One hundred and sixty-six Staphylococcus aureus isolates from mastitic milk samples from different cows on 26 farms were investigated for staphylococcal enterotoxins(SEs) and toxic shock syndrome toxin-1(TSST-1) by polymerase chain reaction(PCR) and reverse passive latex agglutination assay(RPLA). SEs and the TSST-1 gene were detected in thirty-seven isolates based on a multiplex PCR; SEA was detected in 32 isolates, SEB in 3 isolates, SEC in 1 isolate, and SEA and the TSST-1 gene in 1 isolate. Of the 37 enterotoxigenic isolates, thirty-three isolates were enterotoxigenic according to RPLA, where 29 isolates produced SEA, 3 isolates produced SEB, and 1 isolate produced SEC. The enterotoxin-producing S. aureus isolates were further characterized by pulsed-field gel electrophoresis(PFGE). A macrorestriction analysis revealed 11 PFGE patterns. Among the 33 enterotoxigenic S. aureus isolates, 45.4% exhibited the same PFGE pattern I. Accordingly, although the enterotoxin-producing S. aureus isolates from bovine mastitis were genetically diverse, 1 common genotype prevailed on the farms, indicating that PFGE pattern I isolates may be the most disseminated in Korea.

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The prevalence of enterohemorrhagic Escherichia coli (EHEC) O157 was examined in bovine faeces. EHEC O157 was isolated from the faeces of 42 (13.0%) of 324 cattle. Of the 4 farms and the facilities tested, the 3 farms and the facilities were found positive for EHEC O157. The highest isolation rate among the farms was 33.7%. The prevalence of EHEC O157 in heifers was higher than that in calves and other cattle. No cattle positive for EHEC O157 showed any clinical signs except 2 calves with diarrhea in a veterinary hospital. Almost all isolates possessed the stx gene, and Stx-positive strains carrying both stx₁ and stx₂ genes were predominant. These results indicate that EHEC O157 are distributed in bovine faeces, and that dairy and beef farms in selected regions of Japan are heavily contaminated with the organisms.

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The feces, colonic contents, and colonic mucosa from two beagles and a mongrel with and without mucohemorrhagic diarrhea were anaerobically cultured and after 72 hr, lucent and rough colonies with weak β-hemolysis were observed. Small spirochetes with one or two loose waves in the colonies were observed under a phase contrast microscope and the spirochete cells sometimes aggregated. The 16SrDNA sequencing results demonstrated that the canine isolates were B. pilosicoli and the base alignments registered in DDBJ. This is the first report concerning the isolation of B. pilosicoli from dogs in Japan.

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A brief period of ischemia, i.e. preconditioning of the middle cerebral artery territory, induces ischemic tolerance reducing the cerebral infarction volume caused by subsequent lethal ischemia. Nevertheless, little is known about the molecular mechanisms underlying this phenomenon. In the present study, we examined the involvement of the activation of Akt, a serine/threonine kinase, in the cerebral ischemic tolerance. Western blot analysis showed that Akt was activated in both non-preconditioned and preconditioned groups after ischemia for 1 hr, but the activation was long-lasting in the preconditioned rats. Immunohistochemical analysis demonstrated that the preconditioning-induced preventive effect on a rapid decrease in the activation level of Akt was due to the persistent activation of Akt in the penumbra region. In addition, TUNEL staining demonstrated that the preconditioning treatment inhibited the augmentation of neuronal death probably through apoptosis in the penumbra region to prevent the spread of infarction. Since the activation of Akt has been reported to protect cells from stress, the present results suggest that the preconditioning-induced persistent activation of Akt in the penumbra region plays an important role in ischemic tolerance of the brain.

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Thoracic radiographs of fifteen beagles with mild-to-moderate obesity revealed that oppression of the thoracic cavity increased with increasing degree of obesity. Oppression of the thoracic cavity was evaluated based on the length, depth, width and area of the thoracic cavity. To obtain thoracic radiographs at the terminal inspiration and expiration phases, thoracic fluororadiographs were recorded with a digital video camera. Bodyweight and the depth of the back fat layer at the seventh lumbar vertebra (DB, measured by ultrasonography) were used as indicators of the degree of obesity. The length of the thoracic cavity tended to become shorter and the depth and width of the thoracic cavity tended to increase as bodyweight increased and as DB increased. On the other hand, the area of the thoracic cavity was not clearly related to bodyweight or DB. These results suggest that oppression of the thoracic cavity due to the cranial shift of the diaphragm is compensated for by increases in the depth and width of the thoracic cavity in beagles with mild-to-moderate obesity.

