A summary of reptile and anuran amphibian species from Brazilian sandy coastal plains: 31 years of sampling efforts of the "Laboratório de Ecologia de Vertebrados, Universidade do Estado do Rio de Janeiro" / Um resumo das espécies de répteis e anfíbios anuros das planícies costeiras arenosas brasileiras: 31 anos de esforços de amostragem do "Laboratório de Ecologia de Vertebrados da Universidade do Estado do Rio de Janeiro"

Abstract Although currently there is already a set of studies regarding ecological aspects of some particular reptile and amphibian species living in Brazilian sandy coastal plains (including the so-called "restinga" and "campo nativo" habitats), there is comparatively few information on the species composition usually associated to these environments. During 31 years (1988-2019) of herpetological studies carried out in sandy coastal plains environments by our research team of the Laboratory of Vertebrate Ecology (Department of Ecology, Universidade do Estado do Rio de Janeiro, in Rio de Janeiro Brazil) we have surveyed reptile and amphibian communities and performed different studies with similar methods in 70 sites from 10 different states along the Brazilian coast. Our surveys resulted in records of 87 species of reptile (five turtles, two crocodylians, six amphisbaenians, 36 lizards and 39 snakes) from 24 families, and 77 species of anuran amphibians from nine families. We have studied multiple natural history topics for anurans and reptiles which resulted in the publication of some specific ecological studies, especially regarding some species, encompassing population and community ecology, foraging and feeding habits, species activity, thermoregulation, reproduction, use of microhabitats, and parasitism by ecto and endoparasites. Our results along these three decades have also contributed for the description of four new lizard species (Ameivula nativo, Glaucomastix littoralis, G. abaetensis and G. itabaianensis). Our studies constitute an important contribution to the knowledge of the ecology of anuran amphibians and reptiles in these ecosystems, as well as to the conservation of sandy coastal plains environment. The checklist presented in this study, based on our records of sandy coastal plains herpetofauna, provides for many localities along the Brazilian coast, the needed knowledge on species occurrence, including the presence of endemic and/or endangered species, which can be of value for many conservation actions.

Resumo Embora atualmente exista um conjunto de estudos sobre aspectos ecológicos de algumas espécies de répteis e de anfíbios que ocorrem nas planícies costeiras arenosas brasileiras (incluindo os chamados habitats de "restinga" e de "campo nativo"), há relativamente poucas informações sobre a composição de espécies geralmente associada a esses ambientes. Durante 31 anos (1988-2019) de estudos herpetológicos realizados em restingas por nossa equipe de pesquisa do Laboratório de Ecologia de Vertebrados (Departamento de Ecologia, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brasil) nós estudamos comunidades de répteis e de anfíbios e realizamos diferentes estudos com métodos semelhantes em 70 localidades de dez diferentes Estados ao longo da costa brasileira. Nossas pesquisas resultaram em registros de 87 espécies de répteis (cinco tartarugas, dois crocodilianos, seis anfisbênios, 36 lagartos e 39 serpentes) de 24 famílias, e 77 espécies de anfíbios anuros de nove famílias. Estudamos vários tópicos de história natural sobre anuros e répteis, que resultaram na publicação de alguns estudos ecológicos específicos, especialmente em relação a algumas espécies, abrangendo ecologia populacional e de comunidades, forrageamento e dieta, horário de atividade de espécies, termorregulação, reprodução, uso do microhabitat e parasitismo por ecto e endoparasitas. Nossos resultados ao longo dessas três décadas também contribuíram para a descrição de quatro novas espécies de lagartos (Ameivula nativo, Glaucomastix littoralis, G. abaetensis e G. itabaianensis). Nossos estudos constituem uma importante contribuição para o conhecimento da ecologia de répteis e de anfíbios anuros nesses ecossistemas, bem como para a conservação dos ecossistemas de restinga. A lista de espécies apresentada neste estudo, com base em nossos registros de herpetofauna das planícies costeiras arenosas, fornece para muitas localidades ao longo da costa brasileira o conhecimento necessário sobre a ocorrência de espécies, incluindo a presença de espécies endêmicas e/ ou ameaçadas de extinção, que podem ser úteis para muitas ações de conservação.

A summary of reptile and anuran amphibian species from Brazilian sandy coastal plains: 31 years of sampling efforts of the "Laboratório de Ecologia de Vertebrados, Universidade do Estado do Rio de Janeiro".

Although currently there is already a set of studies regarding ecological aspects of some particular reptile and amphibian species living in Brazilian sandy coastal plains (including the so-called "restinga" and "campo nativo" habitats), there is comparatively few information on the species composition usually associated to these environments. During 31 years (1988-2019) of herpetological studies carried out in sandy coastal plains environments by our research team of the Laboratory of Vertebrate Ecology (Department of Ecology, Universidade do Estado do Rio de Janeiro, in Rio de Janeiro Brazil) we have surveyed reptile and amphibian communities and performed different studies with similar methods in 70 sites from 10 different states along the Brazilian coast. Our surveys resulted in records of 87 species of reptile (five turtles, two crocodylians, six amphisbaenians, 36 lizards and 39 snakes) from 24 families, and 77 species of anuran amphibians from nine families. We have studied multiple natural history topics for anurans and reptiles which resulted in the publication of some specific ecological studies, especially regarding some species, encompassing population and community ecology, foraging and feeding habits, species activity, thermoregulation, reproduction, use of microhabitats, and parasitism by ecto and endoparasites. Our results along these three decades have also contributed for the description of four new lizard species (Ameivula nativo, Glaucomastix littoralis, G. abaetensis and G. itabaianensis). Our studies constitute an important contribution to the knowledge of the ecology of anuran amphibians and reptiles in these ecosystems, as well as to the conservation of sandy coastal plains environment. The checklist presented in this study, based on our records of sandy coastal plains herpetofauna, provides for many localities along the Brazilian coast, the needed knowledge on species occurrence, including the presence of endemic and/or endangered species, which can be of value for many conservation actions.

