Identification of a novel Hemoplasma species from pigs in Zhejiang province, China.

Hemoplasmas belong to Mycoplasmataceae (Mollicutes: Mycoplasmatales) and are able to infect a broad range of mammalian species. We investigated prevalence of hemotropic mycoplasma species in pig farms in the region of Zhejiang by a PCR scheme using universal primers targeting 16S rRNA and RNase P RNA gene (rnpB). Representative positive samples from different farms were selected for sequencing of 16S rRNA and the 219bp rnpB gene fragments for phylogenetic analysis. Sequencing analysis of PCR products from first samples identified a novel hemoplasma species present in several pig farms in the region with highest nucleotide identity of 92% to Candidatus Mycoplasma turicensis. A duplex PCR assay was then designed for differential detection of the novel hemoplasma from Mycoplasma parvum/M. suis in field samples. Of 324 blood samples from clinically healthy pigs, 26.5% was positive for this novel hemoplasma species and 50% positive for M. suis/M. parvum, indicating that the novel hemotropic mycoplasma species were of considerably high prevalence in Zhejiang province, China.

A Colpoda aspera isolate from animal faeces: In vitro cultivation and identification.

A colpodean ciliate was found in the faeces of experimental rabbits. It was initially cultivated in medium mixed with 2% (w/v) rabbit faeces. Subsequently, two chemically defined media, designated CA-1 and CA-2, were found to be suitable for axenical cultivation of the ciliate. The maximum abundance of the ciliate isolate in the CA media was 1-2 × 10(5) cells/ml. The ciliate isolate was further identified with silver impregnation and molecular analysis. Features of the left oral polykinetid, somatic dikinetids, and sliverline pattern were similar to those of Colpoda aspera as described by Foissner (1993). The 18S small subunit ribosomal RNA gene of the ciliate isolate shared 99% sequence identity with that of C. aspera, with 100% coverage, and formed a sister clade in the phylogenetic tree with the reference C. aspera isolate. In addition, the trophozoite of C. aspera could proliferate over a temperature range from 25-37°C. When resting cysts were cultivated in CA-1 medium at 30-35°C, 98.2% of the trophozoites were detached from the cyst wall after 7 h.