Baki, Hilal; Bekircan, Çağrı
A new microsporidium was isolated from Subcoccinella vigintiquatuorpunctata L. (Coleoptera: Coccinellidae), a pest of Galega officinalis L. in Turkey. Infection in larval and adult stages was systemic with mature spores produced in the midgut, gonads, Malpighian tubules and, most extensively, fat body tissues. The microsporidium was polymorphic with two sporulation sequences producing two types of spores, binucleate spores with 13-15 coils of the polar tube, and uninucleate spores with 7 coils of the polar tube that developed within a sporophorous vesicle (SPV) to form meiospores. The 16S small subunit rRNA (SSU rRNA) gene of the microsporidium was sequenced and compared with twenty-seven microsporidian sequences from GenBank. Based on the phylogenetic analysis of the SSU rRNA sequence, this microsporidium is unique within the Vairimorpha group. Morphological and genetic characters indicate that the described microsporidium is dissimilar to all known Vairimorpha species, and so is named here as Vairimorpha subcoccinellae n. sp. Copyright © 2017 Elsevier Inc. All rights reserved.
Yaegashi, Hajime; Kanematsu, Satoko
Fungi are an important component of the soil ecosystem. Mycoviruses have numerous potential impacts on soil fungi, including phytopathogenic fungal species. However, the diversity and ecology of mycoviruses in soil fungi is largely unexplored. Our previous work has shown that the soil-borne phytopathogenic fungus Rosellinia necatrix was infected with several novel mycoviruses after growing for 2-3 years in an apple orchard. In this study, we investigated whether natural infection of R. necatrix with mycoviruses occurs under limited conditions. Virus-free R. necatrix isolates were grown in a small bucket containing soil samples for a short time (1.5-4.5 months) under greenhouse conditions. Screening of dsRNA mycoviruses among 365 retrieved isolates showed that four, including 6-31, 6-33, 6-35, and 7-11, harbored virus-like dsRNAs. Molecular characterization of the dsRNAs revealed that three retrieved isolates, 6-31, 6-33, and 6-35 were infected with a novel endornavirus and isolate 7-11 is infected with a novel partitivirus belonging to the genus Alphapartitivirus. These novel mycoviruses had no overt biological impact on R. necatrix. Overall, this study indicates that natural infections of R. necatrix with new mycoviruses can occur under experimental soil conditions. Copyright © 2015 Elsevier B.V. All rights reserved.
Shimizu, Takeo; Ito, Tsutae; Kanematsu, Satoko
Rosellinia necatrix causes white root rot in a wide range of fruit trees and persists for extended periods as pseudosclerotia on root debris. However, the pathogenesis of this disease has yet to be clarified. The functions of endogeneous target genes have not been determined because of the inefficiency in genetic transformation. In this study, the function of a melanin biosynthetic gene was determined to examine its role in morphology and virulence. A polyketide synthase gene (termed as RnPKS1) in the R. necatrix genome is homologous to the 1,8-dihydroxynaphthalene (DHN) melanin biosynthetic gene of Colletotrichum lagenarium. Melanin-deficient strains of R. necatrix were obtained by RNA interference-mediated knockdown of RnPKS1. The virulence of these strains was not significantly reduced compared with the parental melanin-producing strain. However, knockdown strains failed to develop pseudosclerotia and were degraded sooner in soil than the parental strain. Microscopic observations of albino conidiomata produced by knockdown strains revealed that melanization is involved in synnema integrity. These results suggest that melanin is not necessary for R. necatrix pathogenesis but is involved in survival through morphogenesis. This is the first report on the functional analysis of an endogenous target gene in R. necatrix. Copyright © 2014 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.
Yaman, Mustafa; Pınar Güngör, F; Gonca Güner, Beyza; Radek, Renate; Linde, Andreas
The present study describes the first isolation and characterization of Vairimorpha plodiae, a microsporidian pathogen of Plodia interpunctella (Lepidoptera: Pyralidae), from Turkey. We present characteristic light and electron microscopical features of the spores. Fresh binucleate spores are oval and measure 4.48 ± 0.23 (4.01-4.84) µm in length and 2.21 ± 0.15 (1.91-2.48) µm in width. Ultrastructural studies showed that the spore wall measures 150 to 200 nm and consists of a clear endospore (125-150 nm) and an electron-dense, uniform, thin exospore (30-50 nm). The polar filament is isofilar and with 10-12 coils. The well-developed polaroplast consists of two parts with thin lamellae anteriorly and thick, irregularly arranged lamellae posteriorly. The identity of our isolate is discussed.
Yaegashi, Hajime; Shimizu, Takeo; Ito, Tsutae; Kanematsu, Satoko
RNA silencing acts as a defense mechanism against virus infection in a wide variety of organisms. Here, we investigated inductions of RNA silencing against encapsidated double-stranded RNA (dsRNA) fungal viruses (mycoviruses), including a partitivirus (RnPV1), a quadrivirus (RnQV1), a victorivirus (RnVV1), a mycoreovirus (RnMyRV3), and a megabirnavirus (RnMBV1) in the phytopathogenic fungus Rosellinia necatrix Expression profiling of RNA silencing-related genes revealed that a dicer-like gene, an Argonaute-like gene, and two RNA-dependent RNA polymerase genes were upregulated by RnMyRV3 or RnMBV1 infection but not by other virus infections or by constitutive expression of dsRNA in R. necatrix Massive analysis of viral small RNAs (vsRNAs) from the five mycoviruses showed that 19- to 22-nucleotide (nt) vsRNAs were predominant; however, their ability to form duplexes with 3' overhangs and the 5' nucleotide preferences of vsRNAs differed among the five mycoviruses. The abundances of 19- to 22-nt vsRNAs from RnPV1, RnQV1, RnVV1, RnMyRV3, and RnMBV1 were 6.8%, 1.2%, 0.3%, 13.0%, and 24.9%, respectively. Importantly, the vsRNA abundances and accumulation levels of viral RNA were not always correlated, and the origins of the vsRNAs were distinguishable among the five mycoviruses. These data corroborated diverse interactions between encapsidated dsRNA mycoviruses and RNA silencing. Moreover, a green fluorescent protein (GFP)-based sensor assay in R. necatrix revealed that RnMBV1 infection induced silencing of the target sensor gene (GFP gene and the partial RnMBV1 sequence), suggesting that vsRNAs from RnMBV1 activated the RNA-induced silencing complex. Overall, this study provides insights into RNA silencing against encapsidated dsRNA mycoviruses. Encapsidated dsRNA fungal viruses (mycoviruses) are believed to replicate inside their virions; therefore, there is a question of whether they induce RNA silencing. Here, we investigated inductions of RNA silencing against
Full Text Available Conventional and Scorpion primers were designed from the ITS regions to identify Rosellinia necatrix, Phytophthora nicotianae, and P. citrophthora and from the IGS regions to identify Verticillium dahliae and V. alboatrum. Specificity of primers and probes was assessed using genomic DNA from a large number of fungi from several hosts and by means of BLAST analyses, to exclude the presence of similar sequences in other micro-organisms among available DNA databases (GenBank. Simple and rapid procedures for DNA extraction from naturally infected matrices (soils, roots, bark, and/or woody tissues were utilised to yield DNA of a purity and quality suitable for PCR assays. Combining these protocols with a double amplification (nested Scorpion-PCR, the real-time detection of these pathogens was possible from naturally infested soils and from infected citrus roots (P. nicotianae and P. citrophthora, from the roots and bark of stone fruits and olive (R. necatrix and from olive branches (V. dahliae. For target pathogens, the limit of detection was 1 pg µl-1 in Scorpion-PCR and 1 fg µl-1 in nested Scorpion-PCR. High and significant correlations between pathogen propagule concentrations and real-time PCR cycle thresholds (Ct were obtained. Moreover, specific tests with R. necatrix seem to indicate that its DNA is quite rapidly degraded in the soil, excluding the risk of false positives due to the presence of dead cells.