Role of Stenotrophomonas maltophilia isolation in patients with non-CF bronchiectasis.

INTRODUCTION: Stenotrophomonas maltophilia is a bacteria whose role in patients with cystic fibrosis (CF) bronchiectasis has been previously studied; little is known about its role in non-CF bronchiectasis. MATERIALS AND METHODS: Aim of our study is to investigate the risk factors for S. maltophilia acquisition and its clinical impact on bronchiectasis patients. A retrospective observational cohort study enrolling patients attending the Bronchiectasis Clinic at the Royal Infirmary of Edinburgh, Scotland, UK. A total of 167 bronchiectasis patients undergoing intravenous (IV) antibiotic therapy were selected and divided according to single or chronic S. maltophilia isolation in sputum. The risk factors and prognostic impact were studied. RESULTS: Single isolation was independently associated with lower baseline % predicted forced expiratory volume in 1 s [odds ratio (OR) 0.98; 95% confidence interval (CI) 0.970-1.044; P = 0.025] and with less radiological involvement (OR 0.379; 95% CI 0.175-0.819; P = 0.01). Chronic isolation was associated with the number of IV antibiotic courses in the year before and after the first isolation (OR 1.2; 95% CI 1.053-1.398; P = 0.007) and with the absence of Pseudomonas aeruginosa colonization (OR 0.207; 95% CI 0.056-0.764; P = 0.02). In the chronic isolation group, there were more exacerbations and more need of IV antibiotics in the year after the first isolation. CONCLUSIONS: Poor lung function is the main independent risk factor for single isolation of S. maltophilia. For chronic colonization, the main independent risk factor is the number of IV antibiotic courses and the absence of P. aeruginosa chronic colonization. Only when chronically present, S. maltophilia had a clinical impact with more exacerbations.

Co-Culture of Plant Beneficial Microbes as Source of Bioactive Metabolites.

In microbial cultures the production of secondary metabolites is affected by experimental conditions, and the discovery of novel compounds is often prevented by the re-isolation of known metabolites. To limit this, it is possible to cultivate microorganisms by simulating naturally occurring interactions, where microbes co-exist in complex communities. In this work, co-culturing experiments of the biocontrol agent Trichoderma harzianum M10 and the endophyte Talaromyces pinophilus F36CF have been performed to elicit the expression of genes which are not transcribed in standard laboratory assays. Metabolomic analysis revealed that the co-culture induced the accumulation of siderophores for both fungi, while production of M10 harzianic and iso-harzianic acids was not affected by F36CF. Conversely, metabolites of the latter strain, 3-O-methylfunicone and herquline B, were less abundant when M10 was present. A novel compound, hereby named harziaphilic acid, was isolated from fungal co-cultures, and fully characterized. Moreover, harzianic and harziaphilic acids did not affect viability of colorectal cancer and healthy colonic epithelial cells, but selectively reduced cancer cell proliferation. Our results demonstrated that the co-cultivation of plant beneficial fungi may represent an effective strategy to modulate the production of bioactive metabolites and possibly identify novel compounds.

Secondary metabolites from the endophytic fungus Talaromyces pinophilus.

Endophytic fungi have a great influence on plant health and growth, and are an important source of bioactive natural compounds. Organic extracts obtained from the culture filtrate of an endophytic strain of Talaromyces pinophilus isolated from strawberry tree (Arbutus unedo) were studied. Metabolomic analysis revealed the presence of three bioactive metabolites, the siderophore ferrirubin, the platelet-aggregation inhibitor herquline B and the antibiotic 3-O-methylfunicone. The latter was the major metabolite produced by this strain and displayed toxic effects against the pea aphid Acyrthosiphon pisum (Homoptera Aphidiidae). This toxicity represents an additional indication that the widespread endophytic occurrence of T. pinophilus may be related to a possible role in defensive mutualism. Moreover, the toxic activity on aphids could promote further study on 3-O-methylfunicone, or its derivatives, as an alternative to synthetic chemicals in agriculture.

