Mus musculus (M. m.) molossinus has been considered an independent subspecies of Mus musculus. To elucidate the evolutional origin of this subspecies, we carried out double-color FISH using 18s-28s ribosomal DNA and mouse chromosome paint probes. Among eleven rDNA loci detected, five loci on chromosomes 12, 15, 16, 18 and 19 were common to both Mus musculus (M. m.) musculus and M. m.molossinus and the other six loci, on chromosomes 1, 5, 10, 11, 13 and 17, were characteristic in M. m. molossinus. As M. m. molossinus is thought to originate from a hybrid between ancestral colonies of M. m. musculus and Mus musculus castaneus, we supposed that these six rDNA loci might have evolved after geographical isolation of the ancestral hybrid animals from M. m. musculus and M. m. castaneus.
Two commercial flocks of Chinese partridge experienced increased mortality associated with a wasting disease at 120-day old in June 2006. Postmortem examination of dead chickens consistently showed visceral tissues mainly proventriculus, liver and spleen were diffuse enlargement. Microscopic examination revealed masses of immature lymphocytes with frequent mitotic figures were seen in various tissues including proventriculus, liver, spleen, kidney, heart, lung, thymus and intestine. Reticuloendotheliosis virus (REV) was isolated from each of four blood samples. Viral antigens were observed in cultured CEF (SPF embryos came from the Ji-nan poultry institute) inoculated blood samples via on indirect immunofluorescent assay. Three hundred bp fragments of LTR of REV obtained from liver samples of six chickens by PCR. This disease has not previously been reported in Chinese partridge. Chinese partridge may represent a potential reservoir of infection for other Chinese local chickens.
A field isolate of Newcastle disease virus (NDV) isolated from northern pintail (Anas acuta) in Tohoku district, northeast Japan, was characterized. Phylogenetic analysis of the fusion protein indicated that the isolate belonged to genotype I and was closely related to isolates from the Far East corresponded to the migration route for this bird species. The isolate had the typical avirulent cleavage site of the fusion protein 112GKQGR*L117. In addition, pathogenicity tests indicated the isolate to have avirulent characteristics. However, the isolate has been shown to cause fusion cytopathic effects and form plaques on chicken embryo fibroblasts (CEF) in the absence of trypsin. The present results suggest that the CEF-adapted NDV, which is avirulent, is circulating among waterfowl populations.
We have investigated 37 minor serogroup Shiga toxin-producing Eschrichia coli (STEC) strains other than O157, O26, and O111 isolated from human specimens in Osaka prefecture to determine their serological and biochemical characteristics, virulence-associated genes, and clinical signs in patients. The same serotype strains were genotyped by pulsed-field gel electrophoresis (PFGE). The O antigen of 33 strains were typed into 10 serogroups; O28, O63, O65, O91, O103, O119, O121, O126, O165, and O177, and other 4 strains were not agglutinated with any serum. Four different Shiga toxin (Stx) types (1, 2, 2c, and 2f) were distributed in these isolates. The intimin gene was present in 83.8% of the strains and subtyped into intimin α, β, ε, and ζ. STEC O165, O177, and OUT isolated from hemolytic uremic syndrome (HUS) patients revealed atypical biochemical characters; negative reaction for lysine decarboxylase and gas production from glucose. Eleven strains including the isolates from HUS patients generated no colonies on cefixime-tellurite (CT)-sorbitol-MacConkey agar plates, since they showed high sensitivity (MIC ≤ 1.25 μg/ml) to potassium tellurite. The finding show supportive information for use the selective agar plates with and without CT supplement for the isolation of minor serogroup STEC. PFGE analysis revealed that the strains isolated from family cases were closely related within the respective family, and it was useful for epidemiological analysis of minor serogroup STEC.