A Novel Bipartite Double-Stranded RNA Mycovirus from the White Root Rot Fungus Rosellinia necatrix: Molecular and Biological Characterization, Taxonomic Considerations, and Potential for Biological Control▿
Chiba, Sotaro; Salaipeth, Lakha; Lin, Yu-Hsin; Sasaki, Atsuko; Kanematsu, Satoko; Suzuki, Nobuhiro
White root rot, caused by the ascomycete Rosellinia necatrix, is a devastating disease worldwide, particularly in fruit trees in Japan. Here we report on the biological and molecular properties of a novel bipartite double-stranded RNA (dsRNA) virus encompassing dsRNA-1 (8,931 bp) and dsRNA-2 (7,180 bp), which was isolated from a field strain of R. necatrix, W779. Besides the strictly conserved 5′ (24 nt) and 3′ (8 nt) terminal sequences, both segments show high levels of sequence similarity in the long 5′ untranslated region of approximately 1.6 kbp. dsRNA-1 and -2 each possess two open reading frames (ORFs) named ORF1 to -4. Although the protein encoded by 3′-proximal ORF2 on dsRNA-1 shows sequence identities of 22 to 32% with RNA-dependent RNA polymerases from members of the families Totiviridae and Chrysoviridae, the remaining three virus-encoded proteins lack sequence similarities with any reported mycovirus proteins. Phylogenetic analysis showed that the W779 virus belongs to a separate clade distinct from those of other known mycoviruses. Purified virions ∼50 nm in diameter consisted of dsRNA-1 and -2 and a single major capsid protein of 135 kDa, which was shown by peptide mass fingerprinting to be encoded by dsRNA-1 ORF1. We developed a transfection protocol using purified virions to show that the virus was responsible for reduction of virulence and mycelial growth in several host strains. These combined results indicate that the W779 virus is a novel bipartite dsRNA virus with potential for biological control (virocontrol), named Rosellinia necatrix megabirnavirus 1 (RnMBV1), that possibly belongs to a new virus family. PMID:19828620
Desenvolvimento de Pantoea agglomerans em diversas temperaturas, pH e concentrações de carboxi-metil-celulose e o seu impacto no controle de Rosellinia necatrix Development of Pantoea agglomerans under different temperatures, pH and carboximethyl cellulose rates and their effect on Rosellinia necatrix control
Full Text Available Rosellinia necatrix Prill induz a podridão branca da raiz da macieira, doença que causa severa perda em pomares localizados no sul do Brasil. O manejo da doença é principalmente preventivo e inclui o uso de porta-enxertos resistentes e a fumigação do solo. A proteção das mudas de macieiras antes do plantio com um isolado antagonista de Pantoea agglomerans foi recentemente proposto para reduzir a incidência da doença. Os objetivos desta pesquisa foram caracterizar o relacionamento entre o patógeno e a bactéria antagonista; a produção de metabólitos biológicamente ativos pelo isolado bacteriano e a sua ação sobre o patógeno; o efeito do pH, da temperatura e de carboxi-metil-celulose (CMC sobre o crescimento do antagonista e do patógeno, isolados ou não e no controle da doença. Os resultados demonstraram que o crescimento de P. agglomerans foi maior em pH 5,5 e 6,0 e nas temperaturas de 20 °C e 30 °C. O crescimento micelial de R. necatrix foi inibido em meio de cultura e previamente colonizado pelo antagonista e com CMC nas concentrações de 0,25 e 0,5 %. A proteção das macieiras da infecção por R. necatrix foi observada quando utilizadas as concentrações de 10(7; 10(8 e 10(9 cel/mL e nas diferentes concentrações de CMC. Maior desenvolvimento radicular das macieiras foi constatado em todas as concentrações de CMC quando usada a concentração de 10(9 cel/mL. A formulação de P. agglomerans com CMC tornará possível a incorporação desta estratégia de controle ao manejo integrado da podridão branca das raízes da macieira no Sul do Brasil.Rosellinia necatrix Prill induces the white root rot of apple, a disease that causes severe losses in orchards located in Southern Brazil. The management of the disease is mostly preventative and include usage of less susceptible rootstocks and soil fumigation. Protection of nursery apple trees before planting with an antagonistic bacterium Pantoea agglomerans was recently
Zhang, Rui; Liu, Shengxue; Chiba, Sotaro; Kondo, Hideki; Kanematsu, Satoko; Suzuki, Nobuhiro
Here we report a biological and molecular characterization of a novel positive-sense RNA virus isolated from a field isolate (NW10) of a filamentous phytopathogenic fungus, the white root rot fungus that is designated as Rosellinia necatrix fusarivirus 1 (RnFV1). A recently developed technology using zinc ions allowed us to transfer RnFV1 to two mycelially incompatible Rosellinia necatrix strains. A biological comparison of the virus-free and -recipient isogenic fungal strains suggested that RnFV1 infects latently and thus has no potential as a virocontrol agent. The virus has an undivided positive-sense RNA genome of 6286 nucleotides excluding a poly (A) tail. The genome possesses two non-overlapping open reading frames (ORFs): a large ORF1 that encodes polypeptides with RNA replication functions and a smaller ORF2 that encodes polypeptides of unknown function. A lack of coat protein genes was suggested by the failure of virus particles from infected mycelia. No evidence was obtained by Northern analysis or classical 5′-RACE for the presence of subgenomic RNA for the downstream ORF. Sequence similarities were found in amino-acid sequence between RnFV1 putative proteins and counterparts of a previously reported mycovirus, Fusarium graminearum virus 1 (FgV1). Interestingly, several related sequences were detected by BLAST searches of independent transcriptome assembly databases one of which probably represents an entire virus genome. Phylogenetic analysis based on the conserved RNA-dependent RNA polymerase showed that RnFV1, FgV1, and these similar sequences are grouped in a cluster distinct from distantly related hypoviruses. It is proposed that a new taxonomic family termed Fusariviridae be created to include RnFV1 and FgV1. PMID:25101066
Full Text Available Here we report a biological and molecular characterization of a novel positive-sense RNA virus isolated from a field isolate (NW10 of a filamentous phytopathogenic fungus, the white root rot fungus that is designated as Rosellinia necatrix fusarivirus 1 (RnFV1. A recently developed technology using zinc ions allowed us to transfer RnFV1 to two mycelially incompatible Rosellinia necatrix strains. A biological comparison of the virus-free and -recipient isogenic fungal strains suggested that RnFV1 infects latently and thus has no potential as a virocontrol agent. The virus has an undivided positive-sense RNA genome of 6286 nucleotides excluding a poly (A tail. The genome possesses two non-overlapping open reading frames (ORFs: a large ORF1 that encodes polypeptides with RNA replication functions and a smaller ORF2 that encodes polypeptides of unknown function. A lack of coat protein genes was suggested by the failure of virus particles from infected mycelia. No evidence was obtained by Northern analysis or classical 5'-RACE for the presence of subgenomic RNA for the downstream ORF. Sequence similarities were found in amino-acid sequence between RnFV1 putative proteins and counterparts of a previously reported mycovirus, Fusarium graminearum virus 1 (FgV1. Interestingly, several related sequences were detected by BLAST searches of independent transcriptome assembly databases one of which probably represents an entire virus genome. Phylogenetic analysis based on the conserved RNA-dependent RNA polymerase showed that RnFV1, FgV1, and these similar sequences are grouped in a cluster distinct from distantly related hypoviruses. It is proposed that a new taxonomic family termed Fusariviridae be created to include RnFV1and FgV1.
Chiba, Sotaro; Lin, Yu-Hsin; Kondo, Hideki; Kanematsu, Satoko
A novel mycovirus termed Rosellinia necatrix partitivirus 2 (RnPV2), isolated from a phytopathogenic fungus, Rosellinina necatrix strain W57, was molecularly and biologically characterized in both natural and experimental host fungi. Three double-stranded RNA (dsRNA) segments, dsRNA1, dsRNA2, and defective interfering dsRNA1 (DI-dsRNA1), whose sizes were approximately 2.0, 1.8, and 1.7 kbp, respectively, were detected in W57. While the dsRNA2 sequence, encoding the coat protein, was reported previously, dsRNA1 and DI-dsRNA1 were shown to encode competent and defective (truncated) RNA-dependent RNA polymerase, respectively. Artificial introduction of RnPV2 into an RNA silencing-defective, Dicer-like 2 knockout mutant (Δdcl-2) of a nonnatural host, Cryphonectria parasitica (chestnut blight fungus), resulted in successful infection by the DI-dsRNA1-carrying and -free RnPV2. The DI-dsRNA1-free RnPV2 strain was characterized by a higher ratio of accumulation of the intact dsRNA1 to dsRNA2, enhanced replication and severer symptom expression, compared with the DI-carrying strain. These findings confirmed the nature of DI-dsRNA1 as a DI-RNA. Both viral strains replicated to higher levels in a Δdcl-2 mutant than in a wild-type C. parasitica fungal strain (EP155) and induced severe symptoms in the Δdcl-2 mutant but subtle symptoms in EP155, indicating that the host RNA silencing targets the partitivirus. No obvious phenotypic effects of infection by either virus strain were detected in the natural host fungus. These combined results represent the first example of a partitivirus with DI-RNA that alters viral symptom induction in a host-dependent manner. PMID:23236074
... of coccidiosis caused by Eimeria tenella, E. necatrix, and E. acervulina. (ii) Limitations. Not to be... percent). (i) Indications for use. Prevention of coccidiosis caused by Eimeria tenella, E. necatrix, and E...
