Molecular diagnosis and intraspecific genetic variability of root pathogens of arid legumes in Western Rajasthan, India / Diagnóstico molecular y la variabilidad genética intraespecífica de patógenos de raíces en leguminosas resistentes a sequías del oeste de Rajasthan, India

Abstract:The productivity of arid legumes, such as Clusterbean (Cyamopsis tetragonoloba), Cowpea (Vigna unguiculata), Moth bean (Vigna aconitifolia) and Horse gram (Macrotyloma uniflorum), may remain stagnant over decades because of their high susceptibility to root diseases. Besides, there is a limitation on the information about molecular diagnosis and intraspecific genetic variability of root pathogens in arid legumes. To contribute in this field, we assessed a total of 52 isolates from 88 root samples that were found infected with fungal pathogens in Jodhpur, Jaipur and Bikaner Districts of Rajasthan. Diseased roots samples were analyzed following standard microbiological methods for fungus extraction and purification, and for genetic studies. Irrespective of the geographical location from where the diseased samples were collected, all pathogen isolates were clustered in RAPD dendrograms as per their respective genera. Phylogram, based on multiple sequence alignment, revealed that different genera (i.e. Fusarium, Neocosmospora and Syncephalastrum), separated from each other, and species within the same genera, clustered together with their reference sequences with apreciable bootstrap values. Out of 20 representative isolates representing each cluster and all outgroups sequenced, eight were molecularly identified as Neocosmospora vasinfecta, five as Fusarium solani, two as Neocosmospora striata, two as Fusarium acutatum, one as Syncephalastrum monosporum, one as Fusarium oxysporum and one as Fusarium species. The root pathogens of the arid legumes were found neither restricted to a geographical location nor were host specific in nature. Fusarium solani wilt in cowpea and seedling rot in moth bean, F. oxysporum wilt in moth bean, F. acutatum damping off in cowpea and Clusterbean, Fusarium sp. seedling rot in Clusterbean, Neocosmospora striata root rot in cowpea and wilt in Clusterbean and Syncephalastrum monosporum root rot in Clusterbean were molecularly identified as new fungal records as pathogens causing root diseases in arid legumes. Rev. Biol. Trop. 64 (4): 1505-1518. Epub 2016 December 01.

Resumen:La producción de leguminosas resistentes a sequías como Cyamopsis tetragonoloba, Vigna unguiculata, Vigna aconitifolia y Macrotyloma uniflorum, puede permanecer inactiva durante décadas debido a su alta susceptibilidad a enfermedades en las raíces. Además, hay información limitada relacionada con el diagnóstico molecular y la variabilidad genética intraespecífica de patógenos de raíces en estas leguminosas resistentes a sequías. Para contribuir en esta área, evaluamos un total de 52 extractos de 88 raíces infectadas con patógenos fúngicos en los distritos de Jodhpur, Jaipur y Bikaner de Rajastán. Las muestras de raíces infectadas se analizaron siguiendo los métodos estándar de microbiología para extracción y purificación de hongos y para estudios genéticos. Independientemente del sitio donde se recolectaron las muestras contaminadas, todos los extractos patógenicos se agruparon en dendrogramas RAPD en cada uno de sus respectivos géneros. El filograma, basado en alineamiento de secuencias múltiples reveló que distintos géneros (Fusarium, Neocosmospora y Syncephalastrum) separados entre ellos y especies del mismo género se agrupan con sus secuencias de referencia con valores de bootstrap significativos. De cada 20 extractos representantes de cada agrupamiento y todos los grupos externos secuenciados, ocho fueron identificados molecularmente como Neocosmospora vasinfecta, dos como Fusarium acutatum, una como Syncephalastrum monosporum, una como Fusarium oxysporum y una como Fusarium. Los patógenos de estas leguminosas resistentes a sequías no están restringidos por la localidad ni por un hospedero específico. Fusarium solani que marchita el frijol de vaca y pudre la semilla de Vigna aconitifolia, F. oxysporum que marchita a Vigna aconitifolia, F. acutatum que marchita a Vigna unguiculata y Cyamopsis tetragonoloba, Fusarium sp. que pudre la semilla de Cyamopsis tetragonoloba, Neocosmospora striata que pudre la raíz de Vigna unguiculata y marchita a Cyamopsis tetragonoloba y, Syncephalastrum monosporum que pudre la raíz en Cyamopsis tetragonoloba, fueron identificados molecularmente como nuevos registros de patógenos fúngicos que causan daños en las raíces de leguminosas resistentes a sequías.

