First Report of Colletotrichum karstii Causing Glomerella Leaf Spot on Apple in Santa Catarina State, Brazil.

Glomerella leaf spot (GLS) is an emerging disease of apple (Malus domestica Borkh.) that has been reported in regions with a humid subtropical climate, such as southern Brazil, the southeastern United States, and more recently eastern China. GLS is favored by high humidity and temperatures between 23 and 28°C and can result in extensive defoliation when the severity is high. The disease was first reported 1988 in Brazil on cvs. Gala and Golden Delicious in orchards in Paraná State (3), but now is widespread in the country's producing areas. Two Colletotrichum species of different complexes have been associated with GLS, C. gloeosporioides (Penz.) Penz. & Sacc. and its sexual stage Glomerella cingulata (Stoneman) Spaulding & Scherenk, and C. acutatum J. H. Simmonds, although GLS is more commonly associated with the former. In the summer of 2012, necrotic spots were observed on apple leaves (cv. Gala) in Santa Catarina state, Brazil. The first symptoms were reddish-brown spots, evolving to small necrotic lesions 1 to 10 mm long at 7 to 10 days after symptoms were first noted. Pure cultures were obtained by monosporic isolation and grown on PDA at 25°C and with a 12-h photoperiod under fluorescent light. The color of the upper surface of the colony varied from white to gray and the reverse was pink. The conidia length and width ranged from 9.1 to 17.1 µm ( = 12.8) and from 2.9 to 6.8 µm ( = 4.9), respectively, and were cylindrical, hyaline, and straight. After germination, conidia formed oval or circular appressoria measuring between 4.0 and 10.0 ( = 6.3) × 3.0 and 9.0 ( = 5.7). To confirm pathogenicity, susceptible apple seedlings (cv. Gala) were inoculated with a suspension of 1 × 106 conidia.mL-1. Seedlings sprayed with sterile distilled water served as controls. Seedlings were incubated in a moist chamber at 25°C and 100% RH for 48 h. First symptoms appeared 4 days after inoculation and were similar to those observed in the field. The control treatment remained symptomless. The pathogen was reisolated from lesions, confirming Koch's postulates. Fungus was molecularly characterized by sequencing the internal transcribed spacer (ITS) rDNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and the nucleotide sequence was deposited in the GenBank database (KC876638 and KC875408). C. karstii, considered as part of the C. boninense species complex (1), was identified with 100% sequence homology. This species was previously reported in China (4), Thailand, and the United States, affecting Orchidaceae plants (2), and in Brazil it has been reported affecting Carica papaya, Eugenia uniflora, and Bombax aquaticum (1). To our knowledge, this is the first report of C. karstii causing GLS on apple in Brazil. The development of pre-harvest management practices may be warranted to manage this disease. References: (1) U. Damm et al. Stud. Mycol. 73:1, 2012. (2) I. Jadrane. Plant Dis. 96:1227, 2012. (3) T. B. Sutton. Plant Dis. 82:267, 1998. (4) Y. Yang. Cryptogamie Mycologie 32:229, 2011.

First Report of Colletotrichum nymphaeae Causing Apple Bitter Rot in Southern Brazil.

