Cutaneous protothecosis in kidney transplant recipient

Abstract Protothecosis is a rare condition caused by the aclorophylated algae of the genus Prototheca. In humans, protothecosis, caused mainly by P. wickerhamii, manifests itself in three forms: cutaneous, articular and systemic. It can occur in both immunocompetent and immunosuppressed individuals, being much more common in the latter. We present a new case of protothecosis in Brazil in a kidney transplant recipient.

A Case of Protothecosis on Scalp and Face in the Immunocompetent Patient / 대한의진균학회지

Protothecosis is an unusual human infection, caused by the genus prototheca, especially Prototheca wickerhamii. A 80-year-old immunocompetent man presented with a 1-month history of multiple reddish brown lobulated plaques on the scalp and face. He denied any history of trauma and had no evidence of underlying diseases such as diabetes mellitus or malignancy. On histopathological examination, characteristic morula-like sporangia in the dermis was revealed. After treatment with oral itraconazole for 8 weeks, the patient's skin lesions are almost healed.

A Case of Cutaneous Protothecosis / 대한의진균학회지

Protothecosis is a rare skin infection caused by pathogenic algae of the genus Prototheca, especially Prototheca wickerhamii. It is widely distributed throughout the natural world in tree, river, lake, sewage, soil, marsh, household waste, etc. A 79-year-old woman presented with ill-defined edematous and erythematous patches with various sized and shaped deep ulcers on both dorsal hands and left distal forearm for 1 month. The left forearm was initially injured by scratching due to pruritus and insidiously aggravated. Biopsy specimen revealed the characteristic morula-like sporangias containing multiple endospores in the dermis. The GMS and D-PAS stains highlighted the characteristic spores showing multiple internal septations and some unicellular endospore. P. wickerhamii was isolated in culture and identified by biochemical studies. Oral itraconazole 200 mg/day administration for 7 weeks resulted in marked improvement of the skin lesion.

Biological Control of Phytophthora palmivora Causing Root Rot of Pomelo Using Chaetomium spp

Phytophthora diseases have become a major impediment in the citrus production in Thailand. In this study, an isolate of Phytophthora denominated as PHY02 was proven to be causal pathogen of root rot of Pomelo (Citrus maxima) in Thailand. The isolate PHY02 was morphologically characterized and identified as Phytophthora palmivora based on molecular analysis of an internal transcribed spacer rDNA sequence. This work also presents in vitro evaluations of the capacities of Chaetomium spp. to control the P. palmivora PHY02. As antagonists, Chaetomium globosum CG05, Chaetomium cupreum CC3003, Chaetomium lucknowense CL01 inhibited 50~61% mycelial growth, degraded mycelia and reduced 92~99% sporangial production of P. palmivora PHY02 in bi-culture test after 30 days. Fungal metabolites from Chaetomium spp. were tested against PHY02. Results showed that, methanol extract of C. globosum CG05 expressed strongest inhibitory effects on mycelial growth and sporangium formation of P. palmivora PHY02 with effective dose ED50 values of 26.5 microg/mL and 2.3 microg/mL, respectively. It is interesting that C. lucknowense is reported for the first time as an effective antagonist against a species of Phytophthora.

Esporogénesis, esporodermo y ornamentaciónde esporas maduras en Lycopodiaceae / Sporogenesis, sporoderm and mature spore ornamentation in Lycopodiaceae

Studies on reproductive aspects, spore morphology and ultrastructure of Lycopodiaceae are not very common in the scientific literature, and constitute essential information to support taxonomic and systematic relationships among the group. In order to complete existing information, adding new and broader contributions on these topics, a comparative analysis of the sporogenesis ultrastructure, with emphasis on cytological aspects of the sporocyte coat development, tapetum, monoplastidic and polyplastidic meiosis, sporoderm ontogeny and ornamentation of the mature spores, was carried out in 43 taxa of eight genera of the Lycopodiaceae: Austrolycopodium, Diphasium, Diphasiastrum, Huperzia (including Phlegmariurus), Lycopodium, Lycopodiella, Palhinhaea and Pseudolycopodiella growing in the Andes of Colombia and the Neotropics. For this study, the transmission electron microscopy (TEM) samples were collected in Cauca and Valle del Cauca Departments, while most of the spores for scanning electron microscopy (SEM) analysis were obtained from herbarium samples. We followed standard preparation procedures for spore observation by TEM and SEM. Results showed that the sporocyte coat is largely composed by primary wall components; the sporocyte develop much of their metabolic activity in the production of their coat, which is retained until the spores release; protective functions for the diploid cells undergoing meiosis is postulated here for this layer. The abundance of dictyosomes in the sporocyte cytoplasm was related to the formation and development of the sporocyte coat. Besides microtubule activity, the membrane of sporocyte folds, associated with electrodense material, and would early determine the final patterns of spore ornamentation. Monoplastidic condition is common in Lycopodium s.l., whereas polyplastidic condition was observed in species of Huperzia and Lycopodiella s. l.. In monoplastidic species, the tapetum presents abundant multivesicular bodies, while in polyplastidic species, the secretory activity of the tapetum is less intense. Sporoderm development is centripetal, exospore is the first formed layer, then the endospore and, if present, perispore is the final deposited layer. Adult spores of the Lycopodiaceae showed two patterns of ornamentation: negative or caviform (foveolate spores) and positive or muriform ornamentation, the latter with two subtypes (rugate and reticulate spores). The spores of Huperzia are characteristically foveolate, the rugate spores were found in a few species of Huperzia and in all of the Lycopodiella s. l. taxa studied, while Lycopodium s.l. spores bear reticulate ornamentation. Numerous ornamentation traits are diagnostic at the specific level. The types of ornamentation found do not support the recent extreme fragmentation of the family in several genera, but could match, a priori, with the idea of three subfamilies. The findings of sporogenesis, extremely similar in all taxa studied, point more to consider fewer genera, more comprehensive, than the recent, markedsplitting of the family. Rev. Biol. Trop. 62 (3): 1161-1195. Epub 2014 September 01.

