Comparison of polymerase chain reaction-restriction fragment length polymorphism assay and enzyme assay for diagnosis of G(M1)-gangliosidosis in Shiba dogs.

In the present study, diagnostic methods for canine G(M1)-gangliosidosis were examined by comparing a DNA mutation assay with an enzyme assay. Sixty-two Shiba dogs of a pedigree with G(M1)-gangliosidosis were differentiated into 3 genotypes, i.e., normal, heterozygous, and homozygous affected dogs, using a DNA mutation assay, which consists of polymerase chain reaction amplification and the determination of restriction fragment length polymorphisms. The beta-galactosidase activity in leukocytes, umbilical cords, and plasma was measured using 4-methylumbelliferyl beta-D-galactoside and p-nitrophenyl beta-D-galactoside as artificial substrates and compared among the 3 genotypes. The results showed that it was possible to identify homozygous dogs with the enzyme assay using leukocytes and umbilical cords. When using leukocytes, heterozygous carriers could be differentiated from normal dogs in many cases. However, the use of the DNA mutation assay is essential for a complete determination of heterozygous carriers because of the overlap in the distribution of enzyme activity between these 2 groups. When umbilical cords were used, heterozygous carriers could not be differentiated from normal dogs because of no significant difference in enzyme activity between these 2 groups. The beta-galactosidase activity in plasma was not applicable to the diagnosis and genotyping of G(M1)-gangliosidosis in Shiba dogs.     

Comparison of feed intake, digestibility and fattening performance of Brahman grade cattle (Bos indicus) and crossbred water buffalo (Bubalus bubalis)

Twenty growing crossbred cattle and crossbred water buffalo (carabao) with an average age of 22 (18–24 months) months were equally distributed into two treatment groups according to species. The animals were fed with the same ration made up of corn silage (50%) + wet brewer's spent grain (30%) + concentrate mixture (20%), and their fattening performance was monitored. The digestibilities of the different nutrients were likewise determined. The economics of raising the animals under intensive production system was calculated. Species differences did not influence total dry matter intake of the animals, when expressed as percentage of the bodyweight and per metabolic body size. There were no significant differences in digestion coefficients of the different nutrients, except for crude protein in crossbred water buffalo and crossbred cattle, although the digestibility of dry matter, organic matter and nitrogen free extract tended to be high in the former than in the latter. Likewise, average daily gain (ADG) was similar, although crossbred water buffalo had numerically higher ADG (828.6 vs 785.5 g) than crossbred cattle during the 6 months feeding. During the first 3 months of feeding (1–90 days), the ADG of crossbred water buffalo was 1066.1 g compared to 940.1 g for crossbred cattle. From 91 to 180 days, the crossbred cattle had slightly higher ADG (630.1 vs 591.1 g) but also the difference was not significant. The return above feed cost was comparable for crossbred cattle and crossbred water buffalo during the first 90 days of feeding. However, extending the feeding period from 91 to 180 days , income over feed cost was higher (P < 0.05) for crossbred cattle by PhP 5.3/kg gain than crossbred water buffalo. Results showed that crossbred water buffalo could attain similar growth rate with that of crossbred cattle under intensive system, when fed with high quality feed materials.     

Controlled release of estradiol-17β and bisphenol A from a silicone tube for long-term administration in mice

Silicone tubes filled with estradiol‐17β (E2) or the xenestrogen, bisphenol A (BPA) were covered with a polyethylene tube and implanted into mice subcutaneously. The releasing patterns from the tube in vivo were obtained from the volume of estrogen remaining in the tube by measurement with high performance lipid chromatography. E2 in glycerol was released in a few days from a silicone tube, which was not covered with a polyethylene tube (Type A). However, E2 in sesame oil was released very slowly from the silicone tube covered with a polyethylene tube (Type B) and approximately 10% of E2 remained in the tube at 56 days after implantation into the mice. The mean linear regression equation for E2 was Y = 70.967 ? 1.2271X (R² = 0.9268) and the quadratic regression equation was Y = 77.97 ? 2.2773X + 0.0194 × 2 (R² = 0.9864). The remainder rate curves of E2 and BPA were not significantly different throughout the experimental period of 56 days. The Type B tube in the present experiment might be useful for long‐term administration of estrogens.     

