Characterization of aerosols containing microcystin

Toxic blooms of cyanobacteria are ubiquitous in both freshwater and brackish water sources throughout the world. One class of cyanobacterial toxins, called microcystins, is cyclic peptides. In addition to ingestion and dermal, inhalation is a likely route of human exposure. A significant increase in reporting of minor symptoms, particularly respiratory symptoms was associated with exposure to higher levels of cyanobacteria during recreational activities. Algae cells, bacteria, and waterborne toxins can be aerosolized by a bubble-bursting process with a wind-driven white-capped wave mechanism. The purposes of this study were to: evaluate sampling and analysis techniques for microcystin aerosol, produce aerosol droplets containing microcystin in the laboratory, and deploy the sampling instruments in field studies. A high-volume impactor and an IOM filter sampler were tried first in the laboratory to collect droplets containing microcystins. Samples were extracted and analyzed for microcystin using an ELISA method. The laboratory study showed that cyanotoxins in water could be transferred to air via a bubble-bursting process. The droplets containing microcystins showed a bimodal size distribution with the mass median aerodynamic diameter (MMAD) of 1.4 and 27.8 mum. The sampling and analysis methods were successfully used in a pilot field study to measure microcystin aerosol in situ.     

A new Brucella canis species-specific PCR assay for the diagnosis of canine brucellosis

Brucellosis is a zoonotic disease that is transmitted from animals to humans, and the development of a rapid, accurate, and widely available identification method is essential for diagnosing this disease. In this study, we developed a new Brucella canis species-specific (BcSS) PCR assay and evaluated its specificity and sensitivity. A specific PCR primer set was designed based on the BCAN_B0548-0549 region in chromosome II of B. canis. The PCR detection for B. canis included amplification of a 300-bp product that is, not found on other Brucella species or, genetically or serologically related bacteria. The detection limit of BcSS-PCR assay was 6 pg/μl by DNA dilution, or 3 × 10³ colony-forming units (CFU) in the buffy coats separated from whole blood experimentally inoculated with B. canis. Using the buffy coat in this PCR assay resulted in approximately 100-times higher sensitivity for B. canis as compared to detect directly from whole blood. This is the first report of a species-specific PCR assay to detect B. canis, and the new assay will provide a valuable tool for the diagnosis of B. canis infection.     

A Cathepsin B Inhibitor,E-64, Improves the Preimplantation Development of Bovine Somatic Cell Nuclear Transfer Embryos

Bovine somatic cell nuclear transfer (SCNT) is an important and powerful tool for basic research and biomedical and agricultural applications, however, the efficiency of SCNT has remained extremely low. In this study, we investigated the effects of cathepsin B inhibitor (E-64) supplementation of culture medium on in vitro development of bovine SCNT embryos. We initially used three concentrations of E-64 (0.1, 0.5, 1.0 μm), among which 0.5 μm resulted in the highest rate of blastocysts production after in vitro fertilization (IVF), and was therefore used for further experiments. Blastocyst development of SCNT embryos in the E-64 treatment group also increased relative to the control. Moreover, the cryosurvival rates of IVF and SCNT blastocysts were increased in E-64 treatment groups when compared with the control. On the other hand, we found that IVF and SCNT blastocysts derived from E-64-treated groups had increased total cell numbers and decreased apoptotic nuclei. Furthermore, assessment of the expression of apoptosis-related genes (Bax and Bcl-xL) in bovine IVF and SCNT blastocysts treated with E-64 by real-time RT-PCR analysis revealed suppressed expression of the pro-apoptotic gene Bax and stimulated expression of the anti-apoptotic gene Bcl-xL. Taken together, these finding indicate that addition of E-64 to embryo culture medium may have important implications for improving developmental competence and preimplantation quality in bovine IVF and SCNT embryos.     

First discovery of a cave-dwelling Tineid moth （Lepidoptera,Tineidae） from East Asia

In this study, we report Monopis crocicapitella （Clemens, 1859） （Tineidae）, which was collected from bat guano in a cave in the southern region of Korea, for the first time from East Asia. We briefly redescribe and illustrate the external morphology and genital structures of both sexes. Also, we discuss the local habitat characteristics and some of the ecological information that was observed during our field investi-gation.     

Inhibitory effects of a monoclonal antibody (MAb-001) on in vitro oxygen consumption and multiplication of the pathogenic haemoflagellate, Cryptobia salmositica Katz

A monoclonal antibody (MAb-001), against a surface glycoprotein on Cryptobia salmositica inhibited the multiplication and oxygen consumption of both virulent and avirulent strains of the parasite. The classical cysteine proteinase inhibitor (E-64) and a cysteine proteinase activator (EDTA) affected the in vitro multiplication of C. salmositica . Concentrations of E-64 higher than 10 μ M reduced the multiplication of C. salmositica while 5 m M of EDTA enhanced its multiplication. We propose that the cysteine proteinase is an important metabolic enzyme in C. salmositica and that binding of MAb-001 to the enzyme inhibited parasite multiplication and reduced oxygen consumption.