Bitter rot (BR) is a common disease on apples (Malus domestica Borkh.) worldwide. Fruit infection often occurs in warmer weather (25 to 32°C) where high temperatures and humidity strongly favor disease development. Three species causing BR have been reported: Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. and its sexual stage Glomerella cingulata (Stoneman) Spaulding & Schrenk, C. acutatum J. H. Simmonds, and recently C. fragariae in Uruguay (1). In February 2012, typical BR symptoms were observed on apple fruits (cv. Gala) in two orchards located in Santa Catarina, Brazil. Initial symptoms were light brown, slightly sunken lesions that enlarged over time, becoming dark brown and forming sunken or saucer-shaped depressions that extended toward the fruit core in a V-shaped pattern. Acervuli were sparse and the conidia masses varied from dark gray to pale orange in color. To attempt to identify the causal organism, isolations were made from lesions on fully ripened fruits. Pure cultures were obtained following monosporic isolation and grown on PDA at 25°C with a 12-h photoperiod under fluorescent light for 7 to 15 days. The color of the upper surface of the colony varied from white to gray and was orange on the underside. One hundred conidia for each isolate were measured and each was one-celled, hyaline, fusiform, and with a length and width ranging between 8.0 and 25.9 µm (avg. 14.7), and between 2.4 and 9.9 µm (avg. 5.6), respectively. After germination, conidia formed oval appressoria between 5.1 and 9.9 µm (avg. 7.3) × 3.4 and 7.8 µm (avg. 5.2). These morphological characters are consistent with the description of C. nymphaeae (Pass.) Aa (2). To examine pathogenicity, eight 'Pink Lady' apples were inoculated with isolates MANE 25, 137, 143, and 144. A isolate treatments were arranged in a completely randomized design with three replications. Fruits were inoculated with two drops of 10 µl (1 × 106 conidia per ml) using wounded and non-wounded sterile fruit surfaces. Sterile distilled water drops served as controls. Fruit were incubated in a moist chamber at 25°C with a 12-h photoperiod for 10 days. Symptoms were observed at the inoculation site 3 to 4 days after inoculation (DAI) on wounded and 5 to 6 DAI on non-wounded fruits. All of the isolates produced symptoms identical to those observed in Santa Catarina, and each isolate was re-isolated from the apple lesions, confirming Koch's postulates. Fungal isolates were also characterized by sequencing of the internal transcribed spacer (ITS) rDNA using ITS1/IT4 primers and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) using GDF/GDR primers. Isolate sequences had 100% homology to C. nymphaeae. The nucleotide sequences were deposited in GenBank (KC840351, KC840352, KC840353, KC840354, KC875404, KC875405, KC875406, and KC875407). Species identified as C. nymphaeae are considered as part of the C. acutatum species complex and have been reported to occur on water lilies (Nymphaea alba) (3) and causing bitter rot on apples in Korea (2,4). To our knowledge, this is the first report of C. nymphaeae causing bitter rot of apples in southern Brazil. References: (1) S. Alaniz et al. Plant Dis 96:458, 2012 (2) U. Damm et al. Stud. Mycol. 73:37, 2012. (3) D. A. Johnson et al. Mycol. Res. 101:641, 1997. (4) D. H. Lee et al. Plant Pathol. J. 23:37, 2007.

First Report of Colletotrichum acutatum and C. fragariae Causing Bitter Rot of Apple in Uruguay.

Almost 50% of deciduous fruit produced in Uruguay are apples and bitter rot is ubiquitous in the apple-production regions in Uruguay. In rainy and hot seasons (25 to 32°C by day), severe outbreaks of bitter rot occur. In summer 2010, when apple rot incidence reached 70% in some orchards, fruit with typical symptoms of bitter rot were collected from orchards in the south-central region, the main apple-production area. Symptoms included 0.5 to 6.0 cm in diameter, circular, sunken, light brown-to-brown lesions on the fruit surface that contained black, pinhead-sized fruiting structures that produced orange-to-brown conidial masses under high relative humidity. Each lesion progressed to the core of the fruit in a V-shaped pattern. Single-conidial isolates from lesions were examined morphologically (3), and based on sequences of the internal transcribed spacer (ITS) rDNA determined using ITS1/ITS4 primers (4), three species were identified: Colletotrichum acutatum with white-to-pale orange colonies and one-celled, hyaline, fusiform to cylindrical conidia that averaged 14.5 (9.3 to 17.8) × 5.0 (6.9 to 4.0) µm (isolates C11 and C18, GenBank Nos. JN413081 and JN413082, respectively); C. fragariae with white-to-pale gray and/or dark gray colonies and one-celled, hyaline, cylindrical to fusiform conidia that averaged 20.5 (14.3 to 22.9) × 6.0 (4.6 to 7.6) µm (isolate C15 and C37, GenBank Nos. JN413083 and JN413084, respectively); and C. gloeosporioides with white-to-pale gray or gray colonies and one-celled, hyaline, cylindrical to fusiform conidia that averaged 16.5 (13.1 to 20.3) × 6.5 (3.7 to 7.6) µm (isolates C5 and C29, GenBank Nos. JN413079 and JN413080) when grown on potato dextrose agar (PDA) at 25°C. To confirm pathogenicity, two isolates of each Colletotrichum spp. were inoculated onto mature, asymptomatic fruit of cv. Pink Lady (eight fruit per isolate). Each fruit was surface disinfested with 70% ethanol, wounded with a sterile needle, and inoculated with 10 µl of a spore suspension (5 × 105 conidia/ml) of the appropriate isolate. Eight control fruit were each inoculated with 10 µl of sterile water. Inoculated fruit and the control fruit were placed in plastic bags (eight fruit per bag) and incubated at 25°C. Symptoms (sunken, brown lesions each with a V-shaped pattern extending to the core) developed on all inoculated fruit 2 to 4 days after inoculation. No lesions were observed on control fruit. When fungi were reisolated from lesions of inoculated fruit onto PDA and incubated at 25°C, colony and conidial morphology were identical to those of the original isolates, confirming Koch's postulates. This study confirms a previous report of C. gloeosporioides causing bitter rot on apple in Uruguay (1). C. acutaum and C. gloeosporioides are known to cause bitter rot on apple. C. fragariae has traditionally been associated with strawberry and recently with other host plants (2), but not with bitter rot of apple. To our knowledge, this is the first report of bitter rot of apple caused by C. fragariae, and the first report of this disease caused by C. acutatum and C. fragariae in Uruguay. References: (1) S. García. Page 49 in: Guía Para el Manejo Integrado de Plagas y Enfermedades en Frutales. INIA Las Brujas, Canelones, 1998. (2) S. J. MacKenzie et al. Plant Dis. 92:1432, 2008. (3) B. C. Sutton. The Coelomycetes. CAB International Publishing, New York, 1980. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

