First Report of Penicillium polonicum Causing Blue Mold on Stored Onion (Allium cepa) in Serbia.

Penicillium polonicum K. Zaleski is an economically important airborne fungus with a broad host range including cereals, peanuts, onions, dried meats, citrus fruits, and yam tubers (2,4). Secondary metabolites produced by this species include harmful mycotoxins penicillic acid, verucosidin, and nephrotoxic glycopeptides, which may play a role in Balkan Endemic Nephropathy (2,5). In January 2013, decayed onion bulbs (Allium cepa L. cv. Meranto) with blue mold symptoms were found causing significant economic losses at a storage facility in Stara Pazova, Serbia, and were collected. The decayed area of the bulbs was pale yellow to light brown, and tissue was soft and watery. Bluish green sporulation was abundant on the surface and inside the bulb, between decayed scales. Two isolates (designated L1a and L4p) were obtained and further characterized using morphological and molecular methods. Colonies on potato dextrose agar (PDA), Czapek yeast autolysate agar (CYA), malt extract agar (MEA), and yeast extract sucrose agar (YES) media at 25°C after 7 days were blue green, velutinous, with clear exudate present on CYA. Colony reverse color on CYA and YES for both isolates were cream to yellow brown. The mean colony diameter on PDA for L1a was 29.89 ± 0.96 mm, and for L4p was 26 ± 0.37 mm; on CYA 32.56 ± 0.53 mm for L1a and 30.11 ± 2.42 mm for L4p; and on YES 33.86 ± 1.59 mm for L1a and 31.17 ± 1.83 mm for L4p. No growth was observed on CYA when isolates were incubated at 37°C. Conidiophores of both isolates were terverticillate, stipes were septate with smooth to finely roughened walls, and phialides were ampulliform. Conidia were globose to subglobose, smooth-walled, and borne in columns. Conidial dimensions for L4p were 2.72 to 3.82 (3.26) × 2.36 to 3.42 (2.95) µm, and for L1a were 2.87 to 4.39 (3.58) × 2.53 to 3.79 (3.16) µm (n = 50). Both isolates tested positive for the production of cyclopiazonic acid and other alkaloids, as indicated by a violet reaction for the Ehrlich test. Morphological characters of L1a and L4p were in accordance with those described for P. polonicum K. Zaleski (2). Genomic DNA was isolated using CTAB extraction method (1) and molecular identification was completed using gene specific primers for the ß-tubulin locus (Bt-LEV-Up4/Bt-LEV-Lo1) via conventional PCR (3). The nucleotide sequences of amplified products (~800 bp) have been assigned to GenBank (KJ570971 and 72). MegaBLAST of obtained sequences showed a 99% similarity with several sequences of P. polonicum deposited in GenBank, which confirmed the morphological identification. Pathogenicity was tested by wound inoculation of 10 surface sanitized onion bulbs cv. Meranto with 50 µl of a 105/ml conidial suspension from isolates grown on PDA. Ten control onion bulbs were wound-inoculated with Tween-treated sterile distilled water. After 30 days incubation in plastic containers, under high humidity at 22°C, typical symptoms of blue mold developed on inoculated bulbs, while non-inoculated controls remained symptomless. Isolates recovered from inoculated bulbs showed the same morphological characteristics as the original isolates, thus completing Koch's postulates. To our knowledge, this is the first report of P. polonicum on stored onion in Serbia. Results from this study indicate that a holistic approach to control this fungus should be implemented that may include one or all of the following: increased sanitation methods to eliminate inoculum, breeding for resistant onion cultivars, and integration of additional control methods to maintain onion quality during storage. References: (1) J. P. Day and R. C. Shattock. Eur. J. Plant Pathol 103:379, 1997. (2) J. C. Frisvad and R. A. Samson. Stud. Mycol. 49:1, 2004. (3) S. N. de Jong et al. Mycol. Res. 105:658, 2001. (4) W. K. Kim et al. Mycobiology 36:217, 2008. (5) P. G. Mantle. Facta Univ. Ser. Med. Biol. 9:64, 2002.

First Report of Penicillium crustosum Causing Blue Mold on Stored Apple Fruit in Serbia.