A summary of reptile and anuran amphibian species from Brazilian sandy coastal plains: 31 years of sampling efforts of the Laboratório de Ecologia de Vertebrados, Universidade do Estado do Rio de Janeiro

Abstract Although currently there is already a set of studies regarding ecological aspects of some particular reptile and amphibian species living in Brazilian sandy coastal plains (including the so-called restinga and campo nativo habitats), there is comparatively few information on the species composition usually associated to these environments. During 31 years (1988-2019) of herpetological studies carried out in sandy coastal plains environments by our research team of the Laboratory of Vertebrate Ecology (Department of Ecology, Universidade do Estado do Rio de Janeiro, in Rio de Janeiro Brazil) we have surveyed reptile and amphibian communities and performed different studies with similar methods in 70 sites from 10 different states along the Brazilian coast. Our surveys resulted in records of 87 species of reptile (five turtles, two crocodylians, six amphisbaenians, 36 lizards and 39 snakes) from 24 families, and 77 species of anuran amphibians from nine families. We have studied multiple natural history topics for anurans and reptiles which resulted in the publication of some specific ecological studies, especially regarding some species, encompassing population and community ecology, foraging and feeding habits, species activity, thermoregulation, reproduction, use of microhabitats, and parasitism by ecto and endoparasites. Our results along these three decades have also contributed for the description of four new lizard species (Ameivula nativo, Glaucomastix littoralis, G. abaetensis and G. itabaianensis). Our studies constitute an important contribution to the knowledge of the ecology of anuran amphibians and reptiles in these ecosystems, as well as to the conservation of sandy coastal plains environment. The checklist presented in this study, based on our records of sandy coastal plains herpetofauna, provides for many localities along the Brazilian coast, the needed knowledge on species occurrence, including the presence of endemic and/or endangered species, which can be of value for many conservation actions.

Resumo Embora atualmente exista um conjunto de estudos sobre aspectos ecológicos de algumas espécies de répteis e de anfíbios que ocorrem nas planícies costeiras arenosas brasileiras (incluindo os chamados habitats de restinga e de campo nativo), há relativamente poucas informações sobre a composição de espécies geralmente associada a esses ambientes. Durante 31 anos (1988-2019) de estudos herpetológicos realizados em restingas por nossa equipe de pesquisa do Laboratório de Ecologia de Vertebrados (Departamento de Ecologia, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brasil) nós estudamos comunidades de répteis e de anfíbios e realizamos diferentes estudos com métodos semelhantes em 70 localidades de dez diferentes Estados ao longo da costa brasileira. Nossas pesquisas resultaram em registros de 87 espécies de répteis (cinco tartarugas, dois crocodilianos, seis anfisbênios, 36 lagartos e 39 serpentes) de 24 famílias, e 77 espécies de anfíbios anuros de nove famílias. Estudamos vários tópicos de história natural sobre anuros e répteis, que resultaram na publicação de alguns estudos ecológicos específicos, especialmente em relação a algumas espécies, abrangendo ecologia populacional e de comunidades, forrageamento e dieta, horário de atividade de espécies, termorregulação, reprodução, uso do microhabitat e parasitismo por ecto e endoparasitas. Nossos resultados ao longo dessas três décadas também contribuíram para a descrição de quatro novas espécies de lagartos (Ameivula nativo, Glaucomastix littoralis, G. abaetensis e G. itabaianensis). Nossos estudos constituem uma importante contribuição para o conhecimento da ecologia de répteis e de anfíbios anuros nesses ecossistemas, bem como para a conservação dos ecossistemas de restinga. A lista de espécies apresentada neste estudo, com base em nossos registros de herpetofauna das planícies costeiras arenosas, fornece para muitas localidades ao longo da costa brasileira o conhecimento necessário sobre a ocorrência de espécies, incluindo a presença de espécies endêmicas e/ ou ameaçadas de extinção, que podem ser úteis para muitas ações de conservação.

Sudden Vegetation Dieback in Atlantic and Gulf Coast Salt Marshes.

Salt marshes rank as the most productive ecosystems on the planet. Biomass production can be greater than 3 kg dry matter/m2/year, which is 40% more biomass than tropical rainforests produce. Salt marshes provide multiple benefits to mankind. For example, coastal communities receive protection from storm surges and wave erosion. Salt marshes absorb excess nitrogen and phosphorus from sewage and fertilizer run-off into rivers, which, in turn, prevents algal blooms and hypoxia in coastal waters. In addition, these unique ecosystems provide habitat and shelter for many hundreds of species of shellfish, finfish, migratory and sedentary birds, and other marine animals. Despite the richness in animal species, the intertidal marshes of the salt marsh ecosystem are dominated by only a few plant species. Of these, the most prevalent plant species in a marsh are the tall and short forms of smooth cordgrass (Spartina alterniflora). The first recorded account of a dieback in a U.S. salt marsh was in the early 1990s in the Florida panhandle where patches of Sp. alterniflora as large as 1 ha died. This article explores possible causes of Sudden Vegetation Dieback.

First Report of Boxwood Blight Caused by Cylindrocladium pseudonaviculatum in the United States.