Mucorales from the semiarid of Pernambuco, Brazil

Nineteen taxa of Mucorales, belonging to Absidi, Apophysomyces, Cunninghamella, Fennellomyces, Lichtheimi, Mucor, Mycotypha, Rhizopus and Syncephalastrum were isolated from 36 composite soil samples in three semiarid areas in the State of Pernambuco (Triunfo, Cabrobó and Belém de São Francisco), Northeast Brazil, which are characterized by Caatinga vegetation. Triunfo is preserved, whereas Cabroró and Belém de São Francisco are experiencing low and severe desertification processes, respectively. Mucorales were isolated in Petri dishes in triplicate from 5 mg samples of soil placed on the surface of wheat germ agar plus chloramphenicol and Cercobin [Dimethyl 4,49-(103 phenylene) bis (3-thioallophanate)] medium. The plates were left on a bench at room temperature (28 ± 2 ºC) for 72 h of alternating dark and light periods. Absidia cylindrospora presented the highest amount of CFU/g of soil, followed by L. hyalospor, C. phaeospora and C. echinulata var. echinulata. The latter, and R. microsporus var. microsporus, presented the highest frequencies of occurrence. Soils from Triunfo showed higher diversity of Mucorales than the samples from the other areas, although without differing statistically in relation to species richness. The communities of Mucorales from the degraded areas were more similar, while that from the preserved area was quite different. Most of the identified specimens have been commonly isolated from soil in other Brazilian regions, which indicates that they are not endemic of the semiarid. Eleven taxa are registered for the first time in this ecosystem, while F. heterothallicus is reported for the first time in Brazil.

Primary cutaneous zygomycosis from a tertiary care centre in north-west India.

Background & objectives: Zygomycosis is highly invasive fungal infection, with high mortality rate. In most of patients, diabetes mellitus is an underlying factor but in primary cutaneous zygomycosis, presentation may be different. Here we present the description of clinical presentation, fungi isolated and management of cases with cutaneous zygomycosis seen in a tertiary care hospital in north India during 2001-2007. Methods: All patients diagnosed with primary cutaneous zygomycosis between November 2001 and September 2007 presenting with clinical diagnosis of necrotizing fasciitis were included. Detailed history of each patient was taken, clinical presentation, site of involvement, underlying illness and risk factor, if any were noted. The diagnosis was established by direct microscopic evidence of broad, aseptate or sparsely septate ribbon-like hyphae with right angle branching in KOH wet mount and histopathological examination of stained sections. Cultures were put up for fungal isolation and species identification. Outcome of the therapy was analysed. Results: Of the nine patients reviewed, only one had diabetes mellitus. Commonest risk factor was injection abscess (33.3%). Apophysomyces elegans was isolated in four cases, Saksenaea vasiformis and Absidia corymbifera in one each. The fungal culture was sterile in three cases. Mortality rate was high with only four patients responded well to surgical and/or medical therapy. Interpretation & conclusion: Primary cutaneous zygomycosis appears to be on rise in India that calls for high index of clinical suspicion and an early biopsy of the affected area for timely diagnosis. The standard treatment is a combination of amphotericin B therapy, surgical debridement, and reversal of the underlying disease or immunosuppression.