Estudios sobre aspectos reproductivos, morfología y ultraestructura de las esporas de Lycopodiaceae no son abundantes en la literatura científica y constituyen información esencial para apoyar las relaciones taxonómicas y sistemáticas en el grupo. Con el fin de completar la información existente, añadiendo contribuciones nuevas y más amplias sobre estos temas, se realizó un análisis comparado de la ultraestructura de la esporogénesis, con énfasis en aspectos citológicos que tienen que ver con la formación de la cubierta de los esporocitos, el tapete, las meiosis monoplastidial y poliplastidial, la ontogenia del esporodermo y la ornamentación de las esporas maduras en 43 táxones de ocho géneros de Lycopodiaceae: Austrolycopodium, Diphasium, Diphasiastrum, Huperzia (incluyendo Phlegmariurus), Lycopodium, Lycopodiella, Palhinhaea y Pseudolycopodiella que crecen en los Andes de Colombia y el Neotrópico. Para estudios con microscopía electrónica de trasmisión (MET) las muestras se recolectaron en los departamentos de Cauca y Valle del Cauca, mientras que la mayoría de las muestras para microscopía electrónica de barrido (MEB) provienen de material herborizado de colecciones. Para la observación de las muestras con MET y MEB se utilizaron protocolos estándar para el procesamiento de esporas. La cubierta de los esporocitos está formada por pared primaria; los esporocitos invierten gran parte de su actividad metabólica en la producción de esa cubierta, que es mantenida hasta la liberación de las esporas y tiene funciones de protección de las células que harán meiosis. La abundancia de dictiosomas en los esporocitos se relacionó con la formación y desarrollo de la cubierta. Además de la actividad de los microtúbulos, la presencia de sinuosidades y plegamientos asociados con material electro denso en la membrana de los esporocitos determinarían tempranamente los patrones de ornamentación de las esporas. La condición monoplastidial es común en Lycopodium s.l.y la poliplastidial se observó en Huperzia y Lycopodiella s. l. En especies monoplastidiales el tapete presenta abundantes cuerpos plurivesiculares, en las poliplastidiales la actividad secretora del tapete es menos intensa. El desarrollo del esporodermo es centrípeto, el exosporio se forma primero, seguido del endosporio y el perisporio, si está presente, se deposita de último. En las esporas adultas de Lycopodiaceae se encontraron dos patrones de ornamentación: negativo o caviforme (esporas foveoladas) y positivo o muriforme (esporas rugadas y reticuladas). Las esporas foveoladas son características de Huperzia; las rugadas de unas pocas especies de Huperzia y las especies de Lycopodiella s. l., mientras que las reticulada son típicas de Lycopodium s. l.. Numerosos caracteres de la ornamentación resultan diagnósticos en el nivel específico. Los tipos principales no apoyan la extrema fragmentación reciente de la familia en varios géneros, aunque podría coincidir, a priori, con la idea de tres subfamilias. Los hallazgos de la esporogénesis, extremadamente similar en todos los táxones estudiados, apuntan más a la unificación de los géneros en la familia que a su segregación.

Morfología y anatomía de ejes caulinares, licofilos y esporangios de Phlegmariurus phylicifolius: un aporte a la sistemática de las Lycopodiaceae neotropicales / Morphology and anatomy of caulinar axes, lycophylls and sporangia of Phlegmariurus phylicifolius: a contribution to the systematics of Neotropical Lycopodiaceae

