Ml Of Suspension Research Articles

FIRST REPORT OF PECTOBACTERIUM CAROTOVORUM subsp. CAROTOVORUM CAUSING SOFT ROT ON WHITE HEAD CABBAGE IN TURKEY

In October of 2016, soft rotted tissue on white head cabbage (Brassica oleraceaL. var.capitata subvar. Alba) were observed in commercial fields of Samsun province. Infected tissue was macerated in Bioreba extraction bags and extract was plated on Crystal Violet Pectate (CVP) medium. Cavity forming strains on CVP were facultative anaerobic with pectinolytic ability on potato tuber slices, grew at 37°C, produced acid from α-methyl glucoside, were negative for producing indole and reducing substances from sucrose. The sequence of 1400 bp of 16S rDNA gene (Weisburget al., 1991) of strain BTC2 (GenBank accession No. MF314823) showed 99% identity to P. carotovorum subsp. carotovorumstrain ICMP 13550 (KY446059). Further confirmation of strain BTC2 was done with a 713 bp recA gene sequence (Waleron et al., 2002) (MF314822), which showed 99% similarity to P. carotovorum subsp. carotovorum strains in Genbank. Phylogenetic analysis based on 16S rDNA and recA sequences grouped our strain in the same cluster together with type strain ATCC 15713T and respective P. carotovorum subsp. carotovorum strains derived from GenBank. Pathogenicity was performed on cabbage plants (cv. Beyaz Bursa) by inoculation of 10 µl of suspension (108 cfu ml-1 ) using a syringe and hypodermic needle at the leaf axil. Inoculated plants were incubated in humid conditions at 28°C. Inky-black lesions and slimy decay developed within 72 h. To our knowledge,this is the first report of P. carotovorum subsp. carotovorum causing soft rot of white head cabbage in Turkey.

Abstract 5430: GAPDH loss in a tumorigenic human glioblastoma cell line

Abstract Background: Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is a ubiquitous, multifunctional 37 kDa enzyme best known for catalyzing the sixth step in glycolysis. GAPDH expression is so consistent that it has been granted “housekeeping” status, and is used as a standard (along with β-actin) for normalizing Western blots. While increased GAPDH expression has been described in hypoxia, diabetes, and some cancers, decreased expression is rare. Here we describe a new human glioblastoma cell line that exhibits GAPDH loss, while maintaining a high degree of malignancy. Methods: Primary cultures of human glioblastoma cells were initiated and maintained in DMEM with 10% FBS. This particular cell line (designated E297) grew readily, and was passaged more than 40 times. E297 cells were confirmed to be negative for HIV, HTLV, hepatitis B and C, CMV, and mycoplasma contamination. Cells in exponential growth were stained for routine markers, and studied by DNA flow cytometry. GAPDH expression was studied by Western blot analysis using polyclonal rabbit anti-human GAPDH IgG (Abcam, Rosemont, IL), with HeLa, U138, U87 and U251 cells (as well as β-actin staining) as positive controls. For intracranial implantation into Wistar-Furth rats (male, 10-11 weeks), cells were suspended at 20 x 106 cells/ml. Using aseptic technique, rats were anesthetized, and placed into a stereotactic frame. A burr hole was drilled 3 mm right of the bregma and 25 µl of suspension injected (5 mm depth) over 10 min. by Hamilton syringe. Rats were studied by MRI and euthanized when symptomatic. Results: E297 cells have glial/epithelioid morphology, with a doubling time of 24 + 2 hours in logarithmic growth phase, with a single DNA subpopulation with 28% S phase. They stain positively for GFAP, vimentin, bFGF, c-myc and p53, but fail to express GAPDH. 10/10 animals implanted intracerebrally developed tumors, becoming symptomatic and requiring sacrifice at 25 days (mean). Loss of GAPDH expression was confirmed by Western blot analysis. Conclusions: The E297 human glioblastoma cell line is highly aggressive and proliferates rapidly. It is tumorigenic in nonimmunosuppressed rats but lacks GAPDH protein. We conclude that GAPDH expression is not essential for glioblastoma cell proliferation under routine culture conditions, and care needs to be taken when using GAPDH as a normalization standard for Western blots of cancer cells. Citation Format: Zachary Gaertner, Yi Lu, Jinsuh Kim, Gayatry Mohapatra, Ankit Mehta, Sanjani Lakka, Herbert Engelhard. GAPDH loss in a tumorigenic human glioblastoma cell line [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5430. doi:10.1158/1538-7445.AM2017-5430

Benefits of Bifidobacterium animalis subsp. lactis Probiotic in Experimental Periodontitis.