Antimicrobial activities of 139 Enterococcus isolates (48 E. faecium and 91 E. faecalis) obtained from canine feces, boiler meat samples, swine feces, wild waterfowl feces, and human feces were examined against respective bacteria, including Streptococcuspyogenes, Staphylococcus aureus, Bacillus subtilis, Listeria monocytogenes, Salmonella Enteritidis, and Escherichia coli. Bacteriocin (BAC) production assay revealed that the antimicrobial activity against at least one of 6 indicator strains (BAC+ phenotype) was found in 51 (37%) isolates (29 E. faecium and 22 E. faecalis). Twenty-four of 46 isolates positive for at least one of the enterocin structural genes (entA, entB, entL50AB, and cylL) showed a BAC+ phenotype. The existence of other enterocins or nonenterocin factors was implied because the BAC+ phenotype was detected in a total of 27 Enterococcus isolates that had none of the enterocin genes tested. The antimicrobial activity against Gram-negative strains (Salmonella Enteritidis and E. coli) was detected in the 6 Enterococcus isolates that had either the entA, entB, entL50AB or cylL genes. Moreover, the proportion of the antimicrobial activity against L. monocytogenes among the cylL-positive E. faecalis isolates showing beta-hemolysis (10/16) was significantly (p<0.01) higher than among those lacking beta-hemolysis (2/15). The results suggested that certain characteristics are likely to be associated with the antimicrobial activity against specific organisms.
A total of 355 Murrah cross buffaloes, consisting of 23 subclinical and 332 clinical mastitis cases brought to the Veterinary Teaching Hospital, Chitwan, Nepal from 2002 to 2005, were analyzed to determine the organisms involved, the seasonal occurrence of mastitis, and antibiotic susceptibility of mastitis pathogens. Coagulase negative Staphylococci (CNS) such as Staphylococcus albus and S. epidermidis were the predominant organisms associated with subclinical cases, and CNS and Coliforms in clinical cases. The maximum number (16%) of clinical cases of mastitis were observed in the month of July, when temperature and humidity are highest. The incidence of clinical mastitis was higher in animals during 1st calving and during the 1st month of parturition. Resistance to antibiotics was determined for 55, 23 and 149 isolates of Staphylococcus spp., Streptococcus spp. and Coliforms, respectively. In vitro drug sensitivity testing revealed that enrofloxacin had the highest average sensitivity (91%) for all types of bacteria. The effectiveness of other drugs detected were gentamicin (87%), tetracycline (83%) and chloramphenicol (82%). The antibiogram showed that both gentamicin and enrofloxacin are slowly becoming resistant. Mastitis pathogens have developed resistance to ampicillin and penicillin.
Epidemiologic indicators associated with within-farm infection of Johne's disease in dairy farms in Japan were determined through a nationwide investigation of infected farms. We assumed that subsequent detection of the disease within one year after the first detection could represent the occurrence of within-farm spread occurring before the first detection. Of 594 infected farms, 158 farms (27%) had at least one additional detection. Logistic regression analysis using epidemiologic information obtained from infected farms at the time of the first detection revealed three epidemiologic indicators associated with subsequent detection. Farms at which the first cases included cattle with clinical signs were 3.8 (95% confidence interval: 2.2, 6.8) times more likely to have additional detections than those with cattle without clinical signs. Similarly, farms where two or more cattle were detected at the time of first detection and where cattle were held in a loose housing system were 2.8 (95% CI: 1.8, 4.5) and 2.0 (95% CI: 1.1, 3.6) times more likely to have additional detections than those where only one animal was detected and a tied-up housing system was used, respectively. These epidemiologic indicators are likely important determinants in the selection of farms requiring more intensive on-farm control measures.
The purpose of this study was to determine if Mycoplasma haemofelis, `Candidatus Mycoplasma haemominutum' exist in Korea. Three hundreds and thirty one feral cats were evaluated by using PCR assay targeting 16S rRNA gene sequence. Fourteen cats (4.2%) were positive for M. haemofelis, 34 cats (10.3%) were positive for `Candidatus M. haemominutum' and 18 cats (5.4%) were positive for both species. Partial 16S rRNA gene sequences were closely (>98%) related to those from other countries. This is the first molecular detection of feline hemoplasmas in Korea.
Supplementation with both cystine and glutamic acid increases the synthesis of glutathione (GSH), which has a marked effect on immune cell function, as compared with supplementation with either amino acid alone in human macrophages in vitro. As dietary glutamic acid is metabolized during intestinal transport, oral administration of L-theanine (γ-glutamylethylamide), which is metabolized to glutamic acid mainly in the liver, may act as a glutamic acid donor in vivo. The present study was performed to investigate the effects of oral administration of L-cystine and/or L-theanine on GSH levels and immune responses. Co-administration of L-cystine (200 mg/kg) and L-theanine (80 mg/kg) for 11 days before immunization significantly increased the levels of total GSH in the liver 6 hr after immunization as compared with the levels in control mice. To examine the effects of administration of L-cystine and/or L-theanine on the balance of T helper (Th) 1/Th2 cell responses, the serum ratios of the Th1 cytokine, interferon (IFN)-γ, and the Th2 cytokine, interleukin (IL)-10, were investigated. At 24 hr after immunization, co-administration significantly increased the IL-10/IFN-γ ratio compared with the ratios of the control and single-administration mice. Furthermore, co-administration before primary immunization significantly enhanced serum antigen-specific IgG levels. Taken together, these findings suggest that co-administration of L-cystine and L-theanine enhances antigen-specific IgG production partly through augmentation of GSH levels and Th2-mediated responses.
