Laboratory diagnosis of canine GM2-gangliosidosis using blood and cerebrospinal fluid.

In the present study, laboratory techniques were used to diagnose canine GM2-gangliosidosis using blood and cerebrospinal fluid (CSF) that can be collected noninvasively from living individuals. Lysosomal acid beta-hexosaminidase (Hex) was measured spectrofluorometrically using 4-methylumbelliferyl N-acetyl-beta-D-glucosaminide and 4-methylumbelliferyl 7-(6-sulfo-2-acetamido-2-deoxy-beta-D-glucopyranoside) as substrates. Main isoenzymes A and B of Hex in leukocytes were also analyzed using cellulose acetate membrane electrophoresis. GM2-ganglioside in CSF was detected and determined quantitatively by using thin-layer chromatography/enzyme-immunostaining method with anti-GM2-ganglioside antibody. In normal dogs, Hex activities could be determined in leukocytes, serum, and CSF and the total activities were markedly reduced in all the enzyme sources in a dog with Sandhoff disease. Electrophoresis of a leukocyte lysate from a normal dog showed that the Hex A and Hex B were not separated distinctively with formation of a broad band, whereas there were no bands in electrophoresis of a lysate from a dog with Sandhoff disease, showing a deficiency in the total enzyme activity. GM2-ganglioside could be detected and determined quantitatively in as little as 100 microl of canine CSE GM2-ganglioside in CSF in a dog with Sandhoff disease increased to 46 times the normal level. In conclusion, the methods in the present study are useful for diagnosis of canine GM2-gangliosidosis. These techniques enable definitive and early diagnosis of canine GM2-gangliosidosis even if tissues and organs cannot be obtained.     

Involvement of a host erythrocyte sialic acid content in Babesia bovis infection

Host sialic acid (SA) has recently been suggested to play an important role in erythrocyte (RBC) infection by Babesia spp. The present study attempted to further determine the specific type of SAs important in the RBC invasion. Bovine RBC was found to bear abundant alpha2-3-linked SA residues but not alpha2-6-linked SA in nature, confirmed by flow cytometric analysis of the neuraminidase (Nm)-treated RBCs. Lectin-blot analyses revealed the removal of alpha2-3-linked SAs from the 97-, 33-, and 31-kDa bands by the Nm treatment. Addition of the Nm-treated RBCs into an in vitro culture of B. bovis resulted in a decreased population of the parasitized RBCs. The thin smear samples from the cultures were then observed under a confocal laser scanning microscope after staining with the alpha2-3-linked SA-specific lectin: a selective invasion of B. bovis was found only in the intact RBCs bearing the SAs, but not in the desialylated RBCs. Furthermore, a significant reduction of the parasitized RBCs was also observed in the culture supplemented with exogenous 3'-sialyllactose containing the alpha2-3-linked SAs. However, the complete inhibition of parasite proliferation was not achieved in the culture. These findings indicate that while the alpha2-3-linked SA-dependent pathway is needed for highly efficient invasion of host RBCs by B. bovis, there might also be other potential alternative pathways.     

Daily growth rate model of Japanese anchovy larvae Engraulis japonicus in Hiuchi-nada Sea, central Seto Inland Sea

In the present study, we developed a larval anchovy growth model in relation to sea temperature and food availability via food consumption and metabolic process terms, based on biological data from previous laboratory experiments and field surveys from 2003 to 2006 in Hiuchi-nada Sea, central part of Seto Inland Sea, Japan. To investigate when food shortage for larval anchovy and then recruitment failure occur in Hiuchi-nada Sea, anchovy food requirements were estimated by using the growth model, and we compared the food requirement with anchovy food availability. We applied an estimation method for growth model parameters, Hewett?CJohnson p and Q ₁₀, by minimizing the sum of squares of difference between mass-specific growth rates estimated by the models and those by otolith growth analysis. Parameter p was 0.86, slightly higher than typical values, and Q ₁₀ was 2.11, close to the value used for the biological model of larval northern anchovy. Food shortage for anchovy larvae did not occur in Hiuchi-nada Sea, although it was indicated that low food availability led to a low reproductive success rate. The newly developed growth model is considered optimal at present and useful to link environmental conditions and larval growth.     