Prevalence of blood parasites in Japanese wild birds

The prevalence of blood parasites was investigated in 701 Japanese wild birds for 13 years from January, 1988 to March, 2001. Most of the injured or sick birds were caught in the suburbs of Kobe City, Hyogo Prefecture and brought to the zoo for clinical care. Among all the birds examined, 10.6% were infected with hematozoa belonging to three genera as Plasmodium (1.7% of the samples), Haemoproteus (5.1% of the samples) and Leucocytozoon (4.6% of the samples), and two birds (0.29% of the samples), a Japanese grosbeak (Coccothraustes personatus) and a dusky thrush (Turdus naumanni), were infected with microfilariae. Mixed infection with Leucocytozoon sp. and Haemoproteus sp. was observed in 6 individuals of 4 species and that with Leucocytozoon sp. and microfilariae was observed in 2 individuals of 2 species of bird. Relatively high positive rates were 75%(3/4) in the scops owl (Otus scops), 71.4% (10/14) in the ural owl (Strix uralensis), 57.7% (15/26) in the jungle crow (Corvus macrorhynchos), 57.1% (4/7) in the black-tailed gull (Larus crassirostris), 55.6% (5/9) in the brown hawk owl (Ninox scutulata), 41% (16/39) in the carrion crow (Corvus corone) and 24.1% (7/29) in the night heron (Nycticorax nicticorax).     

Different factors affect developmental competence and cryotolerance in in vitro produced bovine embryo

The present study was conducted to examine the effects of culture systems and culture media on developmental competence and freezability of bovine embryos obtained by in vitro culture of in vitro matured and fertilized (IVM-IVF) oocytes. No significant difference was observed in the proportions of oocytes developed to blastocysts, the speed at which the oocytes reached the blastocyst stage and the number of cells, when the IVM-IVF oocytes were cultured in CR1aa with or without cumulus cells. Nevertheless, more of the IVM-IVF oocytes cultured either with or without cumulus cells in CR1aa were seen to reach the blastocyst stage much sooner than those cultured with cumulus cells in TCM199 (P<0.05). The proportion of embryos developed to the blastocyst stage by day 7 in CR1aa culture was significantly higher than embryos cultured in TCM199. Viability after frozen-thawed blastocysts were obtained in vitro, was seen in a significantly higher percentage of embryos cultured in TCM199 and developed to the hatched blastocysts than in those cultured in CR1aa (P<0.05). These results indicate that CR1aa was superior to TCM199 for the potential developmental of IVM-IVF oocytes to blastocysts during in vitro culture regardless of co-culture with or without cumulus cells. But the freezability of blastocysts developed in CR1aa was inferior to those developed in TCM199.     

New race of <Emphasis Type="Italic">Phytophthora vignae</Emphasis> f. sp. <Emphasis Type="Italic">adzukicola</Emphasis>, the causal agent of Phytophthora stem rot of the adzuki bean

 A new race of Phytophthora vignae f. sp. adzukicola, designated race 4, is reported from central and western Hokkaido, Japan. The isolates obtained from diseased plants of a new cultivar, cv. Syumari, which is resistant to races 1, 2, and 3, were determined to be a new race by the pathogenic reaction on a set of differential adzuki bean cultivars (cv. Erimo-shozu, cv. Kotobuki-shozu, cv. Noto-shozu, cv. Urasa-shimane, and cv. Syumari). Received: March 7, 2002 / Accepted: August 13, 2002     

The antiproliferative effect of bovine lactoferrin on canine mammary gland tumor cells

Lactoferrin has several biological activities, including antitumor activities in some human and animal tumor cells. Clinical trials have been carried out in human medicine based on these effects. However, the antitumor effects of lactoferrin in veterinary medicine remain unknown. In this in vitro study, we demonstrated that co-incubation of canine mammary gland tumor cells (CIPp and CHMp) and bovine lactoferrin induced growth arrest of tumor cells. This growth arrest was associated with induction of G1 arrest. Furthermore, this effect was stronger in tumor cells than in normal cells. These findings demonstrate that bovine lactoferrin has anti-tumor activity in canine mammary tumors and has the potential for use in tumor-bearing dogs.     

