First Report of Lasiodiplodia pseudotheobromae Causing Collar Rot of Peanut in Shandong Province, China

In August 2019, a collar rot of peanut was observed in several fields in Qingdao, Shandong province, China. Disease survey was conducted in several peanut fields. Less than 5% plants exhibited various symptoms, including brown or black stem rot, pod rot, leaf chlorotic, wilted, and even dead. Symptomatic stems were cut into small pieces, surface disinfested with 70% ethanol for 1 min, 1% NaClO for 2 minutes, rinsed three times with sterile water, and dried on sterile filter papers. Pieces then were plated on potato dextrose agar (PDA) media and incubated at 25°C in darkness. Fungal cultures were initially white, then turned gray, and eventually turned black, and aerial hyphae were dense, fluffy. Conidia were ellipsoidal, initially hyaline, unicellular, 14.3 to 21.1 × 8.7 to 13.2 µm (n = 50), and mature conidia showed dark brown, with a central septum, and longitudinal stripes. Molecular identification was performed by sequencing ITS with ITS1/ITS4 (White et al., 1990) and beta tubulin gene with Bt2a/Bt2b (Glass and Donaldson, 1995) of a representative isolate ZHX9. ITS and beta tubulin regions (OK427342 and OK489788) of ZHX9 obtained 99.62 and 100% similar to L. pseudotheobromae (KF766193 and EU673111), respectively. Phylogenetic analysis was done using Neighbor-Joining (NJ) analysis based on those gene sequences. The microorganism we have isolated was identified as L. pseudotheobromae based on molecular analysis and morphological characteristics. For pathogenicity assay, twelve ten-days-old peanut (Zhonghua No.12) seedlings were each inoculated with one mycelial plug (8 mm in diameter) by placing the inoculum on the base of the stem. Twelve plants were each inoculated with a plug of non-colonized PDA as controls. Plants were incubated in a growth chamber (30°C in the day and 25°C at night, a 12-h photoperiod and 80% RH). Necrotic lesions were observed on stems of all inoculated seedlings 5 days after inoculation, whereas control plants remained asymptomatic, and L. peudotheobromae was consistently re-isolated from symptomatic stem. In Asia, peanut collar rot caused by L. teudotheobromae has been reported in India, Indonesia, North Vietnam (Nguyen, et al., 2006) and China (Guo, et al., 2014), but collar rot caused by L. pseudotheobromae has not been reported. To our knowledge, this is the first report of L. peudotheobromae causing collar rot on peanut in China. These results will provide crucial information for studying on epidemiology and management of this disease.

Sample collection and eDNA extraction from Sterivex filter units v1

The following workflow covers several steps in the DNA analysis of environmental samples, from the water collection to the analysis back in the lab. The samples can be taken from several water systems (i.e. sea, lakes, rivers, streams) and collected in triplicate (1 L) in Sterivex sterile filter units (Merck, cat. no. SVGP01050). The DNA extraction protocol modifies the Dneasy PowerWater Sterivex kit (Qiagen, cat. no. 14600-50-nf).

Textile fibers resistant to biodestruction is a way to improve indoor ecological conditions

The aim of research was to observe and to describe biodestructions of fabrics that usually used inside a home, to estimate the destruction (total number of destruction “N” and destruction index “K”). The samples of fabrics were placed into a thermostat in sterile Petri dishes on moistened sterile filter paper in order to stimulate the microflora occurred on fibers. Incubation carried out in a thermostat at a +24-28 °C, humidity of 90-100% and exposed for 8 month. The samples examined with a microscope. As a result of the study of fiber’s damage, the types of damage were identified as follow: layering, fretting, mottling, fouling, spotting, swelling, granular disintegration, delamination, thinning, and damage of the fiber wall. Initially the most destructed was detected sample of natural silk and hemp according to the highest value of destruction index K. However, after 8 months of exposure, this sample turned out to be the most resistant to biodegradation. The data obtained based on the evaluation of the biostability of fabrics made from various textile fibers showed that the usage of natural silk, hemp and polyester able to supply textile with resistance to biodegradation by spontaneous microflora.

