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Nagai S.,Japan National Research Institute of Fisheries And Environment of Inland Sea | Yamamoto K.,Research Institute of Environment Agriculture and Fisheries | Hata N.,Mie Prefectural Science and Technology Promotion Center | Itakura S.,Japan National Research Institute of Fisheries And Environment of Inland Sea
Marine Genomics | Year: 2012

In a previous study, we experienced instable amplification and a low amplification success in loop-mediated isothermal amplification (LAMP) reactions from naturally occurring vegetative cells or resting cysts of the toxic dinoflagellates Alexandrium tamarense and Alexandrium catenella. In this study, we examined 4 methods for extracting DNA from single resting cysts of A. tamarense and A. catenella to obtain more stable and better amplification success and to facilitate unambiguous detection using the LAMP method. Apart from comparing the 4 different DNA extraction methods, namely, (1) boiling in Tris-EDTA (TE) buffer, (2) heating at 65. °C in hexadecyltrimethylammonium bromide buffer, (3) boiling in 0.5% Chelex buffer, and (4) boiling in 5% Chelex buffer, we also examined the need for homogenization to crush the resting cysts before DNA extraction in each method. Homogenization of resting cysts was found to be essential for DNA extraction in all 4 methods. The detection time was significantly shorter in 5% Chelex buffer than in the other buffers and the amplification success was 100% (65/65), indicating the importance of DNA extraction and the effectiveness of 5% Chelex buffer in the Alexandrium LAMP. © 2012 Elsevier B.V.

Yasumatsuya K.,Research Institute of Environment Agriculture and Fisheries | Kasai K.,Research Institute of Environment Agriculture and Fisheries | Yamanaka K.,Shiga Prefectural Livestock Technology Promotion Center | Nishino O.,Nara Prefectural Livestock Technology Center | And 4 more authors.
Livestock Science | Year: 2012

Data from 63 Japanese Black calves were collected to clarify the effects of feeding whey protein on the growth rate and mucosal IgA induction in calves. Dietary treatments in milk replacers were 1) 26% CP as in skim milk (control), 2) 26% CP as whey and skim milk and 3) 26% CP as whey. Diets were offered from 3 to 63. days of age in calves. Feeding whey protein had no effects on growth rate, fecal consistency and fecal water in calves. Compared with 2. days of age, fecal IgA concentration in calves decreased at 14. days of age, while fecal water increased. Feeding whey protein increased fecal IgA in calves after 14. days of age, which was thought to be the increased mucosal IgA induction in the gut. Serum cholesterol concentration tended to be lower in calves fed whey than in control group, but feeding whey protein had no clear effects on serum glucose, NEFA, total protein and urea-N concentrations. These results suggest that feeding whey protein enhances mucosal IgA induction in calves, but feeding whey protein has little effect on growth rate and fecal consistency in calves. © 2011 Elsevier B.V.

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