Previously, we observed that electroacupuncture (EA) at ST36 (Zusanli) and GV20 (Baihui) enhanced cell proliferation and neuroblast differentiation in the rat dentate gyrus. In this study, we investigated the possible mechanisms of EA in this effect. For this, we applied EA at ST36 and GV20 of Wistar rats (13-week-old) once a day for 3 weeks. Application of EA at these acupoints significantly increased the number of phosphorylated cyclic AMP response element-binding protein (pCREB)-immunoreactive cells in the dentate gyrus. In addition, EA significantly increased the levels of brain-derived neurotrophic factor (BDNF) and pCREB protein in the dentate gyrus. The administration of K252a, an inhibitor of BDNF receptor, significantly reduced cell proliferation in the subgranular zone of dentate gyrus. These results suggest that EA significantly increased neuroblast plasticity via pCREB and BDNF activation in the dentate gyrus.
We previously reported the development of an inactivated oil-adjuvanted avian influenza vaccine using an apathogenic H5N1 strain of the same lineage as the Eurasian lineage viruses currently epidemic in Asia. In this study, we confirmed the safety and evaluated the efficacy of this vaccine in layer chicken farms by field trials. No problematic adverse reactions occurred in the safety test. In addition, no adverse effects were observed in the field trial, and the antibody titer exceeded a protective level (hemagglutination inhibition (HI) antibody titer of 16) at 3 weeks after a single injection. Based on the above findings, this vaccine was confirmed to be safe and induced a protective level of antibody titer with a single injection in the chickens at the farms.
The aim of this study was to evaluate the genotypic characteristics of Staphylococcus aureus isolates (n=170) from bovine milk collected from seven dairy farms in Italy. On the basis of cultural and biochemical properties and by amplification of the 23S rRNA specific to S. aureus, all isolates were identified as S. aureus. To genotypically characterize S. aureus isolates, genes encoding virulence determinants (nuc, clfA, spa-IgG-binding, spa-X-region, fnbA and fnbB, cap5 and cap8) and staphylococcal enterotoxins (sea, seb, sec, sed, see, seg, seh, sei, sej) were investigated using a PCR technique. The results showed that the isolates of S. aureus in each farm had the same genotypic characteristics, while the isolates genotipically differed between the different farms. The present study might help to understand the distribution of prevalent S. aureus strains in dairy farms.
Molecular epidemiology analyses of the 36 clinical isolates of Pasteurella multocida from various avian hosts in Japan between 1976 to 2007 including 5 reference strains from the U.S.A., Taiwan and Indonesia were performed by employing the single-enzyme amplified fragment length polymorphism (SE-AFLP) comparison with the classical ApaI-based pulsed-field gel electrophoresis (PFGE). As the results, SE-AFLP gave 21 profiles while PFGE gave 20 profiles. The Simpson's index of diversity analysis indicated that SE-AFLP gave a high discrimination power than PFGE. This concluded that SE-AFLP is a higher discrimination power than PFGE to differentiate avian P. multocida isolates in Japan. In addition, the genetical profiles suggested that there is the evolution of somatic serotype 3 strain in the indigenous host of Japan.
Avian pathogenic Escherichia coli (APEC) serogroup O78 isolates in Japan were characterized using multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE). Thirty-four of the serotype O78 isolates were clustered into 11 PFGE types with 80% similarity and were classified into 5 sequence types (STs), ST23, ST117, ST155, ST369 and 1 novel ST (ST1645). The most dominant ST was ST23 (54.3%). Fluoroquinolone-resistant strains belonged to ST23 (9 strains), ST155 (3 strains) and ST117 (1 strain). The combined findings of MLST and PFGE suggested that 1 genotype belonging to ST23 was the genotype specific to fluoroquinolone-resistant strains. These results indicate that the combination of MLST and PFGE is useful for conducting epidemiological studies on APEC O78 strains.
