Neospora caninum is a recognized protozoan parasite of a wide range of mammalian hosts, and was reported for the first time in 1988. The isolation of its oocysts in dog's faeces in 1998 led to its establishment as a parasitic species undergoing typical coccidian life cycle. Infection with N. caninum causes paralysis and death in young livestock and companion animals, and is associated with abortions and stillbirth in cattle, and neurologic disease in calves. Considering the economic and agricultural importance of neosporosis, there is the urgent need to develop biological control measures aimed at preventing its transmission, infection, as well as reducing severity of the disease. In this paper, we have reviewed the progress made to date on the parasite-host immunology and on vaccine development including its prospects, and discussed possible strategies in the formulation of vaccine(s) against neosporosis.

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The protective effect of an alpha-toxoid vaccine of Clostridium septicum purified alpha-toxin was investigated in guinea pigs. Purified alpha-toxin was treated with formalin to make toxoid, and alpha-toxoid vaccine was prepared by mixing alpha-toxoid (4 to 64 μg/dose) with an aluminum phosphate gel as adjuvant. Guinea pigs were immunized twice with different doses of alpha-toxoid vaccine, and challenged with spores of C. septicum. The guinea pigs surviving after challenge had been immunized with 8 μg/dose or more of alpha-toxoid. All these animals produced titers of 20 units or higher of antitoxin at the challenge. The results suggest that C. septicum alpha-toxin plays an important role in protection against challenge with spores in guinea pigs.

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The in vitro susceptibilities of 285 isolates of Escherichia coli from preweaned and postweaned pigs with diarrhea and edema disease were tested with the 15 commonly used antimicrobial drugs by an agar dilution minimal inhibitory concentration procedure according to National Committee for Clinical Laboratory Standards (NCCLS) guidelines. All E. coli isolates tested in this study belonged to enterotoxigenic E. coli, attaching and effacing E. coli, or Shiga toxin-producing E. coli. Field isolates had low MIC₉₀ for ceftiofur (1 μg/ml). No correlation in antimicrobial resistance was found in three types of E. coli.

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Reduced feed intake near parturition is suggested to be one of the major causal factors for the development of fatty liver in cows, and nonfeeding has been used as an experimental model for fatty liver. In cows with fatty liver, concentrations of lipoprotein lipids and proteins are decreased. In addition, the acute-phase protein haptoglobin is induced. The purpose of the present study was to examine whether the decrease of lipoprotein concentrations and the induction of acute-phase proteins were similarly reproduced by nonfeeding. Holstein female calves (n=5) were nonfed for 3 days and thereafter refed. Serum concentrations of nonesterified fatty acids and β-hydroxybutyric acid were initially increased by the nonfeeding, and followed by decreases in concentrations of cholesteryl esters, phospholipids, apolipoprotein (apo) B-100 and apoA-I. The apoC-III concentration was not distinctly decreased. Haptoglobin and serum amyloid A were induced during the nonfeeding and refeeding process. Haptoglobin was distributed in different proportions in the high-density lipoprotein, very high-density lipoprotein and the lipoprotein-deficient fractions, whereas almost all serum amyloid A was associated with the high-density lipoprotein fraction. These results suggest that the decreases in lipoprotein concentrations and induction of acute-phase proteins found in cows with fatty liver and those with fatty liver-related diseases such as ketosis are primarily due to the reduced feed intake near parturition.

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Lactoferrin with a molecular mass of 80 kDa was purified from equine seminal plasma by heparin-Agarose affinity chromatography and Sephacryl S-200 gel filtration. Purified lactoferrin was found to be highly homogeneous on the bases of its migration as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and of the monospecificity of rabbit antibodies to the purified protein in immunoblotting of seminal plasma proteins. A sandwich enzyme-linked immunosorbent assay was developed for quantifying lactoferrin in equine seminal plasma. Seminal plasma lactoferrin concentrations in 23 normal stallions ranged from 42 to 453 μg/ml, with a mean value of 157 ± 118 μg/ml (S.D.).

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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.