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The expression of constitutive endothelial nitric oxide synthase (eNOS) and inducible NOS (iNOS) in the brains of cattle with natural rabies was studied. Increased expression of eNOS was detected in neurons of the brain stem and Purkinje cells of cerebellum. By contrast, iNOS was diffusely localized in the cytoplasm of affected neurons, and some inflammatory cells were positive. eNOS and rabies antigen were co-localized in inclusion bodies (Negri bodies) in neurons. The specific localization of eNOS, but not iNOS, in the Negri bodies suggests that eNOS is involved in the formation of rabies virus inclusion bodies.

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Effectiveness of vaccine that used recombinant R7 protein (rR7) as antigen that is derived from second-generation schizont (2GS) of Leucocytozoon caulleryi was verified under a field condition against chicken leucocytozoonosis. Chickens reared in a poultry farm where the chickens are attacked by leucocytozoonosis in every year were inoculated with oil-adjuvanted rR7 vaccine (O-rR7), and the immunized chickens were found to have production of antibodies against 2GS at a high level by one shot. Leucocytozoonosis was observed at post-injection. During the epidemic period of leucocytozoonosis, the unique clinical signs of the disease such as discharge of green feces and anemia, and also parasitemia were observed, however, compared to chickens in control group, those in O-rR7 vaccinated group had significantly slight symptoms (P<0.05). In addition to this, immunized chickens had better result of egg production than the unvaccinated chickens did, and the maximum difference of egg production rate, 22%, was observed at the peak of the disease. In conclusion, it is verified that O-rR7 vaccine has efficacy against leucocytozoonosis under field condition, and this vaccine can be put into practical use.

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A three-month-old female Siberian tiger cub with hindlimb ataxia was referred to the veterinary teaching hospital of Konkuk University. The patient was fed only beef without supplementation of calcium and vitamins after weaning. The tiger was presented with ataxia and back pain on digital palpation. In addition, abnormal gait, reluctance to move, and depressed withdrawal reflex were noted at the neurological examination. The overall osteodystrophic change of the lumbosacral vertebrae was observed on the lateral and ventrodorsal view of radiographic examination. And also PTH level was increased in hormonal assay when compared to that of cat reference range. Based on the results of examinations, nutritional secondary hyperparathyroidism was diagnosed. Clinical signs of this patient were improved after administration of vitamin D and calcium. This case demonstrates that nutritional hyperparathyroidism could be occurred in wild animals raised on a meat diet containing imbalanced calcium and phosphate.

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The canine caspase-3 gene was cloned and sequenced. The canine caspase-3 cDNA clone was 1473 base pairs in length and encoded 277 amino acids. The predicted amino acid sequence showed 88.4%, 88.0%, 85.9%, 65.9% and 60.6% homology with that of human, pig, mouse, chicken and zebra fish caspase-3, respectively. The caspase-3 mRNA was confirmed to express in skin, lymph node, bone marrow, small intestine and lung from a healthy dog by RT-PCR analysis.

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The incidence of DIC in 208 dogs with a malignant tumor was evaluated. The incidence of DIC was 9.6% in dogs with a malignant tumor which was a solid tumor in all. In 164 dogs with a malignant solid tumor, the incidence of DIC was 12.2%. The incidence of DIC in dogs with hemangiosarcoma, mammary gland carcinoma and adenocarcinoma of the lung was significantly higher than that in dogs with other malignant tumors. These results suggested that special care in looking for DIC should be taken in dogs with a malignant solid tumor.

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Alimentary lymphoma was evaluated endoscopically in 7 dogs and a histopathological examination was made to detect the origin of neoplastic lymphocytes in 5 dogs. A solitary mass in the rectum (1 case), irregular cobblestone appearance in the duodenum (4 cases) and a moderate irregular appearance resembling lymphocytic-plasmacytic enteritis (2 cases) were endoscopically detected. Endoscopic ultrasonography demonstrated increased thickness of the duodenal wall in 2 cases examined. Neoplastic lymphocytes of alimentary lymphoma proved to originate in T cells in all 5 cases examined by immunohistochemical analysis.

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A 10-year-old spayed mongrel dog was referred with repeated intercurrent hematochezia and anal bleeding. The dog was vigorous and had a normal appetite, and the fecal test showed no abnormal signs. Despite treatment primarily with sulfasalazine, the condition did not improve and unilateral blindness developed. A Prototheca zopfii infection was identified by further examination with bowel culture on Sabouraud's agar without cyclohexane and antibiotics. Subsequent to a vision loss in the other eye, the dog died showing signs of neurological disorder.