Phlegmariurus is the only genus of Lycopodiaceae with the species grouped in 22 informal groups. Species level relationships within Phlegmariurus are poorly understood and their circumscriptions require a thorough molecular and morphological review. A detailed study of morphology and anatomy of caulinar axes, lycophylls and sporangia of Phlegmariurus phylicifolius was carried out in order to contribute to the elucidation of species circumscription in the informal group Phlegmariurus phlegmaria. Small pieces of caulinar axes bearing trophophylls, sporophylls and sporangia were fixed, dehydrated, Histowax (paraffin) embedded, sectioned in a rotatory microtome, and stained using the common Safranin O-Fast Green technique; handmade cross sections were also made and stained with the same technique. P. phylicifolius includes slender, pendulous plants up to 40cm long. Shoots heterophyllous, in the basal divisions ca. 10-20(-25)mm in diameter including the trophophylls, then abruptly constricted to (l-) 1.5-2(-2.5)mm in diameter including the imbricate, reduced sporophylls. Trophophylls are borne in alternating whorls of three, or decussate, subdecussate, or alternate, widely spaced in alternate leaved caulinar axes portions, perpendicular to the caulinar axes to falcately ascending, lanceolate to linear-lanceolate, with flat to slightly revolute margins. Each lycophyll is supplied by a single central vascular bundle, connected to a protoxylem pole in the stele. At the site of leaf-trace departure, no leaf (lycophyll) gap is present. Caulinar axes excluding leaves 0.7-1.2mm thick at the base, upward tapering to ca. 0.5mm. Caulinar axes present unistratified epidermis and endodermis, the cortex is characterized by the presence of a trabecular structure of lisigenous origin formed in the parenchimatous tissue next to the endodermis. The vascular tissue occupies the central part of the caulinar axes, forming a plectostele of subradiate organization, with five poles of protoxylem. The epidermal cells present sinuous anticlinal walls; invaginations in the inner side of external periclinal wall of the epidermal cells could be probably adaptive morphological feature of a water deficient environment. Leaves of constricted terminal divisions are decussate, or subdecussate, continuously or discontinuously sporangiate, appressed, abaxially rounded to carinate, widely lanceolate to widely ovate or subcordate, acute to mucronate or cuspidate, shorter than the sporangia. Each sporangium originates from a group of epidermal cells, axilar to the sporophylls. The cell walls of epidermal cell of the sporangia are Huperzioideae type. The morphological studies of trophophylls contribute to confirm the differences between P. phylicifolius and P. subulatus. Rev. Biol. Trop. 62 (3): 1217-1227. Epub 2014 September 01.

Phlegmariurus es el único género de Lycopodiaceae con las especies reunidas en 22 grupos informales. Las relaciones a nivel de especie dentro de Phlegmariurus están pobremente estudiadas y la circunscripción de las mismas requiere profundos exámenes moleculares y morfológicos. Se ha llevado a cabo un estudio detallado de la morfología y la anatomía de ejes caulinares, licofilos y esporangios de P. phylicifolius, con el fin de contribuir al esclarecimiento en la delimitación de las especies en el grupo Phlegmariurus phlegmaria. Segmentos de ejes caulinares con trofofilos, esporofilos y esporangios fueron fijados, deshidratados, incluidos en Histowax (parafina), cortados con un micrótomo rotatorio y coloreados usando la técnica tradicional Safranina O-Verde Rápido; además se hicieron cortes a mano alzada y se colorearon con la misma técnica. P. phylicifolius incluye plantas colgantes y péndulas de hasta 40cm de longitud. Los ejes son heterofilos, de aproximadamente 10-20(-25)mm de diámetro en las divisiones basales incluyendo los trofofilos, luego abruptamente reducidos a (l-) 1.5-2(-2.5)mm de diámetro incluyendo los esporofilos reducidos e imbricados. Los trofofilos están dispuestos en anillos alternantes de a tres, o decusados, subdecusados o alternos, dispuestos en forma espaciada en los ejes caulinares, perpendiculares al tallo hasta falcadamente ascendentes, lanceolados a lineal-lanceolados, con márgenes lisos o levemente revolutos. Cada licofilo está provisto de un haz vascular simple y central, conectado a un polo de protoxilema de la estela y sin laguna foliar. Los tallos poseen un ancho de 0.7-1.2mm en la base, excluyendo los licofilos, estrechándose hasta cerca de 0.5mm hacia el ápice. Los ejes caulinares presentan una epidermis uniestratificada y endodermis, la corteza se caracteriza por la presencia de una estructura trabecular de origen lisígeno formada en el tejido parenquimático próximo a la endodermis. El tejido vascular ocupa la parte central del eje caulinar, formando una plectostela de organización subradiada, con cinco polos de protoxilema. Las células epidérmicas presentan paredes anticlinales sinuosas; las invaginaciones en la cara interna de la pared periclinal externa podrían ser probablemente un característica morfológica adaptativa a un ambiente con períodos de sequía. Las hojas de las porciones apicales son decusadas o subdecusadas, con esporangio de disposición continua o discontinua, adpresas, abaxialmente redondeadas a carinadas, ampliamente lanceoladas a ovadas o subcordadas, ápice agudo a mucronado o cuspidado, más corto que el esporangio. Cada esporangio se origina de un grupo de células epidérmicas, en la axila de los esporofilos con el eje caulinar. Las paredes celulares de las células epidérmicas del esporangio son de tipo Huperzioideae. El estudio de la morfología de los trofofilos contribuye a confirmar las diferencias entre P. phylicifolius y P. subulatus.

