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The dry bubble disease, caused by Lecanicillium fungicola, is an important fungal disease of white button mushroom in Iranian mushroom production farms. Twenty-three isolates of the pathogen collected in Iran and identified as L. fungicola var. fungicola, were compared for genetic polymorphism, diversity in growth rate and virulence. Ten Universal Rice Primers (URP) were used to evaluate the genetic diversity of L. fungicola var. fungicola. URP analysis showed that the genetic diversity of Iranian isolates was low (average 10 % over the 10 primers used) and that they were almost clonal. Relative correlations between geographical origins of isolates and molecular grouping were observed but there was no correlation between mycelial growth rate, virulence assays and URP patterns. Significant differences were observed between isolates based on mycelial growth rate and virulence assays. The high level of genetic homogeneity is attributed to the effect of fungicides used for control of the mushroom diseases which might have imposed a significant selection pressure on the fungal populations.  

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The genus Pseudomonas consists of more than 120 species that are ubiquitous in moist environments such as water and soil ecosystems and are pathogenic to animals and humans. Within the genus of Pseudomonas, P. aeruginosa is most frequently associated with human infections. The bacterium is regarded as an opportunistic pathogen, primarily causing nosocomial infections in immunocompromised patients. The existing knowledge regarding the pathogenesis of P. aeruginosa has mainly been obtained through studying clinical isolates; particularly those involved in causing chronic lung infection in cystic fibrosis patients. Nosocomial infections commonly associated with P. aeruginosa include ventilator-associated pneumonia, catheter-associated urinary tract infections, wound infections in severe burn patients and septicaemia with their pathogenesis shown to be multifactorial. The bacterium is also capable of producing a number of toxins via the type III secretion system, as well as secreting enzymes and proteins including elastase, phospholipase C and siderophores. However, P. aeruginosa is also a waterborne pathogen, commonly found in environmental waters as well as in other sources such as sewage treatment plants. The public health implication of these bacteria whilst in the environment has not been fully investigated. Here we review our present knowledge about the pathogenesis of P. aeruginosa in clinical settings and the environment. 

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Background: Masske is a traditional Iranian butter made from yoghurt. The first aim of this study was to isolate and identify the nonlactic pathogenic microflora by culture and molecular methods of identification, and the second purpose was to identify genetic similarity of the isolated bacteria in Masske.
Materials and Methods: In order to detect pathogenic dominant indicator microorganisms, a number of 150 bacterial isolates from three Masske samples, which may comprise the repetitive isolates and could grow on appropriate media for Staphylococci and E.coli, were classified into 8 groups according to their phenotypic characterization followed by chemical tests. Then 2approximately similar isolates from each group were chosen (total 18 isolates; we selected 3 isolates from 2 groups of eight), and the sequencing of 16S rRNA gene was done for subsequent analysis.
Results: Among 18 bacterial isolates, Staphylococcus hominis was the most frequently isolated species during the manufacture of Masske as the presence of this bacterium was confirmed in 14 out of 18 samples. Also, the presence of Staphylococcus epidermidis and Escherichia coli was identical across the samples (for each one, 2 out of 18).
Conclusion: Our results based on cultural and molecular methods suggest making some improvements to the hygiene of Masske manufacture due to the high population of minor pathogens.

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Background: Salmonella typhimurium is one of the most important species of Salmonella that is intracellular parasite and attacks host mucus membrane. These bacteria can cause gastroenteritis, and their main transmission route is water, poultry, meat, egg, and raw food. The aim of this study was to detect three virulence genes associated with S. typhimurium named invA, STM4497, and fliC183 genes by Multiplex PCR method.
Materials and Methods: 183 samples of poultry were collected from food products in Zanjan (Iran) and cultured in BPW (Buffered Peptone Water) for 18 hr and at 37°C, and in RVS broth (Rappaport Vassiliadis Soya) for 6 hr at 41.5°C. After amplification of genomic DNA by Multiplex PCR method, occurrence of pathogen contamination was checked and compared with standard strain.
Results: From the total of 183 collected samples, 52(28.4%) samples were positive for S. typhimurium. The frequency of STM4497, fliC183, and invA genes were 49 (27%), 3 (2%), and 53 (29%), respectively.
Conclusion: Simultaneous detection of invA, STM4497, and fliC183 genes were recognized as a key for detection of S. typhimurium by Multiplex PCR method. 

