First Report of Leaf Spot of Spinach Caused by Pleospora betae in Spain.

The production of spinach (Spinacia oleracea L.) in Spain has increased 50% since 2009, mainly due to the commercialization of fresh-cut spinach leaves packaged in modified atmosphere containers. In October 2012, light brown leaf spots 1 to 2 cm in diameter with dark concentric rings were observed in a commercial spinach production area in Valencia Province, Spain. The initial outbreak comprised an area of about 3 ha with a 20% disease incidence. Symptomatic leaves from spinach cv. Apollo were collected in the affected area and were surface disinfected with 0.5% NaOCl for 2 min. Small fragments from lesions were placed onto potato dextrose agar (PDA) amended with 0.5 g streptomycin sulfate/liter. Fungal colonies developed after 3 days of incubation at 23°C from about 90% of the infected tissues plated. Isolates were transferred to oatmeal agar (OA) (1) and water agar (WA) amended with autoclaved pea seeds (2). Plates were incubated for 30 days at 24°C with 13 h of fluorescent light and 11 h of dark for morphological examination. Colonies were olivaceous grey in OA and pycnidia developed in WA were globose to subglobose, olivaceous black, and 100 to 200 µm in diameter. Conidia were globose to ellipsoidal, hyaline, aseptate, and 3.8 to 7.7 × 2.4 to 3.9 µm. Swollen cells were observed. Isolates showed a positive reaction to NaOH (1). Partial 18S, ITS1, 5.8S, ITS2, and partial 28S ribosomal RNA (rRNA) regions were amplified using the primers ITS1 and ITS4 (4) and sequenced from DNA extracted from the isolate designated as IVIA-V004 (GenBank Accession No. KF321782). The sequence had 100% identity (e-value 0.0) with that of Pleospora betae (Berl.) Nevod. (syn. Phoma betae A.B. Frank) representative strain CBS 523.66 (1). Pathogenicity tests were performed twice by inoculating 4-month-old plants of spinach cv. Apollo, table beet (Beta vulgaris L.) cv. Detroit, and Swiss chard (B. vulgaris subsp. cicla) cv. Verde de Penca Blanca. Plants were inoculated by spraying a conidial suspension of isolate IVIA-V004 (10 ml/plant, 105 conidia/ml water) using a manual pressure sprayer. Plants were immediately covered with black plastic bags and incubated in a growth chamber at 23°C. In each experiment, four plants of each host were inoculated with the fungus and four additional plants sprayed with sterile distilled water were used as controls. Plastic bags were removed after 48 h and leaf spots similar to those observed in affected spinach plants in the field were visible on all spinach, table beets, and Swiss chard plants 3 to 5 days after inoculation. No symptoms were observed on control plants. Fungal colonies morphologically identified as P. betae were re-isolated from leaf lesions on inoculated plants, but not from asymptomatic leaves of control plants. To our knowledge, this is the first report of leaf spot caused by P. betae on spinach in Spain, where it was previously described affecting sugar beet (3). The disease reduces the quality of spinach leaves and proper control measures should be implemented. References: (1) G. H. Boerema et al. Phoma Identification Manual, Differentiation of Specific and Infra-Specific Taxa in Culture. CABI Publishing, Wallingford, UK, 2004. (2) O. D. Dhingra and J. B. Sinclair. Basic Plant Pathology Methods, 2nd ed. CRC Press, Boca Raton, FL, 1995. (3) P. Melgarejo et al. Patógenos de Plantas Descritos en España. MARM-SEF, Madrid, 2010. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

Inoculum Sources, Infection Periods, and Effects of Environmental Factors on Alternaria Brown Spot of Mandarin in Mediterranean Climate Conditions.

Alternaria brown spot (ABS), caused by Alternaria alternata, is a serious disease affecting mandarin in humid and in semi-arid regions. The information available from Florida cannot be easily extrapolated to Mediterranean regions; thus, epidemiological studies were conducted during two consecutive years in Spain. Pathogenic isolates were found in the canopy and leaf litter and on weeds of the genus Sonchus. The pathogen survived in fallen immature leaves for up to 76 days, until complete leaf decay. Conidia of Alternaria spp. were captured continuously and pathogenic isolates were detected in all sampling dates. However, the number of pathogenic isolates was not correlated with the total captured, indicating that morphological identification is not sufficient for airborne inoculum monitoring. In contrast to humid areas, infections occurred mainly in spring and autumn. Classification tree analysis indicated that virtually all infections occurred on weeks with rainfall ≥2.5 mm and average temperature ≥12.5°C. Based on regression quantiles, the amount of rainfall, number of rain days, and total wetness duration were considered important factors increasing ABS incidence during infection periods. The development of decision support systems for ABS control in Mediterranean conditions may benefit from the restricted periods of infection and the strong influence of weather factors in disease onset.