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Several peripheral blood microfilaria concentration methods that use Acetone (Acetone test), 2% formalin (modified Knott method), 5% Tween 20 solution, distilled water, 1% or 0.1% SDS were compared for their efficacy in detecting Setaria digitata microfilaria in cattle. The Acetone test was found to be more efficacious than the modified Knott method or the 5% Tween 20 solution test for detecting the S. digitata microfilaria in bovine blood. However, besides the Acetone test, the modified Knott method was also found to be suitable for Dirofilaria immitis microfilaria detection in dogs. SDS and distilled water were found not to be effective as hemolytic agent for the disruption of the red blood cell of both the cattle and dogs. Thus, the Acetone test is recommended for the primary screening of microfilaremia of S. digitata in cattle.

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The aim of this study was to detect immunohistochemically means growth hormone (GH) in 24 hepatoid gland adenomas and 5 hepatoid gland carcinomas and to compare the difference of immunoreactivity between types of tumors. The tumors were classified according to the WHO standards. Tissue sections which were prepared from formalin-fixed, paraffin wax-embedded tissues from 25 male and 4 female dogs were carried out immunostaining using polyclonal primary anti-hGH and EnVision method. Of 24 hepatoid gland adenomas (perianal gland adenomas) 23 (95.8%) were positive. All 5 hepatoid gland carcinomas (perianal gland carcinomas) were positive. No statistically significant differences in percentage of labelled cells between malignant and benign tumors were seen. The present demonstration of GH in hepatoid gland tumors adds new data on GH in extra-pituitary tissues and hormon-dependent tumors.

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A survey of proliferative enteritis (PE) in pigs at a meat processing plant was conducted using polymerase chain reaction (PCR) testing methods. During the investigation period, 227 of 83,717 pigs brought to the meat processing plant from Iwate, Fukushima, Miyagi, Niigata, and Yamagata Prefectures displayed characteristic general pathological features in terminal ileum, including mucosal hypertrophy and reticulation of serosal surface. Of these, 179 cases were further examined in the laboratory. All cases displayed characteristic histopathological features, and the specific band of the Lawsonia intracellularis (Li) causative agent of PE in pigs was detected in 155 cases by PCR testing methods. These results suggested a general infiltration of Li in the Tohoku district.

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The effects of xylazine or medetomidine on tiletamine/zolazepam (Zoletile^®) anesthesia were evaluated by changes in the canine electroencephalogram (EEG). Experimental groups were three: the group treated with 10 mg/kg of Zoletile intravenously (TZ), the group treated with 1.1 mg/kg of xylazine intramuscularly and 10 mg/kg of Zoletile intravenously (XTZ), and the group treated with 30 μg/kg of medetomidine intramuscularly and 10 mg/kg of Zoletile intravenously (MTZ). EEG recording electrode was positioned at Cz, which was applied to International 10-20 system. For all recording times, the powers of each band (band 1: 1-2.5 Hz, band 2: 2.5-4.5 Hz, band 3: 4.5-8 Hz, band 4: 8-13 Hz, band 5: 13-20 Hz, band 6: 20-30 Hz, band 7: 30-50 Hz, band 8: 1-50 Hz) were calculated. In TZ, at 10 min after Zoletile injection, the powers of bands 3, 4, 5 and 8 significantly decreased. At 20 min after Zoletile injection, the powers of band 1 and 8 were significantly decreased. After Zoletile injection, there were significant decreases in bands 1, 4, 5, 6, 7 and 8 in XTZ, and in bands 1, 3, 4, 5, 6, 7 and 8 in MTZ. These significant changes disappeared in all band powers in TZ and MTZ in the last 1 min just prior to the dogs showing head-up movement. But, in XTZ, all band powers, except band 2, were significantly reduced. EEG power spectral analysis revealed the differences in band powers on awakening, even though the same kind of drugs were used. We thought that was a useful method to compare the effect of xylazine and medetomidine on Zoletile-induced anesthesia in dogs.

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A three-month-old male Golden Retriever had symptoms including exercise intolerance, dyspnea and syncope and was diagnosed with subvalvular aortic stenosis. Cardiac catheterization revealed a left ventricular-aortic systolic pressure gradient of 90 mm Hg. Surgical correction of the condition was achieved using cardiopulmonary bypass. The subvalvular fibrous lesion was resected through an aortotomy. The stenosis was dilated from 8.5 mm to 12.0 mm in diameter. Postoperatively the dog was asymptomatic. Seven months after surgery, the pressure gradient decreased to 44 mm Hg. However, after another three months, the dog died suddenly without any premonitory signs. Postmortem examination revealed that pathologic changes caused by increased left ventricular pressure overload were not severe.