In September and October 2011, a new disease was observed on Buxus spp. in North Carolina and Connecticut, respectively. In North Carolina, over 10,000 containerized Buxus sempervirens (American boxwood) were affected at one location. A few weeks later, the disease was found in Connecticut on entire plantings of B. sempervirens 'Suffruticosa' (English boxwood) at two residential properties, and shortly thereafter on over 150,000 plants at two production nurseries. Initial foliar symptoms appeared as light to dark brown spots, often with dark borders. Spots enlarged and coalesced, often with a concentric pattern, and black streaks or cankers developed on stems. Infected leaves became brown or straw colored and dropped quickly after foliar symptoms were visible. Branch dieback and plant death were also observed in Connecticut. Cultures were isolated from symptomatic leaves and stems and identified as Cylindrocladium pseudonaviculatum Crous, Groenewald & Hill 2002 (1) (syn. Cylindrocladium buxicola Henricot 2002 [2]) on the basis of morphological characteristics. Macroconidiophores were single or in groups of up to three and comprised a stipe, stipe extension, and a penicillate arrangement of fertile branches. The stipe extension was septate, hyaline (89 to 170 × 2 to 4.5 µm), and terminated in an ellipsoidal vesicle (6 to 11 µm diameter) with a papillate or pointed apex. Conidia were cylindrical, straight, hyaline, and one septate (48 to 62 × 4 to 6 µm), occurring in slimy clusters. No microconidiophores were observed. Chlamydospores were medium to dark brown, thick walled, and smooth to rough. Microsclerotia were observed on PDA (1). A portion of ß-tubulin gene sequence from two Connecticut (Genbank Accession Nos. JQ866628 and JQ866629) and two North Carolina isolates showed 100% similarity with only C. pseudonaviculatum strains. USDA-APHIS-PPQ confirmed this new United States record on October 24, 2011. Pathogenicity was confirmed by inoculating three 1-gallon container plants of B. sempervirens 'Suffruticosa' in North Carolina and four liners of B. sinica var. insularis × B. sempervirens 'Green Velvet' in Connecticut with a spore suspension of approximately 5.0 × 106 conidia (North Carolina) or 1.0 × 106 conidia (Connecticut) on the foliage of each plant; untreated control plants were sprayed with water. After incubation at ambient temperature, all inoculated plants developed foliar and stem lesions within 3 to 4 days and blighting occurred within 2 weeks; control plants remained asymptomatic. C. pseudonaviculatum was reisolated from inoculated plants. To our knowledge, this is the first report of C. pseudonaviculatum in the United States. C. pseudonaviculatum causes a serious disease of Buxus spp. in the United Kingdom and several other European countries as well as New Zealand (1). Confirmation of boxwood blight in the United States is significant because of the popularity of boxwood as a landscape plant, and because of the potential economic impact this disease may have on commercial growers; boxwood production in the United States has an annual wholesale market value of approximately $103 million (3). References: (1) P. Crous, et al. Sydowia 54:23, 2002. (2) B. Henricot and A. Culham Mycologia 94: 980, 2002. (3) USDA-NASS, Census of Horticulture, 2010.

First Report of Cylindrocladium pseudonaviculatum Causing Leaf Spot of Pachysandra terminalis.

Cylindrocladium pseudonaviculatum Crous, J.Z., Groenew. & C.F. Hill 2002 was recently reported infecting common boxwood, Buxus sempervirens L., in Connecticut (2). We isolated the pathogen from leaf and stem lesions of B. sempervirens and obtained single-spored cultures on half-strength potato dextrose agar (½PDA). The pathogen was identified as C. pseudonaviculatum by morphological characteristics (1). Colony size reached 71 mm in diameter after 14 days at room temperature on ½PDA, and was fluffy with white aerial hyphae, mars brown, and reverse color chestnut brown at the center fading to pale brown forming concentric bands. Macroconidiophores were solitary or in a group of up to three, comprised a stipe, a sterile elongation, and one to three penicillate fertile branches. The stipe was up to nine septate, 90 to 250 µm long, colorless, smooth, terminating in a naviculate or broadly ellipsoidal vesicle with a pointed or papillate apex, and 27 to 50 × 6.5 to 9 µm. Primary branches were zero- to one-septate, 20 to 36 × 4 to 5 µm; secondary branches were aseptate and 11 to 20 × 3 to 4.5 µm; tertiary branches were rare, each terminal branch producing two to five phialides; phialides were doliiform or reniform, colorless, 12 to 18 µm. Conidia were cylindrical, rounded at both ends, straight, smooth, colorless, two-celled, 48 to 55 × 4.5 to 5.5 µm, and in colorless slimy cylindrical clusters. Microconidiophores were not observed. Chlamydospores were golden to dark brown, thick-walled, and smooth or rough. Microsclerotia were present on ½PDA. Primers T1 and T22 (3) were used to amplify a portion of the ß-tubulin gene from isolates Cps-CT-L1 and Cps-CT-S1. Amplified sequences were used in a BLAST search against the GenBank database to demonstrate 100% sequence identity only with other C. pseudonaviculatum strains. Both sequences were deposited in GenBank (Accession Nos. JQ866628 and JQ866629), using corresponding gene data from C. pseudonaviculatum strain STE-U 3399 (GenBank Accession No. AF449455) to distinguish coding from noncoding regions. Healthy plants of Japanese spurge, Pachysandra terminalis, with three plants per 10 cm diameter pot, were inoculated with water alone or a conidial suspension of C. pseudonaviculatum isolate Cps-CT-L1 (ATCC MYA-4891) (1.0 × 106 conidia/plant) with a handheld sprayer until runoff. Plants were kept moist in a plastic bag for 48 h at laboratory temperature and then transferred to the greenhouse. Circular lesions (1- to 4-mm diameter) were evident on leaves after 10 days. All 12 inoculated plants developed lesions, and no lesions were observed on noninoculated plants. Leaves with lesions were surface sterilized in 0.5% NaOCl for 30 s, rinsed twice in sterile water, and lesion margins plated onto water agar or ½PDA. The pathogen was reisolated from at least one leaf per plant. Koch's postulates were performed again with isolate Cps-CT-S1 (ATCC MYA-4890). After 3 weeks, many of the leaves with lesions yellowed and dropped to the soil surface and heavy sporulation of C. pseudonaviculatum and microsclerotia were observed. To our knowledge, this is the first report of C. pseudonaviculatum causing a leaf spot disease on P. terminalis. Pachysandra is a widely grown ground cover suitable for shady, humid environmental conditions that may be conducive for the development of disease. References: (1) P. Crous, et al. Sydowia 54:23, 2002. (2) K. Ivors et al. Plant Disease. 96:X, 2012. (3) K. O'Donnell and E. Cigelnik Mol. Phylogenet. Evol. 7:103, 1997.