...) Indications for use. For the prevention of coccidiosis caused by Eimeria tenella, E. necatrix, E. acervulina...) Indications for use. For the prevention of coccidiosis caused by Eimeria tenella, E. necatrix, E. acervulina...) Indications for use. For the prevention of coccidiosis caused by Eimeria tenella, E. necatrix, E. acervulina...
... coryza (Haemophilus gallinarum), coccidiosis (Eimeria tenella, E. necatrix), acute fowl cholera... per day (ii) Indications for use. For control of coccidiosis (E. meleagrimitis, E. adenoeides). (iii...
... (Avibacterium paragallinarum), coccidiosis (Eimeria tenella, Eimeria necatrix), acute fowl cholera (Pasteurella...) * * * (ii) Indications for use. For control of infectious coryza (Avibacterium paragallinarum), coccidiosis...
...) Indications for use. For prevention of coccidiosis caused by Eimeria necatrix, E. tenella, E. acervulina, E... grams (0.005 percent). (A) Indications for use. For the prevention of coccidiosis in broiler chickens.... For the prevention of coccidiosis caused by Eimeria necatrix, E. tenella, E. acervulina, E. brunetti...
...) Indications for use. As an aid in the prevention of coccidiosis caused by E. tenella and E. necatrix. (ii... aid in the prevention of coccidiosis caused by E. tenella, E. necatrix, and E. acervulina. (ii... grams (0.0025-0.005 percent). (i) Indications for use. As an aid in the prevention of coccidiosis caused...
Thomas Kolling; Andreas Linde
The quantification of the transmission of entomopathogens is important for the evaluation of their establishment and potential as biological control agents, however, only few field or semi-field studies were performed. The microsporidium Vairimorpha sp. was isolated from a gypsy moth (Lymantria dispar) population in Bulgaria and is...
... use—(A) As an aid in the control of coccidiosis caused by Eimeria tenella and E. necatrix susceptible... consumption. (2) Turkeys—(i) Amounts and indications for use—(A) As an aid in the control of coccidiosis...
...) Indications for use. An aid in the prevention of coccidiosis caused by E. tenella, E. necatrix, E. acervulina....002 percent). (ii) Indications for use. An aid in the prevention of coccidiosis caused by E. tenella...
... million (ppm)) Broiler chickens: For the prevention of coccidiosis caused by Eimeria tenella, E. necatrix... Indications for use Limitations Sponsor (i) 0.91 (1 ppm) Growing turkeys: For the prevention of coccidiosis...
Awais, Mian Muhammad; Akhtar, Masood; Iqbal, Zafar; Muhammad, Faqir; Anwar, Muhammad Irfan
The present paper reports the prevalence of coccidiosis in industrial broiler chickens in Faisalabad, Punjab, Pakistan to determine the occurrence of different species of Eimeria in the area and to assess their correlation with the environmental conditions including temperature, humidity and rainfall. The study was conducted from January 2009 to December 2010. Examination of chicken guts revealed 43.89% (n = 3,283/7,480) prevalence of coccidiosis. The highest prevalence (27.04%) was recorded of Eimeria tenella followed by Eimeria maxima (22.42%), Eimeria acervulina (19.89%) and Eimeria necatrix (4.02%). The prevalence of disease was significantly higher (P coccidiosis was recorded except E. necatrix.
... Indications for use Limitations Sponsor 27 to 45 Narasin 27 to 45 Broiler chickens; prevention of coccidiosis...; prevention of coccidiosis caused by Eimeria tenella, E. necatrix, E. acervulina, E. maxima, E. brunetti, E... 45 and bacitracin methylene disalicylate 50. Broiler chickens: For prevention of coccidiosis caused...
The study indicated that 16.92% (44/260) of the chickens were infected with coccidia oocysts. Four Eimeria species were identified, namely, Eimeria maxima, Eimeria acervulina, Eimeria necatrix and Eimeria tenella with the prevalence estimate of 36.3%, 29.5%, 18.2%, and 15.9%, respectively. Age unlike sex was found to ...
...) Indications for use. For the prevention of coccidiosis caused by Eimeria tenella, E. necatrix, E. acervulina... bambermycins 1 to 2 grams. (A) Indications for use. For the prevention of coccidiosis caused by Eimeria tenella...) plus virginiamycin 5 grams. (A) Indications for use. For the prevention of coccidiosis caused by...
Forty brooder houses were sampled from (our States in the southern part of Nigeria, namely: Oyo, Lagos, Imo and Anambra States, to identify the species of coccidia. Five species of Eimeria were isolated including Eimeria tenella,· E. necatrix; E. maxima and E. mivati/E. mitis. Using the McMaster count, the number of oocyst ...
... coccidiosis caused by Eimeria tenella, E. necatrix, E. acervulina, E. maxima, and E. brunetti under average... continue past the age when coccidiosis is ordinarily a hazard. If death losses exceed 0.5 percent in a 2-day period, obtain a laboratory diagnosis. If coccidiosis is the cause, use the sulfaquinoxaline...
... Fusarium solani Mart. and Sphaerostilbe repens B. et Br. The latter is the most widespread in the Congo; (ii) dry root rots caused by Rigidoporus lignosus (Klot.) Imazeki, Rosellinia necatrix (Hart.) Berk., Botriodiplodia theobromae Pat., Armillaria mellea (Vahn: Fr.) P. Kumm., Armillaria heimii and Phaeolus manihotis Heim.
Polonio, Álvaro; Vida, Carmen; de Vicente, Antonio; Cazorla, Francisco M
The biocontrol rhizobacterium Pseudomonas chlororaphis PCL1606 has the ability to protect avocado plants against white root rot produced by the phytopathogenic fungus Rosellinia necatrix. Moreover, PCL1606 displayed direct interactions with avocado roots and the pathogenic fungus. Thus, nonmotile (flgK mutant) and non-chemotactic (cheA mutant) derivatives of PCL1606 were constructed to emphasize the importance of motility and chemotaxis in the biological behaviour of PCL1606 during the biocontrol interaction. Plate chemotaxis assay showed that PCL1606 was attracted to the single compounds tested, such as glucose, glutamate, succinate, aspartate and malate, but no chemotaxis was observed to avocado or R. necatrix exudates. Using the more sensitive capillary assay, it was reported that smaller concentrations (1 mM) of single compounds elicited high chemotactic responses, and strong attraction was confirmed to avocado and R. necatrix exudates. Finally, biocontrol experiments revealed that the cheA and fglK derivative mutants reduced root protection against R. necatrix, suggesting an important role for these biological traits in biocontrol by P. chlororaphis PCL1606. [Int Microbiol 20(2):94-104 (2017)]. Copyright© by the Spanish Society for Microbiology and Institute for Catalan Studies.
Arianne Brown Jordan
Full Text Available Coccidiosis is an intestinal disease of chickens of major economic importance to broiler industries worldwide. Species of coccidia found in chickens include Eimeria acervulina, Eimeria brunetti, Eimeria maxima, Eimeria mitis, Eimeria necatrix, Eimeria praecox, and Eimeria tenella. In recent years, polymerase chain reaction (PCR has been developed to provide accurate and rapid identification of the seven known Eimeria species of chickens. The aim of this study was to use species-specific real-time PCR (qPCR to identify which of the seven Eimeria species are present in Trinidad poultry. Seventeen pooled fecal samples were collected from 6 broiler farms (2–5 pens per farm across Trinidad. Feces were also collected from birds showing clinical signs of coccidiosis in two live bird markets (pluck shops. qPCR revealed the presence of five species of Eimeria (E. acervulina, E. maxima, E. mitis, E. necatrix, and E. tenella, but not E. brunetti or E. praecox. Mixed infections were detected on all broiler farms, and DNA of two highly pathogenic Eimeria species (E. tenella and E. necatrix was detected in feces taken from clinically sick birds sampled from the two pluck shops.
Vida, Carmen; Cazorla, Francisco M; de Vicente, Antonio
The improvement in soil quality of avocado crops through organic amendments with composted almond shells has a positive effect on crop yield and plant health, and enhances soil suppressiveness against the phytopathogenic fungus Rosellinia necatrix. In previous studies, induced soil suppressiveness against this pathogen was related to stimulation of Gammaproteobacteria, especially some members of Pseudomonas spp. with biocontrol-related activities. In this work, we isolated bacteria from this suppressiveness-induced amended soil using a selective medium for Pseudomonas-like microorganisms. We characterized the obtained bacterial collection to aid in identification, including metabolic profiles, antagonistic responses, hybridization to biosynthetic genes of antifungal compounds, production of lytic exoenzymatic activities and plant growth-promotion-related traits, and sequenced and compared amplified 16S rDNA genes from representative bacteria. The final selection of representative strains mainly belonged to the genus Pseudomonas, but also included the genera Serratia and Stenotrophomonas. Their biocontrol-related activities were assayed using the experimental avocado model, and results showed that all selected strains protected the avocado roots against R. necatrix. This work confirmed the biocontrol activity of these Gammaproteobacteria-related members against R. necatrix following specific stimulation in a suppressiveness-induced soil after a composted almond shell application. Copyright © 2017 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.
