Ligand-binding characteristics of feline insulin-binding immunoglobulin G

Polyclonal immunoglobulin (Ig) G autoantibodies against insulin have been identified in sera of healthy cats. We purified and fractionated insulin-binding IgGs from cat sera by affinity chromatography and analyzed affinity of insulin-binding IgGs for insulin and their epitopes. Following the passing of fraction A, which did not bind to insulin, insulin-binding IgGs were eluted into two fractions, B and C, by affinity chromatography using a column fixed with bovine insulin. Dissociation constant (KD) values between insulin-binding IgGs and insulin, determined by surface plasmon resonance analysis (Biacore™system), were 1.64e⁻⁴ M for fraction B (low affinity IgGs) and 2e⁻⁵ M for fraction C (high affinity IgGs). Epitope analysis was conducted using 16 peptide fragments synthesized in concord with the amino acid sequence of feline insulin by an enzyme-linked immunosorbent assay. Fractions B and C showed higher absorbance (affinity) of the peptide fragment of 10 amino acid residues at the carboxyl-terminal of the B chain (peptide No. 19), followed by peptide fragments of 6 to 15 amino acid residues of the B chain (peptide No. 8). Fraction C showed a higher absorbance to 7 to 16 amino acid residues of the B chain (peptide No. 5) compared with the absorbance of fraction B. Polyclonal insulin-binding IgGs may form a macromolecule complex with insulin through the multiple affinity sites of IgG molecules. Feline insulin-binding IgGs are multifocal and may be composed of multiple IgG components and insulin.     

Triploid Induction in the Yellowtail Tetra,Astyanax altiparanae,Using Temperature Shock: Tools for Conservation and Aquaculture

Triploidization is an interesting tool to produce sterile fish. In the yellowtail tetra, Astyanax altiparanae, this can be applied for aquaculture and surrogate technologies. In this study, we compared the efficacy of cold (2 C) or heat shock (38 C, 40 C, and 42 C) on triploid induction in the yellowtail tetra. The eggs were treated with cold or heat shock, 2 min postfertilization (30 min in cold shock or 2 min in heat shock). Intact embryos served as the control group. Ploidy status was confirmed by karyotyping, flow cytometry, and nuclear diameter of erythrocytes. The hatching rate decreased after cold shock (12.69 ± 15.76%) and heat shock at 42 C (0.35 ± 0.69%) in comparison with the control group (63.19 ± 16.82%). At 38 C and 40 C, hatching rates (61.29 ± 17.73% and 61.75 ± 22.1%, respectively) were not decreased. Only one triploid arose at 38 C (1/80). At 40 C, a high number of triploids arose (72/78). At 42 C, very few embryos developed into the hatching stage. A large number of haploid individuals arose after cold shock (61/75), with only one triploid. Our results indicate that heat shocking of embryos at 40 C is optimum for triploid production in the yellowtail tetra.     

Bioavailability of amino acids chelated and glass embedded zinc to rainbow trout, Oncorhynchus mykiss, fingerlings

A 2 × 4 factorial experiment was conducted to determine the bioavailability of zinc (Zn) from amino acids chelated (Zn–Am) and glass embedded Zn (Zn–Gl) as sources for rainbow trout, Oncorhynchus mykiss , fed practical type diets. Two levels of Zn (20 and 40 mg kg⁻¹) were supplemented to the diets using either zinc sulphate (Zn–Sf), zinc methionine (Zn–Mt), Zn–Am or Zn–Gl. Rainbow trout with an average weight of 2 g were fed the experimental diets for 15 weeks. Growth and feed gain ratio (FGR) were not significantly influenced by the dietary Zn content and forms. Alkaline phosphatase (ALP) activity for both levels of Zn–Am was significantly higher than that of Zn–Sf and Zn–Gl at 20 mg supplementation. In another experiment, fish of about 95 g were fed the same experimental diets to determine the absorption of Zn and it was found to be significantly higher from Zn–Am compared with the rest. Retention from Zn–Am at 20 mg was significantly higher than the rest, excluding Zn–Sf. The results suggest that the availability of Zn from Zn–Am might be superior among the sources compared.     