Hemoplasma infections in wild ungulates have not been reported yet in Japan. We examined presence of hemoplasmas in blood samples collected from 147 sika deer (Cervus nippon) in the Iwate prefecture by real-time PCR, and found 13 (9%) were positive. Almost entire region of the 16S rRNA gene of the representative strains from positive samples was amplified by conventional PCR. The nucleotide sequences of the 16S rRNA gene were further determined and compared with those of other hemoplasmas. Our examinations 1st revealed the presence of 2 distinct hemoplasma species in sika deer, which are previously not described. One of them was closely related to M. ovis by the 16S rRNA sequence analysis, but was found distinct by comparison of the RNase P RNA gene sequences. Pathogenicity of these two hemoplasma species in sika deer is currently unknown.
The effects of aflatoxin B1 (AFB1), aflatoxin M1 (AFM1), deoxynivalenol (DON) and zearalenone (ZEA) on the viability, chemiluminescent (CL) response and expression of cytokine mRNA of bovine neutrophils (PMNs) were evaluated. The opsonized zymosan (OPZ)-stimulated CL response of PMNs was significantly (P<0.05) decreased by AFB1 (>50 pg/ml), AFM1 (>50 pg/ml) and ZEA (>50 pg/ml). The phorbol myristate acetate (PMA)-stimulated CL response PMNs was significantly (P<0.05) decreased by AFB1 (>0.5 pg/ml), AFM1 (>50 pg/ml), ZEA (>500 pg/ml) and DON (>5 pg/ml). Treatment with AFB1 resulted in reduction in the mRNA expression of interleukin-1β and tumor necrosis factor-α of PMNs stimulated with OPZ and PMA. These results suggest that these four mycotoxins have inhibitory effects on the function of bovine PMNs.
Interleukin-1beta (IL-1β) plays a significant role in the onset and pathogenesis of inflammation in mammalian hosts. Although well characterized in a range of vertebrate species, little is known about this important cytokine in marsupial mammals. We report here the molecular cloning and characterization of IL-1β in the tammar wallaby (Macropus eugenii). M. eugenii IL-1β has an open-reading frame of 813 nucleotides, coding for a putative protein of 270 amino acids to the termination codon. The IL-1 family motif and potential caspase cleavage site (necessary for production of the mature protein) is also present in the sequence. Molecular characterization of tammar wallaby IL-1β provides fundamental information necessary to progress the study of functional immune responses in this unique group of mammals.
The present study aimed to objectively evaluate the adverse events after the administration of chemotherapeutic agents used in the University of Wisconsin (UW)-Madison chemotherapy protocol (UW-25) for canine lymphoma, using the Veterinary Co-operative Oncology Group common terminology criteria for adverse events (VCOG-CTCAE). The medical records of 40 dogs with multicentric high-grade lymphoma that underwent UW-25 were reviewed. Gastrointestinal adverse events of grade 2 and above and blood/bone marrow adverse events of all grades were evaluated. Gastrointestinal adverse events occurring at least once during the entire period of UW-25 were observed in 50% (20/40), 17.9% (7/39), and 8.1% (3/37) of the dogs after the administration of vincristine (VCR), cyclophosphamide (CPA), and doxorubicin (DXR), respectively. Blood/bone marrow adverse events occurring at least once during UW-25 were observed in 57.5% (23/40), 41% (16/39), and 8.1% (3/37) of the dogs after the administration of VCR, CPA, and DXR, respectively. The rate of patients that experienced gastrointestinal adverse events was higher after the first administration of VCR than after the first administration of DXR. Findings obtained in this study will be helpful in predicting the adverse events that could occur when dogs with lymphoma are treated with UW-25.
There has been a need for improvement of the elimination diet used for diagnosis of adverse food reaction (AFR) in dogs. Recently, a novel elimination diet composed of a mixture of amino acids and potatoes was developed. We evaluated the efficacy of the elimination diet for diagnosis of AFR in dogs. Twenty dogs that were suspected to have allergic dermatitis were enrolled in a 2-month food elimination trial using the diet. Before and after the trial, the clinical symptoms were evaluated based on the change in canine atopic dermatitis extent and severity index (CADESI), pruritus score and medication score. Of the 20 dogs, 15 completed the food elimination trial. The remaining 5 dogs were removed from the trial because of diet unpalatability, skin disease progression or diarrhea. On the basis of evaluation of the clinical scores, we observed that the clinical symptoms improved in 11 of the 15 dogs that completed the food elimination trial. Provocative challenge was performed in 10 of the 11 dogs that showed improvement in their clinical symptoms. Of the 10 dogs, 7 were diagnosed as having AFR against food ingredients such as pork, beef, chicken and wheat because their skin symptoms reappeared after intake of these ingredients. The results of the food elimination trial and the provocative challenge indicated the usefulness of the novel elimination diet for diagnosis of AFR.