First Report of Alternaria japonica Causing Black Spot of Turnip in Spain.

Turnip (Brassica rapa subsp. rapa L.) is an annual vegetable crop cultivated for consumption of its succulent root. In July 2011, symptoms consisting of leaf spots 1 to 8 mm in diameter with a dark brown color surrounded by a yellow halo and black sunken lesions in the swollen storage root were observed in production areas in Alicante Province in east-central Spain. Disease incidence was approximately 20% in fields of about 3 ha where infection was highest. Symptomatic leaves and roots collected from turnip cv. Virtudes-Martillo in three affected fields were surface disinfected with 0.5% NaOCl for 2 min, and small fragments from necrotic lesions were plated on potato dextrose agar (PDA) amended with 0.5 g streptomycin sulfate per liter. Alternaria colonies were consistently isolated from affected leaves and roots after 7 days of incubation at 24°C, and were transferred to V-8 with autoclaved turnip cv. Virtudes-Martillo leaves. Two isolates from leaves and two isolates from roots were included in the study. Plates were incubated for 15 days at 24°C with an 8-h fluorescent light period and a 12-h dark period for morphological examination. Conidia produced in culture were mostly solitary or in short chains of 2 to 3 spores, beakless, ovoid to ellipsoid, and light brown. Conidia were 32 to 78 × 13 to 24 µm, with 3 to 7 transverse septa and 1 to 2 longisepta. Aggregated hyphal chains of dark, thick-walled ornamented cells distinctive of Alternaria japonica Yoshii (3) were observed. The 5.8S, ITS2, and 28S ribosomal RNA (rRNA) regions were amplified using the primers ITS3 and ITS4 (4) and sequenced from DNA extracted from the isolate designated as IVIA-A070, obtained from turnip leaves cv. Virtudes-Martillo in Alicante Province (GenBank Accession No. JX983044). The sequence had 100% identity (total score 302, 73% coverage) with that of A. japonica strain ATCC 13618 (2) (AY376639). Pathogenicity tests were performed twice on two 3-month-old plants of turnip cv. Virtudes-Martillo and cv. Blanco-Globo, and cabbage (B. oleracea var. capitata L.) cv. Brunswick. Plants were inoculated by spraying a conidial suspension of the isolate IVIA-A070 (10 ml/plant, 104 conidia/ml water) using manual pressure sprayer. Two plants of each host sprayed with sterile distilled water were used as controls in each experiment. Plants were covered with black plastic bags and incubated in a growth chamber for 48 h at 25°C. Leaf spots similar to those observed in affected plants in the field were visible on all turnip and cabbage plants 4 days after inoculation with the fungus. No symptoms were observed on control plants. Fungal colonies morphologically identified as A. japonica were reisolated from leaf lesions on inoculated turnip and cabbage plants, but not from asymptomatic leaves of control plants. Based on these results, the disease was identified as black spot of turnip caused by A. japonica. In Spain, black spot of brassicas was previously associated only with A. brassicae (Berkeley) Saccardo and A. brassicicola (Schw.) Wiltshire (1). References: (1) P. Melgarejo et al. Patógenos de Plantas Descritos en España. MARM-SEF, Madrid, 2010. (2) B. M. Pryor and R. L. Gilbertson. Mycol. Res. 104:1312, 2000. (3) E. G. Simmons. Alternaria: An Identification Manual. CBS Fungal Biodiversity Centre, Utrecht, 2007. (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 Alternaria petroselini Causing Leaf Blight of Fennel in Spain.