Polinéstor Aguilar Celi
Full Text Available 1.- Se. ha. estudiado 152 casos de Tiña del cuero cabelludo, han sido identificadas y pertenecen a las siguientes especies: Genera Species N° human de cases Trichophyton T. tonsurans 120 T. mentagrophytes 7 T. violaceum 1 Microsporum M. canis 24 2.- El contagio se hace de preferencia en las escuelas, especialmente las tricofitias que dan pequeñas placas que son fácilmente disimuladas por el peinado, pasando inadvertidas. 3.- El mayor porcentaje de Tiñas lo hemos encontrado entre los 7 y 12 años de edad, predominando el género Trichophyton, como, se puede apreciar en la gráfica 1. 4.- En casi todos los casos en que hemos aislado Trichophyton metagrophytes los niños han vivido en el campo o estado en contacto con perros,gatos o caballos. Lo mismo sucede con las microsporias. 5.- En dos cultivos de Trichophyton tonsurans hemos observado, variaciones en la forma de la colonia. En todas las variaciones se hizo el estudio micológico no encontrándose diferencias. Actualmente no se toma el aspecto de la colonia gigante como signo fundamental para establecer variedades. 6.- No hemos encontrado el Microsporum Audouini, como en otras partes; en su lugar se encuentra el M. canis. 7.- Hemos encontrado 8 casos de Kerion causado por el Trichophyton tonsurans.
M. Q. KHAN, H. IRSHAD, R. ANJUM, M. JAHANGIR AND U. NASIR
Full Text Available This study was designed to determine the prevalence of eimeriosis in poultry and identify potential risk factors for its spread in Rawalpindi/Islamabad area of Pakistan. Of 359 gut samples (suspected for harbouring eimeriosis examined, 258 (71.86% were found infected. Four species of Eimeria (E. maxima, 34.10%, E. tenella, 30.62%, E. mitis, 13.95% and E. necatrix, 7.75% were recorded. The prevalence of eimeriosis was highest in the month of September (89.74%, while lowest during June (28.57%. The disease was more common at the farms where the litter was wet and not managed properly.
M. M. Ayaz, M. Akhtar, C.S. Hayat, M.A. Hafeez and A. Haq1
Full Text Available A study was conducted during the period from October, 2000 to June, 2001 to record the prevalence of coccidiosis in broiler chickens in Faisalabad district. A total of 930 guts were collected and processed at Immunoparasitology Laboratory, Department of Veterinary Parasitology, University of Agriculture, Faisalabad. Seven species of Eimeria (E viz. E. tenella (50%, E. maxima (40%, E. mitis (2%, E. praecox (0.8%, E. acervulina (4%; E. necatrix (2% and E. brunetti (1.2% were recorded. The overall prevalence of coccidiosis was found to be 37.95%.
Esteban B Sir
Full Text Available Se reporta por primera vez a Rosellinia canzacotoana en el Cono Sur de América y a Rosellinia arcuata para la Argentina. Rosellinia necatrix se cita creciendo sobre árboles en descomposición de la flora autóctona de la Argentina. Se realiza una revisión de tres holotipos de Spegazzini: Rosellinia australis, Rosellinia bonaerensis, Rosellinia smilacina, detallando su estado nomenclatural y taxonómico actual. También se presenta una clave dicotómica de las especies con aspecto rosellinoide del Noroeste del país y un mapa de distribución.Novelties for the genus Rosellinia (Ascomycota-Xylariaceae from Northwest of Argentina. Rosellinia canzacotoana is reported for the first time in the southern Cone of America and Rosellinia arcuata is a new record from Argentina. Rosellinia necatrix is reported growing on a native host of the Argentine flora. Three Spegazzini's holotypes: Rosellinia australis, Rosellinia bonaerensis and Rosellinia smilacina were studied and their taxonomic and nomenclatural positions are discussed. A dichotomous key to rosellinoid taxa so far known from Northwest Argentina and distributions maps is presented.
Shirzad, Mohammad Reza; Seifi, Saeed; Gheisari, Hamid Reza; Hachesoo, Bahman Abdi; Habibi, Hasan; Bujmehrani, Hossein
Coccidiosis is a disease of almost universal importance in Poultry production. The subclinical form of coccidiosis has most significant economic impact due to impaired growth rate and feed conversion. Some factors such as: age, size of flock, season, etc., may effect on severity of this disease. In this study, the relationships between some major risk factors and prevalence of subclinical coccidiosis were investigated. This study was done in 120 broiler farms in five different cities of Mazandaran province, north of Iran. Five chicks (3-8 weeks of ages) were taken randomly from every 120 farms and post-mortem and parasitological examinations were performed. Five Eimeria spp. were recognized: Eimeria tenella, Eimeria maxima, Eimeria acervulina, Eimeria brunetti, and Eimeria necatrix. The prevalence rate of subclinical coccidiosis among them is 75% (90 farms out of 120). E. acervulina was the most prevalent species (65.5%) followed by E. maxima (17.7%), E. tenella (15.5%), E. brunetti (10%), and E. necatrix (5.5%). According to what the results approve, the occurrence of subclinical coccidiosis is significantly related to the age and size of flock, whereas the other factors such as the season of year, industrial strains, chicken's keeping system, and anticoccidial drugs do not affect this phenomenon remarkably.
M. E. M. Habib
Full Text Available Some biological effects of feeding larvae of the armyworm Spodoptera latifascia with leaves of three plant species (cotton, soybean and lettuce have been studied. Some indicative measurements were utilized to determine the host suitability of these three plant species. Cotton leaves (Gossypium hirsutum were found to be better than soybean (Glycine max for the development and growth of this insect. Lettuce leaves (Lactuca sativa were incapable of sustaining this insect. Moreover, the stress of the latter diet was associated with a latent microbial disease, a mixed infection caused by Vairimorpha sp. and a nuclear polyhedrosis virus (NPV. Cotton leaves can be recommended as a suitable diet for mass rearing of this insect species. Due to recent outbreks of S. latifascia observed in the States of São Paulo, Santa Catarina and Paraná and because of the frequent applications of wide spectrum chemical insecticides, it can be expected that this insect will soon become one of the important pests threatening cotton and other economic plantations in Brazil.
Gharekhani, Jamal; Sadeghi-Dehkordi, Zivar; Bahrami, Mohammadali
The main goal of current study was to investigate the prevalence of coccidiosis in broiler farms in Hamedan province, western Iran. Chicks and fecal samples were collected in all of the 220 broiler farms in this region. All viscera were examined for gross pathological changes. The mucosa of small intestine and the caeca were examined for the presence and identification of parasitic forms using parasitology methods. The overall rate of coccidiosis was 31.8%; E. acervulina (75.7%), E. tenella (54.3%), E. necatrix (28.6%), and E. maxima (20%) were determined. Mixed infections were observed in all of the positive farms. There was a statistical significant difference (P 0.05). This is the first report of coccidiosis rate in broiler farms in this region. Further additional researches and design control strategies for improving management in farms are necessary. PMID:26464948
Full Text Available The main goal of current study was to investigate the prevalence of coccidiosis in broiler farms in Hamedan province, western Iran. Chicks and fecal samples were collected in all of the 220 broiler farms in this region. All viscera were examined for gross pathological changes. The mucosa of small intestine and the caeca were examined for the presence and identification of parasitic forms using parasitology methods. The overall rate of coccidiosis was 31.8%; E. acervulina (75.7%, E. tenella (54.3%, E. necatrix (28.6%, and E. maxima (20% were determined. Mixed infections were observed in all of the positive farms. There was a statistical significant difference (P0.05. This is the first report of coccidiosis rate in broiler farms in this region. Further additional researches and design control strategies for improving management in farms are necessary.
Vida, Carmen; Bonilla, Nuria; de Vicente, Antonio; Cazorla, Francisco M
This study focused on the microbial profile present in an agricultural soil that becomes suppressive after the application of composted almond shells (AS) as organic amendments. For this purpose, we analyzed the functions and composition of the complex communities present in an experimental orchard of 40-year-old avocado trees, many of them historically amended with composted almond shells. The role of microbes in the suppression of Rosellinia necatrix, the causative agent of avocado white root rot, was determined after heat-treatment and complementation experiments with different types of soil. Bacterial and fungal profiles obtained from natural soil samples based on the 16S rRNA gene and ITS sequencing revealed slight differences among the amended (AS) and unamended (CT) soils. When the soil was under the influence of composted almond shells as organic amendments, an increase in Proteobacteria and Ascomycota groups was observed, as well as a reduction in Acidobacteria and Mortierellales. Complementary to these findings, functional analysis by GeoChip 4.6 confirmed these subtle differences, mainly present in the relative abundance of genes involved in the carbon cycle. Interestingly, a group of specific probes included in the "soil benefit" category was present only in AS-amended soils, corresponding to specific microorganisms previously described as potential biocontrol agents, such as Pseudomonas spp., Burkholderia spp., or Actinobacteria. Considering the results of both analyses, we determined that AS-amendments to the soil led to an increase in some orders of Gammaproteobacteria, Betaproteobacteria, and Dothideomycetes, as well as a reduction in the abundance of Xylariales fungi (where R. necatrix is allocated). The combination of microbial action and substrate properties of suppressiveness are discussed.