This study evaluates effects of topical administration of probiotic bacteria of the genus Bifidobacterium on experimental periodontitis (EP) in rats. Thirty-two rats were divided into groups C (control; without EP), EP (EP only), C-HN019 (control+probiotic), and EP-HN019 (EP+probiotic). On day 0 of the experiment, animals of groups EP and EP-HN019 received cotton ligatures around mandibular first molars (MFMs). In groups C-HN019 and EP-HN019, 1 mL of suspensions containing Bifidobacterium animalis subsp. lactis (B. lactis) HN019 was topically administered in the subgingival region of MFMs on days 0, 3, and 7. In groups C and EP, topical administrations were performed using a sham suspension (without probiotic). All animals were euthanized at day 14. Gingival tissue, hemimandibles, and oral biofilm were collected. Data were statistically analyzed (P <0.05). Group EP presented greater bone porosity, trabecular separation, and connective tissue attachment loss (CTAL) as well as reduced bone volume than all other groups (P <0.05). In group EP-HN019, there were greater proportions of Actinomyces and Streptococcus-like species and lower proportions of Veillonella parvula, Capnocytophaga sputigena, Eikenella corrodens, and Prevotella intermedia-like species than group EP. Group EP-HN019 presented greater expressions of osteoprotegerin and β-defensins than group EP (P <0.05). Group EP presented greater levels of interleukin-1β and receptor activator of nuclear factor-kappa B ligand than group EP-HN019 (P <0.05). Topical use of B. lactis HN019 promotes a protective effect against alveolar bone loss and CTALs attributable to EP in rats, modifying immunoinflammatory and microbiologic parameters.

MAST CELLS CONTENT IN THE STRUCTURES OF THE ISCHEMIC MODEL WOUND ON THE 12TH DAY OF REGENERATIVE POTENTIAL STIMULATION BY AUTO- AND HETEROFIBROBLASTS AND DERMAL EQUIVALENT

Mast cell is a traditional cell for wound healing, but their content in the wound after transplantation of autoand heterofibroblasts and dermal equivalent on the basis of these cells remains poorly studied. Aim. To study the morphological structure, collagen formation, angiogenesis and content of mast cells in biopsy materials of the newly formed epidermis and dermis on the 12th day of their recovery in a model ischemic wound after administration of auto- and heterofibroblasts and after transplantation of dermal equivalent with heterofibroblasts. Materials and methods . The study was performed on 28 white mature mice of the C57 / B1 line aged up to 1 year. 0.4 ml of suspension with 1.33 million fibroblasts in growth medium DMEM F12 (Lonza) and in dermal equivalent were administered around and into the bottom of skin surgical model wound in the scapular region. The scar was embedded in paraffin and stained with hematoxylin and eosin, according to Weigert-Van Gieson, and kit for mast cells staining (Biovitrum). Results . On the 12th day epidermis is thicker after introduction of autofibroblasts, but it is more differentiated after transplantation of dermal equivalent with heterofibroblasts. Angiogenesis is also most active after the administration of autofibroblasts. Fibroblasts production of collagen fibers in granulation tissue, angiogenesis and the content of functionally active mast cells indicates the most favorable influence of autofibroblasts transplantation on the wound healing. Conclusion. The effect of dermal equivalent with heterofibroblasts differs from exposure autofibroblasts by several percent: the area of collagen fibers – by 2%, the area of blood vessels − by 5%, the index of degranulation of mast cells – by 4%.

Identification of a Facultative Bacterium Strain with the Ability to Methylate Mercury Under Both Aerobic and Anaerobic Conditions

A strain with the ability to methylate mercury under both the aerobic and anaerobic conditions was isolated from soil of the water-level-fluctuation-zone in the Three Gorge Reservoir in Shibaozhai Village, Zhongxian Country, Chongqing, China (E108°12'3″ and N30°24'36″). The soil was classified as Purple soil with a pH of 7.97 (0-20 cm depth). The isolation was performed under 1.0 mg·L-1 HgCl2 conditions. After its morphological and physiological characterization, and its phylogenetic analysis using 16S rDNA gene sequence, the strain was identified as Pseudomonas fluorescens sp., and named as Pseudomonas fluorescens XD-MeHg-B2 (GenBank accession number: KU954349). On one hand, at 30℃ under aerobic condition, the concentration of methylmercury (MeHg) in the PBS (phosphate buffer saline) solution, which was inoculated with 1×1011 cfu·mL-1 suspension of P. fluorescens XD-MeHg-B2 and an initial Hg2+ of 200 ng·L-1, was exponentially increased to 1.22 ng·L-1±0.15 ng·L-1 after 60 min incubation and then approached to the maximum of 3.85 ng·L-1±0.33 ng·L-1 160 min after incubation. The largest mercury methylation rate was 1.93%. On the other hand, at 30℃ under anaerobic condition, the concentration of MeHg in the PBS solution, which was also inoculated with 1×1011 cfu·mL-1 suspension of P. fluorescens XD-MeHg-B2 and an initial Hg2+ of 200 ng·L-1, was 2.86 ng·L-1±0.73 ng·L-1 and the largest mercury methylation rate was 1.43% 180 min after incubation. As a result, P. fluorescens XD-MeHg-B2 showed its ability to methylate mercury under both aerobic and anaerobic conditions while with a comparatively hysteretic and lower ability of mercury methylation. These results demonstrated that P. fluorescens XD-MeHg-B2 could be a promising candidate for further studies on mercury biogeochemical cycle, particularly under dry-wet alternative conditions.