Isolation of Mucorales from processed maize (Zea mays L. ) and screening for protease activity / Isolamento de Mucorales de milho processado (Zea mays L. ) e seleção quanto à atividade proteásica

Mucoraleswere isolated from maize flour, corn meal and cooked cornflakes using surface and depth plate methods. Rhizopus oryzae, Circinella muscae, Mucor subtilissimus,Mucor hiemalis f. hiemalis, Syncephalastrum racemosum, Rhizopus microsporus var. chinensis and Absidia cylindrospora showed protease activity.

Mucorales foram isolados da farinha de milho, fubá e flocos de milho pré-cozidos pelos métodos de plaqueamento em superfície e em profundidade. Rhizopus oryzae, Circinella muscae, Mucor subtilissimus,Mucor hiemalis f. hiemalis, Syncephalastrum racemosum, Rhizopus microsporus var. chinensis e Absidia cylindrospora exibiram atividade proteásica.

Fatal rhino-orbito-cerebral infection caused by Saksenaea vasiformis in an immunocompetent individual: first case report from India.

A 56-year-old female patient presented with history of nasal blockage, nasal bleeding, headache, since one month. On admission the patient developed severe headache, decreased vision of eyes and blindness. Clinical diagnosis of rhino-orbital involvement was confirmed by computerized tomography of the paranasal sinuses. The diagnosis of fungal infection was confirmed by KOH examination, haematoxylin and eosin examination and Gomori's methanamine silver stain of the biopsy material. Diagnosis was confirmed by culture on Sabouraud Dextrose agar, slide culture on Czapek Dox medium and the isolate was identified as Saksenaea vasiformis. The patient was started on intravenous amphotericin B (received only one dose before succumbing), but she did not respond to the treatment, developed hemiparesis, slurred speech, diminished reflexes and ultimately died. The involvement of the brain was confirmed by computerised tomographic scan. We believe this case to be the first case of rhino-orbito-cerebral infection due to Saksenaea vasiformis and that of second case in an immunocompetent patient in the world.

Apophysomyces elegans - renal mucormycosis in a healthy host: a case report from south India.

Zygomycosis is an opportunistic fungal infection that seldom occurs in individuals with a competent immune system. Isolated involvement of any organ is rare and only a few cases of renal zygomycosis have been reported. We present an unusual case of renal zygomycosis caused by Apophysomyces elegans in a patient with no known predisposing factor. He presented with flakes in the urine and was found to have a poorly functioning right kidney. Ureterorenoscopy was performed, fungal elements removed and pathological confirmation obtained. The patient subsequently underwent nephrectomy after treatment with amphotericin B. He made an uneventful recovery.

Primary cutaneous mucormycosis (zygomycosis) caused by Apophysomyces elegans.

A 53 year-old male diabetic presented with a month-old, painful ulcer with necrotic margins over the right thigh. Wound debridement was done twice and the ulcer showed recurrent growth of a white, cottony filamentous structure. Cutaneous mucormycosis was suspected and confirmed by histopathology and a culture isolate of Apophysomyces elegans . The patient was treated with liposomal amphotericin-B and itraconazole followed by partial thickness skin grafting, and then discharged after being prescribed posaconazole syrup for three weeks. Regular follow-up was done and during the last visit after six months following discharge, the ulcer was found to have healed well with no recurrence of the fungus.

Fatal cellulitis caused by Apophysomyces elegans.

A case of cellulitis of the left lateral side of the face caused by the zygomycete Apophysomyces elegans in a healthy male following a road traffic accident is reported. The contaminated soil was the source of fungus. Broad aseptate fungal hyphae were seen in the necrosed tissues. Extensive tissue debridement and treatment with amphotericin B were not successful in controlling the rapid invasion of the tissues by the fungus. Patient developed angioinvasion, severe cellulitis and finally succumbed to the infection three weeks after admission.