First Report of Crown Rot of Bloodroot (Sanguinaria canadensis) Caused by Fusarium oxysporum in the United States.

Bloodroot (Sanguinaria canadensis L [Papaveraceae]) is a native herbaceous perennial in eastern North America, found from Nova Scotia to Florida. Although it is a common wildflower, rhizomes of double-flowered forms are sold commercially. Rhizomes planted into a wooded area in Guilford, CT produced healthy stems and flowers for a few years and then began to collapse and die in 2008. The same symptoms were observed with a new planting in 2011. Initially, leaves were dull green and were more leathery than healthy leaves. Eventually, the leaves collapsed at the junction of the petioles and the rhizomes. Vascular discoloration, if present, was obscured by the red pigmentation in the rhizome. A Fusarium sp. sporulated on the discolored tissue at the junction between healthy and rotted tissue. Stem pieces were surface disinfested (0.53% NaClO for 1 min), rinsed, and placed on Peptone-PCNB agar (2) at room temperature for 5 days. Colonies originating from single spores were subcultured on carnation leaf agar (2) and identified as Fusarium oxysporum based on falcate, thin-walled, three-septate macroconida borne in monophialides on doliform conidiophores (2). Four rhizomes of double-flowered bloodroot were planted in potting mix in the greenhouse in October 2010; sprouts were observed in March 2011. Two plants were inoculated in March 2011 by drenching the soil with 100 ml of a conidial suspension (106 spores/ml) and two control plants were treated with deionized water. Two months later, the inoculated plants were smaller than the controls. The treated plants subsequently collapsed and F. oxysporum was reisolated. Control plants remained healthy and F. oxysporum was not isolated. DNA extracted from the F. oxysporum isolates was used to obtain partial sequences of the translational elongation factor 1-alpha (tef1) gene, which were then blasted against the GenBank database. We observed a 100% similarity with F. oxysporum f. sp. lycopersici. The bloodroot isolates were compared with a known F. oxysporum f. sp. lycopersici isolate for their ability to cause disease on 2-week-old tomato seedlings (cv. Brandywine), using pathogenicity tests as described above. The known F. oxysporum f. sp. lycopersici isolate caused severe wilt and stunting of the tomato seedlings, but the bloodroot isolate caused no symptoms in inoculated seedling compared with those not inoculated. These results suggest that there may be more hosts for isolates in the F. oxysporum f. sp. lycopersici species complex than previously thought (1). An isolate (O-2603) has been deposited at the Fusarium Research Laboratory at Pennsylvania State University, University Park. Since bloodroot is now being sold commercially as an ornamental, disease management strategies may be needed. To our knowledge, this is the first report of a Fusarium crown rot of bloodroot. References: (1).V. Edel-Hermann et al. Online publication. doi:10.1111/j.1365-3059.2011.02551.x. Plant Pathology, 2011. (2) J. Leslie and B. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006.

Secondary metabolites produced by a root-inhabiting sterile fungus antagonistic towards pathogenic fungi.

AIMS: A sterile red fungus (SRF) isolated from cortices of roots of both wheat (Triticum aestivum cv. Gamenya) and ryegrass (Lolium rigidum cv. Wimmera) was found to protect the hosts from phytopathogens and promote plant growth. In this work, the major secondary metabolites produced by this SRF were analysed, and their antibiotic and plant-growth-promoting activities investigated. METHODS AND RESULTS: Two main compounds, veratryl alcohol (VA) and 4-(hydroxymethyl)-quinoline, were isolated from the culture filtrate of the fungus. In antifungal assays, VA inhibited the growth of Sclerotinia sclerotiorum and Pythium irregulare even at low amounts, while high concentrations (>100 microg per plug) of 4-(hydroxymethyl)-quinoline were needed. Both metabolites revealed weak inhibition of Rhizoctonia solani. Furthermore, both compounds showed a growth promotion activity on canola (Brassica napus) seedlings used as bioassays. CONCLUSIONS: Isolation and characterization of the main secondary metabolites from culture filtrates of a root-inhabiting sterile fungus are reported. The biological assays indicate that these secondary metabolites may have a role in both plant growth regulation and antifungal activity. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides a better understanding of the metabolism of a cortical fungus that may have a useful role in the biological suppression of root-infecting soil-borne plant pathogens.