Calderón, Claudia E; de Vicente, Antonio; Cazorla, Francisco M
Different bacterial traits can contribute to the biocontrol of soilborne phytopathogenic fungus. Among others, (1) antagonism, (2) competition for nutrients and niches, (3) induction of systemic resistance of the plants and (4) predation and parasitism are the most studied. Pseudomonas chlororaphis PCL1606 is an antagonistic rhizobacterium that produces the antifungal metabolite 2-hexyl, 5-propyl resorcinol (HPR). This bacterium can biologically control the avocado white root rot caused by Rosellinia necatrix. Confocal laser scanning microscopy of the avocado rhizosphere revealed that this biocontrol bacterium and the fungal pathogen compete for the same niche and presumably also for root exudate nutrients. The use of derivative mutants in the geners related to HPR biosynthesis (dar genes) revealed that the lack of HPR production by P. chlororaphis PCL1606 negatively influences the bacterial colonisation of the avocado root surface. Microscopical analysis showed that P. chlororaphis PCL1606 closely interacts and colonises the fungal hyphae, which may represent a novel biocontrol mechanism in this pseudomonad. Additionally, the presence of HPR-producing biocontrol bacteria negatively affects the ability of the fungi to infect the avocado root. HPR production negatively affects hyphal growth, leading to alterations in the R. necatrix physiology visible under microscopy, including the curling, vacuolisation and branching of hyphae, which presumably affects the colonisation and infection abilities of the fungus. This study provides the first report of multitrophic interactions in the avocado rhizosphere, advancing our understanding of the role of HPR production in those interactions. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.
Krebes, Lukas; Zeidler, Lisza; Frankowski, Jens; Bastrop, Ralf
Microsporidia are single-celled, intracellular eukaryotes that parasitise a wide range of animals. The Nosema/Vairimorpha group includes some putative asexual species, and asexuality is proposed to have originated multiple times from sexual ancestors. Here, we studied the variation in the ribosomal DNA (rDNA) of 14 isolates of the presumed apomictic and vertically transmitted Nosema granulosis to evaluate its sexual status. The analysed DNA fragment contained a part of the small-subunit ribosomal gene (SSU) and the entire intergenic spacer (IGS). The mitochondrial cox1 gene of the host Gammarus duebeni (Crustacea) was analysed to temporally calibrate the system and to test the expectation of cophylogeny of host and parasite genealogies. Genetic variability of the SSU gene was very low within and between the isolates. In contrast, intraisolate (within a single host) variability of the IGS felt in two categories, because 12 isolates possess a very high IGS genetic diversity and two isolates were almost invariable in the IGS. This difference suggests variable models of rDNA evolution involving birth-and-death and unexpectedly concerted evolution. An alternative explanation could be a likewise unattended mixed infection of host individuals by more than one parasite strain. Despite considerable genetic divergence between associated host mitochondrial haplotypes, some N. granulosis 'IGS populations' seem not to belong to different gene pools; the relevant tests failed to show significant differences between populations. A set of recombinant IGS sequences made our data incompatible with the model of a solely maternally inherited, asexual species. In line with recent reports, our study supports the hypothesis that some assumed apomictic Microsporidia did not entirely abstain from the evolutionary advantages of sex. In addition, the presented data indicate that horizontal transmission may occur occasionally. This transmission mode could be a survival strategy of N
Cruz, Andre Freire; Ishii, Takaaki
The aim of this research was to isolate and characterize bacteria from spores of arbuscular mycorrhizal fungi (AMF). We designated these bacteria 'probable endobacteria' (PE). Three bacterial strains were isolated from approximately 500 spores of Gigaspora margarita (Becker and Hall) using a hypodermic needle (diameter, 200 μm). The bacteria were identified by morphological methods and on the basis of ribosomal gene sequences as Bacillus sp. (KTCIGM01), Bacillus thuringiensis (KTCIGM02), and Paenibacillus rhizospherae (KTCIGM03). We evaluated the effect of these probable endobacteria on antagonistic activity to the soil-borne plant pathogens (SBPPs) Fusarium oxysporum f. sp. lactucae MAFF 744088, Rosellinia necatrix, Rhizoctonia solani MAFF 237426, and Pythium ultimum NBRC 100123. We also tested whether these probable endobacteria affected phosphorus solubilization, ethylene production, nitrogenase activity (NA), and stimulation of AMF hyphal growth. In addition, fresh samples of spores and hyphae were photographed using an in situ scanning electron microscope (SEM) (Quanta 250FEG; FEI Co., Japan). Bacterial aggregates (BAs), structures similar to biofilms, could be detected on the surface of hyphae and spores. We demonstrate that using extraction with an ultrathin needle, it is possible to isolate AMF-associated bacterial species that are likely derived from inside the fungal spores.
Su, L; Dong, L; Bughio, S; Guo, M; Wang, L
Breast cancer resistance protein (BCRP, ABCG2) is a member of ABC (ATP-binding cassette) transporter superfamily that occurs in a variety of tissues including liver and small intestine of animals. As BCRP is involved in drug absorption, distribution, and elimination, modulation of its expression may affect the clinical efficacy of drugs. However, little is known about the effects of coccidiosis or colibacillosis infection on the levels of BCRP expression in chickens. Here, we studied the effect of infection with Escherichia coli (E. coli) or Eimeriida mixture (E. necatrix and E. tenella) on the expression levels of ABCG2 mRNA and BCRP in the different segments of small intestine and liver in chickens. Expression of ABCG2 mRNA or BCRP was detected in the entire small intestine and liver of healthy chickens, and the expression levels in liver and ileum were significantly higher than duodenum and jejunum. Infection with E. coli or Eimeriida mixture resulted in significant decrease in ABCG2 mRNA and BCRP expression in liver, ileum, and jejunum, but not in duodenum, in comparison with noninfection control. The results indicate that coccidiosis or colibacillosis infection inhibits BCRP expression in chickens, which may consequently influence drug distribution and therapeutic efficacy. © 2013 John Wiley & Sons Ltd.
Full Text Available Recent studies have demonstrated that a diverse array of mycoviruses infect the plant pathogenic fungus Sclerotinia sclerotiorum. Here, we report the molecular characterization of a newly identified mycovirus, Sclerotinia sclerotiorum fusarivirus 1 (SsFV1, which was isolated from a sclerotia-defective strain JMTJ14 of S. sclerotiorum. Excluding a poly (A tail, the genome of SsFV1 comprises 7754 nucleotides (nts in length with 83 and 418 nts for 5'- and 3'-untranslated regions, respectively. SsFV1 has four non-overlapping open reading frames (ORFs: ORF1 encodes a 191 kDa polyprotein (1664 amino acid residues in length containing conserved RNA-dependent RNA polymerase (RdRp and helicase domains; the other three ORFs encode three putative hypothetical proteins of unknown function. Phylogenetic analysis, based on RdRp and Helicase domains, indicated that SsFV1 is phylogenetically related to Rosellinia necatrix fusarivirus 1 (RnFV1, Fusarium graminearum virus-DK21 (FgV1, and Penicillium roqueforti RNA mycovirus 1 (PrRV1, a cluster of an independent group belonging to a newly proposed family Fusarividae. However, SsFV1 is markedly different from FgV1 and RnFV1 in genome organization and nucleotide sequence. SsFV1 was transmitted successfully to two vegetatively incompatible virus-free strains. SsFV1 is not responsible for the abnormal phenotype of strain JMTJ14.