Mucormicosis: infección necrotizante como complicación de dermolipectomía abdominal / Mucormycosis: necrotizing infection as a complication of abdominal dermolipectomy

Las infecciones necrotizantes de los tejidos blandos varían en su presentación clínica, pero en general son de curso grave y alta mortalidad. El cuadro clínico incluye: fiebre, celulitis, edema, crepitación, necrosis y sepsis; con frecuencia existe un antecedente o traumático quirúrgico. Como hallazgo operatorio se encuentra necrosis de la piel y tejido subcutáneo, con o sin mionecrosis. El tratamiento es una inmediata y amplia debridación acompañada con antibioticoterapia de amplio espectro. Se presenta un caso clínico de infección necrotizante de tejidos blandos por mucor, en una mujer con antecedente de abdominoplastia y lipoescultura con progresión crítica y evolución fatal. La mucormicosis es una patología agresiva, cuyo diagnostico clínico no es fácil por lo cual debe ser considerada seriamente por el cirujano plástico.

Cutaneous zygomycosis: necrotising fascitis due to Saksenaea vasiformis.

Saksenaea vasiformis is an emerging human pathogen, most often associated with cutaneous or subcutaneous lesions following trauma. This is the report of a case of subcutaneous zygomycosis from which Saksenaea vasiformis was isolated on culture. As the patient developed acute interstitial nephritis, amphotericin B could not be administered in full dose. Surgical debridement was carried out, but the patient deteriorated gradually and died. To the best of our knowledge, this is the first reported case of Zygomycosis due to Saksenaea vasiformis from Visakhapatnam.

Rhino-orbito-cerebral mucormycosis. A retrospective analysis of clinical features and treatment outcomes.

OBJECTIVE: The conventional management of rhino-orbito-cerebral (ROC) mucormycosis includes control of metabolic abnormality, administration of amphotericin B and surgery that spans simple sinus clearance, radical debridement and orbital exenteration. Recent literature includes anecdotal descriptions of successful treatment with conservative management of involved orbits. We evaluated the clinical features and outcome of treatment for the different stages of ROC mucormycosis. METHOD: In this retrospective case series, 34 case records of patients with a histopathological diagnosis of ROC mucormycosis treated between 1992 and 2000 were reviewed. Three clinical stages and three treatment groups were identified. Patients with limited sino-nasal disease (Clinical stage I) underwent sino-nasal debridement (Treatment group A). Patients with limited rhino-orbital disease (Clinical stage II) underwent either sino-nasal debridement alone (Treatment group A) or orbital exenteration in addition to sino-nasal debridement (Treatment group B). Patients with rhino-orbito-cerebral disease (Clinical stage III) did not undergo any surgical procedure (Treatment group C). Thirty-three patients received intravenous amphotericin B. Outcome for each group was measured as "Treatment success" (disease free, stable patient with metabolic abnormality under control) and "Treatment failure" (progression of disease with worsening general condition or mortality due to the disease). RESULTS: Uncontrolled diabetes in 30 (88.2%) of 34 patients was the commonest underlying disease and 16 (53.3%) of 30 diabetics had ketoacidosis. Chronic renal failure (n = 4), hepatic disease (n = 3) and idiopathic thrombocytopenia (n = 1) were the other underlying diseases. Eleven patients had stage I disease, 16 patients had stage II disease and seven patients had stage III disease. All 11 patients with stage I disease received treatment A; of 16 patients with stage II disease, 7 received treatment A and the remaining with stage III disease received treatment B; 7 patients with stage II disease received treatment C. Ten of 11 patients (91%) with stage I disease had treatment success. In patients with stage II disease, 7 of 7 (100%) with treatment A and 1 of 9 (11.1%) with treatment B had treatment success. All seven patients with stage III disease had treatment failure. CONCLUSION: Debridement of the sinuses is necessary in all cases of rhino-orbito-cerebral mucormycosis. Diagnosis in the early stage needs a high degree of suspicion. There is a definite role for retention of orbits in patients whose metabolic derangement is rapidly controlled and orbital involvement is non-progressive.

