Grouped, stacked rods and tapeta lucida in the retina of Japanese anchovy Engraulis japonicus

ABSTRACT: Morphology of the photoreceptor cells and tapetum of the Japanese anchovy Engraulis japonicus was studied by histologically and by chemical analysis. The Japanese anchovy has a duplex retina. The cones form parallel rows consisting of alternately placed long cones and bifid cones. Both types of cones are intimately associated and form triple units that are regularly spaced along the row of cones. The rods are grouped and stacked. This fish has a retinal tapetum lucidum composed of guanine and hypoxanthine. Three structures of the tapetum lucidum were recognized: platelet, diamond and rod types. Photomechanical changes include movements of the photoreceptor cells and the retinal tapetum. The retina of the Japanese anchovy is thought to be highly sensitive and well adapted to a dim light environment.     

Competitive enzyme-linked immunosorbent assay for the detection of the antibodies specific to akabane virus.

A competitive enzyme-linked immunosorbent assay (C-ELISA) using neutralizing monoclonal antibodies (MAbs) against Akabane virus (AKAV) was developed to detect antibodies to AKAV in cattle sera. The performance of the test using 7 different competitor MAbs was evaluated in sequential serum samples and sera from cattle infected with various bovine arboviruses. The dynamics of the antibody response expressed by percentage of inhibition (PI) in C-ELISA coincided with those of neutralizing antibody titers in sequential serum samples from 2 cattle experimentally infected with AKAV. The value of PI in C-ELISA for convalescent sera from cattle infected with arboviruses correlated with the neutralizing antibody titer to AKAV but was unaffected by the antibodies to other arboviruses. In the validation experiment of C-ELISA using 286 bovine sera previously examined for the AKAV antibody by serum neutralization (SN) test, the relative specificity of C-ELISA was more than 98%, whereas the relative sensitivities of individual MAbs ranged from 49% to 82.2%. Overall agreement between C-ELISA and the SN test varied from 72% to 90% depending on the MAb. These results suggest that the C-ELISA is acceptable as a rapid and specific method for detecting antibodies to AKAV and is a potential alternative to the SN test.     

Vulnerability of feline T-lymphocytes to charged particles

An analysis of ionizing radiation-induced damage in peripheral lymphocytes has been employed to predict the prognosis of radiotherapy in terms of toxicity in normal tissues. Therefore, understanding the sensitivity of lymphocytes to high linear energy transfer (LET)-charged particles would be indispensable for utilizing charged particle therapy in veterinary medicine. However, the availability of such information is very limited. This study aimed to compare the radiosensitivity of feline T lymphocytes to gamma-rays (0.2 keV/microm) and 4 different types of charged particles with LET values ranging from 2.8 to 114 keV/microm. It was observed that the relative biological effectiveness, inactivation cross-section, and isodose-induced apoptosis increased in an LET-dependent manner. On the other hand, no difference in apoptosis frequency was observed in the cells exposed to an isosurvival dose of all the radiation types tested. This is the first study that demonstrates the LET dependence of cell killing and apoptosis induction in feline T lymphocytes. Our results suggest that lymphocytes can be effectively used to predict the prognosis of charged-particle therapy in cat patients.     

Clinical evaluation of growth hormone secretion in cattle using insulin tolerance test

Growth hormone secretion was evaluated in cattle. Clinically healthy bovine growth hormone (bGH) concentrations were 10.7 +/- 1.6 ng/ml in Holstein and 7.8 +/- 3.9 ng/ml in Japanese black cattle. The bGH concentration alternated at three-hour intervals, and tended to be higher at midnight and lower in the morning and before feeding. Insulin tolerance test (ITT) at an insulin dosage of 0.25 U/kg showed a significant increase of bGH concentration to 331 +/- 153% at 60 to 90 min after injection. In ITT applied to five under-growth calves of Japanese black cattle, the basal bGH concentrations were lower and peak values after insulin injection were shown to be significantly low. The ITT is useful for the clinical examination of bGH secretion in cattle.     

Isolation and serological survey of Salmonella in pigs in Japan

A total of 267 fecal and serum samples collected from individual pigs reared on a Salmonella-positive farm were subjected to bacteriological and serological examinations of Salmonella. Salmonella was isolated from 47 pigs (17.6%) and prevalence of antibody to lipopolysaccharide (LPS) of S. Typhimurium, which was partly common to S. O4, 12: d: -, was observed in 90 pigs (33.7%). Salmonella was isolated from 26 (28.9%) of 90 antibody-positive pigs and 21 (11.9%) of 177 antibody-negative pigs. Twenty-one of 36 pigs (58.3%) positive for S. O4, 12: d: -, five of 10 pigs (50.0%) positive for S. Havana, and none for S. Anatum had antibodies. Thus, seropositive rates were higher than isolation-positive rates, and antibody prevalence was associated with serovars of the isolates. Then, we analyzed antibody prevalence among pigs on Japanese pig farms. The antibodies to LPS of S. Typhimurium were found in 195 of 1,498 pigs (13.0%) and in at least one serum sample on 35 of 52 farms (67.3%). Our results indicate that Salmonella does not seem to be so prevalent in pigs though it is widely prevalent among pig farms.     