Liu, Rong; Cheng, Jiasen; Fu, Yanping; Jiang, Daohong; Xie, Jiatao
Recent studies have demonstrated that a diverse array of mycoviruses infect the plant pathogenic fungus Sclerotinia sclerotiorum. Here, we report the molecular characterization of a newly identified mycovirus, Sclerotinia sclerotiorum fusarivirus 1 (SsFV1), which was isolated from a sclerotia-defective strain JMTJ14 of S. sclerotiorum. Excluding a poly (A) tail, the genome of SsFV1 comprises 7754 nucleotides (nts) in length with 83 and 418 nts for 5'- and 3'-untranslated regions, respectively. SsFV1 has four non-overlapping open reading frames (ORFs): ORF1 encodes a 191 kDa polyprotein (1664 amino acid residues in length) containing conserved RNA-dependent RNA polymerase (RdRp) and helicase domains; the other three ORFs encode three putative hypothetical proteins of unknown function. Phylogenetic analysis, based on RdRp and Helicase domains, indicated that SsFV1 is phylogenetically related to Rosellinia necatrix fusarivirus 1 (RnFV1), Fusarium graminearum virus-DK21 (FgV1), and Penicillium roqueforti RNA mycovirus 1 (PrRV1), a cluster of an independent group belonging to a newly proposed family Fusarividae. However, SsFV1 is markedly different from FgV1 and RnFV1 in genome organization and nucleotide sequence. SsFV1 was transmitted successfully to two vegetatively incompatible virus-free strains. SsFV1 is not responsible for the abnormal phenotype of strain JMTJ14.
There appear to be over a million of fungal species including those that have been unidentified and unreported, where a variety of viruses make a world as well. Studies on a very small number of them conducted during the last two decades demonstrated the infectivity of fungal viruses that had previously been assumed to be inheritable, indigenus and non-infectious. Also, great technical advances were achieved. The chest blight fungus (Cryphonectria parasitica), a phytopathogenic ascomycetous fungus, has emerged as a model filamentous fungus for fungal virology. The genome sequence with annotations, albeit not thorough, many useful research tools, and gene manipulation technologies are available for this fungus. Importantly, C. parasitica can support replication of homologous viruses naturally infecting it, in addition to heterologous viruses infecting another plant pathogenic fungus, Rosellinia necatrix taxonomically belonging to a different order. In this article, I overview general properties of fungal viruses and advantages of the chestnut blight fungus as a mycovirus host. Furthermore, I introduce two recent studies carried out using this fungal host:''Defective interfering RNA and RNA silencing that regulate the replication of a partitivirus'' and'' RNA silencing and RNA recombination''.
Hema, Srinivasan; Arun, Thangavel; Senthilkumar, Balakrishnan; Senbagam, Duraisamy; Sureshkumar, Muthusamy
A total of thirty suspected broiler chicks were screened for coccidiosis, of them 25 chicks were found to be infected with coccidiosis viz. Eimeria tenella (15) Eimeria maxima (5) Eimeria necatrix (6) and Eimeria mitis (4). The anticoccidial efficacy of Azadirachta indica and Carica papaya with Salinomycin as a dietary feed supplement on the representative E. tenella (25 x 10³ oocyst) infection challenged in broiler chicks was studied in six groups for the period of six weeks. A. indica and C. papaya leaves were administered in powder form at the concentration of 0.1% and 0.2% respectively. The Oocysts per gram (OPG) count were observed on 8th, 9th, 10th, 11th day of post inoculation (DPI). The challenged experimental chicks revealed haemorrhage, thickening of mucosa, cores of blood and ballooning of caecum. The experimental group T5 chicks treated with A. indica were analyzed to possess the maximum weight gain (2.003), better feed conversion ratio (FCR) (2.32), OPG count (5.87), livability percentage (88) and the lesion score (3.33). Chi-square test analysis revealed no significant differences among the treated groups and the performance parameters. Therefore, this study concludes that plant sources used as a remedial curate for coccidiosis is a perforated growth in the commercial broiler industries.
Full Text Available Reuse of olive mill wastewaters (OMWWs in agriculture represents a significant challenge for health and safety of our planet. Phytotoxic compounds in OMWW generally prohibit use of untreated OMWWs for agricultural irrigation or direct discharge into surface waters. However, pretreated OMWW can have positive effects on chemical and microbiological soil characteristics, to fight against fungal soil-borne pathogens. Low amounts of OMWW following thermal (TT-OMWW and high-pressure homogenization (HPH-OMWW pretreatments counteracted growth of some of 12 soil-borne and/or pathogenic fungi examined. With fungal growth measured as standardized change in time to half maximum colony diameter, Δτ, overall, HPH-OMWW showed increased bioactivity, as increased mean Δτ from 3.0 to 4.8 days. Principal component analysis highlighted two fungal groups: Colletotrichum gloeosporioides, Alternaria alternata, Sclerotium rolfsii, and Rosellinia necatrix, with growth strongly inhibited by the treated OMWWs; and Aspergillus ochraceus and Phaeoacremonium parasiticum, with stimulated growth by the treated OMWWs. As a non-thermal treatment, HPH-OMWW generally shows improved positive effects, which potentially arise from preservation of the phenols.
Zhong, Jie; Shang, Hong Hong; Zhu, Chuan Xia; Zhu, Jun Zi; Zhu, Hong Jian; Hu, Yan; Gao, Bi Da
The alternaria blackspot of rapeseed is one of the most prominent diseases of rapeseed. It is caused by three species of the genus Alternaria: Alternaria brassicicola, Alternaria brassicae, and Alternaria raphanin. Here we report a novel positive-sense RNA virus from an A. brassicicola strain 817-14. The virus has a 6639 nucleotide (nt) long genome, excluding a poly (A)-tail, and was predicted to contain three putative open reading frames (ORF1, ORF2, and ORF3). The large ORF1 encoded a 174-kDa polyprotein (composed of 1522 amino acid residues) containing a conserved RNA-dependent RNA polymerase (RdRp) domain and a helicase domain. The other two smaller ORFs encoded polypeptides with unknown function. Homology search and phylogenetic analysis, based on the RdRp and helicase domains, suggest that this virus is related to and grouped with Sclerotinia sclerotiorum fusarivirus 1 (SsFV1), Rosellinia necatrix fusarivirus 1 (RnFV1), Fusarium graminearum virus-DK21 (FgV1), and Penicillium roqueforti RNA mycovirus 1 (PrRV1), all of which belong to a newly proposed family Fusariviridae. For this study, we designed the virus as "Alternaria brassicicola fusarivirus 1" (AbFV1). Virus elimination revealed that AbFV1 has no conspicuous impact on the biological properties of its host. Copyright © 2016. Published by Elsevier B.V.
Kostadinović Ljiljana M.
Full Text Available The effect of Artemisia absinthium essential oil (AAEO on enzymatic activity of super-oxide-dismutase (SOD, glutathione-peroxidase (GSHPx, glutathione-reductase (GR, peroxidase (POD, xantine-oxidase (XOD and non-enzymatic (content of lipid peroxides (LPx and gluthathione (GSH antioxidative status of broilers infected with mixture of oocysts of Eimeria tenella, Eimeria mitis and Eimeria necatrix in comparison to coccidiostat salinomycin was investigated. The in vivo investigation were carried out on 120 Arbor acres broilers of both sexes. Broilers were randomly distributed into four groups. Group A was uninfected and untreated; group B was infected and was kept untreated; group C preventively received coccidiostatic salinomycin in quantity of 60 mg/kg of feed and was inoculated with coccidia species at 21st day-of-age and group D received in feed AAEO in quantity of 3 g/kg and was infected with Eimeria oocysts at 21st day-of-age. Livers were collected for the subsequent evaluation of antioxidative status. It was concluded that AAEO added in feed for broilers prevented the development of coccidia oocysts and therefore it can be used as prophylactic feed additive.
Andre Freire Cruz
The aim of this research was to isolate and characterize bacteria from spores of arbuscular mycorrhizal fungi (AMF. We designated these bacteria ‘probable endobacteria’ (PE. Three bacterial strains were isolated from approximately 500 spores of Gigaspora margarita (Becker and Hall using a hypodermic needle (diameter, 200 μm. The bacteria were identified by morphological methods and on the basis of ribosomal gene sequences as Bacillus sp. (KTCIGM01, Bacillus thuringiensis (KTCIGM02, and Paenibacillus rhizospherae (KTCIGM03. We evaluated the effect of these probable endobacteria on antagonistic activity to the soil-borne plant pathogens (SBPPs Fusarium oxysporum f. sp. lactucae MAFF 744088, Rosellinia necatrix, Rhizoctonia solani MAFF 237426, and Pythium ultimum NBRC 100123. We also tested whether these probable endobacteria affected phosphorus solubilization, ethylene production, nitrogenase activity (NA, and stimulation of AMF hyphal growth. In addition, fresh samples of spores and hyphae were photographed using an in situ scanning electron microscope (SEM (Quanta 250FEG; FEI Co., Japan. Bacterial aggregates (BAs, structures similar to biofilms, could be detected on the surface of hyphae and spores. We demonstrate that using extraction with an ultrathin needle, it is possible to isolate AMF-associated bacterial species that are likely derived from inside the fungal spores.