Penicillium crustosum Thom (1930) causes blue mold on pome fruits and is also regularly found on cheese, nuts, and soil (1,3). The fungus produces a wide range of mycotoxins such as penitrem A, roquefortine C, terrestric acid, and cyclopenol, which impact human health (1). In January and February 2013, 20 decayed apples, 'Golden Delicious' and 'Jonagold' (Malus × domestica Borkh.) with blue mold symptoms were collected from cold storages in Svilajnac and Bela Crkva, Serbia. Decayed areas were light to medium brown with blue green sporulation on the surface of the lesion. Decayed tissue was soft and watery with a sharp margin between the diseased and healthy areas. One isolate from each cultivar was designated JP2 ('Golden Delicious') and JBC7 ('Jonagold') and further characterized. Conidiophores of both isolates were terverticillate, stipes were septate with rough walls, and phialides were ampulliform. Conidia were smooth, borne in columns, and were spherical to subglobose. Conidial dimensions for JP2 were 3.2 to 4.56 (3.73) × 2.64 to 4.3 (3.32) µm and for JBC7 were 3.1 to 4.46 (3.65) × 2.81 to 4.27 (3.31) µm (n = 50). The isolates were cultured on Czapek yeast autolysate agar (CYA), malt extract agar (MEA), and yeast extract sucrose agar (YES) media and incubated at 25°C for 7 days. Mycelia were white with heavy sporulation yielding grayish green colonies on all media. Colonies were radially sulcate and velutinous, with clear exudate, and produced a yellow to orange reverse on CYA and YES. On MEA, colonies were plane, low, and mycelia subsurface with conidia having a dry powdery appearance. Crusts of conidial masses formed after 10 or more days. No growth was observed on CYA when these isolates were incubated at 37°C. Both isolates were identified as P. crustosum Thom using morphological characters according to (2) and (1). Species level identification was confirmed by isolating genomic DNA followed by amplification of the ß-tubulin locus using gene specific primers via conventional PCR (4). MegaBLAST analysis of the 2X consensus nucleotide sequences revealed that JP2 and JBC7 (GenBank KJ433984 and 85) were 99% identical to P. crustosum culture collection isolate IBT 21518 (JN112030.1). Koch's postulates were examined using two apple cvs. Idared and Kolacara. Ten fruit per cultivar per isolate were inoculated on two sides of each fruit; 20 fruit were used as water-only inoculated controls. Fruit were washed with soap and water, surface sanitized with 70% ethanol, and placed into polyethylene boxes. Using a finishing nail, 4-mm wounds were created and inoculated with 50 µl of a 3 × 105/ml conidial suspension or Tween-treated sterile distilled water. Boxes with inoculated and control fruit were stored at 25°C for 10 days. The inoculated fruit developed small, soft, watery lesions, which enlarged into decayed areas with defined edges and abundant sporulation on the surface. Symptoms were identical to the original ones, while the control fruit remained symptomless. The fungus was re-isolated from infected tissue and showed the same morphological characteristics as the original isolates, thus completing Koch's postulates. Blue mold occurs during long term storage of apples and is predominantly caused by P. expansum. This is the first report of P. crustosum causing postharvest blue mold decay on apple fruit obtained from storage in Serbia and indicates that P. crustosum is an emerging pathogen for the Serbian pome fruit growing and packing industry. References: (1) J. C. Frisvad and R. A. Samson. Stud. Mycol. 49:1, 2004. (2) J. I. Pitt and A. D. Hocking. Fungi and Food Spoilage, 239. Springer, 2009. (3) P. G. Sanderson and R. A. Spotts. Phytopathology 85:103. 1995. (4) P. L. Sholberg et al. Postharvest Biol. Technol. 36:41, 2005.

First Report of Penicillium expansum Isolates Resistant to Pyrimethanil from Stored Apple Fruit in Pennsylvania.