First Report of Pestalotiopsis clavispora Causing Dieback on Blueberry in Uruguay.

During the last 10 years, blueberry (Vaccinium corymbosum) production in Uruguay has increased to more than 850 ha. From 2005, symptoms of dieback characterized by the death of twigs and branches have been frequently observed in blueberry plants cv. O'Neal in orchards located in Uruguay. Symptomatic 4-year-old plants (cv. O'Neal) were collected and small pieces of necrotic tissues were surface disinfected and plated onto potato dextrose agar (PDA) with 0.2 g liter-1 of streptomycin sulfate. Plates were incubated at 25°C in the dark. All affected tissues consistently developed colonies with white and cottony mycelium, turning slightly yellow after 7 to 10 days. Black acervuli distributed in concentric circles were observed after 10 days. Conidia were fusiform, straight, and had five cells. Basal and apical cells were colorless while the three median cells were dark brown. Conidia (n = 50) had an average of 22.1 (16.5 to 28.2) × 6.6 (5.6 to 7.7) µm. All conidia had one basal appendage of 6.1 (3.9 to 14.3) µm and two to four (usually three) apical appendages of 22.8 (17.4 to 42.9) µm. According to colony and conidia morphology, the isolates were initially identified as Pestalotiopsis clavispora (G.F. Atk.) Steyaert (1). To identify, the internal transcribed spacers (ITS1, 5.8S, ITS2) region of rDNA of a representative isolate (Ara-1) was amplified with ITS1/ITS4 primers (4), sequenced, and compared with those deposited in GenBank. The isolate Ara-1 (Accession No. JQ008944) had 100% sequence identity with P. clavispora (Accession Nos. FJ517545 and EU342214). To confirm pathogenicity, isolate Ara-1 was inoculated onto asymptomatic 1-year-old blueberry plants (cv. O'Neal). Mycelial plugs (4 mm in diameter) from an actively growing colony on PDA were applied to same-size bark wounds made with a cork borer in the center of the stems previously disinfected with 70% ethanol and covered with Parafilm. Control plants were inoculated with sterile PDA plugs. Inoculated plants (five per treatment) were randomly distributed in a greenhouse and watered as needed. After 2 weeks, all stems inoculated with P. clavispora showed brown necrotic lesions 2 to 3 cm in length and 1 to 2 mm deep. White mycelium was observed over lesions. Control plants remained symptomless. The pathogen was reisolated from all necrotic lesions, thus fulfilling Koch's postulates. P. clavispora has been reported as associated with blueberry in Hawaii (3) and Chile (2). To our knowledge, this is the first report of P. clavispora causing dieback disease on blueberry in Uruguay. References: (1) E. F. Guba, Monograph of Pestalotia and Monocheatia. Harvard University Press, Cambridge, MA, 1961. (2) J. G. Espinoza et al. Plant Dis. 92:1407, 2008. (3) L. M. Keith et al. Plant Dis. 90:16, 2006. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

Iliac artery-ureteral fistula: a fatal hemorrhage in a man with ureterocutaneostomy.

Primary and secondary artery-ureteral fistulas are a rare condition. In this case report we describe a case of misunderstood iliac artery-ureteral fistula, with negative radiological preoperative imaging, which occurred in a man as late complication of ureterocutaneostomy.

[Update on epidemiology and risk factors of prostate cancer].

Prostate cancer is one of the most common malignancies in the world; more and more men are being diagnosed with prostate cancer worldwide. According to epidemiological studies, prostate cancer will become the most common male cancer by 2010, with 900,000 cases per year. Nevertheless, although the constant increase in incidence, knowledge about the aetiology and risk factors of this tumour is still poor. Several important issues could foster the understanding and prevention of this disease, such as the variation in incidence of prostate cancer between ethnic populations, studies on migrants, dietary and genetic factors. Here we provide an update on epidemiology and risk factors of prostate cancer.

