ULTRASONOGRAPHIC FEATURES OF DIFFUSE HEPATOCELLULAR DISORDERS IN DAIRY CATTLE

Ultrasonography of the liver was performed in 200 Holstein-Friesian dairy cattle using a 3.5 MHz transducer with a linear array electronic scanner. Liver samples were taken, processed and examined microscopically and the fat occupying rate was calculated. The hepatic ultrasonograms were evaluated according to the presence of bright pattern, dark pattern, deep attenuation, vascular blurring and blurring of edges. Of the 200 animals, 96 had a normal liver, 63 had hydropic degeneration of the liver, 37 had fatty infiltration of the liver, 3 had liver dystrophy and I had hepatic amyloidosis, diagnosed through histopathological examination. Amyloidosis was characterized by bright pattern and blurring of edges. Liver dystrophy had higher percentages of bright pattern and blurring of edges than normal liver. Hydropic degeneration had higher percentages of dark pattern and blurring of edges than normal liver. Fatty infiltration had higher percentages of bright pattern, deep attenuation, vascular blurring and blurring of edges than normal liver. The present results suggest that different ultrasonographic patterns can be observed in various diffuse hepatocellular disorders in dairy cattle     

Quantitative analysis of the optic nerve of the horse (Thoroughbred)

Three optic nerves (L1, R2, R3) 12-18 mm behind the eyeball of the horse (Thoroughbred) were investigated quantitatively under light and electron microscopes. Thin sections at the thickness of 0.35 microm were cut, stained by toluidine blue and observed under the light microscope. The areas of the optic nerve and the axon bundles were 20.03 +/- 1.04 and 16.59 +/- 0.79 mm2 (mean +/- SD, n=3), respectively. The axon numbers for optic nerve L1, R2 and R3, estimated from light micrographs, were about 481 x 10(3), 543 x 10(3), and 494 x 10(3), respectively. Axons of optic nerve L1 were also counted from electron micrographs and the total number of 488 x 10(3) was received. Furthemore, axon diameters of optic nerve L1 were also measured from electron micrographs. The diameter of a circle with the same peripheral length as an axon, was regarded as its diameter. The medullary sheath of the axon was not included during measuring. Altogether 5,744 axons were measured and axon diameters were in a range of 0.23-12.69 microm, with a mean of 2.56 +/- 1.45 microm (mean +/- SD). A regional difference of axonal diameters was found across the optic nerve: the mean diameter of axons in the centrodorsal region (2.28 microm) was the smallest, and had significant difference with those in several peripheral regions (P<0.05).     

Flow cytometric analysis of chicken NK activity and its use on the effect of restraint stress

Immune system is organized by the influence of both neural and endocrine systems. NK activity plays an important role in the innate immunity. In this study, we observed the effects of restraint stress on chicken peripheral blood NK activity. Viability of FITC-labeled RP9 was measured with PI after treatment with the effector cells. Chicken peripheral blood CD8alpha+ cells expressed strong cytotoxic activity, in contrast to thrombocytes, while peripheral blood CD3+ CD8alpha+ cells and CD4+ cells had little cytotoxic activity. Con A supernatant enhanced the cytotoxic activity of CD8alpha+ cells. Therefore, it is considered that these cytotoxic activities measured by flow cytometry (FCM) analysis are NK activity. When chickens were exposed to restraint stress, the levels of serum corticosterone increased transiently over a short period of time while the NK activity decreased. The decreased NK activity, however, did not recover to the intact levels for a long time, even once the serum corticosterone levels had recovered. These data indicate that chicken NK activity is able to be measured by flow cytometric analysis and that restraint stress causes severe damage to the chicken NK activity.     

An immunohistochemical study of endocrine cells in the abomasum of vagotomized calf

The effect of thoraco-vagotomy on the distribution and frequency of chromogranin-, serotonin-, somatostatin- and gastrin-immunoreactive cells in the abomasum of the calf were investigated by immunohistochemistry. Calves were vagotomized at 1 week old and sampled 2 and 4 weeks later. The endocrine cells generally decreased in number in vagotomized calves as compared to non-operated control calves. However, the detailed responses of endocrine cells to vagotomy varied depending on the endocrine cell type, region of gastric mucosa, and period after vagotomy. The present result suggests that the vagus nerve has an influence on the intrinsic regulatory system by endocrine cell control in the ruminant abomasum.     

Topography of ganglion cells in the retina of the horse

Topography of ganglion cells in the retina of the horse (Thoroughbred) was analyzed in the wholemount retina stained with cresyl violet, and a total number of ganglion cells were estimated. Sizes of ganglion cells were also measured and size spectra were analysed. The main results showed that: (1) a common point in 4 wholemount retinae, based on cell densities and retinal locations, was that a retina could be divided into 5 regions, namely visual streak, nasal, temporal, dorsal and ventral region to the visual streak. A maximum cell density of 4,000 cells/mm2 was found in the visual streak. And a total number of ganglion cells was estimated in a range of 398 x 10(3) - 469 x 10(3), with a mean of 441 x 10(3) +/- 31 x 10(3) (n=4). (2) cell sizes were measured as the mean lengths of the major and minor axes of the somas, and were in a range of 5-53.8 microm. The lowest mean diameter was 14.0 microm (+/- 3.7) in the visual streak and the highest was 25.9 microm (+/- 7.6) in the ventral region. Cell size spectra were unimodal and positively skewed. It is expected that these analyses will provide an anatomical and physiological background for further study of the visual system in the horse.     