In-Situ TEM Analysis of Ink-Jet Printed MnO2-Graphene for Supercapacitor Electrodes

Supercapacitors composed of nano-layered material electrodes represent a new generation of energy storage technology. Understanding and analyzing the mechanism of how the electrode material changes and degrades during the charge and discharge process is fundamental to maximizing the efficiency of supercapacitors. The primary objective of this body of work was to develop a method whereby the electrode-electrolyte interface of a supercapacitor could be analyzed by real time in-situ TEM imaging during the charge-discharge process. This experimental approach combines advanced microscopy techniques and electrochemical testing. This approach is in the preliminary stages of experimental development, with very few major published works exploring the specific use of this technique. First, an effective method for repeatable deposition of our layered electrode material was developed, using inkjet printing methods. Our selected active material, MnO2-Graphene nanoflakes suspended in Isopropanol was used as our material ink for printing. This suspension was printed onto a specialized electrochemistry TEM chip, featuring Au electrodes and a transparent SiN2 window for TEM viewing. A low concentration dispersion of 0.5 mg mL-1 was first used to optimize the printing process and to find the printing parameters that minimized the spreading of the printed line. Having fully optimized the printing process, a high concentration 10 mg mL-1 suspension of MnO2-Graphene was used to deposit the working electrodes of our micro electrochemical cell. This cell was tested with a range of potential windows and multiple material deposition thicknesses. In most potential ranges, responses from the active material were difficult to observe due to the noise created by the inherent double layer capacitance of the Au electrode. Higher amounts of deposited material produced slightly higher responses but were for the most part unobservable above the aforementioned noise. These high layer depositions were also electron opaque. During in-situ TEM CV testing of these cells, prominent dendritic growth outward from our electrodes were observed. Higher CV responses ex-situ could be achieved with higher amounts of deposited active material, overcoming the noise generated by the Au-electrolyte reactions. For in-situ tests, morphological changes could be observed with lower amounts of material, so that the individual flakes of the Solid-Electrolyte interface can be resolved via TEM analysis.

English

Silicon is the main objective of many researchers whose goal is to see its use as an inductor resistance. The action of the inductor happens in the plants, where it does not affect the pathogen directly, but the effect will contribute for its control. Therefore, the main objective of this report was to evaluate the effect of silicon on the embryonic development and the hatching of Meloidogyne javanica. For this, three sources of silicon (Silifort&reg;, Rocksil&reg; and wollastonite) in the dosage 0 (distilled water), &frac12;, 1 and 2x the producer indication, resulting in a factorial 3 x 4, were evaluated in two&nbsp; different times. In Petri dishes were add 1 ml of suspension with 1500 eggs, and eventually juveniles of M. javanica, and 9 ml of treatment, which were evaluated, in Peters chambers, the percentage of eggs one, two, tetra and multicellular, eggs containing&nbsp;juvenile&nbsp;formed, and the hatched in a period of three days and seven days after the incubation (DAI). The wollastonite had stimulated the embryonic developments, as the increase of the dosage, were not seen with the Silifort&reg; and Rocksil&reg; experiments. The increase in the dosage of wollastonite caused stimulus to the hatching in 3 DAI, while Silifort&reg; caused the reduction of hatching in 3 DAI in both experiments. In 7 DAI, there was a reduction of hatching of juveniles exposed to Silifort&reg; and Rocksil&reg;, in the experiments 1 and 2, respectively. &nbsp; Key words: Egg, silicon, root-knot nematode, juvenile.

Clinicopathological Studies on Neem and Ginger Effects as Feed Additives in Normal and E. coli Infected Weaned Rabbits

The present study was performed to investigate the clinicopathological effects of neem and ginger as feed additives. One hundred and twenty weaned White New Zealand rabbits were divided into 6 equal groups. Group (1) kept as control. Groups (2 and 5) received ration contained neem leaves daily (5% of diet). Groups (3 and 6) received ration contained ginger powder daily (2% of diet). Groups (4, 5 and 6) were experimentally infected by E. coli (O103 once orally with a dose of 3 ml of suspension containing 3x10 7 CFU at the end of the 4 th week of experiment. The results revealed normal parameters in none infected as well as neem and ginger treated groups (1, 2 and 3). However a significant decrease in the serum total protein, albumin, globulin and catalase levels (CAT) on the 1 st , 3 rd and 15 th day PI was observed in infected non treated animals (Group 4). On the other hand a significant increase in alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), malondialdehyde level (MDA), phagocytic percentage and phagocytic index were observed in the same group. In Groups 5 and 6; animals showed a significant increase in total protein, albumin, globulin, CAT and a significant decrease in the other parameters comparing with the infected group. It could be concluded that both neem and ginger can be used as feed additives in rabbit ration to enhance hepatic, renal and antioxidant activities beside cell mediated immunity .Moreover, Neem was better than ginger in amelioration of the harmful effects of E. coli infection .