Ontogenia de los esporangios, formación y citoquímica de esporas en licopodios (Lycopodiaceae) colombianos / Ontogeny of the sporangium, spore formation and cytochemistry in Colombian Lycopodials (Lycopodiaceae)

Studies on reproductive aspects of Lycopodiaceae are not very abundant in the scientific literature, and constitute essential information to support taxonomic and systematic relationships among the group. Here we present a detailed study of the ontogeny of sporangia and sporogenesis, and the chemical determination of several compounds generated during spore formation. The analyses were performed in 14 taxa of six genera of the family, Diphasiastrum, Diphasium, Huperzia (a genus which is treated here including Phlegmariurus), Lycopodiella, Lycopodium and Palhinhaea. Specimens were collected in three departments from the Colombian Andes between 1 454-3 677m altitude. Ontogeny was studied in small, 1cm long pieces of strobili and axis, which were fixed in glutaraldehyde or FAA, dehydrated in alcohol, embedded in LR White, sectioned in 0.2-0.5μm and stained with toluidine blue (TBO), a metachromatic dye that allows to detect both sporopollenin and lignin or its precursors, during these processes. For other studies, paraplast plus-embedded sections (3-5μm) were stained with safranin-fast green and alcian blue-hematoxylin. Chemical tests were also conducted in sections of fresh sporangia at different stages of maturity using alcian blue (mucopolysaccharides), Lugol solution (starch), Sudan III (lipids), phloroglucinol (lignin) and orcein (chromosomes). Sections were observed with photonic microscope equipped with differential interference contrast (DIC) and fluorescence microscopy (for spore and sporangium walls unstained). Strobili and sporangia were dehydrated with 2.2 dimethoxypropane, critical point dried and coated with gold for scanning electron microscopy (SEM). Our results indicated that the ontogeny of sporangia and sporogenesis were very similar to the previously observed in Huperzia brevifolia. Cutinisation occurs in early stages of development of sporangium cell walls, but in their final stages walls become lignified. As for the sporoderm development, the exospore is the first layer formed, composed by sporopollenin. The endospore deposits as a thin inner layer composed of cellulose, pectin and carboxylated polysaccharides. The perispore, if present, deposits at last. Mucopolysaccharides were found on the sporocyte coat and its abundance in sporangial cavity persists up to the immature tetrads stage, and then disappears. The lipids were abundant in the sporocytes, tetrads and spores, representing the main source of energy of the latter. In contrast, starch is not detected in the spores, but is abundant in premeiotic sporocytes and immature tetrads, developmental stages of high cellular metabolic activity. Intrinsic fluorescence corroborates the presence of lignin and cutin in the sporangium wall, while the sporopollenin is restricted to the exospore. The transfusion cells and the perispore are not always present. However, the processes of ontogeny and sporogenesis are extremely similar throughout the taxa studied, suggesting that they represent conservative family traits, nonspecific or generic.

Los estudios sobre aspectos reproductivos no son muy abundantes en la literatura científica sobre los taxones de Lycopodiaceae y constituyen información esencial para apoyar la taxonomía y relaciones sistemáticas en el grupo. Por lo tanto, se presenta aquí un análisis detallado de la ontogenia de los esporangios y esporogénesis, así como determinaciones químicas de varios compuestos generados durante la formación de las esporas. Los análisis se llevaron a cabo en 14 taxones de seis géneros de la familia: Diphasiastrum, Diphasium, Huperzia (un género que se trata aquí, incluyendo Phlegmariurus), Lycopodiella, Lycopodium y Palhinhaea. Las muestras fueron recolectadas en tres departamentos de los Andes de Colombia entre 1 454-3 677m de altitud. La ontogenia se estudió en trozos de estróbilos y ejes, de 1cm de largo, que se fijaron en glutaraldehido o FAA, se deshidrataron en alcohol, se incluyeron en LR White, se seccionaron en cortes de 0.2-0.5μm y se colorearon con azul de toluidina (TBO), un colorante metacromático que permite detectar tanto esporopolenina como lignina o sus precursores. Para estudios adicionales, secciones de 3-5μm de material incluido en paraplast plus se colorearon con safranina-verde rápido y azul alciánhematoxilina. Las pruebas químicas se llevaron a cabo en secciones de esporangios sin fijar en diferentes etapas de madurez utilizando azul alcián (mucopolisacáridos), solución de Lugol (almidón), Sudán III (lípidos), fluoroglucinol (lignina) y orceína (cromosomas). Las observaciones se efectuaron con microscopio fotónico equipado con contraste diferencial de interferencia (DIC) y microscopía de fluorescencia (para esporas y pared de los esporangios sin colorear). Para observaciones con microscopía electrónica de barrido (MEB), los estróbilos y esporangios se deshidrataron con 2,2 dimetoxipropano, se desecaron a punto crítico y se metalizaron con oro. Los resultados indican que la ontogenia de los esporangios y esporogénesis es muy similar a la observada previamente en Huperzia brevifolia. En las primeras etapas de desarrollo, las paredes celulares de la epidermis del esporangio se cutinizan y en las finales se lignifican. En el desarrollo del esporodermo, la primera capa que se forma es el exosporio, compuesto por esporopolenina. El endosporio es una capa interna delgada compuesta de celulosa, pectina y polisacáridos carboxilados. El perisporio, si está presente, es la última capa que se deposita. Los mucopolisacáridos se encontraron en la cubierta del esporocito, son abundantes en la cavidad esporangial hasta la etapa de tétradas inmaduras y luego desaparecen. Los lípidos son abundantes en esporocitos, tétradas y esporas, y representan la principal fuente de energía de estas. En contraste, el almidón no se detecta en las esporas pero es abundante en esporocitos premeióticos y tétradas inmaduras, ambos con gran actividad metabólica. La fluorescencia intrínseca corrobora la presencia de lignina y cutina en la pared del esporangio, mientras que la esporopolenina se limita al exosporio. Las células de transfusión y el perisporio no siempre están presentes. Sin embargo, los procesos de la ontogenia y esporogénesis son extremadamente similares en todos los taxones estudiados, lo que sugiere que representan rasgos típicos de familia, no específicos ni genéricos.