Evaluation of cytokine messenger RNA expression in peripheral blood mononuclear cells from dogs with canine demodicosis

Using RT-PCR and semi-quantitative PCR, mRNA expression for canine interferon (IFN)-gamma, interleukin (IL)-2, IL-4, IL-5, IL-10, tumor necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta in peripheral blood mononuclear cells (PBMCs) was examined in dogs with or without demodicosis. mRNA expression for IFN-gamma as well as TNF-alpha in dogs with demodicosis (localized (LD) and generalized (GD)) was slightly lower than those in dogs without demodicosis (healthy controls). Expression of IL-5 mRNA in dogs with demodicosis was higher than that in control dogs, but there were no significant differences in IL-4 and IL-10 mRNA expression levels among the three groups. On the other hand, expression levels of TGF-beta mRNA in dogs with GD were higher than those in control dogs and dogs with LD. The expression levels of IL-5 and TGF-beta mRNA decreased in all three dogs with GD which showed resolution of the clinical signs. Taken together, these results suggest that the Th2-like response in PBMCs from dogs with demodicosis is up-regulated, and that subsequent increased expression of IL-5 and TGF-beta mRNA in dogs with GD is reversible after treatment. Therefore, these cytokines, particularly IL-5, might be a useful clinical index of the clinical course in demodicosis. Also, increased TGF-beta mRNA expression might be a key factor for revealing the difference in the mechanism of onset between LD and GD.     

Comparison of Ni/Zn Accumulation Ability of Thlaspi japonicum from Three Different Areas in Hokkaido

Thlaspi japonicum is a Japanese Ni-hyperaccumulator plant growing in areas with serpentine soil containing a high level of Ni. In the present study, we investigated the ability of wild Thlaspi japonicum plants inhabiting non-ultramafic soil areas of Hokkaido to accumulate Ni/Zn. Wild T. japonicum plants growing on Rebun Island (Rebun-T) accumulated small amounts of Ni, while the plants cultivated on ultramafic rock soil accumulated large amounts of Ni, like T. japonicum growing on Mt. Yubari (Yubari-T), and this change was also observed for T. japonicum growing on Mt. Tengu (Tengu-T). All the T. japonicum plants transplanted on both ultramaflc soil or Andosol accumulated over 100 mg kg⁻¹ Zn, especially the Ni/Zn content of Tengu-T plants tended to be higher than that of the plants growing in the other two areas. In contrast, transplanting of Yubari-T plants from ultramafic soil with a high level of Ni to soil with a lower-Ni content decrease of the Ni concentration in the leaves. Wild Rebun-T plants showed a smaller biomass than the Yubari-T and Tengu-T plants, and some individuals of the Rebun-T population were smaller than those of the other populations, with tiny radical leaves. Zinc content of the small leaves was not significantly different from that in large leaves. These results suggested that T. japonicum plants are able to accumulate high Ni/Zn accumulation, regardless ecotype, habitat and leaf size.     

Histochemical properties of skeletal muscles in Japanese cattle and their meat production ability

