a correlation between SEN-V infection with cervicitis, especially in chronic cases with unknown etiology which needs more studies to be proved.
Publication Date: 2011
New Microbiologica (11217138)34(3)pp. 263-274
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.
Publication Date: 2011
International Endodontic Journal (13652591)44(6)pp. 499-504
Aim To determine mRNA expression levels of Nav 1.8 in inflamed pulps of rats. Methodology Inflammation was induced by creating pulp exposures in rat incisors. Histopathological changes in the induced pulpitis were evaluated 1, 3, 7 and 10days after exposure. Using real-time PCR, the relative mRNA expression levels of Nav 1.8 in the inflamed rat dental pulp was determined. Results At day 1, no inflammation was evident in the pulp tissue, whereas increased levels of inflammatory responses were identified at day 3 and day 7. No pulpal inflammation was evident in day 10 or in the control group. Nav 1.8 was expressed in the rat dental pulp and increased at day 3 and day 7. Time course study of dental pulp inflammation indicated that differences in relative mRNA expression levels of Nav 1.8 were correlated with the severity of inflammation. Conclusions Nav 1.8 channels seem to be expressed significantly more under a temporal control so as to be associated with a severity of inflammation during pulpitis. As Nav 1.8 has been considered to have a role in neuropathic pain, its expression within dental pulp may contribute to the pathophysiology of tooth pain. © 2011 International Endodontic Journal.
Publication Date: 2011
International Journal of Neuroscience (15635279)121(8)pp. 437-441
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 2,860 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. © 2011 Informa Healthcare USA, Inc.
Publication Date: 2010
Virology Journal (1743422X)7pp. 1-1
BACKGROUND: SEN virus is a blood-borne, circular ssDNA virus and possessing nine genotypes (A to I). Among nine genotypes, SENV-D and SENV-H genotypes have the strong link with patients with unknown (none-A to E) hepatitis infections. Infection with blood-borne viruses is the second important cause of death in thalassemic patients. The aim of this study was to determine the frequency of SENV-D and SENV-H genotypes viremia by performing nested-PCR in 120 and 100 sera from healthy blood donors and thalassemic patients in Guilan Province, North of Iran respectively. Also, to explicate a possible role of SEN virus in liver disease and established changes in blood factors, the serum aminotransferases (ALT and AST) and some of the blood factors were measured. RESULTS: Frequency of SENV-D, SENV (SENV-H or SENV-D) and co-infection (both SENV-D and SENV-H) viremia was significantly higher among thalassemic patients than healthy individuals. Frequency of SENV-H viremia was significantly higher than SENV-D among healthy individuals. In comparison to SENV-D negative patients, the mean of mean corpuscular hemoglobin was significantly higher in SENV-D positive and co-infection cases (P < 0.05). The means of AST and ALT were significantly higher in thalassemic patients than healthy blood donors, but there were not any significant differences in the means of the liver levels between SENV-positive and -negative individuals in healthy blood donors and thalassemic patients. High nucleotide homology observed among PCR amplicon's sequences in healthy blood donors and thalassemic patients. CONCLUSIONS: The high rate of co-infection shows that different genotypes of SENV have no negative effects on each other. The high frequency of SENV infection among thalassemic patients suggests blood transfusion as main route of transmission. High frequency of SENV infection in healthy individuals indicates that other routes rather than blood transfusion also are important. Frequency of 90.8% of SENV infection among healthy blood donors as well as high nucleotide homology of sequenced amplicons between two groups can probably suggest that healthy blood donors infected by SENV act partly as a source of SENV transmission to the thalassemic patients. In conclusion, SENV-D isolate in Guilan Province may be having a pathogenic agent for thalassemic patients.
Publication Date: 2009
Iranian Journal of Clinical Infectious Diseases (20081081)4(3)pp. 143-150
Background: Staphylococcus aureus is a major pathogen in hospital setting and in the community and causes a wide range of diseases. MRSA infection has recently become a serious problem in anti-microbial chemotherapy. The aim of the study was to detect and analyze the antibiotic diversity and isolation of methicillin resistance gene (mecA) of S. aureus isolated from Tehran hospitals as a rapid and reliable method. Patients and methods: We studied 585 isolates of staphylococcus spp. recovered from patients at 3 clinical centers in Tehran from October 2005 to October 2006. Antibiotic susceptibility test of isolates was achieved with 13 antibiotics by disc diffusion. The MIC of methicillin was also performed by broth micro dilution assay. PCR was used for detection of mecA gene. Results: Totally, 321 (54.7%) isolates were identified as S. aureus. 66, 65, 88, 88, 100, 41, 38, 41, 0, 40, 93, 20 and 64% of S. aureus isolates were resistant to kanamycin, cephotaxim, methicillin, oxacillin, ampicillin, erythromycin, clindamycin, sulphamethoxazole-trimethoprime, vancomycin, chloramphenicol, ciprofloxacin, gentamicin and tetracycline, respectively. All MRSA and 63% of intermediate isolates carried mecA gene. Conclusion: In contrary to other studies in Iran, the prevalence of methicillin resistance is rising up in Tehran and most of MRSA isolates were resistance to 5 antibiotics at least. Vancomycin, chloramphenicol, gentamicin and clindamycin are the most effective antibiotics. All MRSA isolates had mecA gene with different expression. Detection of mecA gene is a rapid and reliable method for identification of MRSA isolates. ©2009 IDTMRC, Infectious Diseases and Tropical Medicine Research Center.
Publication Date: 2006
Journal of Poultry Science (13467395)43(4)pp. 408-414
Heat-resistant strains of Newcastle disease virus, such as strain V4, are being used as vaccines to protect flocks of rural chickens in developing countries. Sometimes these vaccines are administered on food, a procedure that has yet to be optimized. Experiments were undertaken to determine the anatomical sites of initial infection with orally administered V4 virus, and the sites of initial viral replication. During the first 24 hours after introduction of V4 into the mouth of 3-weeks-old chickens, virus was isolated by egg inoculation from oesophagus, crop and trachea and less frequently from proventriculus. V4 virus was not isolated from other parts of the digestive tract, nor from blood. Evidence for viral replication was sought by egg inoculation and by immunohistochemistry 4 to 10 days after V4 strain was placed in the crop. Virus was detected most frequently and at highest titre from jejunum, ileum and caecum 6 days after infection. There were also isolations from blood, 4 days after infection. Viral antigens were detected in epithelial cells in most parts of the digestive tract and in some cells, possibly lymphocytes or macrophages, in lamina propria. It is suggested that V4 vaccine virus, given orally, attaches to cells in the upper digestive tract. Viral replication occurs mainly in cells of the lower digestive tract, probably reaching these sites as the result of a viraemia. © 2006, Japan Poultry Science Association. All rights reserved.