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The objectives of the present study were (1) to establish and maintain catheters non-surgically in the portal and hepatic veins of cattle for long-term monitoring of postprandially absorbed metabolites, and (2) to assess the safety of the technique used. Aspiration and catheterization were not performed if there was risk of bleeding, as determined by hemostatic data and enlargement of portal and hepatic veins. In five Holstein cattle, before and 60-min after feeding, percutaneous ultrasound-guided aspiration of portal and hepatic blood, together with jugular blood, was performed utilizing 17G × 17.5 cm needles. Concentration of ammonia nitrogen tended to be higher after feeding, but that of urea nitrogen and glucose were not significantly changed. In five other Holstein cows liver veins were catheterized with 14 × 16.5 cm guide-needles attached with 50 cm polyethylene tubes and blood samples were collected from portal, hepatic and jugular veins at 30-min intervals for 4.5 hr. The concentration of plasma acetate and 3-hydroxybutyrate (3-HB) were significantly increased after feeding, while that of lactate was not changed. Catheters in the portal and hepatic veins enabled the collection of blood samples up to 15 days (mean, 7 days). After a 7-day observation period, five cows were euthanised and necropsied. Changes in the intercostal musculature, peritoneum and liver were negligible. During the observation period, clinical profiles, aspartate aminotransferase, alanine aminotransferase and total and differential WBC counts were within normal ranges.

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Serum alkaline phosphatase (ALP) isoenzymes were studied in normal dogs using a commercially available polyacrylamide gel disk electrophoresis kit (PAG/disk kit). Serum samples taken from the dogs were incubated with neuraminidase, after which most showed ALP isoenzymes as two characteristic stained bands. To determine the origin of each band, ALP isoenzymes of serum and tissue extracts (liver, intestine and bone) were characterized by heating, wheat germ agglutinin (WGA) and levamisole treatments. The results suggested that the band detected on the anode was liver ALP (LALP) and that the band detected on the cathode represented bone ALP (BALP), and both were corticosteroid-induced ALP (CALP). The percentage of each ALP isoenzyme to total ALP activity was estimated by densitometry. The percentage of BALP was the highest in young dogs (age<1 year, 64.7% ), and this value decreased with age. In contrast, the percentage of LALP in young dogs (22.2%) was much lower than that in middle-aged dogs (ages 1 year to 7 years, 59.3%) and old dogs (ages>7 years, 50.4%). The present results suggested that a commercially available PAG/disk kit is capable of detecting three serum ALP isoenzymes in dogs, and further that it may have clinical applications in the evaluation of ALP isoenzymes in veterinary medicine.

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In veterinary medicine, dopamine is currently being administered clinically by infusion for treatment of kidney disorders at low doses (≤3 μg/kg/min) and for assessment of hemodynamics at high doses (≥5 μg/kg/min). However, since high doses of dopamine cause peripheral vasoconstriction due to its effect on alpha adrenoceptors, high doses have no longer been recommended. The present study was conducted to explore possible regimens for the use of dopamine infusion in dogs. The regional (renal and cardiac) blood flow for 60 min was measured by using colored microspheres at three doses (3, 10 and 20 μg/kg/min) of dopamine infusion in healthy anesthetized mongrel dogs. The effects on kidney and peripheral hemodynamics at each dose and the resultant cardiac output, mean arterial blood pressure and total peripheral resistance were determined. Renal blood flow increased markedly at 3 μg/kg/min dopamine. Improvement in hemodynamics indicated by marked increase in cardiac blood flow, cardiac output and mean arterial blood pressure and decreased total peripheral resistance was observed at higher doses (10 and 20 μg/kg/min). At 10 μg/kg/min, in addition to the satisfactory increase in cardiac blood flow, there was also a stable satisfactory increase in renal blood flow. However, at 20 μg/kg/min, increased myocardial oxygen consumption (manifested by marked increased in cardiac output), arrythmia and irregular increase in renal blood flow were detected. This study suggests that the clinical use of dopamine infusion in dogs could be safely expanded to moderately higher doses.

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The effects of non-specific immunotherapy with anti-allergic peptides extracted from the urine of human allergic patients (MS-antigen^TM), in two dogs with allergic dermatitis (AD) have been described. Clinically, severe pruritus accompanied by secondary bacterial pyoderma did not respond to conventional therapy with systemic antibiotics. The first clinical change appeared as a significant reduction in pruritus within 3 months, around the time of the 15th injection in both cases. The clinical condition was stabilized after 5 months, allowing the gradual withdrawal of concurrent therapies and an increase of injection intervals. The correlation between the results of intradermal skin tests before and after treatment and the improvement of clinical signs was not obvious.