Does the exposure mode to ENPs influence their toxicity to aquatic species? A case study with TiO2 nanoparticles and Daphnia magna.

Recent studies suggest that the ecotoxicity of engineered nanoparticles (ENPs) is dependent upon the treatment of ENPs in suspensions (e.g. sonication or use of solvents) and on the mode of exposure to test organisms. We conducted several bioassays with Daphnia magna in order to determine how adverse effects of TiO2 nanoparticles (n-TiO2) are influenced by experimental set-up. Several treatments were applied, including three test media, several treatments of n-TiO2 suspensions (stirring, sonication) and different exposure modes (exposure duration and volume of test suspension). No adverse effects were observed when D. magna were exposed to 50mL of suspension, regardless of TiO2 concentration (up to 250mg/L) and exposure duration. Conversely, adverse effects were observed when D. magna were exposed to 2 mL of suspension for 96 h with a 50 % effect concentration EC50 values ranging from 32 mg/L to 82 mg/L. Test media had no significant influence on the outcome of all treatments. For a better mechanistic understanding of the experimental set-up at which adverse effects were observed, the particle size of n-TiO2 in the test media was characterized throughout the test duration. These measurements revealed a fast and strong agglomeration with a secondary particle size in the order of magnitude of micrometers. Our study describes how the effects of n-TiO2 on D .magna are influenced by the duration of exposure and volume of media, highlighting the need for standardization of experimental methods.

Efficiency of treatments for controlling Trichoderma spp during spawning in cultivation of lignicolous mushrooms.

Trichoderma spp is the cause of the green mold disease in mushroom cultivation production. Many disinfection treatments are commonly applied to lignocellulose substrates to prevent contamination. Mushroom growers are usually worried about the contaminations that may occur after these treatments during handling or spawning. The aim of this paper is to estimate the growth of the green mold Trichoderma sp on lignocellulose substrates after different disinfection treatments to know which of them is more effective to avoid contamination during spawning phase. Three different treatments were assayed: sterilization (121 °C), immersion in hot water (60 and 80 °C), and immersion in alkalinized water. Wheat straw, wheat seeds and Eucalyptus or Populus sawdust were used separately as substrates. After the disinfection treatments, bagged substrates were sprayed with 3 mL of suspension of conidia of Trichoderma sp (10(5) conidia/mL) and then separately spawned with Pleurotus ostreatus or Gymnopilus pampeanus. The growth of Trichoderma sp was evaluated based on a qualitative scale. Trichoderma sp could not grow on non-sterilized substrates. Immersions in hot water treatments and immersion in alkalinized water were also unfavorable treatments for its growth. Co- cultivation with mushrooms favored Trichoderma sp growth. Mushroom cultivation disinfection treatments of lignocellulose substrates influence on the growth of Trichoderma sp when contaminations occur during spawning phase. The immersion in hot water at 60 °C for 30 min or in alkalinized water for 36 h, are treatments which better reduced the contaminations with Trichoderma sp during spawning phase for the cultivation of lignicolous species.

Evaluation of Saudi Fluorescent Pseudomonads Isolates as a Biocontrol Agent against Citrus Canker Disease Caused by Xanthomonas citri subsp citri A*

The goal of this study was to determine whetherbacterial antagonists could be used to control Xanthomonas citri subspcitri (Xcc), the causal agent of bacterial citrus canker disease. Atotal of 22 potentially bacterial antagonists isolated as epiphytes from thephylloplane of healthy citrus trees were screened for their in vitro biologicalcontrol capability against Xcc. These strains were identified as Pseudomonas fluorescens onthe basis of biochemical and physiological tests and 16S rDNA. Out of these 22potentially bacterial antagonists, five strains (KSA1, KSA9, KSA14, KSA17, andKSA20) showed high potential growth inhibition capabilities against Xcc.The KSA1 strain was selected for further studies to test its in vivocapability to control bacterial citrus canker pathogen. It was sprayed in asuspension of 107CFU ml-1on citrus leaves which were subsequently inoculated after 72 h with 108 CFU ml-1 suspension of Xcc strain JQ890095.According to the in vivo biocontrol tests, the putative antagonist KSA1significantly reduced the symptoms on the leaves of Mexican lime seedlingscompared with untreated inoculated plants.