Hermosa, M R; Grondona, I; Iturriaga, E A; Diaz-Minguez, J M; Castro, C; Monte, E; Garcia-Acha, I
The most common biological control agents (BCAs) of the genus Trichoderma have been reported to be strains of Trichoderma virens, T. harzianum, and T. viride. Since Trichoderma BCAs use different mechanisms of biocontrol, it is very important to explore the synergistic effects expressed by different genotypes for their practical use in agriculture. Characterization of 16 biocontrol strains, previously identified as "Trichoderma harzianum" Rifai and one biocontrol strain recognized as T. viride, was carried out using several molecular techniques. A certain degree of polymorphism was detected in hybridizations using a probe of mitochondrial DNA. Sequencing of internal transcribed spacers 1 and 2 (ITS1 and ITS2) revealed three different ITS lengths and four different sequence types. Phylogenetic analysis based on ITS1 sequences, including type strains of different species, clustered the 17 biocontrol strains into four groups: T. harzianum-T. inhamatum complex, T. longibrachiatum, T. asperellum, and T. atroviride-T. koningii complex. ITS2 sequences were also useful for locating the biocontrol strains in T. atroviride within the complex T. atroviride-T. koningii. None of the biocontrol strains studied corresponded to biotypes Th2 or Th4 of T. harzianum, which cause mushroom green mold. Correlation between different genotypes and potential biocontrol activity was studied under dual culturing of 17 BCAs in the presence of the phytopathogenic fungi Phoma betae, Rosellinia necatrix, Botrytis cinerea, and Fusarium oxysporum f. sp. dianthi in three different media.
Cruz, A F; Horii, S; Ochiai, S; Yasuda, A; Ishii, T
The aim of this work was to observe bacteria associated with the spores of Gigaspora margarita, an arbuscular mycorrhizal fungus (AMF). First, a direct analysis of DNA from sterilized spores indicated the bacteria belonging to the genus Janthinobacterium. In the second assay, two bacterial strains were isolated by osmosis from protoplasts, which were derived from spores by using two particular enzymes: lysing enzymes and yatalase. After isolation, cultivation and identification by their DNA as performed in the first experiment, the species with the closest relation were Janthinobacterium lividum (KCIGM01) and Paenibacillus polymyxa (KCIGM04) isolated with lysing enzymes and yatalase respectively. Morphologically, J. lividum was Gram negative and oval, while P. polymyxa was also oval, but Gram positive. Both strains had antagonistic effects to the pathogenic fungi Rosellimia necatrix, Pythium ultimum, Fusarium oxysporum and Rhizoctonia solani. In particular, J. lividum was much stronger in this role. However, in phosphorus (P) solubilization P. polymyxa functioned better than J. lividum. This experiment had revealed two new bacteria species (P. polymyxa and J. lividum), associated with AMF spores, which functioned to suppress diseases and to solubilize P. AMF spores could be a useful source for bacterial antagonists to soil-borne diseases and P solubilization.
Full Text Available Abstract Aneurinibacillus aneurinilyticus strain CKMV1 was isolated from rhizosphere of Valeriana jatamansi and possessed multiple plant growth promoting traits like production of phosphate solubilization (260 mg/L, nitrogen fixation (202.91 nmol ethylene mL-1 h-1, indole-3-acetic acid (IAA (8.1 µg/mL, siderophores (61.60%, HCN (hydrogen cyanide production and antifungal activity. We investigated the ability of isolate CKMV1 to solubilize insoluble P via mechanism of organic acid production. High-performance liquid chromatography (HPLC study showed that isolate CKMV1 produced mainly gluconic (1.34% and oxalic acids. However, genetic evidences for nitrogen fixation and phosphate solubilization by organic acid production have been reported first time for A. aneurinilyticus strain CKMV1. A unique combination of glucose dehydrogenase (gdh gene and pyrroloquinoline quinone synthase (pqq gene, a cofactor of gdh involved in phosphate solubilization has been elucidated. Nitrogenase (nif H gene for nitrogen fixation was reported from A. aneurinilyticus. It was notable that isolate CKMV1 exhibited highest antifungal against Sclerotium rolfsii (93.58% followed by Fusarium oxysporum (64.3%, Dematophora necatrix (52.71%, Rhizoctonia solani (91.58%, Alternaria sp. (71.08% and Phytophthora sp. (71.37%. Remarkable increase was observed in seed germination (27.07%, shoot length (42.33%, root length (52.6%, shoot dry weight (62.01% and root dry weight (45.7% along with NPK (0.74, 0.36, 1.82% content of tomato under net house condition. Isolate CKMV1 possessed traits related to plant growth promotion, therefore, could be a potential candidate for the development of biofertiliser or biocontrol agent and this is the first study to include the Aneurinibacillus as PGPR.
Pirali Kheirabadi, Khodadad; Kaboutari Katadj, Jahangir; Bahadoran, Shahab; Teixeira da Silva, Jaime A; Dehghani Samani, Amir; Cheraghchi Bashi, Mehdi
Coccidiosis is the most important parasitic disease of the poultry production industry. Due to increasing resistance to conventional anticoccidial agents, it is necessary to find new anticoccidial compounds. Herbal compounds such as those from Artemisia species are promising weapons in this regard since preliminary studies have shown its anticoccidial effects. To compare the anticoccidial effect of a granulated extract of Artemisia sieberi (GEAS) versus monensin in experimental broiler coccidiosis, 120 one-day old Ross 308 broiler chickens were divided in four groups, each with three replicates (n=10). Group 1 was separated as an uninfected negative control and received no treatment. At 21days of age, groups 2, 3 and 4 were inoculated with a mixed suspension of 2×10(5) oocysts of Eimeria tenella, Eimeria maxima, Eimeria acervulina and Eimeria necatrix. Group 2 was maintained as an infected positive control and received no treatment while groups 3 and 4 received GEAS (5mg/kg feed), and monensin (110mg/kg feed) from the first day until 42days of age as a feed additive, respectively. Five days after inoculation, the number of oocycts per gram (OPG) of feces for 7 successive days was measured. Also, mean body weight (MBW), weight gain (WG), feed intake (FI) and feed conversion ratio (FCR) were determined weekly in all groups and replicates. The results showed that GEAS and monensin improved performance attributes (FI, MBW, WG, FCR) and significantly (P<0.05) decreased OPG in inoculated broiler chickens. Copyright © 2014 Elsevier Inc. All rights reserved.
Chiba, Sotaro; Suzuki, Nobuhiro
Viruses often coinfect single host organisms in nature. Depending on the combination of viruses in such coinfections, the interplay between them may be synergistic, apparently neutral with no effect on each other, or antagonistic. RNA silencing is responsible for many cases of interference or cross-protection between viruses, but such antagonistic interactions are usually restricted to closely related strains of the same viral species. In this study, we present an unprecedented example of RNA silencing-mediated one-way interference between unrelated viruses in a filamentous model fungus, Cryphonectria parasitica. The replication of Rosellinia necatrix victorivirus 1 (RnVV1; Totiviridae) was strongly impaired by coinfection with the prototypic member of the genus Mycoreovirus (MyRV1) or a mutant of the prototype hypovirus (Cryphonectria hypovirus 1, CHV1) lacking the RNA silencing suppressor (CHV1-Δp69). This interference was associated with marked transcriptional induction of key genes in antiviral RNA silencing, dicer-like 2 (dcl2) and argonaute-like 2 (agl2), following MyRV1 or CHV1-Δp69 infection. Interestingly, the inhibition of RnVV1 replication was reproduced when the levels of dcl2 and agl2 transcripts were elevated by transgenic expression of a hairpin construct of an endogenous C. parasitica gene. Disruption of dcl2 completely abolished the interference, whereas that of agl2 did not always lead to its abolishment, suggesting more crucial roles of dcl2 in antiviral defense. Taken altogether, these results demonstrated the susceptible nature of RnVV1 to the antiviral silencing in C. parasitica activated by distinct viruses or transgene-derived double-stranded RNAs and provide insight into the potential for broad-spectrum virus control mediated by RNA silencing. PMID:26283371
Mercado, José A; Barceló, Marta; Pliego, Clara; Rey, Manuel; Caballero, José L; Muñoz-Blanco, Juan; Ruano-Rosa, David; López-Herrera, Carlos; de Los Santos, Berta; Romero-Muñoz, Fernando; Pliego-Alfaro, Fernando
The expression of antifungal genes from Trichoderma harzianum, mainly chitinases, has been used to confer plant resistance to fungal diseases. However, the biotechnological potential of glucanase genes from Trichoderma has been scarcely assessed. In this research, transgenic strawberry plants expressing the β-1,3-glucanase gene bgn13.1 from T. harzianum, under the control of the CaMV35S promoter, have been generated. After acclimatization, five out of 12 independent lines analysed showed a stunted phenotype when growing in the greenhouse. Moreover, most of the lines displayed a reduced yield due to both a reduction in the number of fruit per plant and a lower fruit size. Several transgenic lines showing higher glucanase activity in leaves than control plants were selected for pathogenicity tests. When inoculated with Colletotrichum acutatum, one of the most important strawberry pathogens, transgenic lines showed lower anthracnose symptoms in leaf and crown than control. In the three lines selected, the percentage of plants showing anthracnose symptoms in crown decreased from 61 % to a mean value of 16.5 %, in control and transgenic lines, respectively. Some transgenic lines also showed an enhanced resistance to Rosellinia necatrix, a soil-borne pathogen causing root and crown rot in strawberry. These results indicate that bgn13.1 from T. harzianum can be used to increase strawberry tolerance to crown rot diseases, although its constitutive expression affects plant growth and fruit yield. Alternative strategies such as the use of tissue specific promoters might avoid the negative effects of bgn13.1 expression in plant performance.