Mucormicosis rinocerebral: caso clínico / Rhinocerebral mucormycosis: clinical case

La mucormicosis es una enfermedad poco frecuente, causada por hongos del orden de los mucorales. Suele presentarse en pacientes con algún grado de inmunodepresión. La forma rinocerebral afecta principalmente a pacientes con cetoacidosis diabética. El cuadro clínico corresponde al de una sinusitis invasiva, con un curso generalmente fatal y el tratamiento es médico-quirúrgico. Se presenta un caso clínico con revisión de la literatura

Zigomicose subcutânea causada pelo basidiobolus haptosporus / Subcutaneous zygomycosis caused by basidiobolus haptosporus

Os autores registram caso de zigomicose subcutânea em paciente do sexo feminino, com dois anos de idade, branca, natural de Porto Franco, Estado do Maranhäo - Brasil, provocada por Basidiobolus haptosporus. Os exames histopatológicos e micológicos confirmaram o diagnóstico. Em face da experiência adquirida com o tratamento desses doentes, indicam o iodeto de potássio como a droga de escolha para o tratamento dessa micose

Mucormicosis rinocerebral. Informe de cuatro casos / Rhinocerebral mucormycosis. Four cases report

Se informa de cuatro casos de mucormicosis rinocerebral, de su evolución clínica, antecedentes, métodos diagnósticos y tratamiento, con el objetivo de presentar al personal médico una semblanza de esta patología que orienta al establecimiento del diagnóstico temprano y tratamiento oportuno, que permitan mejorar el pronóstico para los pacientes, ya que en estudios tempranos los índices de curación alcanzan casi 50 por ciento. En aquellos que no reciben atención médica oportuna la evolución lleva invariablemente a la muerte

Micobiota de la rizosfera y rizoplano de suelos contaminados con hidrocarburos / Mycobiota from rhizosphere and rhizoplane in hidrocarbon polluted soils

Se estudió la micobiota saprótrofa de suelo rizosférico y de rizoplano, de plantas que crecen en áreas contaminadas con hidrocarburos y en áreas control. Se aislaron e identificaron 52 especies fúngicas, de las cuales 39 pertenecen a los deuteromycetes, 3 a los ascomycetes, 8 a los zygomycetes (orden mucorales) y 2 son micelios estériles. Mediante el uso de métodos de ordenación, utilizándose el análisis de componentes principales, pudo diferenciarse las muestras obtenidas de áreas contaminadas de las no contaminadas. A pesar que las especies son las mismas en ambas áreas, sus frecuencias relativas permiten una discriminación de los muestreos

Microhongos del suelo y asociados a pastos silvestres senescentes en una localidad precordillerana de la 9ª región, Chile / Microfungi of soil and associated to senescent wild grass in a preandine locality of the IX region, Chile

Mediante métodos de dilución y cámara húmeda, se analizó cualitativamente y cuantitativamente la micota presente en el suelo y en su cobertura de pastos senescentes (gramíneas principalmente) en una localidad precordillerana de pradera en la 9§ región. Nuestra finalidad fue la obtención de datos preliminares sobre la diversidad y composición de especies en suelos subtropicales. La diversidad de géneros fue mayor en los pastos (38), que en el suelo (27), mientras el número de especies en ambos habitat fue similar (50 y48). Solo 14 especies fueron comunes en ambos sustratos (28,9 porciento), representadas mayoritariamente por Hyphomycetes (71,4 porciento) y Coelomycetes (21,45 porciento). Los taxa dominantes en los suelos fueron: Cladosporium cladosporioides, penicillium spp. y las levaduras blancas. En los pastos senescentes: Acremonium spp. Alternaria alternata, A tenuissima, Botrytis cinerea, Cladosporium cladosporioides, C. herbarum, Dactylaria candidula, Eppicoccum purpurascens, Fusarium poae, Gliocladium catenulatum, G. roseum, Heteroconium chaestospira, Leptosphaeria ogilviensis, Myrothecium verrucaria, Papulaspora inmersa, Penicillium spp. Periconia minutissima, Phoma levellei, y Torula herbarum. En los suelos, a diferencia de los pastos, se presentó una gran diversidad de taxa esporádicos (40)