Ability of salmon growth hormone to accelerate somatic growth of juvenil abalone Haliotis discus hannai

ABSTRACT:   The ability of salmon growth hormone (sGH) to accelerate the somatic growth of juvenile abalone Haliotis discus hannai by immersion was examined. After immersion for one hour into a sGH-rich solution at a concentration of 30 mg/L, the sGH immunoreactivity in the body fluid of abalone was maximal after one day, and levels were still detectable at two days. No immunoreactivity was observed in the control group. Following immersion for one hour at one or two-week intervals for 120 weeks into the sGH-rich solution at 30 mg/L, the sGH-immersed abalone exhibited a significant increase in shell length and body weight. On the other hand, abalone immersed into a salmon prolactin (sPRL)-rich solution at 30 mg/L for one hour showed a lower increase in both shell length and body weight than the sGH-immersed abalone. No increases in shell length and body weight were observed in either the bovine serum albumin-immersed abalone or untreated controls. These results provide evidence that sGH can be transported from ambient water into the circulatory system of abalone, and can subsequently improve the somatic growth of juveniles.     

Calcium ions are involved in egress of Babesia bovis merozoites from bovine erythrocytes

Bovine babesiosis is a livestock disease known to cause economic losses in endemic areas. The apicomplexan parasite Babesia bovis is able to invade and destroy the host’s erythrocytes leading to the serious pathologies of the disease, such as anemia and hemoglobinuria. Understanding the egress mechanisms of this parasite is therefore a key step to develop new therapeutic strategies. In this study, the possible involvement of Ca²⁺ in the egress of B. bovis merozoites from infected erythrocytes was investigated. Egress was artificially induced in vitro using calcium ionophore A23187 and thapsigargin to increase Ca²⁺ concentration in the cytosol of the parasite cells. The increased intracellular Ca²⁺ concentration following these treatments was confirmed using live cell Ca²⁺ imaging with confocal laser scanning microscopy. Based on our findings, we suggest a Ca²⁺ signalling pathway in the egress of B. bovis merozoites.     

Synovial fluid total protein concentration as a possible marker for canine idiopathic polyarthritis

Idiopathic polyarthritis (IPA) is a very common inflammatory arthropathy in the dog. Canine IPA is diagnosed mainly by detecting increased number of leukocytes in the synovial fluid (SF), which is easily influenced by glucocorticoid therapy. We obtained 31 SF samples from 24 IPA dogs prior to (n=19) and/or after (n=12) 1 to 10 weeks of glucocorticoid therapy. The SF total protein concentrations of IPA dogs were significantly higher than those of dogs with non-arthritis diseases (n=34) and healthy controls (n=10). Our data revealed that the SF total protein concentrations are not influenced by several weeks of glucocorticoid therapy. Hence, the SF total protein concentration is applicable as a diagnostic marker of canine IPA even when the patients are receiving glucocorticoid therapy.     

Seed productivity test of CMS lines of Japanese bunching onion (Allium fistulosum L.) possessing the cytoplasm of a wild species, A. galanthum Kar. et Kir.

In a previous study, we developed male sterile lines of Japanese bunching onion (Allium fistulosum L.) possessing the cytoplasm of a wild species, A. galanthum Kar. et Kir., by backcrossing. To evaluate seed productivity of the male sterile lines in practise, they were crossed with the male fertile line, cultivar 'Kujyo', using honeybees as pollinators under field conditions. The number of florets and seeds per inflorescence, seed set and seed germination of the material were investigated. Although variation was observed among the male sterile lines, there were several lines having seed productivity equal to cultivar 'Kujyo'. Our data demonstrate that the male sterile lines of A. fistulosum possessing the cytoplasm of A. galanthum are useful as seed parents for the commercial F seed production of A. fistulosum.     

PCDD and PCDF in air,soil, vegetation and oily runoff from a tire fire

In early February 1990, vandals set fire to a used tire storage facility near Hagersville, Ontario, Canada, During the fire, the Ontario Ministry of Environment and Energy (MOEE) monitored the site and surrounding impact zone for numerous organic contaminants. Samples of air, soil, vegetation, runoff water, and oily residue were collected and analyzed for polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDF). Air samples produced complex incineration patterns with a large number of isomers detected. Total toxicity equivalents (TEQ) at 1 km downwind of the fire were an order of magnitude higher than those at 3 km downwind. Soil samples collected during the fire did not show elevated PCDD or PCDF concentrations, however, low concentrations were detected in vegetation collected at 100 m and 200 m from the site. PCDD and PCDF concentrations in the foliage decreased with time but were still detectable for at least 200 days after the fire started. As the tires burned, water used to extinguish the fire caused runoff of oil and oily water. Unique patterns of PCDDs and PCDFs were detected in these samples.