The compositional characteristics of the three basic types of myofiber, namely type I (slow‐twitch oxidative), type IIA (fast‐twitch oxidative glycolytic) and type IIB (fast‐twitch glycolytic), are clarified in the skeletal muscles of Japanese Black cattle. The myofiber composition, which is characteristic of the muscles of Japanese Black cattle, markedly changes during their growth, when some type IIA myofibers are transformed into type I or IIB, depending on the different muscles. Independent of these changes with growth, inter‐ and intramuscular variations of myofiber type distribution is evident. The small extensor muscles in deep regions around bone contain a lot of type I myofibers, whereas the large muscles at surface regions have many type II myofibers. Japanese Black cattle have typical white muscles such as the Longissimus thoracis and Semitendinosus, containing half the myofibers as red (type I + IIA). The muscles of Japanese Black cattle show a tendency to contain a higher percentage of type I myofibers than other breeds over an intrabreed variation of the myofiber type composition. In the big muscles such as the Longissimus thoracis and Biceps femoris, a great diversity of myofiber type composition is observed among the different regions. When fattened, heifers produce Longissimus thoracis and Biceps femoris muscles of smaller weight than steers, but in heifers the myofiber size in each type is rather larger. In the Psoas major, Vastus lateralis and Serratus ventralis muscles, heifers contain a higher frequency of red (type I + IIA) myofibers with no differences in myofiber size. Among the several muscles of fattened Japanese Black steers, the percentage distribution of type I myofibers has a positive correlation with the percentage amount of intramuscular fat. From these results, the high potential of Japanese Black cattle to produce marbled beef could be based on the histochemical properties of myofibers in their skeletal muscles.     

Sr isotopic compositions of ayu otolith and its ambient water

Sr isotopic compositions of ayu otolith and water collected from Japanese rivers were measured to validate the relationship between otolith and ambient water for this species. Micromilling and rostrum-cut methods employed for otolith sampling were compared and yielded almost indistinguishable otolith Sr isotopic composition data. A good correlation observed between data on otolith and those on ambient water (river water and sea water) confirms the usefulness of Sr isotopic composition for elucidating migratory behavior of ayu, as already verified in other species.     

Proportion of abortions due to neosporosis among dairy cattle in Japan

A retrospective cohort study was conducted to clarify the association between seropositive reactions to Neospora caninum and subsequent reproductive disorders among dairy cattle in Japan. A statistically significant association between Neospora seropositive reactions and abortions was observed (P=0.016), and seropositive cattle were 6.1 times more likely to abort compared to seronegative cows. No significant differences were observed between seropositive reactions and other reproductive disorders such as conception failure, perinatal death and calf mortality. As indicated by estimation of the attributable fraction, 83.6% of abortions in Neospora seropositive animals may be attributed to N. caninum. Considering seroprevalence of N. caninum in the cattle which aborted in Japan, 21.8% of abortions were estimated to be caused by neosporosis in Japan.     

Molecular survey of Babesia infection in dogs in Okinawa, Japan

A total of 80 free-roaming dogs on Okinawa Island, Japan, were examined for Babesia infection using the polymerase chain reaction (PCR) and sequence analysis. Of 80 samples, 12 were positive in a Babesia genus-specific PCR. Consequent species-specific PCR for B. canis and B. gibsoni revealed that 5 (6.3%) and 7 (8.8%) dogs were infected with B. canis and B. gibsoni, respectively. Sequence analysis of the PCR products revealed that the 18S rRNA gene sequence of B. canis detected from dogs in Okinawa was very close to B. canis vogeli with sequence similarity of 99.94%.     

Effect of high intensity training on anaerobic capacity of middle gluteal muscle in Thoroughbred horses

We hypothesize that high intensity training for Thoroughbred horses that have been subjected to conventional training could further improve the metabolic properties of the middle gluteal muscle. Nine well-trained horses were subjected to high intensity (80-100% Vdot;O(2)max, 5 minx2) training for 12 weeks. Biopsy samples were obtained from the muscle before and after 4 and 12 weeks of training. Three of the 9 horses did not complete the training programme. In the remaining 6 horses, activities of succinic dehydrogenase (SDH), phosphofructokinase (PFK) and 3-hydroxy acyl CoA dehydrogenase (HAD), and the composition of myosin heavy chain isoforms were analyzed by biochemical techniques. After 12 weeks of training, a significant increase was found in PFK activity but not in the SDH and HAD activities. There were no significant changes in the composition of myosin heavy chain isoforms. The high intensity training in this study was effective at increasing glycolytic enzyme activity, indicating the possibility to improve anaerobic capacity, which potentially could contribute greatly to performance in Thoroughbred horses. This study also highlighted a fact that high intensity training should be given with the great care to prevent the skeletal muscle injuries.