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The aphid species, Cinara pini (Linnaeus, 1758) reported in our previous work as a new aphid on pinus trees for Iran, was described using the classic method and through analysis of COI gene sequence. In the next step, we addressed the efficiency of the entomopathogenic fungus, Lecanicillium longisporum (Zimm.) Zare and Gams strain LRC 190, on the aphid. The fungus was administered to the second instar nymphs and adults using topical application procedure. The results indicated that the entomopathogen caused 90% mortality in adults over seven days at a concentration of 10⁸ spores/ml, while the same control level was achieved for nymphs by 8 × 10⁷ spores/ml. The LC₅₀ values were obtained as 1.2 × 10⁶ and 6.9 × 10⁵ spores/ml for adults and nymphs, respectively. The present study suggests that the entomopathogenic fungus, L. longisporum could be considered as a potential candidate in biocontrol programs of C. pini. This is the first report on the pathogenicity of L. longisporum on C. pini.

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Samples of leaf, twig and fruit from ‘Mexican’ lime (Citrus aurantifolia) and grapefruit (Citrus paradisi) with symptoms of bacterial canker were collected from different provinces throughout Iran during spring and summers of 2010 and 2011. Yellow, gram-negative colonies were isolated from infected tissue samples. Results of pathogenicity assays indicated that some isolates incited tissue hyperplasia, hypertrophy and raised callus-like lesions typical of canker in hosts while other isolates stimulated flat necrotic and water-soaked lesions on leaves. Candidate samples of each group were identified according to morphological and physiological characteristics. Detections were also made using specific primers and partial sequencing of 16SrDNA for Pantoea group and gyrB for Xanthomonas group. Results showed that one group was characterized as the typical Xanthomonas citri subsp. citri strain while the other group containing most of the isolates was identified as Pantoea agglomerans. Samplings done frequently in different seasons revealed the presence of high populations of P. agglomerans with bacterial canker, especially in warmer and drier regions. These bacteria were able to incite canker-like symptoms on grapefruit seedlings and could be reisolated after two months.

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Entomopathogenic fungi produce a variety of degrading enzymes, including proteases, chitinases and lipases, to facilitate their entry through the massive barriers of insect cuticle. Isolates of the entomopathogenic fungi vary considerably in their proteolytic activity and virulence. The proteolytic activity of different isolates has been hypothesized to reflect their virulence toward the host. In this study, we evaluated the virulence and proteolytic activity of 17 Beauveria bassiana sensu lato isolates collected from different geographical regions in Iran. The selective medium D0C2 was used for isolating B. bassiana from soil samples. Casein substrate was used for protease assay. Total mortalities caused by different B. bassiana isolates through the dipping method, ranged from 25 to 60% with the highest and lowest rates for isolates BA and MITE, respectively. Our results revealed a wide variation in both proteolytic activity and virulence among the studied isolates. Additionally, we found a strong positive correlation between the proteolytic activity on Casein substrate and virulence of the isolates against the Khapra beetle, Trogoderma granarium. This finding will facilitate the screening and selection process of virulent fungal isolates as efficient agents for use in biological control programs of insect pests.  

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The antibacterial and antioxidant activities of methanol and acetone extracts of three marine algae, including Hypnea hamulosa, Gracilaria corticata and Enteromorpha intestinalis wereinvestigated.Antioxidant activities were determined by means of total antioxidant capacity, total phenolic compounds, DPPH radical scavenging activity and ferric reducing antioxidant power. Antibacterial activity was determined using a paper disc diffusion method against pathogenic bacteria, including Listeria monocytogenes, Escherichia coli and Bacillus subtilis. Acetone extract of E. intestinalis showed the highest antioxidant activity and contained the highest phenolic compounds. The highest percentage of DPPH radical scavenging activity was observed in the methanol extract of H. hamulosa (p<0.05). The highest ferric reducing antioxidant power was observed in the methanol extract of Glacilaria (p<0.05). The strongest inhibition (p<0.05) against L. monocytogenes was shown by the methanol extract of E. intestinalis and the highest inhibition against B. subtilis and E. coli was observed in the acetone extract (p<0.05). In conclusion, E. intestinalis extracts showed favorable antioxidant and antibacterial activity suggesting its application in food and pharmacological industries.