Rainbow trout (Oncorhynchus mykiss) head kidney phagocytes precultured with a synthetic cytidine-phosphate-guanosine (CpG) oligodeoxynucleotide (ODN) displayed significantly higher phagocytic activity agaist Vibrio ordalii than phagocytes precultured with non-CpG ODN. However, head kidney phagocytes precultured with CpG ODN did not show enhanced phagocytic activity against polystyrene particles.
Ferret polymorphonuclear cells (PMNs) and peripheral blood mononuclear cells (PBMCs) were separated from whole blood by density gradient centrifugation. Using a 50% Percoll solution (density=1.066), PMNs and PBMCs were successfully isolated after centrifugation; the purities of the PMNs and PBMCs were 94.2% and 95.6%, respectively. To evaluate the function of isolated ferret PMNs, we measured the superoxide generation with a MCLA-dependent chemiluminescence assay. The isolated ferret PMNs responded to phorbol 12-myristate 13-acetate (PMA) with kinetics similar to that of human PMNs. The ferret PMNs did not respond to N-formyl-Met-Leu-Phe (fMLF), unlike human PMNs, which rapidly responded. Thus, authors established a method for the rapid separation of highly purified populations of functional PMNs from the whole blood of ferrets.
An 11-month-old Japanese Black steer with chronic bloat underwent clinical and histological analyses. During the observation period, it showed normal appetite and fecal volume but persistent chronic bloat symptoms. Compared to controls, the steer's feces contained undigested large straws. Necropsy revealed normal rumen, reticulum, and abomasum but a small omasum. The rumen, reticulum, and abomasum mucosa was normal, with well-developed ruminal papillae. However, severe hypoplasia of the omasal laminae was observed along with hypoplasia reticular groove and ruminoreticular fold. The contents of the reticulum, omasum, and abomasums comprised undigested large sized hay particles. The omasum papillae showed no pathological abnormalities. This is a rare case of a steer with chronic bloat probably caused by severe hypoplasia of the omasal laminae.
Administration of immunosuppressive doses of glucocorticosteroids is the traditional primary treatment in necrotizing meningoencephalitis (NME) in dogs. However, response is variable and clinical signs often recur quickly with tapering dosage. Prognosis is poor and long-term therapy causes many complications. In the present study, we compared the long-term effects of combination (cyclosporine plus prednisolone) therapy with sole prednisolone therapy in management in dogs with NME. All NME cases in this study were examined with magnetic resonance imaging and cerebrospinal fluid analysis, and confirmed by histopathologic examination. The mean survival time of combination therapy group was 305.7 ± 94.7 days. The mean survival time of sole prednisolone therapy group was 58.3 ± 30.5 days. This case report demonstrates that combination treatment of cyclosporine with prednisolone is more effective in survival time than administration of only prednisolone in NME cases.
An 11-year-old female miniature schnauzer was tentatively diagnosed with the skull base meningioma, based on several examinations. Because surgical treatment was difficult, and outpatient radiation therapy was not available in the local area, chemotherapy with hydroxyurea combined with dexamethasone was selected. The patient's clinical symptoms improved after one week of treatment, and the tumor size was obviously reduced on MRI performed 37 days after treatment began. The patient received hydroxyurea for 7 months, with symptoms remaining stable, and the tumor re-increased to almost the same size at 7 months as that at the initial examination. At that time, hydroxyurea was discontinued. The patient died from pulmonary edema 14 months after treatment began. Pathologically, the tumor was diagnosed as a meningioma.