Reação de genótipos de Capsicum ao nematoide-das-galhas

O objetivo desta pesquisa foi avaliar a reação de híbridos porta-enxerto experimentais de Capsicum annuum ['Hibrido Experimental (HE)' e seu Recíproco (HER)] e genótipos elite do programa de melhoramento da Embrapa Hortaliças de C. chinense, C. frutescense C. baccatumà Meloidogyne incognita raça 1, M. javanica e M. enterolobii. Como testemunha suscetível a M. incognita raça 1 e resistente a M. javanicafoi utilizado o híbrido de pimentão 'Magali R', e como testemunhas resistentes a M. incognita raça 1 e M. javanicaforam utilizados os porta-enxertos 'Silver' e 'Snooker'. Para confirmar a viabilidade do inóculo foi utilizado o tomateiro 'Rutgers'. Dois experimentos foram conduzidos em casa de vegetação, em delineamento inteiramente casualizado com seis repetições. Plântulas com 30 dias de idade foram transplantadas para vasos de 2,0 L contendo substrato composto de solo, areia, esterco de gado e palha de arroz carbonizada (1:1:1:1). As plantas foram inoculadas após o transplantio com a deposição de 5,0 mL de suspensão, contendo 5.000 ovos e J2 de cada espécie. Setenta dias após foram avaliados o índice de massa de ovos e de galhas, o número de ovos por grama de raiz e o fator de reprodução. Os híbridos de C. annuum apresentaram resistência a M. incognita e M. javanica e suscetibilidade a M. enterolobii, assim como as testemunhas 'Silver' e 'Snooker'. O híbrido 'Magali R' apresentou resistência somente a M. javanica. Os genótipos de C. chinense e C. baccatumapresentaram suscetibilidade às três espécies, enquanto genótipos de C. frutescens apresentaram suscetibilidade somente a M. enterolobii.Os híbridos experimentais de pimentão e a maioria dos genótipos de C. frutescens avaliados poderão ser utilizados em áreas de cultivo protegido infestadas com M. incognitaraça 1 e M. javanica. Entretanto, a busca por genótipos resistentes a M. enterolobii, bem como o desenvolvimento e a adoção de estratégias de controle é de suma importância.

The Influence Of Fusarium Ear Infection On The Maize Yield And Quality In Some Maize Hybrids Created At ARDS Turda

Abstract. Maize is a host plant for a large number of pathogens, over 50, that invade all of the plant organs from the moment of germination to the moment of harvesting, and cobs and seeds infections continue most of the times during crops storage. Pathogen agents contributes to deterioration and decrease quantity and quality of yield contributes to productions decreasing, both quantitative and qualitatively, with an average percentage of 20 - 25% in our country. The most damaging in quantitative and qualitative point of view are diseases caused by genus Fusarium spp.. The objectives of this paper are determining the productivity and the quality of corn hybrids created at ARDS Turda, in natural and artificial infection with Fusarium spp, and identification of resistant hybrids to Fusarium spp attack. In 2011-2012 the hybrids were tested in natural and artificial infection conditions. The inoculation was made on each cob using 4 ml of suspension. Following research we can assert the following conclusions: in terms of natural infections in both 2011 and 2012 the most productive hybrids were Turda Favorit (10.74 t / ha, 9.87 t / ha) and Turda Star (10.72 t / ha 8.78 t / ha). Yields hybrids in the study decreased by 0.3 t / ha - 1.4 t / ha under artificial infection. Artificial infections produce changes in biochemical composition of the grain. Hybrids analyzed starch content and oil are lower and protein percentage increased after artificial inoculation.

First Report of "Cylindrocarpon" pauciseptatum Associated with Black Foot Disease of Grapevine in Brazil.

