The nucleotide sequences of nucleocapsid protein (N); phosphoprotein (P); matrix protein (M); hemagglutinin-neuraminidase (HN); and large polymerase protein (L) genes, 3′-end leader, 5′-end trailer and intergenic regions of the avian paramyxovirus (APMV) strain goose/Shimane/67/2000 (APMV/Shimane67) were determined. Together with previously reported data on fusion protein (F) gene sequence [46], the determination of the genome sequence of APMV/Shimane67 has been completed in this study. The genome of APMV/Shimane67 comprised 16,146 nucleotides in length and contains six genes in the order of 3′-N-P-M-F-HN-L-5′. The features of the APMV/Shimane67 genome (e.g., nucleotide length of whole genome and each of the six genes, and predicted amino acid length of each of the six genes) were distinct from those of other APMV serotypes. Phylogenetic analysis indicated that although APMV/Shimane67 was grouped with APMV-1, -9 and -12, the evolutionary distance between APMV/Shimane67 and these viruses was longer than that observed between intra-serotype viruses. These results show that the genome sequence of APMV/Shimane67 contains specific characteristics and is distinguishable from other types of APMV.

The outbreak of Riemerella anatipestifer (RA) infection has been confirmed in meat-type domestic ducks (Anas platyrhynchos) for the first time in 27 years in Japan. In January 2014, increased mortality in a 14- to 21-day-old duck flock was reported to veterinary officials by the owner. The affected ducks exhibited reduced movement, ataxia and dorsal recumbency with leg paddling. Pathological findings were typical for an RA infection. Fibrinous and heterophilic pericarditis, airsacculitis, perihepatitis, ventriculitis and meningitis were observed. The bacterial isolate from duck organs was identified as RA by PCR-based 16S ribosomal RNA sequencing.

Brucella abortus cells were lysed by the N-terminal 24-amino acid fragment (GI24) of the 36-amino acid peptide PMAP-36 (porcine myeloid antimicrobial peptide 36). Next, the protection efficacy of the lysed fragment as a vaccine candidate was evaluated. Group A mice were immunized with sterile PBS, group B mice were intraperitoneally (ip) immunized with 3 × 10⁸ colony-forming units (CFUs) of B. abortus strain RB51, group C mice were immunized ip with 3 × 10⁸ cells of the B. abortus vaccine candidate, and group D mice were orally immunized with 3 × 10⁹ cells of the B. abortus vaccine candidate. Brucella lipopolysaccharide (LPS)-specific serum IgG titers were considerably higher in groups C and D than in group A. The levels of interleukin (IL)-4, IL-10, tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) were significantly higher in groups B–D than in group A. After an ip challenge with B. abortus 544, only group C mice showed a significant level of protection as compared to group A. Overall, these results show that ip immunization with a vaccine candidate lysed by GI24 can effectively protect mice from systemic infection with virulent B. abortus.

Tyrosol (Tyr) is a natural phenolic antioxidant with diverse biological activities. We compared the anti-inflammatory effects of intravenously administered Tyr versus prednisolone (PSL) in an endotoxin-induced uveitis (EIU) rat model. Intravenous administration of 100 mg/kg Tyr was performed 2 hr before, simultaneously and 2 hr after lipopolysaccharide (LPS) injection. Tyr treatment was associated with decreased inflammatory cell number, protein concentration, tumor necrosis factor (TNF)-α, PGE2 and NO levels in AqH and improvements in histopathologic evidence of EIU in ocular tissue at 24 hr after LPS injection. 100 mg/kg Tyr and 1 mg/kg PSL (administered on the same schedule as Tyr) had comparable anti-inflammatory effects. Taken together, Tyr may represent a promising therapeutic agent for the management of intraocular inflammatory diseases.

Pavlovian fear conditioning is an experimental procedure in which a conditioned stimulus (CS) acquires an ability to elicit fear responses. This type of conditioning depends on the basolateral complex of the amygdala (BLA) and/or central amygdala (CeA). We previously found that rats showed reduced fear responses to an auditory CS when they were subjected to a pre-training chemical lesion of the entire right amygdala as compared with the left amygdala. Based on this finding, we hypothesize that the BLA and/or CeA in the right hemisphere will be more strongly activated by an auditory CS than those in the left hemisphere. To test this hypothesis, we re-exposed fear-conditioned and non-conditioned rats to an auditory CS 1 day after fear conditioning. We assessed Fos expression in the BLA and CeA in each hemisphere. We found that fear-conditioned subjects showed fear responses, such as increased freezing and decreased walking, as well as increased Fos expression in the BLA and CeA. When we compared Fos expression between hemispheres, Fos expression in the CeA, but not the BLA, was greater in the right hemisphere compared with the left hemisphere. These results suggest that the right CeA is more strongly activated by the auditory CS.

Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically important disease around the globe. Protection against this virus remains problematic. Here, we evaluated antibody (IgG & IgA) inducibility of a heterologous PRRSV glycoprotein 5 (GP5) expressed in a live attenuated Bordetella bronchisepticaaroA mutant strain (BBS-GP5). Mice and pigs were primed with recombinant GP5 (rGP5) subcutaneously followed by boosting with live BBS-GP5. As a result, anti-GP5 IgG was induced in both mice (P<0.001) and pigs (P<0.1). Pigs were challenged with live PRRSV (VR2332). Viral RNA was found to be significantly (P<0.01) removed in the vaccinated pig group. Overall, BBS-GP5 is a good candidate as a live attenuated vaccine against PRRSV infection.