Fennel (Foeniculum vulgare Mill.) is an aromatic herb widely cultivated in Mediterranean areas for culinary and medicinal uses. In 2010, symptoms consisting of leaf blight and necrosis were observed in commercial organic fennel production areas in Valencia Province in east-central Spain. Disease incidence in affected fields was approximately 20%. Symptomatic leaves from four fields were surface disinfected with 0.5% NaOCl for 2 min, and small fragments from necrotic lesions were then plated on potato dextrose agar (PDA) amended with 0.5 g of streptomycin sulfate/liter. After 7 days at 25°C, isolates of the genus Alternaria were consistently isolated. Single conidium cultures were grown on PDA and V8 agar for morphological examination. On both agar media, colonies were dark olive brown without production of pigments. On V8 agar, conidia were solitary, darkly pigmented, and predominantly ovoid-subsphaeroid. Mature conidia were 25 to 59 × 12 to 23 µm with up to six to seven transepta and one to three longisepta. The 5.8S, ITS2, and 28S ribosomal RNA (rRNA) regions were amplified with the primers ITS3 and ITS4 (3) from DNA extracted from the isolate IVIA-A029, and sequenced (GenBank Accession No. JQ240204). The sequence had 100% identity (total score 399, 97% coverage) with that of Alternaria petroselini (Neergard) Simmons strain EGS 09-159 (GenBank Accession No. AF229454.1) (1). Pathogenicity tests were conducted on four 3-month-old fennel plants (cv. Giotto) by spraying a conidial suspension of the fungus (10 ml/plant, 103 conidia/ml of water). Four control plants were sprayed with sterile, distilled water. Plants were covered with plastic bags and incubated in a growth chamber for 72 h at 25°C. Leaf necrosis was visible on inoculated plants after 4 days, but symptoms were not observed on control plants. The fungus was reisolated from leaf lesions on inoculated plants, but not from leaves of control plants, confirming Koch's postulates. On the basis of the morphological (2), molecular, and pathogenicity data, the disease was identified as Alternaria leaf blight of fennel caused by A. petroselini. To our knowledge, this is the first report of A. petroselini in Spain. References: (1) B. M. Pryor and R. L. Gilbertson. Mycol. Res. 104:1312, 2000. (2) E. G. Simmons. Alternaria: An Identification Manual. CBS Fungal Biodiversity Centre, Utrecht, The Netherlands, 2007. (3) T. J. White et al. Pages 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

Primary gastric choriocarcinoma: a case report and review of the literature.

Choriocarcinoma is a rapidly invasive, widely metastatic human chorionic gonadotropin (HCG)-producing neoplasm, usually intrauterine and gestational. Primary gastric choriocarcinoma is very rare, and its pathogenesis is still uncertain. We report a case of primary gastric choriocarcinoma associated with adenocarcinoma in a 36-year-old woman. The patient presented with gastrointestinal bleeding and a gastric mass clinically suspicious of gastric adenocarcinoma. Histopathologic evaluation proved the tumor to be a choriocarcinoma, with a minor component of a poorly differentiated adenocarcinoma. The patient was treated with a standard nongestational choriocarcinoma chemotherapy regimen. An impressive initial response was evidenced by clinical reduction of the tumor volume and drop of the serum beta-HCG levels after the first cycle. However, the tumor rapidly recurred in the abdomen and disseminated to the lungs, which were documented by new elevation of serum beta-HCG levels and computed tomographic scans despite continuing with 3 more cycles of chemotherapy. The patient died 6 months after diagnosis.

Balloon thermoablation in a woman with complex endometrial hyperplasia with atypia. A case report.

BACKGROUND: Endometrial ablation is an alternative to hysterectomy in patients with menorrhagia refractory to medical therapy. The histologic changes in the hyperplastic endometrium secondary to endometrial ablation are not well understood. CASE REPORT: A 44-year-old woman, gravida 4, para 4, had menorrhagia, was at high risk for medical treatment and underwent thermal balloon ablation. The dilatation and curettage specimen at the time of the procedure revealed complex hyperplasia of the endometrium with atypia. Subsequently a hysterectomy was performed, and the pathology specimen showed scarring, fibrosis and focal, weakly proliferative phase endometrium. There was no evidence of hyperplasia or malignancy. CONCLUSION: Endometrial ablation with a thermal balloon seems to have resulted in resolution of endometrial hyperplasia. However, hyperplasia of the endometrium should still be a contraindication to such a procedure because of the inability to accurately assess the endometrium after ablation.

Diagnosis of malignant granular cell tumor by fine needle aspiration cytology:.

