Molecular identification of Tricula spp. and the parasitized trematode cercariae in schistosomiasis-endemic areas of Yunnan Province / 中国血吸虫病防治杂志

Objective To characterize a species of the genus Tricula and parasitized trematodes in schistosomiasis-endemic areas of Yunnan Province using a molecular analysis, so as to understand their taxonomic positions. Methods Tricula spp. and Oncomelania snails were collected from Xiangyun County, Yunnan Province, and cercaria parasitizing snails were observed using crushing followed by microscopy. Cercaria parasitizing Tricula snails at various morphologies were sampled using a shedding method. Genomic DNA was extracted from snail soft tissues and cercariae, and the 16S rRNA, COI, 28S rDNA genes in snails and the ND1 and 28S rDNA genes in cercariae were amplified using a PCR assay and sequenced. The species of Tricula snails and their parasitized trematodes was characterized using sequence alignment and phylogenetic analysis. Results Among 382 Tricula snails detected, there were three types of trematode cercariae found, including the non-forked (20.94%, 80/382), double-forked (3.40%, 13/382) and swallow shapes (7.07%, 27/382). Sequence and phylogenetic analyses showed that the 16S rRNA, COI and 28S rDNA gene sequences of this species of Tricula had high homology to those in Delavaya dianchiensis, and were clustered in a branch. Sequencing analysis of the ND1 and 28S rDNA genes revealed that the non-forked cercariae belonged to the family Pleu- rogenidae, the swallow-shaped cercariae belonged to the family Opecoelidae, and the double-forked cercariae belonged to another species of the genus Schistosoma that was different from S. sinensium and S. ovuncatum. Conclusion The species and taxonomy of Triculla spp. and their parasitized trematodes are preliminarily determined in schistosomiasis-endemic areas of Yunnan Province; however, further studies are required to investigate the more definite taxonomy and pathogenicity.

Analysis of the genetic diversity of Candida isolates obtained from diabetic patients and kidney transplant recipients

Yeasts of the genus Candida have high genetic variability and are the most common opportunistic pathogenic fungi in humans. In this study, we evaluated the genetic diversity among 120 isolates of Candida spp. obtained from diabetic patients, kidney transplant recipients and patients without any immune deficiencies from Paraná state, Brazil. The analysis was performed using the ITS1-5.8S-ITS2 region and a partial sequence of 28S rDNA. In the phylogenetic analysis, we observed a consistent separation of the species C. albicans, C. dubliniensis, C. glabrata, C. tropicalis, C. parapsilosis, C. metapsilosis and C. orthopsilosis, however with low intraspecific variability. In the analysis of the C. albicans species, two clades were formed. Clade A included the largest number of isolates (91.2%) and the majority of isolates from GenBank (71.4%). The phylogenetic analysis showed low intraspecific genetic diversity, and the genetic polymorphisms between C. albicans isolates were similar to genetic divergence found in other studies performed with isolates from Brazil. This low genetic diversity of isolates can be explained by the geographic proximity of the patients evaluated. It was observed that yeast colonisation was highest in renal transplant recipients and diabetic patients and that C. albicans was the species most frequently isolated.

First Report of Leaf Rust Caused by Puccinia caricis in Farfugium japonicum in Korea

Farfugium japonicum is used in traditional medicine and as an edible herb in China and Korea. In July 2013, leaf spots were observed in F. japonicum seedlings at Ulleung Island, Gyeongsangbuk Province, Korea. Early symptoms on the leaf adaxial surface included roughly circular yellow spots that later developed brown, necrotic centers. The aecia were hypophyllous, cupulate, yellowish, 180~430 microm in diameter, clustered, and erumpent with a peridium with a recurved margin. The aeciospores were globoid, 14~17 x 13~16 microm, light yellow or colorless, and densely verrucose. The 28S rDNA sequence of the isolate was identical to each other and shared 99% identity with Puccinia caricis. This is the first report of rust caused by P. caricis in F. japonicum in Korea or elsewhere in the world.