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The objective of the present study was to retrospectively investigate causes of variation on litter size at birth (total number of piglets born per litter (TB) and number of piglets born alive per litter (BA)) of Landrace (L) and Yorkshire (Y) sows in swine nucleus herds in Thailand. The data included sows farrowed during a four-year period from January 1998 to December 2001. The analyzed data set included observations on 8020 litters from 2199 L sows and 6919 litters from 1680 Y sows. Analysis of variance (ANOVA) was applied for statistical analyses using General Linear Mixed Model (MIXED) procedure of SAS. No breed difference was found for both TB and BA. Farrowing months significantly influenced TB and BA (P<0.001). Sows farrowed in August and September had a lower BA than sows farrowed from November to June (P<0.05). Effect of farrowing months on both TB and BA was more pronounced in primiparous compared with multiparous sows. Average minimum daily temperature during gestation negatively correlated with both TB and BA, average maximum daily temperature during gestation negatively correlated with BA and average daily humidity during gestation negatively correlated with both TB and BA. The correlations were stronger in L than in Y sows.

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Ejaculated semen and cross sections of the cauda epididymides collected from 3 normal dogs were smeared or stamped on glass slides, and the sperm on the slides were stained with 7 different FITC-lectins (Con A, DBA, GS-1, PHA-E, PSA, UEA-1, WGA) to examine the relation between sperm-binding glycoprotein derived from the canine prostate and sperm capacitation. The only lectin that stained the ejaculated sperm but not the cauda epididymal sperm was PHA-E. The sperm ejaculated by 5 other dogs were incubated for 4 hr in fluid flushed from the uterine horns or oviducts of estrous bitches, and then the percentages of actively motile sperm and hyperactivated sperm (HA-sperm) were determined. The percentages of PHA-E-labeled sperm and sperm labeled with fluoresceinated Ca indicator to assess the influx of Ca into the sperm were also evaluated. The mean percentages of actively motile sperm, HA-sperm, and Ca-labeled sperm after 4 hr of incubation in the uterine flush fluid and oviductal flush fluid were significantly higher than in control medium (P<0.05, 0.01), but the mean percentages of PHA-E-labeled sperm were lower (both P<0.01). The percentages of PHA-E-unlabeled sperm correlated with the percentages of both HA-sperm and Ca-labeled sperm (r²=0.787 and 0.812, respectively). The results indicate that loss of the glycoprotein secreted by the canine prostate on the sperm surface induces the influx of Ca into the sperm, and then hyperactivation of the sperm.

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A novel repeated sequence specific to male cattle was identified and named S4. S4 is a highly repetitive sequence and is a 1.5 kb repeating unit that contains various internal repeated sequences. FISH analysis showed that S4 is localized on the whole long arm and the proximal region of the short arm of the Y chromosome. We found that a PCR primer set for S4 amplified a male-specific 178 bp product in addition to a 145 bp product common to both male and female cells. Although the origin of the 145 bp product is unknown, it acts as a positive internal control in practical embryo sexing. Due to the high copy number of S4, PCR required only 0.5 pg purified DNA for accurate amplification. This made it possible to reduce the amount of biopsy sample required for embryo sexing and thus result in less damage to embryos manipulated. These studies indicate that embryo sexing based on the S4 sequence is accurate and sensitive.

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In an effort to understand the genetic diversity of porcine circovirus type 2 (PCV2) and the prevalence of PCV2 infection in Taiwanese herds, we have sequenced the complete genomes from PCV2-infected specimens and individually measured the antibody titer against PCV2 from pigs reared in Taiwan between the years 2000 and 2002. A total of 623 specimens originating from pigs displaying varied clinical signs were screened with the polymerase chain reaction (PCR). Results showed that 309 pigs (49.6%) tested positive for PCV2. Eight of the positive specimens were used for the amplification of the complete viral genome. Sequence comparison of the complete genomes indicated that the 8 Taiwanese PCV2 isolates shared 95-99% similarity. Phylogenetic analysis of all 40 PCV2 isolates from North America, Europe, Asia and Taiwan revealed that those isolates were grouped together in one large group containing two minor subgroups. The Taiwanese PCV2 isolates were classified into the two minor subgroups. The prevalence of serum antibodies to PCV2 in pigs was investigated, and results showed that approximately 83.5% of the pigs in Taiwan were seropositive. Finishing pigs possess the highest titers of antibodies, while 9-week-old pigs contained the lowest titers for specific antibodies. Our results suggest that PCV2 infections have become common in Taiwanese pig farms.