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Ninety one monoconidial Bipolaris isolates were obtained from lesions on different parts of rice in different locations of Mazandaran province during the summer of 2009. Bipolaris species were identified using morphological features such as color and shape of colony and color and size of conidia and conidiophores. The isolates were separated into two species; 85 (93.4%) isolates belonged to Bipolaris oryzae and the remaining 6 (6.6%) isolates to Bipolaris cynodontis. Therefore B. oryzae is regarded as the major cause of rice brown spot disease in Mazandaran province. In order to analyze genetic diversity among B. oryzae isolates, 71 isolates were subjected to fingerprinting analysis by rep-PCR using BOX and REP primers. In cluster analysis, 15 clonal lineages and 54 haplotypes were identified. The largest clonal lineage contained with 36 haplotypes was the most common lineage. These results also indicate a relatively high level of genetic diversity among B. oryzae isolates. Also, pathogenicity test of a few B. oryzae isolates (12 isolates) was conducted under greenhouse condition and showed that those isolates were pathogenic to rice seedlings of cv. Tarom. All isolates produced some leaf spots 24 h after inoculation.

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The main goal of this study was to evaluate the antimicrobial activity of Ziziphora clinopodioides against some food spoilage and pathogenic bacteria and determine Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC). Extract of Ziziphora was tested for its growth inhibitory and bactericidal effect on 6 Gram-negetive (Enterobacter aerogenes, Escherichia coil, Klebsiella pneumoniae, Salmonella enteriticlis, Shigella dysenteria and Pseudomonas aeroginosa) and 3 Gram-positive (Listeria monocytogenes, Staphylococcus aureus and Bacillus cereus) species. Minimum inhibitory Concentration (MIC) was determined using dilution method and minimum bactericidal concentration(MBC) was taken from the concentration of the lowest dosed test tube showing no growth on subcultured. All of microorganisms were inhibited by the extract of Ziziphora clinopodioides except Pseudomonas aeroginosa. The MIC and MBC for Gram-negetive bacteria, including Enterobacter aerogenes, Escherichia coil, Klebsiella pneumoniae, Salmonella enteriticlis and Shigella dysenteria were 1000-2000 μg/L. The MIC and MBC for Gram-positive bacteria, including Listeria monocytogenes, Staphylococcus aureus and Bacillus cereus, were 1000-4000 μg/L. Acording to the results of this study, It is applicable to use extract of Ziziphora as the natural preservatives and flavoring agents in food products.

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 Aims
 Urinary tract infection (UTI) is one of the most common infections worldwide. The aim of this study was to investigate the association between ESBLs genes and quinolone resistance in Uropathogenic Escherichia coli isolated from patients with urinary tract infection .
Materials & Methods
A total of 150 E. coli isolates were collected from patients with urinary tract infection referring to Firouzgar Hospital in Tehran, Iran. Antimicrobial susceptibility of isolates were determined by disk diffusion method. Double-disk diffusion test was performed for phenotypic identification of extended-spectrum β-lactamase- (ESBL) producing isolates. PCR was used for the detection of ESBL-encoding genes in addition to quinolone (qnr) resistance genes.

Findings

 There was a high resistance rate to most of the studied antimicrobial agents. Phenotypically, 75% of the isolates produced an ESBL enzyme and were resistant to different antimicrobial classes. In overall, 83% of the isolates carried ESBL genes, especially bla_TEM and bla_CTX-M  . 75% were positive for the quinolone resistance genes including qnrA , qnrB ,qnrS and qepA. These results indicate the association between the presence of various ESBLs genes and quinolone resistance in uropathogenic E. coli.

Conclusion

Resistance patterns show the increased incidence of antibacterial resistance in E. coli. Results of the current study indicate the high prevalence of ESBL-producing isolates and quinolone resistance genes. Simultaneous presence of genes responsible for antibacterial resistance has made the treatment of UTI more challenging than ever before.

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Blue mold disease caused by Penicillium expansum is a major post-harvest disease of apples. In this research, the biochemical basis of apple resistance to this pathogen was studied in two relatively resistant and susceptible cultivars, Granny smith and Mashhad, respectively. The activities of catalase (CAT), peroxidase (POX), superoxide dismutase (SOD) and polyphenol oxidase (PPO) enzymes and polyphenol content were compared at different time intervals of 0 to 7 days. Based on the results, fruit polyphenol content of Granny smith was higher than that of Mashhad PPO, SOD and CAT activity was higher in Granny smith than Mashhad but CAT activity decreased three days post-treatment. No detectable difference was found in POX activities in the two cultivars. It is concluded that polyphenols contribute in apple resistance to blue mold. Activation of PPO and SOD, lack of POX activity and decrease of CAT activity, all together, could lead to a toxic environment around the blue mold fungus.