Factors affecting the production of Trichoderma harzianum secondary metabolites during the interaction with different plant pathogens.

AIMS: Strains of Trichoderma spp. produce numerous bioactive secondary metabolites. The in vitro production and antibiotic activities of the major compounds synthesized by Trichoderma harzianum strains T22 and T39 against Leptosphaeria maculans, Phytophthora cinnamomi and Botrytis cinerea were evaluated. Moreover, the eliciting effect of viable or nonviable biomasses of Rhizoctonia solani, Pythium ultimum or B. cinerea on the in vitro production of these metabolites was also investigated. METHODS AND RESULTS: T22azaphilone, 1-hydroxy-3-methyl-anthraquinone, 1,8-dihydroxy-3-methyl-anthraquinone, T39butenolide, harzianolide, harzianopyridone were purified, characterized and used as standards. In antifungal assays, T22azaphilone and harzianopyridone inhibited the growth of the pathogens tested even at low doses (1-10 microg per plug), while high concentrations of T39butenolide and harzianolide were needed (>100 microg per plug) for inhibition. The in vitro accumulation of these metabolites was quantified by LC/MS. T22azaphilone production was not enhanced by the presence of the tested pathogens, despite its antibiotic activity. On the other hand, the anthraquinones, which showed no pathogen inhibition, were stimulated by the presence of P. ultimum. The production of T39butenolide was significantly enhanced by co-cultivation with R. solani or B. cinerea. Similarly, viable and nonviable biomasses of R. solani or B. cinerea increased the accumulation of harzianopyridone. Finally, harzianolide was not detected in any of the interactions examined. CONCLUSIONS: The secondary metabolites analysed in this study showed different levels of antibiotic activity. Their production in vitro varied in relation to: (i) the specific compound; (ii) the phytopathogen used for the elicitation; (iii) the viability of the elicitor; and (iv) the balance between elicited biosynthesis and biotransformation rates. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of cultures of phytopathogens to enhance yields of Trichoderma metabolites could improve the production and application of novel biopesticides and biofertilizers based on the active compounds instead of the living microbe. This could have a significant beneficial impact on the management of diseases in crop plants.

First Report of Pestalotiopsis paeoniicola Causing Twig Blight on Paeonia suffruticosa in North America.

Native to China, tree peony (Paeonia suffruticosa Andrews) is a perennial valued for its showy, often fragrant flowers. In May 2007, samples received from two field-grown tree peonies from Torrington, CT exhibited twig blight characterized by small, black spots on the bark of living or dead branches, and associated with subsequent death and loss of the branches. Colonies on potato dextrose agar (PDA) attained 5.5 to 6.5 cm in diameter in 9 days at 25°C and produced abundant, white, aerial mycelium, later turning pink and forming scattered, wet, black, acervular conidiomata containing abundant conidia that were five-celled, fusiform, smooth, straight, or slightly curved, and 24.0 ± 1.8 × 7.1 ± 0.5 µm (n = 20); three intermediate cells were dark brown and end cells were hyaline; two to four hyaline whip-like appendages on the apical cell, 26.6 ± 4.1 µm long, and one appendage on the basal cell, 6.7 ± 1.1 µm long. We identified the fungus as Pestalotiopsis paeoniicola (Tsukam. & T. Hino) J.G. Wei & T. Xu (= Pestalotia paeoniicola Tsukam. & T. Hino) based on morphology and host and have deposited a culture with CBS (CBS 124745). Originally described from Paeonia suffruticosa in Japan (1,3), the fungus is also found in China on Paeonia lactiflora (Pall.) (2). To confirm pathogenicity, two 5-year-old potted plants of Paeonia suffruticosa cv. Shichifukujin were inoculated with the fungus as follows: a 2-week-old PDA culture was used to produce a conidial spore suspension in sterile water; incisions (5 mm) were made with a sterile scalpel; 4 µl of either the conidial suspension or water were applied; and the wounds were wrapped with Parafilm. Each plant received three replicates each of the treatment and the control. Plants were loosely covered with plastic bags, kept on laboratory benches with ambient light for 1 month, and transferred to a greenhouse for an additional 2 months. Subsequent inspection revealed irregular or elongate, grayish brown lesions at the treatment inoculations, while the controls remained symptom free. Lesions gradually girdled the branches and the infected cortex turned dark brown. At advanced stages, small, black acervuli developed. Treatment and control tissues were cut into four pieces, surface sterilized, and placed on malt extract agar in petri dishes, four per dish. These were incubated for 9 days at 25°C under ambient light. Reisolation of P. paeoniicola only from tissues that had been treated with the pathogen, not from control inoculations, confirmed that the causal agent was P. paeoniicola. DNA sequences were obtained from the ß-tubulin gene (1,512 bp) and the internal transcribed spacer (ITS1 and ITS2) and 5.8S regions of the rDNA (550 bp) and were deposited in GenBank (Nos. FJ975603 and FJ997645, respectively). Neither ITS nor ß-tubulin sequences distinguished this P. paeoniicola isolate from Pestalotiopsis spp. whose sequences are deposited in GenBank. Morphological characteristics identify the fungus as P. paeoniicola, constituting, to our knowledge, the first report of this pathogen in North America. Discovery of P. paeoniicola in field-grown plants warrants further monitoring by growers because of the uncertain level of threat this pathogen may pose to the tree peony industry. References: (1) E. F. Guba. Monograph of Monochaetia and Pestalotia: 224. Harvard University Press, Cambridge, MA, 1961. (2) F. Tai. Sylloge Fungorum Sinicorum: 1021. Sci. Acad. Sin, Peking, 1979. (3) E. Tsukamoto et al. Ann. Phytopathol. Soc. Jpn. 4:183, 1956.