Since 1999, the decline of American grapevines (Vitis labrusca L.) has been common in Rio Grande do Sul, Brazil (1). In August 2012, V. labrusca with black foot symptoms were collected in vineyards in the Serra Gaúcha Region. Symptomatic plants had low vigor, vascular lesions, delayed budding, and decline and death of vines. Symptomatic roots had necrotic lesions and reduced biomass. Fungal isolations were made from necrotic root and crown fragments (own-rooted cultivar) on potato dextrose agar (PDA) medium amended with 0.5 g L-1 streptomycin sulfate. Putative colonies of "Cylindrocarpon" pauciseptatum Schroers & Crous were obtained from single macroconidia isolations. Two isolates were used to confirm the identity of isolated colonies: Cy12UFSM and Cy13UFSM. After incubation in the dark for 10 days at 20°C, the isolated mycelial colonies, which were cottony white to felty in texture, became dark orange to brown. Both isolates produced chlamydospores in chains at 40 days. Chlamydospores of Cy12UFSM and Cy13UFSM were 9 to 12 μm and 5 to 11.5 μm in diameter. Sporodochia formation on carnation leaf agar (CLA) medium was observed after 30 days. To encourage development of conidia, the isolates were grown on spezieller nährstoffarmer agar (SNA) medium for five weeks at 20°C with addition of two pieces of 1 cm2 filter paper. Microconidia of Cy12UFSM were 4 to 8.5 × 3.5 to 5 μm and those of Cy13UFSM were 3.5 to 7.5 × 3 to 5 μm. Macroconida were predominantly 3-septate (Cy12UFSM was 36 to 45 × 7.5 to 9 μm and Cy13UFSM was 30 to 38 × 7.5 to 8 μm), but 1-, 2- septate macroconidia were observed. The sizes of the three spore types and colony morphology for our isolates were similar to those described by Schroers et al. (3) for "C." pauciseptatum. To further confirm the identity of Cy12UFSM and Cy13UFSM, multi-gene DNA sequence analysis (rDNA-ITS, β-tubulin, and histone H3) was conducted using primer pairs ITS1 and ITS4 (4), Bt2a and Bt2b, and H3-1a and H3-1b (2), which amplify the ITS1-5.8S rRNA-ITS2 genes, part of the β-tubulin gene, and the histone H3 gene, respectively. Sequences of these three regions had 99, 99, and 97% similarity with references sequences of "C." pauciseptatum (isolate Cy238; accessions ITS [JF735307]; β-tubulin [JF735435], and histone H3 [JF735582], respectively). To evaluate pathogenicity, 4-month-old rooted cuttings of V. labrusca cv. Bordô were inoculated with two isolates by immersing them in a conidial suspension (106 conidia ml-1) for 60 min. Ten single-vine replicates were used for each isolate, and 10 water-inoculated vines were included as controls. Thirty days after inoculation, vines were re-inoculated with 40 ml of a 106 conidia ml-1 suspension to ensure root infection. After 4 months, the inoculated plants had reduced root mass relative to controls (39.18% for Cy12UFSM and 18.27% for Cy13UFSM). Inoculated plants also had root and crown necrosis, vascular lesions, shoot decline, and vine mortality (60 and 80% mortality for Cy12UFSM and Cy13UFSM, respectively). All water-inoculated control plants remained symptomless. The fungi Cy12UFSM and Cy13UFSM were re-isolated from infected woody tissues, confirming Koch's postulates. To our knowledge, this is the first report of "C." pauciseptatum associated with black foot disease of grapevine in Brazil, which may potentially impact the sustainability of grapevine nurseries and vineyard productivity.

The Influence of Fusarium Ear Infection on the Maize Yield And Quality in Some Maize Hybrids Created at ARDS Turda

Maize is a host plant for a large number of pathogens, over 50, that invade all of the plant organs from the moment of germination to the moment of harvesting, and cobs and seeds infections continue most of the times during crops storage. Pathogen agents contributes to deterioration and decrease quantity and quality of yield contributes to productions decreasing, both quantitative and qualitatively, with an average percentage of 20 - 25% in our country. The most damaging in quantitative and qualitative point of view are diseases caused by genus Fusarium spp.. The objectives of this paper are determining the productivity and the quality of corn hybrids created at ARDS Turda, in natural and artificial infection with Fusarium spp, and identification of resistant hybrids to Fusarium spp attack. In 2011-2012 the hybrids were tested in natural and artificial infection conditions. The inoculation was made on each cob using 4 ml of suspension. Following research we can assert the following conclusions: in terms of natural infections in both 2011 and 2012 the most productive hybrids were Turda Favorit (10.74 t/ha, 9.87 t/ha) and Turda Star (10.72 t/ha 8.78 t/ha). Yields hybrids in the study decreased by 0.3 t/ha - 1.4 t/ha under artificial infection. Artificial infections produce changes in biochemical composition of the grain. Hybrids analyzed starch content and oil are lower and protein percentage increased after artificial inoculation.

Investigation of Fungi in Drinking Water Resources as a Source of Contamination Tap Water in Sari, Iran

Background and purpose: One of the most prominent concerns for the water consumers is pathogenic microorganism contamination. Wells and underground water resources are the main resources of drinking water in Sari city, Iran. The main objectives of the research project were to explore the distribution and frequency of mycoflora in wells and underground water resources of the city and their contamination effects on humans. Materials and methods: Three reservoirs and 18 wells or underground water resources were analyzed. Water samples were then filtered and analyzed according to the World Health Organization guidelines. Each filter and 0.2 ml of suspension inoculated on SDA+CG media. For fungal growth, plates were incubated at 27’C for 7-10 days. The fungi were identified by standard mycological techniques. Results: Fungal colonies were isolated from all samples. From total of 160 fungal colonies isolated from wells water, 14 species of fungi were distinguished. Rhodotorula (54.4%), Monilinia (13.7%), Alternaria (6.9%) were the most commonly isolated. Drechslera, Rhizopus, and Exserohilum (0.6%) had the lowest frequency. There was no significant difference between fungal elements isolated from three major reservoirs (P>0.05). Conclusion: This study revealed that resources of drinking water from an area have to monitored and if its fungal CFU be greater than a certain value, medical and health preventive measures should be taken before the water is used by human. In this context, public and private awareness should also be provided through the media, broadcasting, teachers and scholars. [Yousefi Z. *Aghili S.R., Ebrahimzadeh R. Salmanian B. Investigation of Fungi in drinking water resources, as a source of contamination tap water in Sari, Iran, IJHS 2013; 1(1): 84-91] http://jhs.mazums.ac.ir

First Report of Marasmiellus palmivorus Causing Post-Emergence Damping Off on Coconut Seedlings in Malaysia.

