RESUMO
In May 2014, a severe leaf spot disease was observed on walnut tree (Juglans regia L.) in Hechi, Guangxi, China. Leaf spots were circular to semicircular in shape, water-soaked, later becoming grayish white in the center with a dark brown margin and bordered by a tan halo. Necrotic lesions were approximately 3 to 4 mm in diameter. Diseased leaves were collected from 10 trees in each of five commercial orchards. The diseased leaves were cut into 5 × 5 mm slices, dipped in 75% ethanol for 30 s, washed three times in sterilized water, sterilized with 0.1% (w/v) HgCl2 for 3 min, and then rinsed five times with sterile distilled water. These slices were placed on potato dextrose agar (PDA), followed by incubating at 28°C for about 3 to 4 days. Fungal isolates were obtained from these diseased tissues, transferred onto PDA plates, and incubated at 28°C. These isolates produced gray aerial mycelium and then became pinkish gray with age. Moreover, the reverse of the colony was pink. The growth rate was 8.21 to 8.41 mm per day (average = 8.29 ± 0.11, n = 3) at 28°C. The colonies produced pale orange conidial masses and were fusiform with acute ends, hyaline, sometimes guttulate, 4.02 to 5.25 × 13.71 to 15.72 µm (average = 4.56 ± 0.31 × 14.87 ± 1.14 µm, n = 25). The morphological characteristics and measurements of this fungal isolate matched the previous descriptions of Colletotrichum fioriniae (Marcelino & Gouli) R.G. Shivas & Y.P. Tan (2). Meanwhile, these characterizations were further confirmed by analysis of the partial sequence of five genes: the internal transcribed spacer (ITS) of the ribosomal DNA, beta-tubulin (ß-tub) gene, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene, chitin synthase 3(CHS-1) gene, and actin (ACT) gene, with universal primers ITS4/ITS5, T1/ßt2b, GDF1/GDR1, CHS1-79F/CHS1-354R, and ACT-512F/ACT-783R, respectively (1). BLAST of these DNA sequences using the nucleotide database of GenBank showed a high identify (ITS, 99%; ß-tub, 99%; GAPDH, 99%; CHS-1, 99%; and ACT, 100%) with the previously deposited sequences of C. fioriniae (ITS, KF278459.1, NR111747.1; ß-tub, AB744079.1, AB690809.1; GAPDH, KF944355.1, KF944354.1; CHS-1, JQ948987.1, JQ949005.1; and ACT, JQ949625.1, JQ949626.1). Koch's postulates were fulfilled by inoculating six healthy 1-year-old walnut trees in July 2014 with maximum and minimum temperatures of 33 and 26°C. The 6-mm mycelial plug, which was cut from the margin of a 5-day-old colony of the fungus on PDA, was placed onto each pin-wounded leaf, ensuring good contact between the mycelium and the wound. Non-colonized PDA plugs were placed onto pin-wounds as negative controls. Following inoculation, both inoculated and control plants were covered with plastic bags. Leaf spots, similar to those on naturally infected plants, were observed on the leaves inoculated with C. fioriniae within 5 days. No symptoms were observed on the negative control leaves. Finally, C. fioriniae was re-isolated from symptomatic leaves; in contrast, no fungus was isolated from the control, which confirmed Koch's postulates. To our knowledge, this is the first report of leaf disease on walnut caused by C. fioriniae. References: (1) L. Cai et al. Fungal Divers. 39:183, 2009. (2) R. G. Shivas and Y. P. Tan. Fungal Divers. 39:111, 2009.
RESUMO
PURPOSE: To evaluate the clinical efficacy of CT-guided percutaneous huge ilio-psoas abscesses drainage combined with posterior approach surgery for the management of dorsal and lumbar spinal tuberculosis in 16 adult cases. METHODS: Between January 2006 and June 2013, a total of 16 dorsal and lumbar spinal tuberculosis patients with huge ilio-psoas abscesses underwent two-stage CT-guided percutaneous abscesses drainage and posterior debridement, decompression, intervertebral fusion and instrumentation. Standard quadruple antituberculous chemotherapy was performed both before and after surgery. RESULT: The average follow-up period was 26.7 months (range: 18-38 months). There is no severe complication and relapse of spinal tuberculosis. The blood loss was 921.0±141.3mL, operation time was 174.8±15.7minutes. Kyphotic angle improved from 36.6±10.0° preoperatively to 8.1±1.8° postoperatively with 2.2±1.5° loss of correction at final follow-up. The solid bone fusion was achieved in all cases at average 6.6±2.2 months after surgery. Neurologic deficits were recovered in varying degrees except 4 cases remained the same. The postoperative quality of life significantly improved. The Oswestry Disability Index (ODI) decreased from 32.8±10.6 preoperatively to 14.4±7.9 at the final follow-up. CONCLUSION: CT-guided percutaneous drainage combined with posterior approach surgery was proved to be safe and effective for the management of dorsal and lumbar spinal tuberculosis with huge ilio-psoas abscesses in adults. LEVEL OF STUDY: Level IV, retrospective.