BACKGROUND: Malignant granular cell tumors (GCT) are the rarest of all sarcomas, and the histologic differentiation from their benign counterpart may be extremely difficult or impossible unless metastatic disease is demonstrated. To our knowledge, this is the first report of a malignant GCT diagnosed by fine needle aspiration (FNA) cytology. CASE: A 70-year-old, Caucasian female presented with a progressively enlarging left supraclavicular mass. FNA of the mass revealed a metastatic tumor cytologically consistent with GCT. With this diagnosis, a search for other metastatic sites was initiated. Computed tomography (CT) scan revealed several tumor nodules in the lungs and liver. CT-guided FNA and tru-cut needle biopsy of a liver mass confirmed the diagnosis of metastatic GCT. In searching for a primary site, the patient revealed a clinical history of having had a tumor removed from her back two months before; it was reported to be an atypical GCT. Comparison of the three tumors revealed similar histologic, cytologic and immunohistochemical features. CONCLUSION: Evidence of mild to moderate cytologic atypia; increased mitotic activity; locally aggressive growth; increased proliferative activity as demonstrated by immunohistochemical evaluation of proliferation markers; and DNA ploidy analysis, as reported for this case, may be helpful in predicting malignant behavior of GCTs.

Paracoccidioides brasiliensis in a postpartum Pap smear. A case report.

BACKGROUND: There are several reported cases that describe female genital tract infections with opportunistic fungi, such as Blastomyces dermatitidis, Coccidioides immitis, Aspergillus flavus, Cryptococcus neoformans and Mucor. We describe a case of paracoccidiodomycosis limited to the uterine cervix. To the best of our knowledge, no such case has been described before in the English-language literature. CASE: A 27-year-old, healthy female, gravida 3, para 2, abortion 1, presented for a routine gynecologic examination at six weeks' postpartum. Her past medical history was unremarkable. A routine cervical/endocervical smear revealed the presence of multiple fungal forms at different stages of development with a characteristic "pilot's wheel" appearance consistent with Paracoccidioides brasiliensis. Detailed medical examination of the patient did not reveal the presence of the primary infection in any other system. Cultures of the endometrium revealed no growth of the fungal organisms. The patient was asymptomatic, and therefore no therapy was initiated. Repeat Papanicolaou smears were negative for organisms. CONCLUSION: Paracoccidioidomycosis can present as a limited form, involving the cervix only. Identification and recognition of the infection are important in cytopathology.

Concomitant well-differentiated adenocarcinoma and leiomyosarcoma of the uterus.

We describe a case of a concomitant well-differentiated endometrial endometrioid adenocarcinoma and leiomyosarcoma of the uterus in a 66-year-old woman who presented with a 6-month history of vaginal bleeding. The patient underwent total hysterectomy for endometrial carcinoma diagnosed by endometrial biopsy. Gross examination of the specimen revealed an endometrial mass bulging into the endometrial cavity and an underlying well-circumscribed nodule separated from the endometrial mass by a myometrial band. Frozen section performed at the time of the total hysterectomy rendered a diagnosis of malignant mixed-müllerian tumor. Histologic examination of the permanent sections revealed well-differentiated endometrial endometrioid adenocarcinoma clearly separated from a high-grade leiomyosarcoma. Differential diagnosis included malignant mixed-müllerian tumor. However, no admixture of carcinomatous and sarcomatous elements was present. There were no heterologous elements. To the best of our knowledge, no similar case has been described in the English literature.

Metastatic female adnexal tumor of probable Wolffian origin: a case report and review of the literature.

Female adnexal tumor of probable wolffian origin is a rare neoplasm that can present diagnostic difficulties. We report herein a case of a 60-year-old woman with female adnexal tumor of probable wolffian origin arising within the leaves of a broad ligament and, 5 years later, presenting with metastasis to the liver. The morphologic, immunohistochemical, ultrastructural, and DNA ploidy findings of the original and metastatic tumor, differential diagnoses, and the results of the English-language literature review are presented.

Leiomyoma of the male urethra: a case report and review of the literature.