Production and Semi-Automated Processing of 89Zr Using a Commercially Available TRASIS MiniAiO Module

The increased interest in 89Zr-labelled immunoPET imaging probes for use in preclinical and clinical studies has led to a rising demand for the isotope. The highly penetrating 511 and 909 keV photons emitted by 89Zr deliver an undesirably high radiation dose, which makes it difficult to produce large amounts manually. Additionally, there is a growing demand for Good Manufacturing Practices (GMP)-grade radionuclides for clinical applications. In this study, we have adopted the commercially available TRASIS mini AllinOne (miniAiO) automated synthesis unit to achieve efficient and reproducible batches of 89Zr. This automated module is used for the target dissolution and separation of 89Zr from the yttrium target material. The 89Zr is eluted with a very small volume of oxalic acid (1.5 mL) directly over the sterile filter into the final vial. Using this sophisticated automated purification method, we obtained satisfactory amount of 89Zr in high radionuclidic and radiochemical purities in excess of 99.99%. The specific activity of three production batches were calculated and was found to be in the range of 1351–2323 MBq/µmol. ICP-MS analysis of final solutions showed impurity levels always below 1 ppm.

Evaluation of the anti-mycotic activity of Rosemary Oil Against Candida al-bicans

An antifungal medication, also known as an antimycotic medication, is a pharmaceutical fungicide or fungistatic used to treat and prevent mycoses such as athlete's foot, ringworm, candidiasis, serious systemic infections such as Cryptococcal meningitis, and others. In traditional medicine, extracts and essential oil from flowers and leaves are used in the belief they may be useful to treat a variety of fungal disorders. The aim of this study was to analyse the antimycotic properties of rosemary oil and its principal components. The Rosemary oil was screened for antifungal activity by the disc diffusion method. Activated cultures of Candida albicans in Sabouraud’s broth was adjusted to 0.5 McFarland standards [108cfu/ml]. 100 µl of the inoculum was introduced to molten Sabourauds dextrose agar and poured in the sterile Petri plates and allowed to set. Sterile filter paper discs (6.0 mm diameter) impregnated with 25µl, 50µl and 100µl /disc were placed on fungal seeded plates and incubated at 28oC for 48 hrs. Clear zones within which fungal growth was absent were measured and recorded as the diameter (mm) of complete growth inhibition. All the concentrations of the test solution inhibited the fungal species with varying degree of sensitivity. The extract showed good antifungal activity at different concentrations with a maximum zone of inhibition of 38 mm at concentration 100µl. This study provides a sample large enough to determine the antifungal properties of Rosemary oil and suggests further studies for possible therapeutic use.

Thermal Inactivation of a Single Strain Each of Serotype O26:H11, O45:H2, O103:H2, O104:H4, O111:H−, O121:H19, O145:NM, and O157:H7 Cells of Shiga Toxin–Producing Escherichia coli in Wafers of Ground Beef†

For each of two trials, freshly ground beef of variable fat content (higher: 70:30 %lean:%fat; lower: 93:7 %lean:%fat) was separately inoculated with ca. 7.0 log CFU/g of a single strain of Escherichia coli serotypes O26:H11, O45:H2, O103:H2, O104:H4, O111:H−, O121:H19, O145:NM, and O157:H7. Next, ca. 3-g samples of inoculated beef were transferred into sterile filter bags and then flattened (ca. 1.0 mm thick) and vacuum sealed. For each temperature and sampling time, three bags of the inoculated wafers of beef were submerged in a thermostatically controlled water bath and heated to an internal temperature of 54.4°C (130°F) for up to 90 min, to 60°C (140°F) for up to 4 min, or to 65.6°C (150°F) for up to 0.26 min. In lower fat wafers, D-values ranged from 13.5 to 23.6 min, 0.6 to 1.2 min, and 0.05 to 0.08 min at 54.4, 60.0, and 65.6°C, respectively. Heating higher fat wafers to 54.4, 60.0, and 65.6°C generated D-values of 18.7 to 32.6, 0.7 to 1.1, and 0.05 to 0.2 min, respectively. In addition, we observed reductions of ca. 0.7 to 6.7 log CFU/g at 54.4°C after 90 min, ca. 1.1 to 6.1 log CFU/g at 60.0°C after 4 min, and 0.8 to 5.8 log CFU/g at 65.6°C after 0.26 min. Thus, cooking times and temperatures effective for inactivating a serotype O157:H7 strain of E. coli in ground beef were equally effective against the seven non-O157:H7 Shiga toxin–producing strains investigated herein.