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A total of ten isolates of fungi with Rhizoctonia-like mycelia were obtained from infected roots and stems of Pistachio Pistacia vera grown in a commercial nursery in Rafsanjan, Iran, during the autumn of 2011. The infected seedlings showed symptoms of chlorosis that later turned to necrosis. All of the isolates were identified as binucleate Rhizoctonia on the basis of hyphal characteristics and nuclear number. They were tested for detection of the anastomosis group, optimum growth temperature, rDNA-ITS region traits and pathogenicity on pistachio seedlings in vitro and in vivo. The analysis of hyphal anastomosis reaction was carried out with the tester isolates of binucleate Rhizoctonia AG-A, AG-Ba, AG-G and AG-F as well as multinucleate Rhizoctonia AG4 as already detected on pistachio seedlings. The optimum temperature for growth of binucleate Rhizoctonia sp.was 35 °C. In in vivo test, the symptoms of root rot were observed 30 days after inoculation and mortality happened two weeks thereafter. According to molecular characteristics and anastomosis test groups, all isolates showed greatest similarity to anastomosis group AG-F. This finding is the first report of anastomosis group F (AG-F) of binucleate Rhizoctonia, as causal agent of root and stem rot disease of pistachio in the world and Iran.

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The study was conducted to determine the distribution of the common Meloidogyne species in research stations and vegetable farms in Ibadan, south-western Nigeria. Galled roots were collected from inoculum plots of four research stations and two vegetable farms. Identification of species was based on juvenile and female morphological characters and specific SCAR primers for Meloidogyne species. The pathogenicity of M. incognita and M. javanica was evaluated at different inoculum levels on tomato in a screenhouse study. M. incognita was the dominant species encountered in research plots, although it often occurred in mixed population with M. javanica and other unidentified species. Growth parameters such as plant height, number of leaves, and yield responded negatively to increasing inoculum levels for all the cultivars except Small Fry and Celebrity. Both cultivars were categorized as resistant to M. incognita and tolerant to M. javanca. The most popularly grown tomato cultivars, Ibadan Local, Roma (Roma type) and Beske were susceptible to both species of root-knot nematodes.  

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Cercospora leaf spot caused by Cercospora beticola has a great negative impact on yield and quality of sugar beet. In the present study, pathogenic and genotypic variation of 24 C. beticola isolates collected from different regions of Iran were studied using RFLP of the Internal Transcribed Spacer (ITS-RFLP), and Random Amplified Polymorphic DNA (RAPD-PCR). Pathogenic variability and genotype × isolate interaction were evaluated in greenhouse experiments on five sugar beet cultivars (FD0018, HM1836, Puma, Eudora and Monatuna). All of the 24 isolates tested were found to be pathogenic on the cultivars with significant variation in disease severity. Results of RAPD analysis showed wide DNA polymorphism among the Iranian C. beticola isolates. Restriction pattern of the internal transcribed spacer of rDNA (ITS1-5.8-ITS4) was studied using three restriction endonucleases: EcoR1, Taq1, and Busr1. The length of undigested DNA fragment of all isolates was estimated to be 500bp without rDNA polymorphism after digestion with EcoR1 (280, 270 bp), Taq1 (330 bp) and Busr1 (240, 220, 90 bp). RAPD and ITS-RFLP markers showed the highest level of genetic diversity which confirms the variation in C. beticola detection.

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Bioassay results confirmed the role of low molecular weight phytotoxin in the patho-genesis of Verticillium albo-atrum. The metabolites separated from 21-day-old culture fil-trate by adsorption on the resin Amberlite XAD-4, and further chromatographed on Bio-Gel P2 polyacrylamide gel, induced chlorosis and necrosis on the leaflets of tomato and potato cultivars, similar to those caused by the fungus on diseased plants. Leaflets from tolerant cultivars were much less sensitive to the toxin (s) than those from the susceptible ones. In the presence of toxin(s) plant tissues and individual cells showed ion-leakage and cell death to an extent relating to the plants reaction to the fungus. The relative specificity observed during pathogenicity tests between potato and tomato and their related isolates was shown to be related to the action of toxin (s).