Sahar M Gadelhaq
Full Text Available Coccidiosis is a serious protozoal disease of poultry. The identification of Eimeria species has important implications for diagnosis and control as well as for epidemiology. The molecular characterization of Eimeria species infecting Egyptian baladi chickens was investigated.Eimeria species oocysts were harvested from intestines of naturally infected Egyptian baldi chickens. The morphometry characterization of oocysts along with COCCIMORPH software was done. The DNA was extracted initially by freezing and thawing then the prepared samples was subjected to commercial DNA kits. The DNA products were analyzed through conventional polymerase chain reaction by using amplified region (SCAR marker.The PCR results confirmed the presence of 7 Eimeria species in the examined fecal samples of Egyptian baldi breed with their specific ampilicon sizes being E. acervulina (811bp, E. brunette (626bp, E. tenella (539bp, E. maxima (272bp, E. necatrix (200bp, E. mitis (327bp and E. praecopx (354bp. A sequencing of the two most predominant species of Eimeria was done, on E. tenella and E. máxima. Analysis of the obtained sequences revealed high identities 99% between Egyptian isolates and the reference one. Similarly, E. maxima isolated from Egyptian baldi chickens showed 98% nucleotide identities with the reference strain. Only single nucleotide substitution was observed among the Egyptian E. tenella isolates (A181G when compared to the reference one. The Egyptian isolates acquired 4 unique mutations (A68T, C164T, G190A and C227G in compared with the reference sequence.This is the first time to identify the 7 species of Eimeria from Egyptian baladi chickens.
Chengat Prakashbabu, B; Thenmozhi, V; Limon, G; Kundu, K; Kumar, S; Garg, R; Clark, E L; Srinivasa Rao, A S R; Raj, D G; Raman, M; Banerjee, P S; Tomley, F M; Guitian, J; Blake, D P
Coccidiosis is one of the biggest challenges faced by the global poultry industry. Recent studies have highlighted the ubiquitous distribution of all Eimeria species which can cause this disease in chickens, but intriguingly revealed a regional divide in genetic diversity and population structure for at least one species, Eimeria tenella. The drivers associated with such distinct geographic variation are unclear, but may impact on the occurrence and extent of resistance to anticoccidial drugs and future subunit vaccines. India is one of the largest poultry producers in the world and includes a transition between E. tenella populations defined by high and low genetic diversity. The aim of this study was to identify risk factors associated with the prevalence of Eimeria species defined by high and low pathogenicity in northern and southern states of India, and seek to understand factors which vary between the regions as possible drivers for differential genetic variation. Faecal samples and data relating to farm characteristics and management were collected from 107 farms from northern India and 133 farms from southern India. Faecal samples were analysed using microscopy and PCR to identify Eimeria occurrence. Multiple correspondence analysis was applied to transform correlated putative risk factors into a smaller number of synthetic uncorrelated factors. Hierarchical cluster analysis was used to identify poultry farm typologies, revealing three distinct clusters in the studied regions. The association between clusters and presence of Eimeria species was assessed by logistic regression. The study found that large-scale broiler farms in the north were at greatest risk of harbouring any Eimeria species and a larger proportion of such farms were positive for E. necatrix, the most pathogenic species. Comparison revealed a more even distribution for E. tenella across production systems in south India, but with a lower overall occurrence. Such a polarised region- and
Wang, Minghong; Wang, Yong; Sun, Xiangzhong; Cheng, Jiasen; Fu, Yanping; Liu, Huiquan; Jiang, Daohong; Ghabrial, Said A.
ABSTRACT Mycoviruses have been detected in all major groups of filamentous fungi, and their study represents an important branch of virology. Here, we characterized a novel double-stranded RNA (dsRNA) mycovirus, Sclerotinia sclerotiorum megabirnavirus 1 (SsMBV1), in an apparently hypovirulent strain (SX466) of Sclerotinia sclerotiorum. Two similarly sized dsRNA segments (L1- and L2-dsRNA), the genome of SsMBV1, are packaged in rigid spherical particles purified from strain SX466. The full-length cDNA sequence of L1-dsRNA/SsMBV1 comprises two large open reading frames (ORF1 and ORF2), which encode a putative coat protein and an RNA-dependent RNA polymerase (RdRp), respectively. Phylogenetic analysis of the RdRp domain clearly indicates that SsMBV1 is related to Rosellinia necatrix megabirnavirus 1 (RnMBV1). L2-dsRNA/SsMBV1 comprises two nonoverlapping ORFs (ORFA and ORFB) encoding two hypothetical proteins with unknown functions. The 5′-terminal regions of L1- and L2-dsRNA/SsMBV1 share strictly conserved sequences and form stable stem-loop structures. Although L2-dsRNA/SsMBV1 is dispensable for replication, genome packaging, and pathogenicity of SsMBV1, it enhances transcript accumulation of L1-dsRNA/SsMBV1 and stability of virus-like particles (VLPs). Interestingly, a conserved papain-like protease domain similar to a multifunctional protein (p29) of Cryphonectria hypovirus 1 was detected in the ORFA-encoded protein of L2-dsRNA/SsMBV1. Phylogenetic analysis based on the protease domain suggests that horizontal gene transfer may have occurred from a single-stranded RNA (ssRNA) virus (hypovirus) to a dsRNA virus, SsMBV1. Our results reveal that SsMBV1 has a slight impact on the fundamental biological characteristics of its host regardless of the presence or absence of L2-dsRNA/SsMBV1. IMPORTANCE Mycoviruses are widespread in all major fungal groups, and they possess diverse genomes of mostly ssRNA and dsRNA and, recently, circular ssDNA. Here, we have characterized
Wang, Minghong; Wang, Yong; Sun, Xiangzhong; Cheng, Jiasen; Fu, Yanping; Liu, Huiquan; Jiang, Daohong; Ghabrial, Said A; Xie, Jiatao
Mycoviruses have been detected in all major groups of filamentous fungi, and their study represents an important branch of virology. Here, we characterized a novel double-stranded RNA (dsRNA) mycovirus, Sclerotinia sclerotiorum megabirnavirus 1 (SsMBV1), in an apparently hypovirulent strain (SX466) of Sclerotinia sclerotiorum. Two similarly sized dsRNA segments (L1- and L2-dsRNA), the genome of SsMBV1, are packaged in rigid spherical particles purified from strain SX466. The full-length cDNA sequence of L1-dsRNA/SsMBV1 comprises two large open reading frames (ORF1 and ORF2), which encode a putative coat protein and an RNA-dependent RNA polymerase (RdRp), respectively. Phylogenetic analysis of the RdRp domain clearly indicates that SsMBV1 is related to Rosellinia necatrix megabirnavirus 1 (RnMBV1). L2-dsRNA/SsMBV1 comprises two nonoverlapping ORFs (ORFA and ORFB) encoding two hypothetical proteins with unknown functions. The 5'-terminal regions of L1- and L2-dsRNA/SsMBV1 share strictly conserved sequences and form stable stem-loop structures. Although L2-dsRNA/SsMBV1 is dispensable for replication, genome packaging, and pathogenicity of SsMBV1, it enhances transcript accumulation of L1-dsRNA/SsMBV1 and stability of virus-like particles (VLPs). Interestingly, a conserved papain-like protease domain similar to a multifunctional protein (p29) of Cryphonectria hypovirus 1 was detected in the ORFA-encoded protein of L2-dsRNA/SsMBV1. Phylogenetic analysis based on the protease domain suggests that horizontal gene transfer may have occurred from a single-stranded RNA (ssRNA) virus (hypovirus) to a dsRNA virus, SsMBV1. Our results reveal that SsMBV1 has a slight impact on the fundamental biological characteristics of its host regardless of the presence or absence of L2-dsRNA/SsMBV1. Mycoviruses are widespread in all major fungal groups, and they possess diverse genomes of mostly ssRNA and dsRNA and, recently, circular ssDNA. Here, we have characterized a novel dsRNA virus