The aim of this research was to evaluate the changes in the response of pattern-stimulated visual evoked potential (pVEP) with different pattern size, and demonstrate visual acuity from the minimum visual angle. pVEP was recorded from both eyes of six healthy beagles. Prior to pVEP recording, the dogs were sedated, and a traction fiber was used to prevent the eye from rolling down. The stimulator was set 30 cm from the subject's eye. Pattern reversal frequency of the stimulating monitor was 3 rev/sec, and pattern size was set at seven levels; 14-364 arc-min (1.2-31.4 mm). Amplitude of the P100 component was evaluated, and visual acuity was calculated from the minimum visual angle to obtain a pVEP response. A pVEP response (2.3-3.1 μV) was obtained from all subjects. The P100 component was detectable in 3 eyes with a check size of 14 arc-min and 7 eyes with 28 arc-min, and the component was undetectable with 14 arc-min in all subjects in which it was undetectable with 28 arc-min. From the minimum level to obtain the P100 component, the subjects' visual acuity was extrapolated as 0.54-2.14 cycles per degree. We demonstrated the change in P100 component with check size. However, our technique was inadequate to examine visual acuity because the subject's refractive index was ignored. We suggest that, with further study, pVEP with different check sizes would be applicable for canine visual acuity examination.
The effect of sugar supplementation with 1 g/kg BW twice a week for eight weeks on rumen protozoa was determined in ten retarded growth calves. Rumen juice was sampled by abdominal paracentesis during the experiment. Papillae development of rumens excised by experimental laparotomy was macro- and micromorphologically determined before and after sugar supplementation in a selected calf. The numbers of Entodinium, Isotricha, Dasytricha and Epidinium protozoa increased by 3 to 12 folds after 1-3 wk of supplementation and subsequently decreased. The heights of the rumen papillae after sugar supplementation showed marked development compared with before supplementation (Post vs. Pre: 4.44 ± 0.43 vs. 1.36 ± 0.24 mm). Sugar supplementation accommodates the rumen protozoa profile and stimulates papillae development in retarded growth calves.
A novel canine epidermal keratinocyte cell line, MSCEK, was developed from skin of a healthy dog. The aim of this study was to determine its expression of desmosomal components and to evaluate its use as a detection tool for circulating autoantibodies in canine pemphigus. Immunofluorescence and western blotting analyses revealed that MSCEK expresses desmoglein (Dsg) 1, Dsg2, Dsg3, desmoplakin, plakoglobin and cytokeratins. Moreover, positive fluorescent reactions on the surface of MSCEK cells were observed when the cells were incubated with sera obtained from four dogs diagnosed with pemphigus complex. These findings indicate that MSCEK should be a useful tool for future research to characterize circulating autoantibodies that recognize desmosomal components in dogs with pemphigus.
Two dogs of juvenile-onset skin diseases with involvement of extremities were examined by histopathological, immunohistochemical and ultrastructural analyses. Clinically, both cases showed alopecia and crusts on the face and extremities. Case 1 showed histopathology of dermo-epidermal separation. Ultrastructural analysis revealed that the clefts were recognized between hemidesmosomes and lamina densa. In case 2, histopathology showed follicular atrophy, vacuolar degeneration at lower epidermis and masseter muscle degeneration, without remarkable ultrastructural abnormalities in basement membrane zone of the skin. Thus, the findings in case 1 were compatible to those in junctional epidermolysis bullosa, while those in case 2 were compatible to dermatomyositis-like disease. Combination of histopathological and ultrastructural analyses was useful to distinguish the diseases in two dogs.