Apples in the United States are stored in low-temperature controlled atmospheres for 9 to 12 months and are highly susceptible to blue mold decay. Penicillium spp. cause significant economic losses worldwide and produce mycotoxins that contaminate processed apple products. Blue mold is managed by a combination of cultural practices and the application of fungicides. In 2004, a new postharvest fungicide, pyrimethanil (Penbotec 400 SC, Janseen PMP, Beerse, Belgium) was registered for use in the United States to control blue mold on pome fruits (1). In this study, 10 blue mold symptomatic 'Red Delicious' apples were collected in May 2011 from wooden bins at a commercial facility located in Pennsylvania. These fruit had been treated with Penbotec prior to controlled atmosphere storage. Ten single-spore Penicillium spp. isolates were analyzed for growth using 96-well microtiter plates containing Richards minimal medium amended with a range of technical grade pyrimethanil from 0 to 500 µg/ml. Conidial suspensions adjusted to 1 × 105 conidia/ml were added to three 96-well plates for each experiment; all experiments were repeated three times. Nine resistant isolates had prolific mycelial growth at 500 µg/ml, which is 1,000 times the discriminatory dose that inhibited baseline sensitive P. expansum isolates from Washington State (1). However, one isolate (R13) had limited conidial germination and no mycelial proliferation at 0.5 µg/ml and was categorized as sensitive. One resistant (R22) and one sensitive (R13) isolate were selected on the basis of their different sensitivities to pyrimethanil. Both isolates were identified as P. expansum via conventional PCR using ß-tubulin gene-specific primers according to Sholberg et al. (2). Analysis of the 2X consensus amplicon sequences from R13 and R22 matched perfectly (100% identity and 0.0 E value) with other P. expansum accessions in GenBank including JN872743.1, which was isolated from decayed apple fruit from Washington State. To determine if pyrimethanil applied at the labeled rate of 500 µg/ml would control R13 or R22 in vivo, organic 'Gala' apple fruit were wounded, inoculated with 50 µl of a conidial suspension (1 × 104 conidia/ml) of either isolate, dipped in Penbotec fungicide or sterile water, and stored at 25°C for 7 days. Twenty fruit composed a replicate within a treatment and the experiment was performed twice. Non-inoculated water-only controls were symptomless, while water-dipped inoculated fruit had 100% decay with mean lesion diameters of 36.8 ± 2.68 mm for R22 and 38.5 ± 2.61 mm for R13. The R22 isolate caused 30% decay with 21.6 ± 5.44 mm lesions when inoculated onto Penbotec-treated apples, while the R13 isolate had 7.5% decay incidence with mean lesion diameters of 23.1 ± 3.41 mm. The results from this study demonstrate that P. expansum pyrimethanil-resistant strains are virulent on Penbotec-treated apple fruit and have the potential to manifest in decay during storage. To the best of our knowledge, this is the first report of pyrimethanil resistance in P. expansum from Pennsylvania, a major apple growing region for the United States. Moreover, these results illuminate the need to develop additional chemical, cultural, and biological methods to control this fungus. References: (1) H. X. Li and C. L. Xiao. Phytopathology 98:427, 2008. (2) P. L. Sholberg et al. Postharvest Biol. Technol. 36:41, 2005.

First Report of Botryosphaeria dothidea Causing White Rot on Apple Fruit in Maryland.