[Neuroendocrine bladder cancer: oncological emergency?].

Neuroendocrine bladder cancer is extremely rare, with an estimated incidence of 0.5%- 0.7%. In bladder cancers there is no evident connection between the neuroendocrine phenotypic expression and the clinical history. However, prognosis is usually poor and the survival rate at 5 years does not exceed 8%, if untreated. METHODS. We are here describing three case reports of bladder carcinoma with neuroendocrine differentiation, which is extremely aggressive and leads rapidly to death. At the present time, the local control of these tumors is achieved by radical cystectomy and radiotherapy; they can be both associated to chemotherapy. However, since these lesions are fairly rare, there is no gold standard therapy and there are no prospective studies on the management of these tumors. CONCLUSIONS. Considering the quick evolution and progression of any variant of the neuroendocrine tumors of the bladder, urologists and anesthetists should see them as real oncological emergencies. A prompt intervention through radical surgery with cystectomy and linfadenectomia, and the anathomo-pathologist's systematic investigation of the scraps could make the approach therapeutic and not only palliative. Prospective studies on neo-adjuvant chemotherapy as well as experimental studies about target therapies may yield new guidelines on the tumor management.

Molecular characterization and transfer among Staphylococcus strains of a plasmid conferring high-level resistance to mupirocin.

In this work, mupirocin resistance was correlated with the presence of plasmids in methicillin-resistant Staphylococcus aureus (MRSA) strains isolated in the Rio de Janeiro Federal University Hospital in Brazil, where topical mupirocin has been used extensively since 1990. Of 19 strains studied, those exhibiting high-level resistance carried a large and relaxable plasmid of about 35 kb. Mupirocin-sensitive derivatives, obtained by growth at 42 C of a strain exhibiting high-level resistance, were devoid of the large plasmid, which was designated pMG1. Mupirocin resistance was transferred to strain RN8411 during overnight filter-matings at low frequencies (7.0 x 10(-9)/donor). The pMG1 plasmid was shown to be responsible for high-level mupirocin resistance in our isolates and to be incompatible with pGO1. Hybridization experiments suggested that mupirocin resistance in pMG1 is due to the presence of the ileS-2 gene. The pMG1 plasmid was successfully and bidirectionally transferred from Staphylococcus aureus to Staphylococcus epidermidis, suggesting that the latter may be a reservoir of this resistance plasmid. No transfer was detected to Staphylococcus haemolyticus. The development of self-transferable high-level mupirocin resistance should be considered when using mupirocin to control the spread of MRSA in hospitals.

Detection of ileS-2 gene encoding mupirocin resistance in methicillin-resistant Staphylococcus aureus by multiplex PCR.

The presence of the ileS-2 gene, responsible for mupirocin resistance, in clinical isolates of methicillin-resistant Staphylococcus aureus was determined by multiplex polymerase chain reaction. Three pairs of primers were used, which yielded specific fragments of femA (encoding a unique feature of S. aureus), mecA (encoding resistance to methicillin) and ileS-2 genes. The multiplex polymerase chain reaction system is an easy and time-saving technique that, together with a rapid method for DNA extraction by boiling, may be incorporated as a routine analysis in clinical diagnostic laboratories.

[Our experience with a combination of rifampicin and minocycline in the therapy of brucellosis]. / La nostra esperienza con l'associazione rifampicina-minociclina nella terapia della brucellosi.

After a brief review of the literature on the treatment of brucellosis the results obtained with a new protocol (Rifampicin + Minocycline for 20 days) are reported. The combination was well-tolerated and undeniably effective producing a 94% cure rate.

[295 cases of brucellosis treated with minocycline]. / Su 295 casi di brucellosi trattati con minociclina.

295 cases of acute and subacute cases of brucellosis were treated with minocycline. It is concluded that because of its in vitro activity, its penetrability in macrophages and granulomas and its tolerance, minocycline is the antibiotic of choice for brucellosis.

Enterite por cepa invasiva de Escherichia coli. Apresentacao de um caso. / Enteritis due to invasive cepa of Escherichia coli. Report of a case

Os autores apresentam um caso de enterite por cepa invasiva de Escherichia coli. Recomendam a efetuacao rotineira de tipagem, com soro especifico nos casos de diarreias com fezes mucossanguinolentas e purulentas, principalmente quando nao se identificarem Shigella sp., Salmonella sp. ou outros enteropatogenicos de acao reconhecida. Preconizam que o teste de rastreamento seja feito com soro polivalente especifico e usando-se no minimo cinco colonias suspeitas