In mammals with a hemochorial placenta (e.g., primates and rodents), the maternal and fetal bloodstreams are separated by the blood-placenta barrier. However, a few maternal cells in the general circulation pass through the barrier during normal pregnancy. So far, the transfer mechanism has not been investigated. In this study, we established a chemokine (C-C motif) ligand 3 (CCL3)-deficient mouse model to examine the effect of fetus-derived chemokine(s) on the migration of maternal cells through the blood-placenta barrier. Using this model, we obtained CCL3-positive and -negative littermates from a mother expressing both CCL3 and green fluorescent protein (GFP). The numbers of GFP positive maternal cells in the lung, liver, spleen and heart of CCL3-positive and -negative fetuses were compared. A few GFP-positive cells were detected in the lung and liver of both types of fetus. These results indicate that maternal cells can migrate through the blood-placenta barrier even in the absence of fetal CCL3.
An alkaline-based chemical antigen retrieval pretreatment step was used to enhance immunolabeling of disease-associated prion protein (PrPSc) in formalin-fixed and paraffin-embedded tissue sections from cattle naturally affected with bovine spongiform encephalopathy (BSE). The modified chemical method used in this study amplified the PrPSc signal by unmasking PrPSc compared with the normal cellular prion protein. In addition, this method reduced nonspecific background immunolabeling that resulted from the destruction of the residual normal cellular form of prion protein, and reduced the treatment time compared with the usual autoclave pretreatment step. Immunolabeled PrPSc was thereby clearly detected in the myenteric plexus of the ileum in naturally occurring BSE cattle.
A case of intracranial cholesterol granuloma is described in a 4-year-old neutered European male cat presented with a 5-month history of progressive weakness, ataxia and depression. On clinical evaluation, haematological and biochemical profiles revealed only mild hypercholesterolemia and magnetic resonance imaging showed a large space-occupying extra-axial mass in the area of the falx, not homogeneous after contrast enhancement. At post-mortem examination, an orange-yellowish mass of 22 mm in diameter extended from the right frontal lobe to the temporo-parietal region, causing atrophy of the prosencephalic region of the brain. The site of origin of the mass was within the subarachnoid space of the supracallosum sulcus of the right cerebral hemisphere. Histological examination of the lesion revealed abundant deposits of cholesterol clefts, surrounded by clusters of macrophages and multinucleated giant cells. Neither inflammatory lesions, nor cholesterol deposits were detected in other areas of the brain and in other organs. On the basis of the histological examination, a diagnosis of intracranial cholesterol granuloma was made.
Two 60-day-old pigs showing clinical signs of malgrowth and diarrhea were diagnosed as atypical porcine proliferative enteropathy (PPE). The intestinal mucosal lesions in the piglets were characterized by the adenomatous proliferation of the crypt epithelium together with growth of small curved bacteria within the enterocytes. The lesions could be seen in the ileum and other portions of the intestine histologically, although no significant thickening of the gut wall could be observed grossly in the present case. The macroscopic findings are extremely important for the diagnosis of PPE, however, this paper shows that the histopathological and/or immunohistochemical findings were also critical to identify the disease.
The cerebellar lesions of three dogs with canine neuroaxonal dystrophy (NAD), one dog with cerebellar cortical abiotrophy (CCA), and 4 dogs with neuronal ceroid-lipofuscinosis (NCL) were examined to understand their pathogeneses. Purkinje cell loss was most severe in the vermis of a dog with CCA, and granule cell loss was most prominent in the cerebellar hemisphere of dogs with NCL. Immunohistochemically, CD3-and HLA-DR-positive cells were most frequent in the dogs with NCL, and moderate in dogs with NAD, but not in a dog with CCA. The number of cleaved caspase 3-positive cells was prominent in a dog with CCA, but no significant in the dogs with NAD. The results indicate different pathway of neuronal loss of these canine neuronal disorders.
A variety of chemotherapeutic drugs, e.g., etoposide and bleomycin, are widely used in clinical practice to treat many types of animal malignancies. In the clinical situation, cellular resistance to chemotherapy is a significant component of tumor treatment failure. A variety of DNA repair factors, e.g., Ku80, might be a key contributor to chemoresistance to anticancer agents. In both cancer and normal cells, Ku80 plays a key role as a sensor of DNA double-strand break (DSB) induced by treatment with some chemotherapeutic drugs. Although the localization and mobility of Ku80 play a key role in regulating the physiological function of Ku80, it is not clear whether those of Ku80 are affected after treatment with chemotherapeutic drugs. We examined the localization and mobility of Ku80 in living hamster cells with or without DSBs, which were induced by treatment with chemotherapeutic drugs. Our data showed that Ku80, in contrast to H2AX, is highly mobile in the nuclei. We found that before and after the induction of DNA damage by treatment with etoposide or bleomycin, a major portion of Ku80 is exchanged by the same kinetics in the nuclei of interphase cells. These results suggest that the mobility of a major portion of Ku80 is not affected by DNA DSBs in order to find other DSBs. In addition, the information would be worthy to develop some new chemotherapeutic drugs to treat many types of animal malignancies.
