Pueruli of Panulirus longipes bispinosus (Crustacea, Decapoda, Palinuridae) stranded on the beach of Kuroshima Island, Ryukyu Archipelago, southern Japan

ABSTRACT: Stranded organisms were observed on the sandy beach of Kuroshima Island, Ryukyu Archipelago, in the early morning of 28 January 1993. Stranded organisms of 525 individuals were classified into 74 species. These organisms included 13 individuals of Panulirus pueruli, which were identical to P. longipes bispinosus. Kuroshima Island is located near Ishigakijima and Iriomote islands, Ryukyu Archipelago, where coral reefs develop. Coral reefs are common and develop in the Ryukyu Archipelago, southern Japan, with the exception of Yakushima and Tanegashima islands. Panulirus japonicus in Japanese waters has not been found in coral reef regions where P. longipes bispinosus are mainly found. The present study describes and identifies to species level puerulus specimens that were stranded on the beach of Kuroshima Island.     

Hemorrhagic necrotizing splenitis in a slaughter pig infected with Arcanobacterium species

A 6-month-old barrow presented with lethargy, inappetence and dysstasia. At necropsy, multiple coalescing hemorrhagic foci were detected in the margins of the spleen. Gram-positive bacilli were isolated from the spleen, kidney, muscle and liver. Comparative 16S rDNA gene sequencing analysis of the isolates (TO16177) revealed that they would be the same species of unpublished Arcanobacterium species strain HJ57-14E (accession no. gi 18873551) (99.7% similarity based on a comparison of 675 bp). Histologic examination of the splenic tissue sections revealed extensive necrosis and inflammation, and gram-positive bacilli were discernible. Multifocal necrosis was also detected in the liver. Immunohistochemically, the isolates were cross-reacted with polyclonal antibodies against Arcanobacterium pyogenes and Actinomyces naeslundii, and the reaction was strong for the latter. Similar reactions were found in the suppurative lesions of the tonsil, and occasionally in the spleen and lymph nodes. The present results indicate that the unpublished Arcanobacterium species induced multiple organ failure accompanied by acute hemorrhagic necrotizing splenitis in this growing-finishing pig.     

Strong antiviral activity of heated and hydrated dolomite--preliminary investigation

Heated and hydrated naturally occurring dolomite showed very strong antiviral activity. Infectivity of avian and human influenza, avian infectious bronchitis (coronavirus), Newcastle disease (paramyxovirus) and avian laryngotracheitis (herpesvirus) viruses dropped at least 1,000 fold following contact with the dolomite for five minutes at 4 degrees C. Dolomite is expected to be useful to inhibit the incidence of emerging and re-emerging infectious diseases.     

Changes in the microbial community structure in soils treated with a mixture of glucose and peptone with reference to the respiratory quinone profile

Based on the respiratory quinone profile, changes in the structure of microbial communities in the soil samples from Nagoya University Farm were monitored after the treatment with 1% of a mixture of glucose and peptone. Samples of two soils differing in the fertilization history were examined: CF-soil with the application of only chemical fertilizers and FYM-soil with the application of only farmyard manure at a high rate. In the CF-soil, the amount of water-soluble organic carbon (WOC), indicator of the mixture of glucose and peptone, decreased to the original level after 14 d. After 7 d, the soil pH reached the maximum level, then decreased gradually. Changes in the inorganic nitrogen levels in the water extract also reflected the 14-d period of mineralization. The amount of respiratory quinones reached maximum levels after 7 d and gradually decreased, reflecting the changes in the microbial biomass. The quinone composition significantly changed during the 14-d period and returned to a profile similar to the original one after 28 d. Diversity of quinones significantly decreased during the 14-d period due to the predominance of ubiquinone with 9 isoprenoid units. In the FYM-soil, the amount of WOC decreased to the original level after 1 d, and the pH and inorganic nitrogen levels in the water extract reflected the one-day mineralization period, and nitrification started after 3 d. Although the amount of quinones indicated an increase in the microbial biomass for 14 d, the quinone composition did not change. These findings suggested that long-term application of farmyard manure resulted in stable microbial communities in response to the incorporation of organic matter in soil.     

