A sequencing protocol of some DNA regions in nuclear, chloroplastic and mitochondrial genomes with an individual colony of Thalassiosira nordenskioeldii Cleve (Bacillariophyceae)

From cultured cells of micro algal species of Thalassiosira nordenskioeldii Cleve, total DNA was extracted with a buffer containing a chelating agent, Chelex 100. Each DNA fragment for 18s rDNA, 16s rDNA and coxI in nuclear, chloroplastic and mitochondrial genomes, respectively, were amplified from the DNA extracts by polymerase chain reactions, and their sequences were determined. This protocol for determining DNA sequences, was scaled down for the DNA extracts from small amounts of cells. Finally, DNA sequences in genomes were determined with 10 cells collected by a micromanipulation technique. The quantity of cells corresponded to an individual colony of this species. With this protocol, DNA sequences in various regions in genomes of each individual micro algal species living in natural environments can be determined directly, without isolation of strains and cultures

Changes in the numbers of patients with acute gastroenteritis after voluntary introduction of the rotavirus vaccine in a Japanese children’s primary emergency medical center

Abstract Background Acute gastroenteritis (AGE) is a major reason for presentation to pediatric primary emergency medical centers. Because rotavirus vaccines were introduced in November 2011 for voluntary vaccination in Japan, we analyzed the changes in the numbers of AGE patients. Methods The number and proportion of patients visiting Kobe children’s primary emergency medical center from January 2011 to February 2015 due to AGE, out of all visiting children, were investigated retrospectively. The rotavirus and norovirus epidemic periods were defined as the periods from March to June and from November to February, respectively, based on their disease prevalence. Results In patients ≤2 years of age, the numbers and proportions of patients with AGE were significantly decreased from 2464/14098 (17%) in 2011 to 1888/12321 (15%) in 2014 (p < 0.01). In patients ≤2 and 3–5 years of age, significant decreases in AGE patients between 2011 and 2014 were observed during the rotavirus season (from 20% [1090/5329] to 14% [642/4482] in patients aged ≤2 years and from 23% [704/3047] to 20% [572/2807] in patients aged 3–5 years, p < 0.01 and p < 0.05, respectively), but not during the norovirus season (from 19% [834/4436] to 19% [797/4160] in patients aged ≤2 years and from 20% [679/3334] to 25% [710/2852] in patients aged 3–5 years). Conclusions The estimated rotavirus vaccine coverage in our area increased from 1% in 2011 to 49% in 2014; this coverage may have resulted in a reduction in AGE patients, both directly and indirectly, in our Japanese children’s primary emergency medical center