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Proliferation of chondrocytes from nucleus pulposus (NP) and anulus fibrosus (AF) was confirmed in three-dimensional culture using alginate microspheres. Cells isolated from NP and AF were incorporated in microspheres and cultured for 14 days. Round mononuclear cells of 20-25 μm in diameter proliferated and formed aggregates. At day 14, alcian blue positive matrix surrounded the proliferating cells. The cells had cytoplasmic vacuoles stained positively by toluidine blue. On electron microscopy, the cells contained proteoglycan vacuoles and lipid droplets in the cytoplasm and synthesized collagen fibrils and electron dense granules surrounding the cell. These features of the cells were characteristic for chondrocytes. This culture system should be useful to further investigate metabolic activities of intervertebral disk chondrocytes.

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A female Japanese Black cow showed hemorrhage under the mucous membrane of the eye conjunctiva and the vagina, and thrombocytopenia in peripheral blood. The clinical manifestations repeated spontaneously during long-term observation. The clinical findings were stable except for severe periodic bleeding. Prothrombin time, activated partial thromboplastin time and fibrinogen levels were normal. The anti-platelet antibody in the serum was analyzed by indirect platelet coagulation tests. Because the platelets in the micro-plate wells of the subject series showed coagulation up to 1:128, it was judged that anti-platelet antibody was present in the serum. A number of megakaryocytes were recognized in smear samples obtained from bone marrow of the scapula immediately after euthanasia. Therefore, this case was diagnosed as idiopathic thrombocytopenia.

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Age-related changes in bone mineral density (BMD), cross-sectional area and strength strain index (SSI) of the long bones in the limbs and first lumbar vertebra of male Wistar rats were measured by a peripheral quantitative computed tomography (pQCT) method. One hundred and ten rats aged 2-30 months were used. The results indicate that the total (cortical + trabecular), cortical and trabecular BMD values of the metaphysis and cortical BMD values of the diaphysis in the long bones varied for each bone and differed from those of the first lumbar vertebra. The total BMD of long bones showed high values at 6-21 months and then decreased, but these did not always coincide with cortical and trabecular BMD. The values of SSI in the long bones varied. The values of total and cortical BMD and SSI of lumbar vertebra increased for 6-12 months and then decreased, but the trabecular BMD increased after 12 months. The total area in both the long bones and the first lumbar vertebra increased with the decrease in cortical area and the increase in the trabecular area with increasing age. It was concluded that age-related changes in bones, similar to those observed in humans, could be observed in some bones and parameters, although the age in rats when the so-called peak bone mass appears in the whole skeleton could not be clearly determined.

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The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is an animal model for obese type 2 diabetes. We showed that the OLETF rat exhibits higher levels of intramuscular fat content in Musculus longissimus as compared to the Fischer-344 (F344) rat. Our investigation was designed to identify quantitative trait loci (QTLs) contributing to the increased levels of intramuscular fat content by performing a whole-genome search using 108 F₂ intercross obtained by mating the OLETF and the F344 rats. We identified one QTL responsible for intramuscular fat accumulation on rat chromosome 1 with a maximum lod score of 3.4, which accounts for 5% of the total variance. As expected, the OLETF allele corresponds to the increased levels of intramuscular fat content.

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One 5-month-old female native Korean calf and a 2-year-old female Holstein cow raised in two farms about 4 km apart from each other in Korea, were found to have the left eye opaque, which included motile white worms in the aqueous humor. The parasite removed from the left eye of the calf was identified as Setaria digitata based on both light and electron microscopic features. The ocular infection with S. digitata reported herein may document the first aberrant case in Korean cattle.