Esporogénesis y esporas de Equisetum bogotense (Equisetaceae) de las áreas montañosas de Colombia / Sporogenesis and spores of Equisetum bogotense (Equisetaceae) from mountain areas of Colombia

Studies on some reproductive traits in Equisetum species are scarce and valuable to understand species distribution. Therefore, a detailed study of the sporogenesis process and spore development in E. bogotense is presented, with an analysis of the main events during meiosis, maturation of spores, spore wall ultrastructure, orbicules and elaters. Specimens were collected from 500 to 4 500m in Cauca, Colombia. Strobili at different maturation stages were fixed, dehydrated, embedded in resin, and ultra-microtome obtained sections were stained with Toluidine blue. Observations were made with optical microscopy with differential interference contrast illumination technique (DIC), transmission and scanning electron microscopy (TEM and SEM). Ultrathin sections (70-80μm) for TEM observations were stained with uranyl acetate and lead citrate; while samples for SEM observations, were fixed, dehydrated in 2.2-dimethoxypropane and dried at critical point as in standard methods. Strobili have numerous mature sporangiophores, each one with a peltate structure, the scutellum, bearing five-six sessile sporangia attached to the axis of strobilus by the manubrium. Immature sporocytes (spore mother cells) are tightly packed within the young sporangia. The sporocytes quickly undergo meiosis, by passing the stage of archesporium and give origin to tetrads of spores. The tapetum loses histological integrity during early stages of sporogenesis, intrudes as a plasmodial mass into the cavity of the sporangium, partially surrounding premeiotic sporocytes, and then, tetrads and adult spores. The tapetum disintegrates towards the end of the sporogenesis, leaving spores free within the sporangial cavity. Spores present several cytological changes that allow them to achieve greater size and increase the number of plastids, before reaching the adult stage. Sporoderm includes three layers external to the cytoplasmic membrane of the spore cell, and they are pseudoendospore, exospore and perispore. Viewed with SEM, the exospore is smooth to rugulate, with micro perforations, while the perispore is muriform, rugate, with narrow, delicate, discontinuous, randomly distributed folds delimiting incomplete, irregular areolae, externally covered by of different size, densely distributed orbicules. These orbicules are also found all over the external face and margins of the elaters, while the internal face is smooth and lack orbicules. Viewed with TEM, the exospore is a thick layer of fine granular material, while perispore is a thinner layer of dense, separate orbicules. The elaters are composed by two layers of fibrillar material: an inner layer with longitudinally oriented fibrils and an outer, thicker and less dense layer with fibrils transversely fibrils and abundant, external orbicules. It is suggested that the processes of ontogeny and characters of the sporoderm are relatively constant in Equisetum; however, sporogenesis in E. bogotense is synchronous and this condition has been observed so far only in E. giganteum, a tropical genus also found in Colombia.

Los estudios sobre aspectos reproductivos son escasos en Equisetum. Por eso, hemos realizado un análisis detallado del proceso de esporogénesis, desarrollo de las esporas, ultraestructura de procesos que tienen lugar durante la meiosis, formación de la pared esporal, orbículas y eláteres de E. bogotense, en especímenes procedentes del Cauca, Colombia. Los estudios se efectuaron mediante microscopía fotónica, electrónica de transmisión (TEM) y de barrido (SEM). Los estróbilos llevan numerosos esporangióforos maduros, cada uno con un escutelo peltado, unido al eje del estróbilo por el manubrio y portador de 5-6 esporangios sésiles. Los esporocitos experimentan meiosis dando origen a tétradas de esporas. El tapete pierde la integridad histológica en las primeras etapas de esporogénesis y rodea los esporocitos premeióticos, posteriormente a las tétradas y finalmente las esporas inmaduras, que experimentan cambios citológicos y de tamaño antes de alcanzar la etapa adulta. El esporodermo de las esporas adultas de E. bogotense consiste de seudoendosporio, exosporio y perisporio. Vistos con MEB, el exosporio de las esporas adultas es liso a rugulado con microperforaciones y el perisporio es muriforme, rugado, con pliegues delicados, estrechos, discontinuos, que se distribuyen al azar y delimitan aréolas incompletas. Externamente el perisporio está cubierto por orbículas, que se forman también en la cara externa y los márgenes de los eláteres. Vistos con TEM, el exosporio es una capa de material granular fino y el perisporio, una capa mucho más delgada con orbículas discretas. Los eláteres están formados por dos capas de naturaleza fibrilar, orientadas longitudinalmente y transversalmente. La esporogénesis en E. bogotense es sincrónica, similar a la de E. giganteum, otra especie de distribución tropical que también crece en Colombia.