Outbreak of Rust Caused by Coleosporium asterum on Solidago virgaurea var. gigantea in Ulleung-do

From June to October 2012, severe outbreaks of rust disease on Solidago virgaurea var. gigantea were observed in 5 different regions of Ulleung-do, Korea. Typical rust symptoms appeared as yellow spots, uredinia on leaves, and massive urediniospores covering the entire plant. Severely infected plants became discolored and died. On the basis of its morphological and molecular characteristics, the causal agent was identified as Coleosporium asterum. This pathogen has been reported as rust disease on S. virgaurea in the Canada and United States. In Korea, rust disease on S. virgaurea var. gigantea was reported as Uromyces rudbeckia in 1992. To the best of our knowledge, C. asterum is a novel rust pathogen on S. virgaurea var. gigantea in Korea.

Characterization of Species of Cladobotryum which Cause Cobweb Disease in Edible Mushrooms Grown in Korea

Four Cladobotryum isolates were collected from four different commercially grown mushroom types infected with cobweb disease in Cheongdo-gun and Chilgok-gun of Gyeongbuk Province, Korea in 2010. The isolates were identified as C. mycophilum from Agaricus bisporus and Pleurotus eryngii, C. varium from Flammulina velutipes and Hypsizygus marmoreus. The cultural characteristics of the four isolates were investigated using potato dextrose agar (PDA) media under nine different temperatures ranging from 5~32degrees C. Rapid growth of the isolates to colony diameters of 47~82 mm was observed at conditions of 18~22degrees C. No growth was observed at 32degrees C. C. mycophilum produced a yellowish red pigment while C. varium produced a cream colored pigment after cultivation for 25 days on PDA. Phylogenetic analysis of the internal transcribed spacer region and partial 28S rDNA from the four isolates confirmed they were C. mycophilum and C. varium. Cross pathogenicity tests revealed that the two isolates of C. mycophilum were highly pathogenic toward three mushroom types, but not toward H. marmoreus. The two isolates of C. varium were less pathogenic than those of C. mycophilum, but were pathogenic toward all mushroom types evaluated.

A Case of Fungal Keratitis Caused by Colletotrichum Species / 대한임상미생물학회지

Colletotrichum is mainly a fungal pathogen of plants, but sporadic cases of human infection have been reported recently. Most of them are fungal keratitis and only a few cases have been reported worldwide. A 63-year-old female farmer developed foreign body sensation and watering in her left eye following trauma by rice leaves. At presentation, her visual acuity decreased and corneal ulcer and inflammation in anterior chamber were observed on a slit lamp examination. Numerous hyphae were found on Gram stain and a rapidly growing mold with cup-shaped acervuli and falcate and nonseptate conidia was observed on fungal culture. As morphological findings did not lead to definite differentiation of the organism, sequencing of the D1-D2 domain of 28S rDNA was performed. It proved to be Colletotrichum species and the patient was treated with amphotericin and natamycin eye drop, but complicated by acute glaucoma. This is the first report of Colletotrichum keratitis in Korea and suggests that its infection should be considered in patients with fungal keratitis.

The Determination of the Partial 28S Ribosomal DNA Sequences and Rapid Detection of Phellinus linteus and Related species

Species of Phellinus were known to harmful fungi causing white pocket rot and severe plant disease such as canker or heart-rot in living trees in the West, but some species have been used to traditional medicines in the Orient for a long time. In this study the partial D1-D2 nucleotide sequences of 28S ribosomal DNA from 13 Phellinus strains were determined and compared with the sequences of 21 strains obtained from GenBank database. According to the neighbor-joining (NJ) method comparing the sequence data the phylogenetic tree was constructed. The phylogenetic tree displayed the presence of four groups. Group I includes P. ferreus, P. gilvus and P. johnsonianus, Group II contains P. laevigatus, P. conchatus and P. tremulae, Group III possesses P. linteus, P. weirianus, P. baumii, P. rhabarbarinus and P. igniarius, and Group IV comprises P. pini, P. chrysoloma. P. linteus and P. baumii, which were used mainly in traditional medicine, belong to the same group, but exactly speaking both were split into two different subgroups. To detect P. linteus only, we developed the PCR primer, D12HR. The primer showed the specific amplification of P. linteus, which is permitted to medicinal mushroom in the East. The results make a potential to be incorporated in a PCR identification system that could be used for the rapid identification of this species from its related species, P. linteus especially.