Botryosphaeria dothidea (Moug.:Fr.) Ces. De Not. causes perennial cankers on apple trees and causes white rot on apple fruit in the field and during storage (1). Prolonged periods of warm wet weather favor rapid disease outbreaks that result in severe losses, which range from 25 to 50% for the southeastern United States (3). A B. dothidea isolate was obtained from decayed 'Fuji' apple fruit exhibiting white rot symptoms from a local farm market in Beltsville, MD, in May 2010. The fruit had characteristic large dark brown lesions with irregular margins and decay expanded unevenly toward the core and the tissue was soft. The pathogen was isolated from symptomatic tissue by spraying the lesion surface with 70% ethanol. The skin with aseptically removed with a scalpel and small pieces of tissue were placed on potato dextrose agar (PDA) and incubated at 20°C. Once fungal growth was evident, the cultures were hyphal-tip transferred to individual PDA plates and incubated at 20°C. The B. dothidea isolate produced black aerial mycelium with a white margin on PDA and had a black reverse. Conidiomata were evident after 10 to 14 days at 20°C only on oatmeal agar. Conidia were hyaline, smooth and straight, fusiform with an subobtuse apex and a truncate base 20 to 26 (24.33) × 4 to 7 (5) µm (n = 50). Genomic DNA was isolated from the fungus and amplified with gene specific primers (ITS 4 and 5) for the ribosomal DNA internal transcribed spacer region ITSI-5.8S-ITS2 as described by White et al. (4). Both forward and reverse strands of the 542-bp amplicon were sequenced and assembled into a contig. The nucleotide sequence (GenBank Accession No. KC473852) indicated 99% identity to B. dothidea isolate CMM3938 (JX513645.1) and to voucher specimens CMW 25686, 25696, and 25222 (FM955381.1, FM955379.1, and FM955377,1). Koch's postulates were conducted using three 'Golden Delicious' apple fruit that were wound-inoculated with 50 µl of a mycelial suspension of the fungus, obtained from aseptically scraping a 7-day-old PDA culture, and was also repeated using 'Fuji' apple fruit. Large, brown, slightly sunken, soft lesions with undefined edges developed 5 days after inoculation at 20°C and water-only inoculated fruit were symptomless. The fungus was reisolated from infected tissue and was morphologically identical to the original isolate from decayed apple fruit. To determine if the B. dothidea isolate was resistant to postharvest fungicides, the minimum inhibitory concentration (MIC) was conducted using the 96 well plate method with a mycelial suspension of the fungus as described by Pianzzola et al. (2). The MIC for the isolate was >1 ppm for Mertect and Scholar and 50 ppm for Penbotec, which are well below the labeled rates for these postharvest fungicides and the experiment was repeated. To our knowledge, this is the first report of B. dothidea causing white rot on apple fruit in Maryland. References: (1) A. R. Biggs and S. S. Miller. HortScience 38:400, 2003. (2) M. J. Pianzzola et al. Plant Dis. 88:23, 2004. (3) T. B. Sutton. White rot and black rot. Pages 16-20 in: Compendium of Apple and Pear Diseases, A. L. Jones and H. S. Aldwinckle, eds. The American Phytopathological Society, St Paul, MN, 1991. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Application. M. A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

First Report of Neofusicoccum ribis Causing Postharvest Decay of Apple Fruit from Cold Storage in Pennsylvania.

Neofusicoccum ribis (Slippers, Crous & M.J. Wingf.), previously known as Botryosphaeria ribis (Grossenb. & Duggar), is an aggressive fungal plant pathogen that is part of the N. ribis/N. parvum species complex that causes stem cankers on a variety of woody plant species (2). An isolate of N. ribis was obtained from decayed 'Honeycrisp' apple fruit from a commercial cold storage facility located in Pennsylvania in October of 2011. The decayed apple fruit sample had a brownish lesion that was soft, dry, and leathery on the surface while sporulation was not evident. To conduct Koch's postulates, three 'Golden Delicious' apple fruits were wound-inoculated with a 50-µl mycelial suspension, obtained from aseptically scraping a 7-day-old potato dextrose agar (PDA) culture of the fungus, and was repeated using 'Fuji' apple fruit. The inoculated fruit developed lesions, while water-inoculated fruit were symptomless after 5 days at 20°C. N. ribis was reisolated from infected tissue and was morphologically identical to the original isolate. Genomic DNA was isolated, a portion of the ß-tubulin gene was amplified with the gene specific primers, and the amplicon was sequenced and analyzed using BLAST (1). The nucleotide sequence (GenBank Accession No. KC47853) had 99% identity with N. ribis SEGA8 isolate (JN607146.1). The N. ribis isolate produced a grayish-white mycelium with abundant aerial hyphae on PDA and had an olive-colored reverse. Microscopic investigation revealed septate mycelia with right angle branching and conidiomata were not evident on PDA, V8, oatmeal agar (OMA), or water agar (WA). Growth on WA was sparse and transparent, and aerial mycelial growth was not produced. Growth rate analyses were conducted on PDA, V8, and OMA and were 10.1 (±1.39), 20.4 (±1.15), and 17.6 (±0.70) mm/day at 20°C and the experiment was repeated. The minimum inhibitory concentrations (MIC) for the N. ribis isolate was carried out for three postharvest fungicides as described by Pianzzola et al. (3). Briefly, 96 well plates were filled with PDA alone (0 ppm) and PDA amended with 10 fungicide concentrations ranging from 1 to 1,200 ppm for thiabendazole (Mertect), and 1 to 1,000 ppm for fludioxonil (Scholar) and pyrimethanil (Penbotec). A mycelial suspension of the fungus was obtained from pure culture, 50 µl of the mycelial suspension was pipetted into each well, and allowed to grow for 72 h at 25°C. The experiment was conducted twice. The N. ribis isolate displayed MIC values of >1 ppm thiabendazole (Mertect), >1 ppm fludioxonil (Scholar), and 50 ppm pyrimethanil (Penbotec), which are all well below the labeled application rates for these postharvest fungicides. To our knowledge, this is the first report of N. ribis causing postharvest decay on apple fruit obtained from a commercial storage facility in Pennsylvania. References: (1) S. F. Altschul et al. J. Mol. Biol. 215:403, 1990. (2) D. Pavlic et al. Mycologia 101:636, 2009. (3) M. J. Pianzzola et al. Plant Dis. 88:23, 2004.

