Improvement of short straws for sperm cryopreservation: installing an air-permeable filter facilitates handling

Saving space for sperm cryopreservation would aid mouse genetics research. We previously developed the ST (sperm freezing in ShorT STraw to reduce STorage space) method for cryopreserving mouse sperm in a smaller storage space than conventional methods. However, our ST method has two drawbacks: difficulties during freeze-thaw procedures and the potential risk of sperm loss during storage. Here, we refine ST, terming the new method improved ST (iST). In iST, the straw has an air-permeable filter and the straw container (2-ml cryotube) is endowed with air vents. As in our ST method, iST frozen-thawed sperm showed good performance upon in vitro fertilization. Moreover, up to nine straws can be stored in one cryotube, occupying less storage space than conventional methods. This method provides an easy and space-saving cryopreservation method for mouse sperm, and thus will be valuable for mouse genetics researchers.     

Genetic variation detected with random amplified polymorphicDNA markers among isolates of the red rot disease fungus Pythiumporphyrae isolated from Porphyra yezoensis from Koreaand Japan

ABSTRACT:   Genetic variation of the fungal parasite Pythium porphyrae ,causative organism of red rot disease of Porphyra , isolatedfrom Asan, Mokpo, Pusan and Wando in Korea, and from Aichi, Fukuokaand Miyagi in Japan was investigated by random amplified polymorphicDNA (RAPD) cluster analysis. The total 67 RAPD markers were generatedfrom 38 isolates by RAPD-polymerase chain reaction (PCR) using arbitraryprimers consisting of 10 nucleotide sequences and 33 of them indicatedpolymorphisms. The dissimilarity coefficients calculated from theRAPD banding patterns ranged from 0.0010 to 0.6983. The dendrogramgenerated by the unweighted pair-group method using arithmetic averagesshowed that the 38 isolates were classified into three clusters(Groups 1, 2 and 3). Group 1 consisting of two isolates from Miyagiwas separated from all other isolates by a genetic distance of 0.6983.Groups 2 and 3 containing the majority of the isolates were branchedon genetic distance of 0.3957. These two clusters subdivided intofour and three subclusters, respectively, which were apparentlyassociated with geographic origins of the isolates. Interestingly,the isolates from Asan of Korea were close to Japanese isolatesrather than Korean isolates on genetic diversity. In addition, thegenetic distances of intra-isolates from Japan were higher thanthose from Korea.