Choline pivaloyl ester strengthened the benefit effects of Tacrine and Galantamine on electroencephalographic and cognitive performances in nucleus basalis magnocellularis-lesioned and aged rats.

The aim of the present work was the assessment of the effects produced on the electroencephalographic (EEG) activity and the cognitive and memory performances of nucleus basalis magnocellularis (NBM)-lesioned or aged rats by the combined treatment with [2-(2,2-dimethylpropionyloxy)ethyl]trimethylammonium 2,2-dimethylpropionate (choline pivaloyl ester) (CPE) and the Cholinesterase inhibitors (ChEIs) Tacrine (THA) and Galantamine (GAL). Intraperitoneal administration of CPE combined with THA or GAL to both NBM-lesioned or aged rats, produced EEG desynchronisation, and a significant decrease in the energy of the total EEG spectrum and the lower frequency bands (delta 0.25-3 and theta 4-7 Hz) lasting many minutes. Furthermore, drug associations reversed in aged rats the scopolamine (0.2 mg/kg, i.p.)-induced increase in EEG power, slow waves and high-voltage spindle (HVS). Furthermore, the combined administration of CPE and Cholinesterase inhibitors in both NBM-lesioned or aged animals, improved performances in all behavioural tasks, enhancing object discrimination, increasing locomotory activity and alternation choice in T-maze, ameliorating retention in passive avoidance and decreasing escape latency in Morris water maze. In all test, AChEIs and CPE combinations proved to be more effective than CPE, THA or GAL given alone. In conclusion, the present work shows the ability of choline pivaloyl ester in strengthening the positive cerebral activity of THA and GAL.

Major secondary metabolites produced by two commercial Trichoderma strains active against different phytopathogens.

AIMS: Trichoderma harzianum strains T22 and T39 are two micro-organisms used as active agents in a variety of commercial biopesticides and biofertilizers and widely applied amongst field and greenhouse crops. The production, isolation, biological and chemical characterization of the main secondary metabolites produced by these strains are investigated. METHODS AND RESULTS: Of the three major compounds produced by strain T22, one is a new azaphilone that shows marked in vitro inhibition of Rhizoctonia solani, Pythium ultimum and Gaeumannomyces graminis var. tritici. In turn, filtrates from strain T39 were demonstrated to contain two compounds previously isolated from other T. harzianum strains and a new butenolide. The production of the isolated metabolites was also monitored by liquid chromatography/mass spectrometry during in vitro interaction with R. solani. CONCLUSIONS: This paper reports the isolation and characterization of the main secondary metabolites obtained from culture filtrates of two T. harzianum strains and their production during antagonistic interaction with the pathogen R. solani. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first work on secondary metabolites produced by the commercially applied strains T22 and T39. Our results provide a better understanding of the metabolism of these fungi, which are both widely used as biopesticides and/or biofertilizers in biocontrol.

Body size, diet and endoparasites of the microhylid frog Chiasmocleis capixaba in an Atlantic Forest area of southern Bahia state, Brazil.

We analyzed the diet composition, endoparasites and sexual size dimorphism of the microhylid frog Chiasmocleis capixaba (Microhylidae) from a "mussununga" habitat in the municipality of Nova Viçosa, southern Bahia state, Brazil. All the 119 specimens analyzed were collected in a single night of heavy rainfall. Females (mean snout-vent length = 15.7 + 3.0 mm) were significantly larger than males (mean snout-vent length = 13.2 + 2.1 mm), and specimens of both sexes were smaller than those of a conspecific population previously reported in Aracruz, state of Espírito Santo state. The diet of C. capixaba was dominated by mites, ants and collembolans. Seventy-nine frogs (66.4% of the total) were infected by helminths, all belonging to a single species, Cosmocerca ornata, an intestinal nematode parasite.

Body size, diet and endoparasites of the microhylid frog Chiasmocleis capixaba in an Atlantic Forest area of southern Bahia state, Brazil

Analisamos a composição da dieta, endoparasitas e dimorfismo sexual em tamanho no anuro Chiasmocleis capixaba (Microhylidae) em um habitat de "mussununga" no município de Nova Viçosa, no sul do Estado da Bahia, Brasil. Todos os 119 espécimes analisados foram coletados em uma única noite com chuva intensa. As fêmeas (comprimento rostro-cloacal médio = 15,7 + 3,0 mm) foram significativamente maiores que os machos (comprimento rostro-anal médio = 13,2 + 2,1 mm), e indivíduos de ambos sexos foram menores do que os de uma população coespecífica de Aracruz, no Estado do Espírito Santo. A dieta de C. capixaba estava dominada por ácaros, formigas e colêmbolos. Setenta e nove indivíduos (66.4% do total) estavam infectados por helmintos, todos pertencentes a uma única espécie, Cosmocerca ornata, um nematódeo intestinal.