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A retrospective study was performed on natural cases of postweaning multisystemic wasting syndrome (PMWS), recorded from January 1999 to December 2000, to determine the prevalence, microscopic lesions, and other coexisting pathogens associated with PMWS. PMWS is diagnosed based on three criteria: the presence of clinical signs (retardation of growth), characteristic microscopic lesions (granulomatous inflammation and inclusion body), and the presence of porcine circovirus (PCV)-2 within these lesions. One hundred and thirty three (8.1%) of the 1634 pigs submitted from 1243 pig farms were diagnosed for PMWS. The affected pigs were from 25 to 120 days old, the majority (78 cases, 58.6%) being 60 to 80 days old. PMWS occurred each month during the two-year study period, but the incidence peaked in May (38 cases, 28.6%), followed by April (18 cases, 13.5%) and June (13 cases, 9.8%). The most consistent and characteristic lesions were multifocal, granulomatous inflammation in lymph nodes, liver and spleen, characterized by infiltration of epithelioid macrophages and multinucleated giant cells. The majority of cases (113 cases, 85.0%) was dual infection with other pathogens. The combination of PCV-2 and Hemophilus parasuis (43 cases, 32.3%) was shown to be the most prevalent followed by PCV-2 and porcine reproductive and respiratory syndrome virus (39 cases, 29.3%). The consistent presence of PCV-2, but lower prevalence of other viral and bacterial pathogens in all pigs examined with PMWS, has led to the speculation that PCV-2 is the etiological agent causing PMWS.

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For the evaluation of canine gastric motility with ultrasonography, contraction number of pyloric antrum and gastric emptying time (GET) by area and volume method developed by Bolondi et al.'s method were studied in 14 dogs. All experimental dogs were administered with saline and soup solution (10 ml/kg, B.W.). The mean values of contraction number of pyloric antrum in saline and soup group were 4.19 ± 1.30/min and 4.82 ± 0.65/min before feeding, and overall mean values were 4.66 ± 1.37/min and 5.13 ± 1.71/min, respectively. The mean values of the GET by area and volume method were 36.73 ± 11.27, 40.00 ± 8.87 min in saline group and 61.35 ± 17.58, 59.11 ± 14.46 min in soup group. In the GET in saline and soup groups, there was no significant difference between the area and volume method (p>0.05). Therefore, Bolondi et al.'s method by ultrasound can be used to evaluate the antropyloric motility and gastric emptying time with area and volume methods. The area method is easier to determine the GET than the volume method, but the latter is more accurate.

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The induction of estrus during the non-breeding season was investigated in 100 Egyptian Baladi goats (Capra hircus). All animals assigned to treatments had low progesterone concentrations (<0.5 ng/ml) tested 2 times 10 days apart to confirm anestrous condition. Animals were assigned to three experimental groups. A group of animals received subcutaneous norgestomet ear implant for 11 days and a single i.m injection of PGF2α 24 hr before implant removal (group I; n=40). Second group of animals received subcutaneous norgestomet ear implant for 11 days and a single i.m injection of PGF2α 24 hr before implant removal and gonadotropin releasing hormone 24 hr after implant removal (group II; n=40). Third group of animals received no treatment (control group; n=20). The percentage of goats that showed estrous behavior during the first 72 hr after implant removal was 77.5, 85.0% and 10.0% in group I, group II and control group, respectively. The fertility rate was 57.5, 70.0% and 10.0% in group I, group II and control group, respectively. In conclusion, estrus can be induced in seasonally anestrous Egyptian Baladi goats using norgestomet and PGF2α and the injection of GnRH 24 hr after norgestomet implant removal synchronized ovulation in a higher percentage of goats.

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Four outbreaks of foot-and-mouth disease (FMD) occurred from March to May 2000 in Miyazaki and Hokkaido Prefectures, Japan. FMD virus isolation was achieved by sampling probang materials from Japanese Black cattle in the third case found in Miyazaki Prefecture. The probang materials were inoculated to bovine kidney (BK) and bovine thyroid cell cultures. CPE was observed in the BK at two days post-inoculation. Specific amplified DNA segments for FMD virus (FMDV) were detected by reverse transcriptase polymerase chain reaction in the culture fluid. The FMDV was identified as type O by enzyme-linked immunosorbent assay (ELISA) for antigen detection and the nucleotide sequence encoding the VP1 was determined. This FMDV is a strain that is widespread in Pan-Asia and was designated as O/JPN/2000 by the World Reference Laboratory of the Pirbright Institute, England. This report marks the first isolation of FMDV in Japan.

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