Lineage specific detection of the scaly form of the pen shell <Emphasis Type="Italic">Atrina</Emphasis> spp. by a loop-mediated isothermal amplification method

Taxonomy of the pen shell (genus Atrina) in East Asia has been confused because of the plasticity of the shell morphology and lack of unified morphological characteristics. We analyzed the mitochondrial cytochrome c oxidase subunit I (COI) sequence of the pen shell sampled from Ariake Bay and Seto Inland Sea, western Japan. All individuals were identified as the scaly form based on external morphology; however, their COI sequences were comprised of two distinct lineages. The sequence divergence between the two was 8.3%, indicating the high possibility that they are different species. The scaly form of the pen shell in Japan is one of the target species for resource enhancement, therefore COI lineage identification of broodstock individuals is critically important to avoid producing hybrid seed. We developed a loop-mediated isothermal amplification method that enables specific detection of the two lineages. This method can contribute to reducing the cost and time required for genotyping broodstock individuals prior to seed production.     

In situ observation of pinewood nematode in wood

In pine wilt disease, xylem dysfunction occurs in relation to nematode migration and proliferation in host tissue, but the detection accuracy of pinewood nematode (PWN), Bursaphelenchus xylophilus, in pine stem tissue remains unclear. This study describes the use of cryo-scanning electron microscopy (cryo-SEM) and fluorescein-conjugated wheat germ agglutinin (F-WGA) staining to detect PWN. After PWN invasion, the frequency of surface fluorescence in PWN increased in pine stems from the day of inoculation to 3 weeks after inoculation. However, the fluorescence frequency decreased significantly during the advanced disease stage after 5 weeks. Thus, detecting PWN based on fluorescent staining of the nematode body surface coat protein can be misleading when used to examine the correlation between the development of disease symptoms and the nematode population. In contrast, all cut body segments were fluorescent, and their fluorescent components were common in pine-stem cross sections, regardless of the timing after inoculation. In addition, PWN were observed under cryo-SEM only in empty resin canals and this distribution was confirmed by F-WGA staining of PWN cut in a cross section. Thus, PWN detection based on fluorescent staining of surface coat proteins by F-WGA was not reliable in longitudinal sections of pine stems because of changes in nematode stainability during disease progression. To detect PWN in infected plants, we concluded that a combination of both methods is most effective.     

Bilateral patellar agenesis in dog: A case report

A bilateral patellar agenesis is an extremely rare congenital condition in which the patient does not develop both patellas and can present secondary alterations as bone, muscle and postural changes. There are some hypotheses that it has a genetic background presenting dominant characteristic. It is not yet standardized a gold treatment for this affection, but according to rare reports, clinical and surgical treatments are possible. This is a case report based on imagining exams of a mix breed male puppy that was born with bilateral patellar agenesis, an affection not yet reported in canine species.