Comparison of the nucleotide and amino acid sequences of parental and attenuated isolates of Zucchini yellow mosaic virus

To identify possible sites of viral attenuation, the complete nucleotide sequences of two isolates of Zucchini yellow mosaic virus (ZYMV) were determined; a severe isolate Z5-1 and an attenuated isolate from Z5-1 (designated ZYMV-2002). The viral genome of both isolates consisted of 9593 nucleotides in size and contained an open reading frame encoding a single polyprotein of 3080 amino acids. Comparison of the nucleotide sequences for Z5-1 and ZYMV-2002 revealed 14 nucleotide mutations, resulting in seven amino acid substitutions with four in the HC-Pro region, two in the CI region, and one in the NIb region. These results provide a genetic basis for future manipulation of the ZYMV reverse genetics system. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession numbers AB188115 and AB188116     

Effect of low temperature on breaking dormancy and flowering of Arisaema sikokianum (Araceae)

Arisaema sikokianum (Araceae) native to Japan is classified as a vulnerable species in the Red Data Book of Japan. Control of dormancy is essential for efficient corm production and forcing culture. Sprouting of both vegetative and reproductive corms was enhanced by exposure to low temperature. Vegetative corms exposed to low temperatures at 5 °C longer sprouted faster when grown at 20 °C. Effective temperatures for breaking dormancy was 5 °C. Reproductive corms treated at 5 °C longer showed shorter days to flower. Successful forcing culture was achieved; corms treated at 5 °C from November for 30 days flowered on 5th February.     

Comparison of S-RNase,RNase T1, T2, and A effects on growth inhibition and RNA degradation of in vitro-cultured pear pollen

To obtain the basic information on fruit set regulation, effects of several RNases including S-RNase on pollen tube growth and RNA degradation in the tube were studied in the pear. Purified S-RNase from the Japanese pear ‘Kosui’ (S₄S₅) predominantly inhibited the growth of ‘Kosui’ pollen tubes (self) in vitro at 0.28 unit μL⁻¹, but it inhibited ‘Chojuro’ (S₂S₃) pollen (cross) only slightly. The same unit of RNase T₁ (EC 3.1.27.3) clearly inhibited the pollen tube growth, but the action was significantly weaker than that of the S-RNase against the self-pollen. Inhibitory effect of RNase T₂ (EC 3.1.27.1) and RNase A (EC 3.1.27.5) was only slight. The proteins other than the S-RNase extracted from pear style did not have any inhibitory action, though they possessed RNase activity 3.8 times higher than S-RNase. Thus, RNases tested here could not substitute for the S-RNase in specific inhibition against the self-pollen tube growth. Total RNA degradation by each RNase occurred in the pollen tubes as following order; S-RNase (self) ≥T₁ > T₂ ≥ A > S-RNase (cross). Degradation degree of 28S and 18S rRNA was as follows; S-RNase (self) > A > T₁ > T₂ > S-RNase (cross). The degradation of 5.8S and 5S rRNA was; S-RNase (self) > S-RNase (cross) > A > T₂ > T_1. The degree of rRNA degradation was, thus, not always in parallel with the degree of pollen growth inhibition. The S-RNase may degrade not only rRNA but also mRNA essential for pollen tube growth, and may be specifically adapted to inhibit the growth of self-pollen tubes. Therefore, controlling S-RNase amount in the style will produce self-thinning cultivars efficiently, which are unnecessary not only for hand-pollination but fruit-thinning practices in the pear. Practically, cultivar with weak self-incompatibility and small amount of S-RNase, such as ‘Okusankichi’, may be an expecting candidate for breeding self-thinning cultivars.     

Phospholipase A<Subscript>1</Subscript> activity of crude enzyme extracted from the ovaries of skipjack tuna

The phospholipid class composition, fatty acid composition and phospholipase A₁ (PLA₁) activity from the ovaries of skipjack tuna were compared with those of six other species of marine fish. In the skipjack ovaries, the lysophosphatidylcholine (LPC) proportion for the phospholipid, the docosahexaenoic acid (DHA) percentage for the total fatty acids of the phospholipids and the PLA₁ activity of the crude enzyme were the highest among those of the seven species. The optimum pH and temperature for the PLA₁ activity of the crude enzyme from the skipjack ovaries were in the range of pH 6–7 and 20–30°C, respectively, and calcium ions were not required. As a substrate, phosphatidylcholine was more easily hydrolyzed than phosphatidylethanolamine by this enzyme, and the plasmalogen-type phospholipid was much lower than the acyl-type phospholipid. After a 6-h hydrolysis reaction of the purified phospholipid extracted from the mixed ovaries of skipjack and yellowfin tuna by this enzyme, the LPC ratio of the phospholipid increased from 20 to 72.6% and the percentage of DHA for the total fatty acids of the phospholipid also increased. Thus, skipjack ovaries might possibly be used as a source of PLA₁.