Soft Rot of Rhizopus oryzae as a Postharvest Pathogen of Banana Fruit in Korea

Soft rot on banana fruit caused by Rhizopus oryzae was identified for the first time in Korea. Colonies were white to light brown and formed numerous sporangiospores. Optimum temperature for mycelial growth was 30degrees C. Sporangia were globose and 30~200 microm. Sporangiophores were usually straight, 8~20 microm, and rhizoids usually in groups of 3~5. Columella were globose to sub-globose and 90~110 microm. Sporangiospores were sub-globose or oval and 4~10 microm. Based on its mycological characteristics, molecular analysis, and pathogenicity to host plants, this fungus was identified as Rhizopus oryzae Went & Prisen Geerligs. This is the first report of soft rot on banana caused by Rhizopus oryzae in Korea.

Ontogenia de los estróbilos, desarrollo de los esporangios y esporogénesis de Equisetum giganteum (Equisetaceae) en los Andes de Colombia

Ontogeny of strobili, sporangia development and sporogenesis in Equisetum giganteum (Equisetaceae) from the Colombian Andes. Studies on the ontogeny of the strobilus, sporangium and reproductive biology of this group of ferns are scarce. Here we describe the ontogeny of the strobilus and sporangia, and the process of sporogenesis using specimens of E. giganteum from Colombia collected along the Rio Frio, Distrito de Sevilla, Piedecuesta, Santander, at 2 200m altitude. The strobili in different stages of development were fixed, dehydrated, embedded in paraffin, sectioned using a rotatory microtome and stained with the safranin O and fast green technique. Observations were made using differential interference contrast microscopy (DIC) or Nomarski microscopy, an optical microscopy illumination technique that enhances the contrast in unstained, transparent. Strobili arise and begin to develop in the apical meristems of the main axis and lateral branches, with no significant differences in the ontogeny of strobili of one or other axis. Successive processes of cell division and differentiation lead to the growth of the strobilus and the formation of sporangiophores. These are formed by the scutellum, the manubrium or pedicel-like, basal part of the sporangiophore, and initial cells of sporangium, which differentiate to form the sporangium wall, the sporocytes and the tapetum. There is not formation of a characteristic arquesporium, as sporocytes quickly undergo meiosis originating tetrads of spores. The tapetum retains its histological integrity, but subsequently the cell walls break down and form a plasmodium that invades the sporangial cavity, partially surrounding the tetrads, and then the spores. Towards the end of the sporogenesis the tapetum disintegrates leaving spores with elaters free within the sporangial cavity. Two layers finally form the sporangium wall: the sporangium wall itself, with thickened, lignified cell walls and an underlying pyknotic layer. The mature spores are chlorofilous, morphologically similar and have exospore, a thin perispore and two elaters. This study of the ontogeny of the spore-producing structures and spores is the first contribution of this type for a tropical species of the genus. Fluorescence microscopy indicates that elaters and the wall of the sporangium are autofluorescent, while other structures induced fluorescence emitted by the fluorescent dye safranin O. The results were also discussed in relation to what is known so far for other species of Equisetum, suggesting that ontogenetic processes and structure of characters sporoderm are relatively constant in Equisetum, which implies important diagnostic value in the taxonomy of the group. Rev. Biol. Trop. 59 (4): 1845-1858. Epub 2011 December 01.

Estudios sobre la ontogenia del estróbilo, los esporangios y la biología reproductiva de Equisetum son escasos, por lo tanto, para la especie E. giganteum, se estudiaron estos aspectos en especímenes recolectados a orillas del Río Frío, Santander, Colombia (2 200m). Los estróbilos en diferentes etapas de maduración fueron fijados, deshidratados, embebidos en parafina, seccionados en micrótomo rotatorio y teñidos con safranina O-fast green. Las observaciones se efectuaron mediante un microscopio óptico de alta resolución con contraste diferencial de interferencia (DIC) y microscopio de fluorescencia. Los estróbilos se inician a partir del meristemo apical, tanto en el eje principal como en los laterales, sin diferencias en el proceso de ontogenia y esporogénesis entre estróbilos de diferentes ejes. Sucesivas mitosis y diferenciación celular conducen al crecimiento del estróbilo, y a la formación de los esporangióforos peltados, formados por el manubrio, o porción basal con aspecto de pedicelo, el escutelo, o porción apical aplanada y las iniciales del esporangio, los cuales se diferenciarán para formar la pared del esporangio, los esporocitos y el tapete. No se forma arquesporio y los esporocitos experimentan meiosis para formar tétradas de esporas. El tapete mantiene la integridad histológica hasta la formación de las tétradas y en esa etapa forma un plasmodio que invade la cavidad esporangial la cual rodea parcialmente las tétradas y luego las esporas, y aparecen las cámaras plasmodiales, un término propuesto aquí para las formaciones designadas en inglés "tapetal gaps". La pared del esporangio queda reducida a dos capas celulares: una externa con engrosamientos lignificados en todas las paredes celulares y una interna picnótica. Al finalizar la esporogénesis, el tapete degenera, y las esporas, con exosporio, perisporio delgado, casi membranáceo y eláteres quedan libres en la cavidad esporangial. El esporodermo, los núcleos y nucléolos presentan fluorescencia roja, inducida por coloración con safranina O, mientras que los eláteres y las células de la pared del esporangio presentan autofluorescencia amarillo-naranja.