In April and June 2010, coconut seedlings with symptoms of very slow growth, yellowing of leaves, and general abnormal leaf growth were observed in germination beds in Teluk Intan, Perak, Malaysia. The roots were soft, rotten, and brown, extending upward and downward from these lesions. Rhizomorphs and basidiocarps were produced on coconut seeds near the germination eye and identified as Marasmiellus palmivorus according description by Turner (2). Three isolates were obtained by plating surface sterilized symptomatic roots and basidiocarp on malt extract agar (MEA) amended with 85% lactic acid (1 ml added to 11 of the medium). To confirm the identity of the fungus, genomic DNA was extracted from mycelia and basidiocarps of isolates and the large subunit (LSU) region was amplified and sequenced using LR0R/LR7 primers (3). All isolates had identical LSU sequences (GenBank Accession No. JQ654233 to JQ654235). Sequences were identical to each other and 99% similar to a M. palmivorus sequence deposited in the NCBI database (Accession No. AY639434).To confirm pathogenicity, three isolates of M. palmivorus that were obtained from symptomatic plant tissue was inoculated onto seeds of Malaysian Red Dwarf variety. Each isolate was grown in 100 ml of malt extract broth in 250 ml Erlenmeyer flasks and incubated at 27 ± 2°C for 5 days on an orbital shaker (125 rpm). The resulting culture was passed through two layers of sterile cloth. Mycelial suspension was obtained by blending mycelia in 100 ml of sterile water. Seeds were sterilized by soaking in 10% v/v sodium hypochlorite in distilled water for 3 min. The seeds were then rinsed three times over running tap water. The calyx portion of the seed was removed and five holes were made around the germination eye. The seeds were inoculated by injecting 2 ml of suspension into each hole. The control seeds were inoculated with sterile distilled water only. The seeds were transferred to 40-cm diameter plastic pots containing a mixture of sand, soil, and peat in the ratio of 3:2:1, respectively, and steam treated at 100°C for 1.5 h. Pots were placed in the glasshouse with normal exposures to day-night cycles, temperatures of 29 ± 4°C, and high relative humidity (85 to 95%) achieved by spraying water twice daily. After 2 months, 75% of the inoculated seeds failed to germinate. It was speculated that the artificial inoculum was higher than under germination bed conditions. Rhizomorphs and basidiocarps were produced on husk seeds near the germination eye. Seedlings that emerged successfully developed symptoms similar to those observed in the germination bed. No symptoms developed in the noninoculated seeds and seedlings. At 80 days post inoculation, basidiocarps were observed emerging from three diseased seedlings near the germination eye. Three reisolations were made on MEA from root lesions surface sterilized. Pathogenicity tests and LSU sequence analyses indicated that M. palmivorus is the causal agent of the symptoms observed on coconut seedlings. M. palmivorus was first recorded on coconuts and oil palm in the 1920s (1) and attacks the fruit and the petiole on oil palm (2). To our knowledge, this is the first report of M. palmivorus causing post-emergence damping off on coconut seedlings.

First Report of Stem and Foliar Blight of Sunflower Caused by Alternariaster helianthi in Louisiana.

During the fall of 2009, sunflower (Helianthus annuus L.) planted at the LSU AgCenter's Burden Center in Baton Rouge, LA exhibited severe stem and foliar blight symptoms. Symptoms on stems and petioles included elongated, slightly sunken lesions with dark brown margins. Leaf symptoms included irregular to circular, dark brown lesions with white centers and surrounded by a yellow halo. Several spots often coalesced to form large, blighted areas, and severely affected leaves turned yellow, followed by defoliation. The corolla and calyx exhibited similar lesions except for the yellow halo. Disease developed rapidly and the whole (100% disease incidence) field was blighted within a week following a rain (4 mm). Infected leaf and stem tissue was surface disinfested and plated on ¼-strength potato dextrose agar (PDA). Both leaf and stem tissue consistently produced dark olivaceous-to-black fungal colonies at room temperature under 12 h of fluorescent light per day. Conidia were 53 to 128 × 10 to 26 μm, borne singly on the conidiophores, hyaline to dark olivaceous, cylindrical, rounded at both ends, and with 6 to 10 transverse and 0 to 2 longitudinal septa. Conidiophores were single, unbranched, septate, hyaline to dark olivaceous, and measured 77 to 128 × 7 to 13 μm. Morphologically, the fungus was identified as Alternariaster helianthi (Hansf.) E.G. Simmons (= Alternaria helianthi [Hansf.] Tubaki & Nishih) (1). A single-spore isolate (PDC-4291) was obtained from the original culture and DNA from this isolate was extracted with a DNeasy Plant Mini Kit (Qiagen Inc., Valencia, CA). Primers ITS1 and ITS4 were used to amplify and sequence the internal transcribed spacer regions 1 and 2, and NCBI blast analysis of the 552-bp sequence (GenBank Accession No. JN208925) resulted in 100% homology with Alternaria helianthi isolated from sunflower infected with leaf spot and blight disease in India (GenBank Accession No. DQ156343). Pathogenicity was determined by inoculating 20 potted sunflower plants (Full Sun Improved TD, Fred C. Gloeckner and Company, Inc., Harrison, NY) with conidia from a 2-week-old culture of isolate PDC-4291. Each plant was sprayed with 25 ml of suspension containing 106 conidia/ml. Twenty control plants were sprayed with 25 ml of sterile distilled water. Inoculated and control plants were covered with plastic bags and maintained in a greenhouse at 28 ± 2°C. Plastic bags were removed 72 h after inoculation. Leaf spots similar to the original symptoms appeared on all 20 inoculated plants 5 days after inoculation. A few stem lesions were observed on 13 plants. Two weeks after inoculation, infected leaves turned yellow and blighted. Alternariaster helianthi (= Alternaria helianthi) was reisolated from the leaf spots and stem lesions. No symptoms developed on any of the 20 control plants. On the basis of morphology and sequence data, this pathogen was identified as A. helianthi, and to our knowledge, this is the first report of sunflower stem and foliar blight caused by A. helianthi in Louisiana. In Louisiana, sunflower is a popular ornamental that is grown in landscapes and gardens and by commercial flower growers who grow it for cut flower arrangements. Louisiana's hot, humid weather is ideal for disease development, which may discourage gardeners and commercial growers from planting sunflower. Reference: (1) E. G. Simmons. Alternaria: An Identification Manual. CBS Fungal Biodiversity Center, Utrecht, the Netherlands, 2007.