We describe the case of a 48-year-old quadriplegic black man with history of C4-C5 cervical spine and cord injury secondary to a fall, who presented to the University of Cincinnati Medical Center Urology Service with obstructive symptoms at urination. A bulbous urethral stricture was diagnosed and subsequently resected with primary urethral reanastomosis. On pathologic examination, the surgical specimen contained an epithelioid leiomyoma at the site of the urethral stricture. Although leiomyomas of the female urethra are relatively common, we identified only 2 previously reported cases of leiomyomas of the male urethra in the English-language medical literature. To the best of our knowledge, we describe the third case of leiomyoma of the male urethra, the first of the epithelioid type.

First Report of Leaf Blight on Sweet Persimmon Tree by Pestalotiopsis theae in Spain.

During July 1998, a leaf blight caused by Pestalotiopsis theae (Saw.) Stey. was observed at an incidence of 18 to 20% in sweet persimmon (Diospyros kaki L.fil.) orchards in Huelva Province (southwestern Spain). Symptoms appeared on leaves as large grayish brown circular ringspots. Usually, they were solitary, but occasionally, two to three spots occurred on an affected leaf. In severe cases, lesions developed on more than one-third of the leaf, resulting in defoliation. Small black acervular conidiomata were visible in the surface of spots. These conidiomata produced fusiform conidia that were straight or rarely curved, four five-celled euseptate, including three olivaceous or dark brown median cells, and hyaline apical and basal cells with appendages that were slightly constricted at septa. Conidiomata were up to 240 µm in diameter; conidiogenous cells were 6 to 13 × 1.2 to 2.8 µm; conidia were 24.7 × 7.8 µm; three median cells were 16.7 µm long; two to three apical appendages (rarely four) were 28.3 µm long; and straight basal appendage was 5.7 µm. P. theae was consistently isolated on potato dextrose agar from diseased leaves and conidiomata. To confirm pathogenicity, both mycelial plugs and a conidial suspension (1.5 × 106 conidia per ml) of the fungus were used as inocula. Young completely developed leaves from persimmon tree cvs. Sharon and Hanafuyu were inoculated in the laboratory and maintained in a moist chamber for 5 days. Lesions resembling symptoms that occurred in the field were observed on leaves after 5 days. Symptoms were not observed on control leaves inoculated with agar media or sprayed with water. The fungus reisolated from diseased leaves was identical to the original isolates. Based on the morphological characteristics of conidiomata and conidia as well as pathogenicity, the fungus was identified as P. theae (1). This is the first report of this fungus as a pathogen of D. kaki in Europe. Possibly the introduction of P. theae to Spain has been through young imported persimmon plants. Unusual climatic conditions (heavy rainfalls during 1997 in southwestern Spain) have been favorable for disease development. The hot and dry conditions that usually occur during flowering, growing, and maturation of persimmon fruits normally prevent dissemination of inoculum and infection of leaves. For these reasons, the wet areas of southwestern Spain could be more favorable for "grey blight" of persimmon trees. Reference: (1) T.-H. Chang et al. Korean J. Plant Pathol. 12:377, 1996.

Malignant mixed müllerian tumor of the extraovarian secondary müllerian system. Report of two cases and review of the English literature.

OBJECTIVE: To report two cases of malignant mixed müllerian tumor of the extraovarian secondary müllerian system and to identify cases reported in the English literature. DESIGN: Two cases are described and discussed along with cases reported previously in the English literature. SETTING: The University of Cincinnati Medical Center. PATIENTS: (1) A 62-year-old woman with a bilateral ovarian poorly differentiated endometrioid adenocarcinoma with multiple peritoneal metastasis and a malignant mixed müllerian tumor with heterologous elements arising from the pelvic peritoneum. (2) An 83-year-old woman with a malignant mixed müllerian tumor with heterologous elements arising from the cecal peritoneum. RESULTS: Seventeen previously reported cases were identified in the English literature. CONCLUSIONS: The malignant mixed müllerian tumor of the extraovarian secondary müllerian system is a rare disease with only 17 cases reported to date to our knowledge. The prognosis is poor. Of 12 patients with follow-up information available, 10 died within 1 year and 2 within 2 years after diagnosis. The histogenesis of the tumor remains unclear; however, data exist that support transformation of epithelial neoplastic cells into sarcomatous cells (metaplastic theory) and origin from a single totipotential cell. It is possible that both situations may occur.

Lipoleiomyoma of the ovary: report of a case and review of the English literature.

The purpose of this communication is to report a case of lipoleiomyoma of the ovary. It appears to be the first case of this condition reported in the English literature. Reference to its possible histogenesis is made.