Microdroplet In Vitro Fertilization Can Reduce the Number of Spermatozoa Necessary for Fertilizing Oocytes

Successful in vitro fertilization (IVF) in mice has been achieved using spermatozoa at concentrations specifically optimized for the experimental conditions, such as species and source of spermatozoa. Although IVF in mice is mostly performed using about 80–500 µl drops, it is expected that the number of spermatozoa used for insemination can be reduced by decreasing the size of the IVF drops. The present study was undertaken to examine the extent to which the number of spermatozoa used for IVF could be reduced by using small droplets (1 µl). We devised the experimental parameters using frozen–thawed spermatozoa from C57BL/6 mice in anticipation of broader applications to other mouse facilities. We found that as few as 5 spermatozoa per droplet could fertilize oocytes (1 or 3 oocytes per droplet), although the fertilization rates were low (13–15%). Practical fertilization rates (> 40%) could be achieved with frozen-thawed C57BL/6J spermatozoa, which are sensitive to cryopreservation, when 20 sperm per droplet were used to inseminate 3 oocytes. Even with spermatozoa from a very poor quality suspension (10% motility), about 25% of oocytes were fertilized. Our calculations indicate that the number of inseminated spermatozoa per oocyte can be reduced to 1/96–1/240 by this method. In two separate embryo transfer experiments, 60% and 47%, respectively, of embryos developed to term. Our microdroplet IVF method may be particularly advantageous when only a limited number of motile spermatozoa are available because of inadequate freezing-thawing or genetic reasons.     

A retrospective study of inflammatory colorectal polyps in miniature dachshunds

Medical records of dogs with colorectal polyps were retrospectively reviewed, and clinical presentation of inflammatory colorectal polyps in miniature dachshunds was evaluated. Of 33 dogs found to have colorectal polyps, miniature dachshunds were markedly over-represented with 16 dogs (48%), of which 12 (75%) were found to have inflammatory polyps. Multiple polyps localized between the rectum and the descending colon was the most common finding in miniature dachshunds with inflammatory polyps. Twenty dogs (80%) out of 25 miniature dachshunds with inflammatory colorectal polyps responded to immunosuppressive therapy using prednisolone and cyclosporine. The results of this study indicate that miniature dachshunds are predisposed to develop inflammatory colorectal multiple polyps, for which immunosuppressive therapy may be a treatment option.     

The therapeutic effects of SET/I2PP2A inhibitors on canine melanoma

Canine melanoma is one of the most important diseases in small animal medicine. Protein phosphatase 2A (PP2A), a well conserved serine/threonine phosphatase, plays a critical role as a tumor suppressor. SET/I2PP2A is an endogenous inhibitor for PP2A, which directly binds to PP2A and suppresses its phosphatase activity. Elevated SET protein levels have been reported to exacerbate human tumor progression. The role of SET in canine melanoma, however, has not been understood. Here, we investigated the potential therapeutic role for SET inhibitors in canine melanoma. The expression of SET protein was observed in 6 canine melanoma cell lines. We used CMeC-1 cells (primary origin) and CMeC-2 cells (metastatic origin) to generate cell lines stably expressing SET-targeting shRNAs. Knockdown of SET expression in CMeC-2, but not in CMeC-1, leads to decreased cell proliferation, invasion and colony formation. Phosphorylation level of p70 S6 kinase was decreased by SET knockdown in CMeC-2, suggesting the involvement of mTOR (mammalian target of rapamycin)/p70 S6 kinase signaling. The SET inhibitors, OP449 and FTY720, more effectively killed CMeC-2 than CMeC-1. We observed PP2A activation in CMeC-2 treated with OP449 and FTY720. These results demonstrated the potential therapeutic application of SET inhibitors for canine melanoma.     

Glomerular lipidosis accompanied by renal tubular oxalosis in wild and laboratory-reared Japanese rock ptarmigans (Lagopus mutus japonicus)

Glomerular lipidosis is a disease characterized by lipid accumulation in mesangial cells but that has not been fully investigated in avian species. We examined four wild and two laboratory-reared Japanese rock ptarmigans (Lagopus mutus japonicus)--an endangered avian species--presenting vacuolar deposits in the glomeruli. All cases had vacuolar deposits in the glomeruli. In the wild cases, fewer than 30% of all glomeruli were affected, compared with more than 90% in the laboratory-reared cases. In the wild cases, most deposits were mild and restricted to the mesangial areas of glomeruli. In the laboratory-reared cases, nearly all of the deposits covered entire glomeruli. Electron microscopy of mild deposits revealed vacuoles in the cytoplasm of mesangial cells. These vacuoles were positive for Sudan III, Sudan black B, oil red O, Nile blue, periodic acid-Schiff, Schultz test, and digitonin stain and were negative for performaric acid-Schiff stains. Based on these results, we diagnosed the glomerular lesion as glomerular lipidosis caused by uptake of low-density lipoprotein in mesangial cells. Except for one wild case, all cases exhibited renal tubular oxalosis. The severity of tubular oxalosis tended to be related to the severity of glomerular lipidosis: In cases of mild glomerular lipidosis, tubular oxalosis was also mild or absent. We therefore diagnosed the primary lesion as glomerular lipidosis accompanied by tubular oxalosis. The four wild cases came from different zones and therefore had no opportunities to interbreed and no common relatives. We believe these data support the hypothesis that glomerular lipidosis is a disease of the general population ofJapanese rock ptarmigans. This is the first report of glomerular lipidosis accompanied by renal tubular oxalosis in an avian species.