Assessment of immunity against avian colibacillosis induced by an aroA mutant containing increased serum survival gene in broilers

Colibacillosis is an important disease in the poultry industry which causes serious economic damages. As it is suggested that vaccination is one of the means to control colibacillosis, we tried to investigate the vaccine potential of a ∆aroA derivative of an O78:K80 avian pathogenic Escherichia coli containing increased serum survival gene. 490 chicks were selected as follows: For assessment of virulence of ∆aroA mutant, 30 chicks were divided into three groups and injected with 0.5ml of PBS or bacterial suspension containing either10(7) colony forming units (CFU) of mutant or parent strains via subcutaneous route. Macroscopic lesions and mortality rate were recorded in different groups during the week after challenge. For assessment of safety and immunogenicity of the ∆aroA mutant, three groups of 20 chicks were vaccinated by aerosol administration of 250 ml of suspension containing 10(8) CFU of mutant strain at days 1 and 14, while the two other groups received PBS or wild type strain. Macroscopic lesions and mortality rate were recorded in different groups until day 21. To determine whether the vaccination is protective against challenges or not, the chickens were vaccinated at days 1 and 14 and challenged intramuscularly with either a homologous or heterologous strains at day 21. Macroscopic lesions and mortality rate were recorded in different groups during the week after challenge. The results revealed that the ∆aroA mutant was slightly virulent, however it was safe and did not cause mortality, lesions or weight loss after vaccination. Antibody responses were similar in the control and mutant groups and vaccination did not induce a significant humoral immunity. The mutant could not protect chickens against both homologous and heterologous challenges. This could be due to several factors such as the high amount of maternal antibodies in the first two weeks of life, and the vaccination procedure.

Assessment of immunity against avian colibacillosis induced by an aroA mutant containing increased serum survival gene in broilers

Colibacillosis is an important disease in the poultry industry which causes serious economic damages. As it is suggested that vaccination is one of the means to control colibacillosis, we tried to investigate the vaccine potential of a ∆aroA derivative of an O78:K80 avian pathogenic Escherichia coli containing increased serum survival gene. 490 chicks were selected as follows: For assessment of virulence of ∆aroA mutant, 30 chicks were divided into three groups and injected with 0.5ml of PBS or bacterial suspension containing either107 colony forming units (CFU) of mutant or parent strains via subcutaneous route. Macroscopic lesions and mortality rate were recorded in different groups during the week after challenge. For assessment of safety and immunogenicity of the ∆aroA mutant, three groups of 20 chicks were vaccinated by aerosol administration of 250 ml of suspension containing 108 CFU of mutant strain at days 1 and 14, while the two other groups received PBS or wild type strain. Macroscopic lesions and mortality rate were recorded in different groups until day 21. To determine whether the vaccination is protective against challenges or not, the chickens were vaccinated at days 1 and 14 and challenged intramuscularly with either a homologous or heterologous strains at day 21. Macroscopic lesions and mortality rate were recorded in different groups during the week after challenge. The results revealed that the ∆aroA mutant was slightly virulent, however it was safe and did not cause mortality, lesions or weight loss after vaccination. Antibody responses were similar in the control and mutant groups and vaccination did not induce a significant humoral immunity. The mutant could not protect chickens against both homologous and heterologous challenges. This could be due to several factors such as the high amount of maternal antibodies in the first two weeks of life, and the vaccination procedure.