Tectonic corneal graft for conjunctival rhinosporidiosis with scleral melt.

A 16 year old girl presented with irritation and watering of the right eye for 3 months. On examination, the superior perilimbal sclera was ectatic with incarcerated uveal tissue covered by conjunctiva. The conjunctiva showed discreet, yellow white mucoid spots. Excision biopsy of the conjunctiva showed subepithelial spherules of sporangia containing numerous endospores, suggestive of rhinosporidiosis. Diathermy was applied to flatten the staphyloma. The ectatic area was covered with a corneal patch graft. The patient was started on prednisolone acetate eye drops and oral dapsone for 6 months. Corneal graft was well incorporated and conjunctivalized by 3 months. Since the graft was not seen within the palpebral aperture, there was good cosmetic result. The corneal graft had the added advantage of transparency which allowed visualization of the underlying tissue to diagnose early recurrence. There was no recurrence at 6 months.

Identification of Diachea leucopodia on Strawberry from Greenhouse in Korea

We have detected the slime mold, Diachea leucopodia (GNU06-10) in a strawberry greenhouse located in Sancheong-gun, Gyeongnam. Typical fruiting bodies had developed gregariously on the strawberry leaves, petioles, and plant debris on ground soil habitat, and also surprisingly on plastic pipes and a vinyl covering. Field samples were examined via stereomicroscopy, light microscopy, and SEM for the determination of morphological characteristics. Dark-brown to black spores formed gregariously within the stipitate cylindrical sporangium, and were covered by an iridescent peridium, which may be intact at maturity, or may have disintegrated. The upper portion of the peridium generally breaks up to expose the spores, whereas the lower portion was usually persistent. The results of energy dispersive X-ray spectrometer (EDS) analysis showed that lime was present in the stalk and columella but absent from the spores, capillitium, and peridium. The above characteristics confirm its taxonomic position in the genus Diachea. However, this genus is intermediate in character between the Physarales and Stemonitales of the Myxogastromycetidae. Hence, this genus had been classified as a member of the Stemonitales until the mid-1970's, on the basis of its iridescent peridium and noncalcareous capillitial system, similar to Comatricha of the Stemonitaceae. By way of contrast, emphasis on morphological characteristics, most notably the calcareous stalk and typical columella, places Diachea within the order Physarales. The presence of a phaneroplasmodium during the trophic stage and lime deposition in its sporophores, as was confirmed in this work, supported the inclusion of Diachea in the Physarales, and the noncalcareous capillitial system verified its identification as a member of the Didymiaceae. Further characteristics of the species D. leucopodia include the following: phaneroplasmodium, spore globose 7.5 microm in diameter, very minutely roughened; sporangia 500 microm x 1mm, more or less cylindrical, gregarious, stalked 1.2mm; stalk and columella white.

A Case of Cutaneous Protothecosis / 대한의진균학회지

Cutaneous protothecosis is a rare skin infection, caused by the genus prototheca, especially Prototheca wickerhamii. A 73-year-old woman had a painful, ulcerative patch and oozing plaque on left wrist and dorsum of hand for 2 months. Biopsy specimen revealed thick walled morula-like sporangia in the dermis. P. wickerhamii was isolated in the culture. Oral itraconazole therapy for 8 weeks is not satisfactory. New skin lesion is formed. After oral fluconazole therapy, patient's skin lesion is improved.

A Case of Cutaneous Protothecosis / 대한피부과학회지

Protothecosis is an unusual human infection caused by the achlorophyllic algae Prototheca, and most commonly by Prototheca wickerhamii. A 65-year-old man presented with a 2-month history of a painful, erythematous, non-healing, ulcerative plaque on the extensor surface of his left forearm. He was previously diagnosed with diabetes and iatrogenic Cushing syndrome. A skin biopsy specimen revealed characteristic morula-like sporangia in the dermis by PAS positive staining. Prototheca wickerhamii was identified by culture and biochemical studies. Initially, itraconazole was administered for 8 weeks, followed by fluconazole, and the patient's condition was greatly improved.

