Heinz body hemolytic anemia with eccentrocytosis from ingestion of Chinese chive (Allium tuberosum) and garlic (Allium sativum) in a dog

A 4-year-old, intact male miniature schnauzer was presented with anorexia. The dog had ingested some Chinese steamed dumplings 2 days before, which contained Chinese chive (Allium tuberosum) and garlic (Allium sativum). Hematological examinations revealed severe Heinz body hemolytic anemia with eccentrocytosis and an increased concentration of methemoglobin, which was thought to result from oxidative damage to erythrocytes by constituents in these Allium plants. In this case, eccentrocytosis was a hallmark finding and could be detected easily, suggesting that this hematological abnormality is useful in diagnosing Allium plant-induced hemolysis.     

Relation between reticulocyte count and characteristics of erythrocyte 5'-nucleotidase in dogs,cats, cattle and humans

To examine substrate specificity and susceptibility to lead, erythrocyte 5'-nucleotidase was measured in dogs, cats, cattle and humans, and its relationship to the reticulocyte count in these species was determined. The reticulocyte count in dogs was similar to that in humans, but the count in cats was higher than that in humans. Reticulocytes were not observed in cattle. The activities of canine erythrocyte 5'-nucleotidase measured using cytidine and uridine 5'-monophosphates, which are preferentially catalyzed by one of the human pyrimidine 5'-nucleotidase isozymes (P5N-I), were similar to those of the human enzyme. The canine enzyme preferentially catalyzed thymidine 3'-monophosphate, which is catalyzed only by human P5N-II, more strongly than the human enzyme. This suggests that canine erythrocytes have two isozymes similar to human P5N-I and P5N-II, and a higher P5N-II-like activity than human erythrocytes. Feline erythrocytes had the highest level of P5N-I-like activity among the species examined, and the bovine enzymic activities including those of P5N-I and II were the lowest among these species. According to these observations, the reticulocyte count was approximately proportional to the P5N-I-like activity in these species. Therefore, the P5N-I-like activity may be involved in the morphological maturation of mammalian erythrocytes. The canine and feline erythrocytes had markedly high activity and preferentially catalyzed purine 5'-monophosphate suggesting the presence of a purine-specific 5'-nucleotidase as in human erythrocytes. In addition, the canine and feline P5N-I-like activity showed less susceptibility to lead than the human P5N-I. This may be a reason why there are few case reports of lead-induced anemia in dogs and cats.     

An enzyme-linked immunosorbent assay with the recombinant merozoite protein as antigen for detection of antibodies to Eimeria necatrix

A cDNA library was constructed with Eimeria necatrix merozoite mRNA and immunologically screened by chicken sera against this parasite. One of the positive clones containing an insert of 879 nucleotides, pNP19, showed similarity to part of a published gene expressed in E. tenella merozoite by the homology search system. The inserted DNA was subcloned into baculovirus, and a 35-kD protein was expressed, purified, and used for the antigen in enzyme-linked immunosorbent assay (ELISA). Antibodies from the chickens vaccinated with the E. necatrix attenuated strain, Nn-P125, were detected from 14 days after vaccination by ELISA. The mean absorbance increased rapidly to a peak around 21 days after vaccination; thereafter, it began to decline. Even though some of the vaccinated chickens showed very low levels of antibody response to the recombinant protein 56 days after vaccination, they were protected against challenge with virulent strain of E. necatrix. The mean absorbances in sera from both vaccinated and nonvaccinated chickens highly increased 14 days after challenge. On the other hand, the antibody was not detected in ELISA when chickens were exposed to other Eimeria species such as E. tenella, E. acervulina, and E. maxima. These results demonstrate that this recombinant protein is suitable for detecting the specific antibody in chickens infected with both attenuated and virulent strains of E. necatrix.     

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.     

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.     

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.     

Nuclear genome constitution and other characteristics of somatic hybrids between dihaploid Solanum acaule and tetraploid S. tuberosum

Eleven somatic hybrids (2n = 68 to 74) obtained between S. tuberosum ssp. tuberosum cv. Dejima (2n = 48) and ATDH-1 (2n = 24), an anther-culture-derived dihaploid of S. acaule (Yamada et al., 1997), were characterized by nuclear RFLP markers using 49 single-copy DNA probes distributed throughout the potato genome (2 to 6 probes per chromosome). One of the somatic hybrids, DA8-2, had 72 chromosomes and all the Dejima- and ATDH-1-specific markers (124 and 103 bands, respectively), suggesting the presence of a whole set of both parental chromosomes. The other somatic hybrids lost varying numbers of markers up to seventeen. The pattern of the loss of markers indicated the elimination of five chromosomes among four somatic hybrids. A nucleolar organizer region of chromosome 2 was often eliminated in the somatic hybrids. The somatic hybrids studied here had higher frequencies of multivalent formation than the S. tuberosum parent. They had reasonably good seed set when pollinated with S. tuberosum pollen. Hence, homoeologous recombination between S. acaule and S. tuberosum chromosomes is possible and useful traits from S. acaule may be transferred to the S. tuberosum gene pool. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Change in intrarenal Ghrelin expression in immune complex-mediated glomerular disease in dogs

Ghrelin is a peptide hormone that is mainly produced by the stomach. The kidney is a major source of local ghrelin, and maintaining body fluid balance is considered a critical role of renal ghrelin. However, there are no reports on renal ghrelin in small animal medicine. The present study investigated the intrarenal localization of and change in ghrelin expression in dogs with immune complex-mediated glomerulonephritis (ICGN). Ghrelin immunoreactivity (IR) was observed in the distal tubules of normal kidneys. Ghrelin IR was weak in ICGN kidneys, and the quantitative ghrelin IR score was significantly lower in ICGN kidneys than in normal kidneys. In cases of ICGN, plasma creatinine concentrations showed a positive correlation with the ghrelin IR score.