First Report of Botryosphaeria dothidea Causing White Rot of Apple Fruit in Serbia.

Botryosphaeria dothidea (Moug.: Fr.) Ces. & De Not has a worldwide distribution infecting species from over 80 genera of plants (1). Apart from being an important pathogen of apple trees in many countries, B. dothidea can cause pre- and postharvest decay on apple fruit (2). It has been known to cause canker and dieback of forest trees in Serbia (3), but has not been recorded either on apple trees or apple fruit. In December 2010, apple fruit cv. Idared (Malus × domestica Borkh.) with symptoms of white rot were collected from one storage in the area of Svilajnac in Serbia. The incidence of the disease was low but the symptoms were severe. Affected fruit were brown, soft, and almost completely decayed, while the internal decayed tissue appeared watery and brown. A fungus was isolated from symptomatic tissue of one fruit after surface sterilization with 70% ethanol (without rinsing) and aseptic removal of the skin. Small fragments of decayed tissue were placed on potato dextrose agar (PDA) and incubated in a chamber at 22°C under alternating light and dark conditions (12/12 h). Fungal colonies were initially whitish, but started turning dark gray to black after 5 to 6 days. Pycnidia were produced after 20 to 25 days of incubation at 22°C and contained one-celled, elliptical, hyaline conidia. Conidia were 17.19 to 23.74 µm (mean 18.93) × 3.72 to 4.93 µm (mean 4.45) (n = 50). These morphological characteristics are in accordance with those described for the fungus B. dothidea (4). Genomic DNA was isolated from the fungus and internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The nucleotide sequence has been assigned to GenBank Accession No. KC994640. BLAST analysis of the 528-bp segment showed a 100% similarity with several sequences of B. dothidea deposited in NCBI GenBank, which confirmed morphological identification. Pathogenicity was tested by wound inoculation of five surface-sterilized, mature apple fruit cv. Idared with mycelium plugs (5 mm in diameter) of the isolate grown on PDA. Five control fruit were inoculated with sterile PDA plugs. After 5 days of incubation in plastic containers, under high humidity (RH 90 to 95%) at 22°C, typical symptoms of white rot developed on inoculated fruit, while wounded, uninoculated, control fruit remained symptomless. The isolate recovered from symptomatic fruit showed the same morphological features as original isolate. To the best of our knowledge, this is the first report of B. dothidea on apple fruit in Serbia. Apple is widely grown in Serbia and it is important to further investigate the presence of this pathogen in apple storage, as well as in orchards since B. dothidea may cause rapid disease outbreaks that result in severe losses. References: (1) G. H. Hapting Agriculture Handbook 386, USDA, Forest Service, 1971. (2) A. L. Jones and H. S. Aldwinckle Compendium of Apple and Pear Diseases. APS Press, St. Paul, MN, 1990. (3) D. Karadzic et al. Glasnik Sumarskog Fakulteta 83:87, 2000. (4) B. Slippers et al. Mycologia 96:83, 2004.

First Report of Penicillium carneum Causing Blue Mold on Stored Apples in Pennsylvania.