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Surveillance for bovine spongiform encephalopathy (BSE) in fallen stock in Japan is conducted with a commercial enzyme-linked immunosorbent assay (ELISA) for mass screening, with Western blotting (WB) and immunohistochemistry performed for confirmation of the ELISA. All tests are based on immunological detection of an abnormal isoform of the prion protein (PrP^Sc) in brain tissues, which have sometimes deteriorated by the time samples from fallen stock reach a diagnostic laboratory. To evaluate BSE surveillance procedures for fallen stock, we examined PrP^Sc detection from artificially deteriorated BSE-affected bovine brain tissues with a commercial ELISA kit and compared the results with those of WB. The optical density (OD) values of the ELISA decreased with advancing deterioration of the tissues, whereas no reduction in the signal for PrP^Sc was observed in WB, even when performed after 4 days of incubation at 37°C. The progressive decrease in the OD values in the ELISA appear to be caused by a partial loss of the N-terminal moiety of PrP^Sc due to digestion by endogeneous and/or contaminated microbial enzymes, and by the presence of ELISA inhibitors that are generated in deteriorated tissues. These results suggest that WB is the most reliable test for fallen stock, especially for cattle brains within decaying carcasses.

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Herpesvirus papio 2 (HVP2), which infects baboons, is much more closely related genetically and antigenically to monkey B virus (BV) than to human herpes simplex virus 1(HSV1) and other related herpes viruses. The usefulness of HVP2 as an alternative test antigen in immunoblotting assays to detect BV-antibody in macaque monkey sera was assessed. Six HVP2 proteins reacted with BV-positive sera in immunoblotting. No specific bands could be detected with BV-negative sera. These results show the usefulness of HVP2 antigen as an alternative and safer antigen than authentic BV antigen in detecting BV antibody in immunoblotting.

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Four swine facilities located in Northern Luzon, in the Philippines, showed lesions and clinical signs similar to those described in post weaning multisystemic wasting syndrome (PMWS). Post-mortem and histopathological examination revealed the presence of typical lymphoid lesions caused by porcine circovirus type 2 (PCV2). An in situ hybridization (ISH) technique allowed the detection of PCV2 nucleic acid associated with the lesions in target tissues. PCV2 DNA from paraffin embedded tissues was extracted, purified and sequenced. The phylogenetic analysis of the sequences obtained confirmed their identity, and grouped them into a PCV2 subgroup, together with some Canadian, French and Dutch isolates. This report is the first description of the presence of PMWS in the Philippines.

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The complete nucleotide sequences of the S1 and N genes of three Japanese and one Taiwanese field strains of IBV are reported. These Japanese strains were found to have S1 sequences most similar to those of Australian strains and N sequences most similar to those of North American strains. This result suggested that these Japanese strains might all be recombinant viruses derived from recombination of Australia- and North America-related viruses. Moreover, the S1 proteins of all these Japanese and Taiwanese strains exhibit only a limited sequence homology to strains of Massachusetts and Connecticut serotypes that have been commonly used as vaccine strains. This result high lightens the importance of development of vaccines based on the local strains of IBV.

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One of the author names of this article Dr. Seung-Chun PARK, was left out. Wrong:KWON Oh-Deog¹⁾²⁾³⁾, YIM Chang-Yeol⁴⁾, JEONG Kyu-Shik³⁾, JUNG Kyu-Yong⁵⁾, McGREGOR John R.¹⁾²⁾⁶⁾, BASTIAN Neil R.²⁾, SAMLOWSKI Wolfram E.¹⁾²⁾⁶⁾

1) University of Utah Cancer Immunotherapy Program
2) Department of Internal Medicine (Oncology and Infectious Disease) of the University of Utah School of Medicine
3) College of Veterinary Medicine, Kyungpook National University
4) Department of Internal Medicine (Hematology/Oncology) of the Chonbuk National University Medical School
5) Department of Pharmacology, Wonkwang University School of Medicine
6) Huntsman Cancer Institute
Right:KWON Oh-Deog¹⁾²⁾³⁾, YIM Chang-Yeol⁴⁾, JEONG Kyu-Shik³⁾, PARK Seung-Chun³⁾, JUNG Kyu-Yong⁵⁾, McGREGOR John R.¹⁾²⁾⁶⁾, BASTIAN Neil R.²⁾, SAMLOWSKI Wolfram E.¹⁾²⁾⁶⁾

1) University of Utah Cancer Immunotherapy Program
2) Department of Internal Medicine (Oncology and Infectious Disease) of the University of Utah School of Medicine
3) College of Veterinary Medicine, Kyungpook National University
4) Department of Internal Medicine (Hematology/Oncology) of the Chonbuk National University Medical School
5) Department of Pharmacology, Wonkwang University School of Medicine
6) Huntsman Cancer Institute

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