In vitro detection of equine arteritis virus from seminal plasma for identification of carrier stallions

Equine arteritis virus (EAV) was readily isolated in RK-13 cell monolayers by plaque assay from seminal plasma of experimental carrier stallions when they contained high titers of virus regardless of the presence of non-viral cytotoxicity in the seminal plasma. The cytotoxicity interfered with virus isolation from seminal plasma which contained virus at titers less than 10 PFU/ml. However, it was possible to detect the virus in seminal plasma pretreated with PEG (#6000). EAV was consistently identified by RT-PCR from crude seminal plasma which contained virus at titers of more than 10(2.7) PFU/ml. In vitro detection of EAV by virus isolation supplemented with RT-PCR using seminal plasma was proved to be an effective alternative to the standard test mating as a diagnostic method for carrier stallions.     

Species identification method for <Emphasis Type="Italic">Scombrops boops</Emphasis> and <Emphasis Type="Italic">Scombrops gilberti</Emphasis> based on polymerase chain reaction-restriction fragment length polymorphism analysis of mitochondrial DNA

ABSTRACT:   Gnomefish Scombrops boops and Scombrops gilberti are commercially important fishes in Japan, but these species are often confused in the markets because of their morphological similarity. To identify these two species, we performed nucleotide sequencing and restriction fragment length polymorphism (RFLP) analysis on 16S ribosomal RNA (rRNA) gene and the control region in mitochondrial DNA. Five and 12 nucleotide substitutions were observed between species in the 777-bp 16S rRNA gene and 471-bp control region, respectively. Diagnostic restriction sites for discriminating between S. boops and S. gilberti were found in the 16S rRNA gene, but not in the control region. Polymerase chain reaction (PCR)–RFLP analysis using two enzymes, Eco NI and Mva I, clearly discriminated between S. boops and S. gilberti identified by meristic characters. The PCR–RFLP analysis identified most of the 168 Scombrops young caught in the coastal waters of the Izu and Miura peninsulas as S. boops , suggesting that S. gilberti juveniles are rare in this area.     

Supplemental effect of cholyltaurine and soybean lecithin to a soybean meal-based fish meal-free diet on hepatic and intestinal morphology of rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>

The supplemental effects of cholyltaurine and soybean lecithin to a defatted soybean meal (SBM)-based fish meal-free diet on the hepatic and intestinal morphology of rainbow trout Oncorhynchus mykiss were investigated. A fish meal-based diet (FM), three fish meal-free diets supplemented either with soybean lecithin (SL), bovine gall powder (BG) or cholyltaurine (C-tau) and an unsupplemented fish meal-free diet (NFM), were given to fish (−11 g) for 10 weeks. The growth was the lowest in fish fed diet NFM, intermediate in fish fed diet SL and the highest in fish fed diet FM, BG and C-tau. Hepatocytes of fish fed diet NFM were atrophied, and the distal intestine of these fish showed abnormal features: disintegrated microvilli and fatty degeneration in epithelial cells and increased amount of connective tissue in the submucosa. Hepatic and intestinal histological features in fish fed diets SL, BG and C-tau were similar to those in fish fed diet FM. These results indicate that soybean lecithin, bovine bile salts and cholyltaurine have similar effects on normalizing the hepatic and intestinal morphologies of rainbow trout fed the SBM-based diet, although growth promotion effect was limited in the soybean lecithin.     

Toxicokinetics and absolute oral bioavailability of melamine in broiler chickens

The objective of this study was to investigate the toxicokinetic characteristics of melamine in broilers due to the limited information available for livestock. Melamine was then administered to broiler chickens at an intravenous (i.v.) or oral (p.o.) dosage of 5.5 mg/kg of body weight, and plasma samples were collected up to 48 h. The concentration of melamine in each plasma sample was analyzed using liquid chromatography‐tandem mass spectrometry (LC‐MS/MS). Melamine was measurable up to 24 h after i.v. and p.o. administration. A one‐compartment model was developed to describe the toxicokinetics of melamine in broilers. Following i.v. administration, the values for the elimination half‐life (t_1/2β), the volume of distribution (V_d), and the clearance (CL) were 4.42 ± 1.02 h, 00.52 ± 0.18 L/kg, and 0.08 ± 0.01 L/h/kg, respectively. The absolute oral bioavailability (F) was 95.63 ± 3.54%. The results suggest that most of the administered melamine is favorably absorbed from the alimentary tract and rapidly cleared by the kidneys in broiler chickens.