During the period of 2007-2008, a total of 270 pig fecal samples were collected from a meat processing plant located in southern Japan and examined for Salmonella species. A total of 44 Salmonella isolates were recovered, and antimicrobial resistance was detected in serotypes Typhimurium (n=9), Infantis and Choleraesuis (n=2), and Derby, Miyazaki and Schwarzengrund (n=1). Multidrug resistance was seen in serotypes Typhimurium (n=8) and Infantis (n=2). The most commonly observed resistance phenotypes were against streptomycin, oxytetracycline and sulfamethoxazole (100%), ampicillin (90%), chloramphenicol (50%), cephalothin (30%) and cefoxitin, ceftazidime and kanamycin (each 20%). Extended-spectrum cephalosporin-resistant Salmonella Infantis isolates producing plasmid-mediated, blaCMY-2 gene were detected. These AmpC-producing isolates showed resistance to ampicillin and cephems (cephalothin, cefoxitin and ceftazidime). Resistance transfer experiments showed that transconjugants and transformants coexpressed resistance phenotypes similar to the donor isolates. To the best of our knowledge, this is the first report worldwide describing serovar Infantis from pigs capable of producing AmpC β-lactamase. Then, we detected the pentadrug-resistance phenotype in Salmonella Typhimurium isolates, which yielded class 1 integron amplicons of 1.0 and 1.2 kb. Genetic fingerprinting analysis by pulsed-field gel electrophoresis and an assay by polymerase chain reaction confirmed the isolates to be Salmonella Typhimurium DT104. In conclusion, the findings of this survey call for the systematic and comprehensive domestic and international surveillance programs to determine the true rates of occurrence of AmpC-producing Salmonella both in the livestock and public health sectors.
The seroprevalence of Bartonella vinsonii subsp. berkhoffii was investigated in stray urban dogs and shepherd and farm guard dogs from rural areas sampled from 10 provinces of Turkey. Sera from 855 dogs were examined for the presence of anti-B. vinsonii subsp. berkhoffii antibodies by indirect fluorescent antibody test. Overall, 56 (6.6%) of the 855 dogs examined, including 16 (3%) of the 522 stray dogs and 40 (12%) of the 333 rural dogs, were seropositive. This is the first report on prevalence of antibodies to B. vinsonii subsp. berkhoffii in dogs in Turkey.
A 4-year-old beagle had intermittent vomiting and weight loss for 1 month. On plain radiography, an approximately 3.5-cm, radiopaque, linear foreign body was identified in the region of the liver. On ultrasonography, a hyperechoic linear structure with reverberation was identified in the left lobe or quadrate lobe. On computed tomography(CT), a thin hyperattenuating material consistent with a metallic foreign body was identified just medial to the gall bladder. Removal of the foreign body was performed without complications. We present a rare case in which a metallic foreign body was embedded in the liver asymptomatically and found incidentally during a clinical examination performed to ascertain the cause of clinical signs.
The effects of acute restraint stress on sperm motility and reproductive endocrinology were investigated in adult rats. Sperm motility was determined by computer-assisted sperm analysis. Acute restraint stress reduced sperm motility starting after 30 min, and the sperm motility parameters, percentage of motile spermatozoa (%), straight-line velocity, curvilinear velocity, deviation of the sperm head from the mean trajectory and the maximum amplitude of lateral head displacement decreased. It also induced a significant elevation in plasma adrenocorticotropic hormone, prolactin, corticosterone and progesterone and decreased follicle-stimulating hormone, luteinizing hormone, testosterone and immunoreactive (ir-) inhibin. These results clearly demonstrated that the acute restraint stress rapidly suppressed sperm motility and increased the activity of the hypothalamus-pituitary-adrenal axis, whereas it disturbed hypothalamus- pituitary-gonadal axis activity.