Blue mold decay occurs during long term storage of apples and is predominantly caused by Penicillium expansum Link. Apples harvested in 2010 were stored in a controlled atmosphere at a commercial Pennsylvania apple packing and storage facility, and were examined for occurrence of decay in May 2011. Several decayed apples from different cultivars, exhibiting blue mold symptoms with a sporulating fungus were collected. One isolate recovered from a decayed 'Golden Delicious' apple fruit was identified as P. carneum Frisvad. Genomic DNA was isolated, 800 bp of the 3' end of the ß-tubulin locus was amplified using gene specific primers and sequenced (4). The recovered nucleotide sequence (GenBank Accession No. JX127312) indicated 99% sequence identity with P. carneum strain IBT 3472 (GenBank Accession No. JF302650) (3). The P. carneum colonies strongly sporulated and had a blue green color on potato dextrose agar (PDA), Czapek yeast autolysate agar (CYA), malt extract agar (MEA), and yeast extract sucrose agar (YES) media at 25°C after 7 days. The colonies also had a beige color on plate reverse on CYA and YES media. The species tested positive for the production of alkaloids, as indicated by a violet reaction for the Ehrlich test, and grew on CYA at 30°C and on Czapek with 1,000 ppm propionic acid agar at 25°C; all of which are diagnostic characters of this species (2). The conidiophores were hyaline and tetraverticillate with a finely rough stipe. Conida were produced in long columns, blue green, globose, and averaged 2.9 µm in diameter. To prove pathogenicity, Koch's postulates were conducted using 20 'Golden Delicious' apple fruits. Fruits were washed, surface sterilized with 70% ethanol, and placed onto fruit trays. Using a nail, 3-mm wounds were created and inoculated with 50 µl of a 106/ml conidial suspension or water only as a negative control. The fruit trays were placed into boxes and were stored in the laboratory at 20°C for 7 days. The inoculated fruit developed soft watery lesions, with hard defined edges 37 ± 4 mm in diameter. The sporulating fungus was reisolated from infected tissue of all conidia inoculated apples and confirmed to be P. carneum by polymerase chain reaction (PCR) using the ß-tubulin locus as described. Water inoculated control apples were symptomless. Originally grouped with P. roqueforti, P. carneum was reclassified in 1996 as a separate species (1). P. carneum is typically associated with meat products, beverages, and bread spoilage and produces patulin, which is not produced by P. roqueforti (1,2). Our isolate of P. carneum was susceptible to the thiabendazole (TBZ) fungicide at 250 ppm, which is below the recommended labeled application rate of 600 ppm. The susceptibility to TBZ suggests that this P. carneum isolate has been recently introduced because resistance to TBZ has evolved rapidly in P. expansum (4). To the best of our knowledge, P. carneum has not previously been described on apple, and this is the first report of P. carneum causing postharvest decay on apple fruits obtained from storage in Pennsylvania. References: (1) M. Boyson et al. Microbiology 142:541, 1996. (2) J. C. Frisvad and R. A. Samson. Stud. Mycol. 49:1, 2004. (3) B. G. Hansen et al. BMC Microbiology 11:202, 2011. (4) P. L. Sholberg et al. Postharvest Biol. Technol. 36:41, 2005.

Embolización selectiva de las arterias uterinas en el tratamiento de miomas sintomáticos: dos años de seguimiento / Selective uterine artery embolization in the treatment of symptomatic myomas: A two-year follow-up

El leiomioma es el tumor ginecológico más frecuente. Entre las opciones terapéuticas, la embolización de arterias uterinas (EAU) se emplea como alternativa a la miomectomía o a la histerectomía. Resumen El objetivo es conocer las características epidemiológicas y clínicas de las pacientes sometidas a EAU y analizar los resultados a los dos años. Material y métodos Estudio observacional y descriptivo de 93 pacientes que fueron sometidas a EAU para el tratamiento del mioma uterino. A las pacientes se les realizó una anamnesis completa, exploración clínica y analítica. Además un seguimiento en consulta a los 2, a los 6, a los 12 y a los 24 meses. Resultados La media de días de sangrado fue de 6,74 (4–14). La tasa media de hemoglobina fue de 11,94g/dl (7–16). La media de los días de ingreso fue de 4,2 (2–10) y la media de los días de dolor posterior fue de 5,8 (1–15). Hubo 3 casos de amenorrea transitoria (3,2%) y 3 casos de amenorrea definitiva (3,2%).Resultados En el volumen uterino y en el tamaño del mioma se apreciaba una reducción significativa en cada revisión hasta los dos años de la embolización. Respecto a la variación clínica, se aprecia una disminución significativa en la sintomatología, el sangrado y el dolor. Discusión En el estudio, la EAU se revela como una técnica bien tolerada y eficaz con una respuesta estadísticamente significativa de todos los parámetros clínicos y ecográficos. Sus indicaciones deben ser especificadas y respetadas para mantener su eficacia y utilidad (AU)