An 11-day-old Holstein calf presented with a high rectal temperature and tachypnea. Treatment with antibiotics and non-steroidal anti-inflammatory drugs did not improve the clinical signs. Bleeding tendency, with several hemorrhage spots on the body surface, appeared five days after admission. Severe pancytopenia was observed in the blood examination. The calf died on the 11th day after admission with severe bleeding from an injection site. Necropsy findings revealed that the pancytopenia had resulted from severe bone marrow aplasia. A congenital disorder was suspected to be the cause of pancytopenia associated with bone marrow aplasia.
Canine ganglioradiculitis (sensory neuropathy) was examined pathologically in two dogs (dog Nos. 1 and 2). The affected dogs had 1 and 2 years clinical courses from the onset, respectively. As common clinical signs, both cases showed progressive ataxia, difficulty in prehending food, visual deficit, and several sensory abnormalities. Gross observation after tissue fixation revealed whitish discoloration in the dorsal column of the spinal cords. The histological lesions were mainly distributed in the spinal dorsal roots, ganglions, and dorsal columns. In the spinal dorsal roots and ganglions, there were striking myelin loss, mild infiltration of mononuclear cells, and proliferation of small spindle cells. In the dorsal funiculus, there were moderate to severe diffuse myelin-loss and axonal degeneration. Immunohistochemistry for substance P (SP) revealed marked reduction of SP-immunopositive granules in the spinal substantia gelatinosa of affected dogs. By immunohistochemistry, CD3-positive cells were observed in the dorsal roots of dog No. 2, while CD3-positive cells were rare in those of dog No. 1. In the spinal ganglion of dog No. 1 there were many CD3- and MHC class II-positive cells. By indirect immunofluorescence assay using sera from affected dogs, no autoantibodies against canine nerve tissues were detected. The clinicopathological features of the present cases are almost consistent with those in previous reports of canine sensory neuropathies, while the etiology remains unclear.
Colonic mucosal mast cells are implicated in the pathogenesis of visceral hypersensitivity associated with irritable bowel syndromes. This study was designed to investigate the roles of mucosal mast cells in development of an experimental visceral hypersensitivity induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS) in rats. TNBS, when injected into the proximal colon through laparotomy, produced a significant decrease in pain threshold of the distal colon to mechanical distention, indicating a visceral hypersensitivity. In the proximal colon that was directly insulted by TNBS, mucosal necrosis and extensive inflammatory cell infiltration were observed with concomitant increase in tissue myeloperoxide (MPO) activity. In the distal colon where distention stimuli were applied, the number of mucosal mast cells significantly increased following TNBS treatment, although neither mucosal injury nor increase in tissue MPO activity was observed. In an organ culture, spontaneous release of a mucosal mast cell-specific protease (RMCP-2) from the distal colon tissue of TNBS-treated rats was significantly larger than that of sham animals. Furthermore, TNBS-induced visceral hypersensitivity was significantly suppressed by subcutaneous pretreatment with a mast cell stabilizer doxantrazole in a dose-dependent manner. These findings suggest that prominent colonic mast cell infiltration associated with an enhanced spontaneous mediator release is responsible, at least partly, for development of visceral hypersensitivity induced by TNBS in rats.
Beraprost sodium (BPS) is an orally active prostacyclin analogue. The effects of BPS on the heart, including coronary circulation improvement, myocardial and vascular protection and anti-fibrosis effect on myocardium interstitium, have previously been demonstrated. However, the effects of BPS on hemodynamics, cardiac function and myocardial contractility in patients in the hypertrophic phase have not been clarified. Therefore, in the present study, the effects of BPS under long-term administration were investigated using the hypertension model of salt-sensitive Dahl rats. Six-week-old Dahl rats were divided into three groups, an 8% high salt diet group treated with BPS (BPS group), an untreated 8% high salt diet group (HHF group) and an untreated 0.3% low salt diet group (Control group), and observations were conducted until 17 weeks of age. In the BPS and HHF groups, the survival rates after 11 weeks of high salt diet intake were 87.5% and 47.1%, respectively (p<0.05). At 17 weeks of age, the atrial systolic peak velocity/early diastolic peak velocity and heart weight index of the BPS group decreased significantly compared with the HHF group (p<0.05). The HHF group exhibited significantly more severe myocardial fibrosis mainly in the endocardial layer of the left and right ventricles compared with the BPS and Control groups (p<0.05). In the present study, long-term BPS administration preserved diastolic function and prevented myocardial interstitial fibrosis in the non-compensatory phase. The results of the present study suggest that BPS is effective for treatment of hypertensive cardiac hypertrophy.