Rhizopus Soft Rot on Pear (Pyrus serotina) Caused by Rhizopus stolonifer in Korea

Rhizopus soft rot caused by Rhizopus stolonifer occurred on pears (Pyrus serotina) in the Jinju City Agricultural Products Wholesale Market in Korea from 2004 to 2005. The infection usually started from wounds due to cracking at harvest time. The lesions started as water-soaked, rapidly softened, then gradually expanded. The mycelia grew vigorously on the surface of the fruits and formed stolons. Colonies on potato dextrose agar at 25degrees C were white cottony to brownish black. Sporangia were globose, black and 90~120 microm in size. Sporangiophores were light brown and 480~2600 x 12~18 microm in size. Sporangiospores were globose to oval, brownish, streaked, and 8~14 x 6~10 microm in size. Columella were light brownish gray, hemispherical and 70~80 microm in size. On the basis of these symptoms, mycological characteristics and pathogenicity tests on host plants, the fungus was identified as Rhizopus stolonifer (Ehrenb.) Vuill. This is the first report of rhizopus soft rot on pear (P. serotina) caused by R. stolonifer in Korea.

Soft Rot of Eggplant (Solanum melongena) Caused by Choanephora cucurbitarum in Korea

In April 2002 and 2003, soft rot on fruit of eggplant (Solanum melongena) caused by Choanephora cucurbitarum was observed in the experimental fields at Gyeongnam Agricultural Research and Extension Services in Korea. The disease began with water-soaking and dark-green lesions, and then the infected tissues were rapidly rotten. Sporangium was subglobose in shape and sized 40~130 microm. Monosporous sporangiola were elliptic, fusiform or ovoid, brown in color, and measured as 12~20 x 6~14 microm. Sporangiospores having three or more appendages were elliptic, fusiform or ovoid in shape, dark brown or brown in color, and sized 14~20 x 7~16 microm. The fungus grew well on potato dextrose agar between 15 and 40degrees C and its optimum growth temperature was 30degrees C. Based on morphological characteristics, the causal fungus of the fruit soft rot of eggplant was identified as C. cucurbitarum. This is the first report on the soft rot of S. melongena caused by C. cucurbitarum in Korea.

Soft Rot of Tomato Caused by Mucor racemosus in Korea

A soft rot of fruits caused by Mucor racemosus occurred on cherry tomato collected in Agricultural Products Wholesale Market in Jinju, Korea. The disease infection usually occurred wounded areas after cracking of fruits. At first, the lesions started with water soaked and rapidly softened and diseased lesion gradually expanded. Colonies were white to brownish to gray in color. Sporangia were 32~54 microm in size and globose in shape. Sporangiophores were 8~14 microm in width. Sporangiospores were 5~12 x 4~8 microm in size, ellipsoidal to subglobose in shape. Columella was 27~42 microm in size, obovoid, ellipsoidal, cylindrical-ellipsoidal, slightly pyriform in shape. Chlamydospores were numerous in sporangiophores and barrelshaped when young, subglobose in old cultures. Optimum growth temperature was about 25degrees C. The fungus was identified as M. racemosus Fres. This is the first report of soft rot on cherry tomato caused by M. racemosus in Korea.

Fruit Soft Rot of Sweet Persimmon Caused by Mucor piriformis in Korea

A fruit soft rot caused by Mucor piriformis occurred on sweet persimmon storages in Jinju, Changwon and Gimhae, Gyeongnam province, Korea, 2003. The disease infection usually started from wounding after cracking of fruits. At first, the lesions started with water soaked and rapidly softened and diseased lesion gradually expanded. Colonies on potato dextrose agar at 20degrees C were whitish to olivaceous-buff Sporangia were globose, black and 96~153microm in size. Sporangiophores were 26~42microm in width. Sporangiospores were ellipsoid and 5.8~10.6x4.3~7.6microm in size. Columella was obovoid, cylindrical-ellipsoidal, pyriform, subglobose and 80~125microm in size. Optimal temperature for mycelial growth was 20degrees C on PDA. The causal organism was identified as M. piriformis. This is the first report of fruit soft rot on sweet persimmon caused by M. piriformis in Korea.

A Case of Cutaneous Prototheocsis / 대한의진균학회지

Protothecosis is an infection caused by achloric algae of the genus Prototheca. It is found ubiquitously in nature, particularly in the slime flux of trees and sewage. Only two species including Prototheca wickerhamii and Prototheca zopfii have been known to be pathogenic in humans. A 64-year-old man presented intermittent pruritic erythematous confluent grouped papules and nodules on the left upper arm. Histopathologic study revealed some sporangia of morula-like appearance and chronic granulomatous inflammation. The PAS stain demonstrated the characteristic spores showing multiple internal septations and some unicellular endospore. P. wickerhamii was isolated from culture and identified by the biochemical studies. Oral itraconazole 200 mg/day administration resulted in marked regression of the skin lesion after 8 weeks.

A Case of Cutaneous Protothecosis / 대한피부과학회지

Protothecosis is an unusual cutaneous soft tissue infection caused by the Prototheca, which is a genus of the unicelluar, achloric algae. We report a case of cutaneous protothecosis in a 66-year-old female, who showed erythematous, purulent patches and plaques with ulcerations on the right forearm for 2 months. Biopsy specimen revealed the characteristic thick-walled morulalike sporangia in the dermis. Prototheca wickerhamii was isolated in the culture and the biochemical study. Electron microscopic examination showed the thick-walled spores containing dark dense bodies and amyloplasts. After two months of oral itraconazole 200mg/day, skin lesions were improved.