Leiomyoma is the most common gynecologic tumor. Uterine artery embolization (UAE) is used as an alternative to myomectomy or hysterectomy to treat uterine leiomyoma. Objective To determinate the epidemiological and clinical characteristics of patients undergoing UAE and to analyze the results of UAE at 2 years. Material and methods An observational, descriptive study was carried out in 93 patients who underwent UAE for the treatment of uterine myoma. A complete history and physical examination were performed and laboratory data were collected. Follow-up visits were performed at 2, 6, 12 and 24 months after the procedure. Results The mean number of days of bleeding was 6.74 (4–14). The mean hemoglobin was 11.94g/dl (7–16). The mean length of hospital stay was 4.2 days (2–10), and the number of subsequent days of pain was 5.8 (1–15). There were three cases of transient amenorrhea (3.2%) and three of permanent amenorrhea (3.2%).Results The volume and size of uterine myoma were significantly reduced at each follow-up visit until 2 years after the embolization. A significant decrease in symptoms, bleeding and pain was confirmed in this period. Discussion UAE is a well-tolerated and effective technique, with a statistically significant response of all clinical and ultrasonographic parameters. The indications for UAE should be clearly defined and followed to maintain the efficacy and utility of this procedure (AU)

First Report of Zucchini yellow mosaic virus, Watermelon mosaic virus, and Cucumber mosaic virus in Bottlegourd (Lagenaria siceraria) in Serbia.

During a cucurbit disease survey in August 2004, severe symptoms resembling those caused by viruses were observed on bottlegourd (Lagenaria siceraria (Molina) Standl.) in the Vojvodina region of Serbia. Symptoms included stunting, mosaic, green veinbanding, blistering, yellowing, chlorotic spots, leaf deformation, and fruit distortion. Leaf samples from 25 symptomatic plants were collected from two localities for virus identification using mechanical transmission and serological testing. Crude sap extract from leaf samples was mechanically inoculated onto bottlegourd and pumpkin (Cucurbita pepo) under greenhouse conditions. Field-collected bottlegourd and inoculated plants were tested using double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA). Positive reactions were obtained on collected and inoculated plants with polyclonal antiserum (Loewe Biochemica, Sauerlach, Germany) to Zucchini yellow mosaic virus(ZYMV) in 23 samples, with antiserum to Watermelon mosaic virus (WMV) in eight samples, and with antiserum to Cucumber mosaic virus (CMV) in seven samples. Each of the three viruses was detected in single as well as in mixed infections with the other two viruses. Biological characterization of viruses detected in single infections was done on the following indicator plants: Chenopodium amaranticolor, C. quinoa, Cucumis sativus, Cucumis melo, Citrullus lanatus, Nicotiana glutinosa, and N. tabacum cv. Samsun. The symptoms observed on indicator plants for each isolate corresponded to the results of DAS-ELISA (2,3). All three viruses are known to be important pathogens of cucurbit plants and were previously reported in pumpkin in Serbia (1). To our knowledge, this is the first report of ZYMV, WMV, and CMV in bottlegourd in Serbia. References: (1) N. Dukic et al. J. Agric. Sci. 47:149, 2002. (2) D. E. Lesemann et al. Phytopathol. Z. 108:304, 1983. (3) H. Rahimian and K. Izadpanah. Phytopathol. Z. 92:305, 1978.

Two cases of exogenous endophthalmitis due to Fusarium moniliforme and Pseudomonas species as associated aetiological agents.

Perforative eye injuries in two patients were complicated by fulminant endophthalmitis. In both patients the cornea was free of micro-organisms. Vitrectomy of the posterior pars plana was performed and the contents of the vitreous body were examined mycologically and bacteriologically. Fusarium moniliforme var. subglutinans and Pseudomonas species were isolated from both patients. Fungal and bacterial organisms penetrated the vitreous body via metal foreign bodies.