Effect of bovine lactoferrin on functions of activated feline peripheral blood mononuclear cells during chronic feline immunodeficiency virus infection

Feline immunodeficiency virus (FIV) infection is characterized by chronic overactivation of immune and inflammatory system, resulting in anergic state and dysfunction of immune cells. Lactoferrin (LF), a glycoprotein present in exocrine secretions and neutrophils, plays an important role in host defense system. Our previous study showed that oral administration of bovine LF (bLF) suppressed oral inflammation, improved the clinical symptoms and decreased serum gamma-globulin as a marker of inflammation in FIV-infected cats with intractable stomatitis. The anti-inflammatory effect was partly involved in regulation of neutrophil function by bLF. In this study, to clarify the relationship between anti-inflammatory effects of bLF and peripheral blood mononuclear cells (PBMC), we examined the effect of bLF on proliferation, cell cycle progression and cytokine expression in mitogen-activated PBMC. MTT [3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl tetrazolium bromide] assay showed that bLF inhibited the concanavalin A (ConA)-induced cell proliferation in FIV-infected cats with the asymptomatic carrier and AIDS-related complex (ARC) phase. Bovine LF restored ConA-induced cell cycle progression and resulted in suppression of the induced apoptosis in feline PBMC. Real-time RT-PCR showed that bLF suppressed ConA-induced expression of interferon-gamma and interleukin-2 in cells of the ARC group regardless of the time of its addition to the medium. These results suggest the hypothesis that therapy with bLF may have the potential to improve and protect functions of overactivated lymphocytes by modulating the cell proliferation, cell cycle and cytokines expression in cats in terminal stage of FIV infection.     

Morphological analysis of corneal opacity in Shiba dog with GM1 gangliosidosis

GM1 gangliosidosis is one of the inherited metabolic lysosomal storage disorders characterized by neurological symptoms caused by beta-galactosidase deficiency and consequent accumulation of GM1 ganglioside in neuronal cells. Shiba dogs affected with GM1 gangliosidosis have been found to suffer from corneal opacity. In our morphological analysis, keratocyte enlargement was induced by abnormal intracellular accumulation of neutral carbohydrates, resulting in the loss of normal arrangement of collagen fibrils in the opaque cornea was found to be associated with the disorder. We therefore conclude that corneal opacity in this Shiba dog with GM1 gangliosidosis may be caused by neutral carbohydrate accumulation in lysosomes, swelling and dysfunction of keratocytes, and subsequent irregular arrangement of collagen fibrils in the corneal proper substance.     

Evaluation of equine cecal motility by ausculation, ultrasonography and electrointestinography after jejunocecostomy

OBJECTIVE: Horses often suffer reduced intestinal motility after jejunocecostomy. Therefore, accurate evaluation of intestinal motility is important for the prevention, diagnosis and treatment of this condition. The purpose of this study was to evaluate intestinal motility in horses after jejunocecostomy using three different methods, i.e. auscultation, ultrasonography and electrointestinography. ANIMALS: Six healthy thoroughbreds were used in this study. They were subjected to jejunocecostomy. PROCEDURE: Bowel sounds in the right paralumbar fossa were assigned a score of 0-3 for intestinal motility evaluation by auscultation, and the number of cecal contractions during a 3-min period were counted by ultrasonography. Electrointestinography (EIG) was used to measure percutaneous potential of the cecum. RESULTS: We identified three specific postoperative periods: the period of reduced intestinal motility (postoperative day 1 to day 2), in which intestinal motility declined, the unstable period (day 3 to day 7), in which intestinal motility partially recovered, and the full recovery period (day 8 to day 31), in which intestinal motility returned to preoperative state. Careful management was found to be especially important during the period of reduced intestinal motility and the unstable period. We found that, in healthy horses that underwent jejunocecostomy, it takes approximately one month for the cecum to return to normal motility patterns observed before surgery. CONCLUSION: We have shown in this study that evaluation of intestinal motility after jejunocecostomy in horses by EIG is more objective and provides more details than evaluation by auscultation or ultrasonography.