The present study examined factors associated with suckling, eating and postural behaviors (standing, lying and sitting) and the relationships between sow behaviors and reproductive performance. A commercial farrowing-to-finish farm with approximately 300 female pigs was visited eight times for 3 years to record sow behaviors using time-lapse video recorders for 24 hr. The lactational and postweaning reproductive performances of each sow were recorded using a recording software system. Statistical models were built to investigate the factors associated with each behavior. Regression analyses were performed to determine the associations between behavior measurements and adjusted 21-day litter weights (A21d-Wt), weaning-to-first-mating intervals (WMI) and occurrence of ≥ 2 dead piglets during lactation. The means of suckling intervals and frequency of postural changes of the 145 recorded sows were 47.5 ± 0.59 min and 86.8 ± 2.97, respectively. As parity increased from 1 to ≥ 4, the suckling interval and total standing duration increased (P<0.05). The number of nursing piglets and observation days 4 to 21 of lactation were not associated with the suckling intervals. A longer duration of lactation was associated with a longer total standing duration and a higher frequency of postural changes (P<0.05). Shorter suckling intervals were associated with heavier A21d-Wt (P<0.05). Each change in postural behavior increased the likelihood of occurrence of ≥ 2 dead piglets during lactation (P<0.05). No postural or suckling behaviors were associated with WMI. Eating behavior was not associated with any reproductive performance. In conclusion, sow behaviors during lactation were related to lactational performance but were not related to postweaning reproductive performance.
The proportions of Sertoli cell tumor (SCT), seminoma and Leydig cell tumor in 50 dogs with unilateral testicular tumors were 52%, 36% and 12%, respectively. The rate of occurrence of SCT in the cryptorchid testis was very high (71%). The testicular superoxide dismutase (SOD) activity, testicular heat shock protein (HSP) 70 concentration and peripheral blood plasma inhibin (INH)-α concentration of 10 dogs with a unilateral cryptorchid testis and no testicular tumors, 10 dogs with SCT in a unilateral cryptorchid testis and 10 normal dogs, all aged 5-15 years, were measured in order to identify high risk factors for the occurrence of SCT in the canine cryptorchid testis. The mean SOD activity in cryptorchid testes and SCTs was significantly lower and higher, respectively, than in normal testes (both P<0.01). The mean HSP 70 concentration in both cryptorchid testes and SCTs was significantly higher than in normal testes (both P<0.01). The mean plasma INH-α concentration of the cryptorchid and SCT dogs was significantly lower and higher, respectively, than in normal dogs (P<0.05 and 0.01, respectively). The low SOD activity in the cryptorchid testis, low blood plasma INH-α concentration of the cryptorchid dogs and high HSP 70 concentration in the SCTs may be related to the occurrence of SCT and tumor cell proliferation in canine cryptorchid testes.
We validated a time-resolved fluoroimmunoassay (TR-FIA) using the lanthanide element europium as a non-radiological tracer for measuring luteinizing hormone (LH) in porcine peripheral blood. The dose-response curve of the reference standard ranged from 0.2048 to 50 ng/ml. Good parallelism was noted between the LH standard and plasma sample. The profile of LH throughout the estrous cycle, assessed by daily blood sampling, was consistent with the previous findings obtained by radioimmunoassays (RIAs). Moreover, the secretory patterns of pulsatile LH in the follicular phase and the preovulatory LH surge were also similar to those obtained in previous RIAs. We conclude that TR-FIA can be used to measure LH levels in porcine blood, with practical and convenient applications.
The selection of sheep with scrapie-resistant PrP genotypes is one of the control measures for transmissible spongiform encephalopathies in ruminants. In this study, we investigated the frequencies of PrP genotypes in meat breeds in Japan. The nationwide surveillance revealed that nearly half of the Suffolk sheep, a major meat breed in Japan, carried scrapie-susceptible AQ/AQ and AQ/VQ genotypes. In addition, the VQ haplotype, which confers high susceptibility to scrapie within sheep, was also found in Poll Dorset sheep. A trial of selective breeding using sires with scrapie-resistant PrP genotypes AQ/AR and AR/AR could raise the ratio of scrapie-resistant sheep from less than 50% to 80% within 3 years. However, the use of sires with the AR/AR genotype and the selection of ewes would be required to achieve a higher ratio of scrapie-resistant sheep.