Body size, diet and endoparasites of the microhylid frog Chiasmocleis capixaba in an Atlantic Forest area of southern Bahia state, Brazil

We analyzed the diet composition, endoparasites and sexual size dimorphism of the microhylid frog Chiasmocleis capixaba (Microhylidae) from a "mussununga" habitat in the municipality of Nova Viçosa, southern Bahia state, Brazil. All the 119 specimens analyzed were collected in a single night of heavy rainfall. Females (mean snout-vent length = 15.7 + 3.0 mm) were significantly larger than males (mean snout-vent length = 13.2 + 2.1 mm), and specimens of both sexes were smaller than those of a conspecific population previously reported in Aracruz, state of Espírito Santo state. The diet of C. capixaba was dominated by mites, ants and collembolans. Seventy-nine frogs (66.4% of the total) were infected by helminths, all belonging to a single species, Cosmocerca ornata, an intestinal nematode parasite.

Analisamos a composição da dieta, endoparasitas e dimorfismo sexual em tamanho no anuro Chiasmocleis capixaba (Microhylidae) em um habitat de "mussununga" no município de Nova Viçosa, no sul do Estado da Bahia, Brasil. Todos os 119 espécimes analisados foram coletados em uma única noite com chuva intensa. As fêmeas (comprimento rostro-cloacal médio = 15,7 + 3,0 mm) foram significativamente maiores que os machos (comprimento rostro-anal médio = 13,2 + 2,1 mm), e indivíduos de ambos sexos foram menores do que os de uma população coespecífica de Aracruz, no Estado do Espírito Santo. A dieta de C. capixaba estava dominada por ácaros, formigas e colêmbolos. Setenta e nove indivíduos (66.4% do total) estavam infectados por helmintos, todos pertencentes a uma única espécie, Cosmocerca ornata, um nematódeo intestinal.

Relative efficiency of anuran sampling methods in a restinga habitat (Jurubatiba, Rio de Janeiro, Brazil)

São escassos os estudos sobre anuros em áreas de restinga e, conseqüentemente, há pouca informação sobre os métodos mais eficientes de amostragem nesse habitat. De forma geral, são utilizados dez métodos de amostragem em comunidades de anuros em áreas tropicais e subtropicais. Neste estudo, avaliamos quais seriam os métodos mais eficientes para amostrar a comunidade de anfíbios na restinga do Parque Nacional da Restinga de Jurubatiba, norte do Estado do Rio de Janeiro. Analisamos seis métodos usualmente utilizados para a amostragem de anuros. Registramos, para cada método, o tempo total dispendido (em min), o número de pesquisadores envolvidos e o número de espécies e de anuros capturados. Calculamos, então, um índice de eficiência de captura (tempo necessário para um pesquisador capturar um animal), de forma a tornar comparáveis os dados obtidos, e estimamos a riqueza e a abundância totais obtidas com cada método. Dos métodos avaliados, os mais eficientes foram Inventário completo de espécies (9,7 min/amostrador/indivíduo-MAI, com riqueza = 6 e abundância = 23) e Transectos em sítios reprodutivos (9,5 MAI, com riqueza = 4 e abundância = 22). Os métodos Levantamento por encontros visuais (45,0 MAI) e Amostragem em manchas (65,0 MAI) apresentaram eficiência consideravelmente menor, enquanto Amostragem em parcelas (plots) e Armadilhas de queda com direcionadores (Pit-falls) não capturaram nenhum indivíduo. Concluímos, portanto, que há considerável diferença na eficiência dos métodos em ambiente de restinga e que os métodos Inventário completo de espécies e Transectos em sítios reprodutivos foram os mais eficientes para a amostragem de anuros na restinga estudada e, supostamente, em outros ambientes de restinga. Os dados mostram a importância da procura ativa pelos anuros nos microhabitats durante as amostragens nesse ambiente. Alguns métodos usualmente eficientes em áreas florestadas (como pit-falls e plots) apresentaram baixa eficiência no habitat estudado de restinga aberta.

Mixed mating in natural populations of the chestnut blight fungus, Cryphonectria parasitica.

As in plants, fungi exhibit wide variation in reproductive strategies and mating systems. Although most sexually reproducing fungi are either predominantly outcrossing or predominantly selfing, there are some notable exceptions. The haploid, ascomycete chestnut blight pathogen, Cryphonectria parasitica, has previously been shown to have a mixed mating system in one population in USA. In this report, we show that both selfing and outcrossing occur in 10 additional populations of C. parasitica sampled from Japan, Italy, Switzerland and USA. Progeny arrays from each population were assayed for segregation at vegetative incompatibility (vic) and DNA fingerprinting loci. Outcrossing rates (t(m)) were estimated as the proportion of progeny arrays showing segregation at one or more loci, corrected by the probability of nondetection of outcrossing (alpha). Estimates of t(m) varied from 0.74 to 0.97, with the lowest rates consistently detected in USA populations (0.74-0.78). Five populations (four in USA and one in Italy) had t(m) significantly less than 1, supporting the conclusion that these populations exhibit mixed mating. The underlying causes of variation in outcrossing rates among populations of C. parasitica are not known, but we speculate that--as in plants--outcrossing is a function of ecological, demographic and genetic factors.