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Backgrounds: This study aimed to analyse hybrid Entroaggregative/Uropathogenic Escherichia coli (EAEC/UPEC) isolates. To do so, the antibiotic resistance pattern and virulence genes were investigated in E. coli strains isolated from clinical specimens of patients hospitalized in Isfahan, Iran.
Materials & Methods: Disc diffusion method was used to determine the antibiotic susceptibility pattern of EAEC/UPEC isolates. Also, virulence determinants of these isolates were determinated by singleplex and multiplex PCR.
Findings: Overall, a total of 148 E. coli isolates were collected, of which 12 (8.1%) isolates were hybrid EAEC/UPEC strains, then antibiotic susceptibility examination was operated on these strains. The higest antibiotic resistance rate was related to ofloxacin (42%), followed by trimethoprim-sulfamethoxazole (41%), ceftriaxone and cefepime (33%), and cefoxitin (17%). All the isolates showed susceptibility to fosfomycin.
Conclusion: According to the current study, since resistance to fluoroquinolones has increased in hybrid strains, monitoring the drug susceptibility of hybrid strains seems critical in Iran. Fosfomycin is considered to be the drug of choise for infections caused by multidrug-resistant (MDR) Gram-positive and Gram-negative bacteria. Fortunately, 100% of the strains were sensitive to fosfomycin.

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Beauveria bassiana (Balsamo) is one of the promising microbial control agents for the management of Oryzaephilus surinamensis (L.) Death rate, lethal time and survival expectancy were calculated for an infected population of O. surinamensis at 15, 20, 25, 30 and 35 °C. Results showed that the mean death rate under above mentioned temperatures was 0.89, 1.15, 1.40, 1.21, and 1.11 larvae/day, respectively. The values were 0.99, 1.38, 1.47, 1.18 and 1.16 insects/day for adults respectively. LT₅₀s, at the same temperatures, were 7.11, 7.04, 4.82, 6.07 and 6.89 days for larvae and 7.03, 6.31, 4.83, 5.58, and 6.55 for adults, respectively. Survival curves for both larval and adult populations were more similar at 25, 30 and 35 °C compared to 15 and 20 °C. The survival rates in infected populations were low during 3^rd and 4^th days post inoculation and decreased with a sharp slope toward the end of the experiments under different temperatures. In every case, survival curves were of the 2^nd type in which the mortality decreases in a steady linear form.

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Backgrounds: Uropathogenic Escherichia coli is one of the most important etiological agents of UTI. The aim of this study was to investigate the antibacterial effects of zinc oxide nanoparticles (ZnONPs) on aminoglycoside-resistant E. coli isolates from patients with UTI.
Materials & Methods: After identifying E. coli strains in 100 out of 250 urine samples, antibiotic susceptibility was evaluated against six antibiotic classes (with emphasis on aminoglycosides) by disk diffusion method according to CLSI-2020 guidelines. The presence of aac (6')-Ie-aph (2'') gene in isolates was investigated by PCR. Antibacterial properties and minimum inhibitory concentration (MIC) of zinc oxide nanoparticles were evaluated by agar well diffusion and broth microdilution assays, respectively.
Findings: Among 100 E. coli isolates, the highest and lowest antibiotic resistance rates were observed against tetracycline (70%) and ofloxacin (10%), respectively. Of 30 gentamicin-resistant E. coli isolates, 17 (56.5%) isolates harbored the aac (6')-Ie-aph (2'') gene. In agar well diffusion assay, 22 (74%) gentamicin-resistant isolates were eliminated by zinc oxide nanoparticles at a concentration of 150 mg/L, while ZnONPs at 300 mg/L could eliminate all gentamicin-resistant isolates. Furthermore, ZnONPs could inhibit all bacteria at a concentration of 200 μg/mL (MIC₉₀ ≥ 100).
Conclusion: Spread of the aac(6')-Ie-aph(2'') gene could increase gentamicin resistance among E. coli strains causing UTI. Given the favorable antibacterial effects of zinc oxide nanoparticles in vitro, the clinical application of these nanoparticles in the treatment of UTIs caused by multidrug-resistant E. coli could be investigated in future studies.