Effects of a bacterial probiotic (BP) on ruminal fermentation and plasma metabolites were evaluated in four Holstein cattle (body weight, 645 ± 62 kg; mean ± SD) with induced subacute ruminal acidosis (SARA). SARA was induced by feeding a SARA-inducing diet, and thereafter, 20, 50 or 100 g per head of a commercial BP was administered for 7 consecutive days during the morning feeding. Cattle without BP served as the control. The 24-hr mean ruminal pH in the control was lower, whereas those in the BP groups administered 20 or 50 g were significantly higher compared to the control from days 2 to 7. Circadian patterns of the 1-hr mean ruminal pH were identical (6.4–6.8) among all cattle receiving BP. Although the mean minimum pH in the control on day –7 and day 0 was <5.8, the pH in the treatment groups on day 7 was >5.8 and significantly higher than that of the control group ( >5.2). Ruminal volatile fatty acid (VFA) concentrations were not affected by BP treatment; however, the BP groups had lower lactic acid levels compared with the control group at 20:00 on day 7. Additionally, non-esterified fatty acid levels decreased from 8:00 to 20:00 in all BP groups on day 7. These results suggest that administration of 20 to 50 g of a multi-strain BP for 7 days might improve the low pH and high lactic acid level of the ruminal fluid in SARA cattle.

Persistent left cranial vena cava (PLCVC) is an uncommon congenital thoracic venous anomaly in dogs. This study examines the clinical and CT findings of dogs diagnosed with PLCVC incidentally. In this study, complete type of PLCVC was diagnosed in 26 dogs with CT angiography. Shih tzu (17 cases) and Pekingese dogs (3 cases) were overrepresented. There was no gender predisposition, and the average age at presentation was 10.3 years. Of 26 dogs, one dog had a bridging vein connecting right and left cranial vena cavae, and another dog showed azygos vein terminating PLCVC. On the thoracic CT images in the third dog, the right cranial vena cava was absent so that right brachiocephalic vein ended to PLCVC. However, the right costocervical vein drained another vein coursing caudally to the right atrium with azygos vein. In conclusion, CT angiography is a very useful method to diagnose PLCVC and variations of related thoracic vein anomalies in dogs.

This study aimed to investigate the potential effects of sildenafil on pulmonary hypertension (PH) in the monocrotaline (MCT)-induced PH rat. Twenty-four, 12-week-old, male Sprague-Dawley rats were injected with MCT or saline solution. After injection of MCT, rats received oral sildenafil immediately (early-phase treatment group: E group), 4 weeks after injection (late-phase treatment group: L group) or no treatment (MCT group) until 6 weeks after injection. Serial echocardiography and right ventricular systolic pressure (RVSP) measurements via a cardiac catheter were performed. RVSP was reduced in the E and L groups compared with the MCT group. Echocardiography indicated that sildenafil therapy prevents an increase in RVSP and preserves diastolic function, and this effect is not dependent on timing of initiation of therapy.

A 2-year-old female beagle was referred to our hospital for evaluation of anemia. Laboratory tests, including bone marrow cytology, revealed non-regenerative immune-mediated anemia (NRIMA). Although initial immunosuppressive multi-drug therapy was not effective, additional administration of danazol was successful in treating the anemia. However, hepatocellular carcinoma (HCC) developed about 20 months after the administration of danazol. In humans, several cases of development of HCC after the administration of danazol have been reported. The present report describes a case of HCC development in a dog after chronic administration of danazol in addition to other immunosuppressive drugs.

An epidemiological survey of Theileria annulata infection was undertaken in a cattle population in Rajshahi Division, Bangladesh. The local cattle breeds from the area (North Bengal Gray and Deshi) and crosses between the local breeds and Holstein cattle were predominantly screened. In total, 192 cattle serum samples were collected in two areas of Rajshahi Division, the Rajshahi District (n=147) and Natore District (n=45). The samples were screened with an enzyme-linked immunosorbent assay using T. annulata surface protein (TaSP) as the antigen. The seroprevalence was 80.0% (36/45) in Natore and 20.4% (30/147) in Rajshahi. A logistic regression analysis showed that the sampling location was significantly associated with seropositivity, whereas age, sex and breed were not. Although the logistic regression analysis did not show a linear dependence on age, we considered age-specific seroprevalence separately in the two districts. Seroprevalence did not differ significantly among age categories in the Natore District. In contrast, all the cattle <1 year old in the Rajshahi District were seronegative (11/11). Seroprevalence in the 1- and 2-year-old cattle was significantly lower in the Rajshahi District than in the Natore District. In the older age categories (3, 4 and >5 years), seroprevalence did not differ significantly between the Natore and Rajshahi Districts. These results suggest that the cattle in the Rajshahi District were sporadically exposed to T. annulata, whereas most cattle in the Natore District became infected during an early phase of life.

Antibodies to Encephalitozoon cuniculi (E. cuniculi) were examined by enzyme-linked immunosorbent assay using E. cuniculi PTP2 recombinant protein from serum samples that had been collected from a total of 295 cats in Japan. Of these samples, 6.1% (18/295) had antibodies against E. cuniculi, which included 6.3% (6/96) of the male cats and 6.0% (12/199) of the female cats. The incidence was slightly higher in feral cats (8.3%, 11/132) compared to domesticated cats (4.3%, 7/163). This suggests the possibility that the cats of our country have become a reservoir of E. cuniculi. This study is the first to demonstrate the prevalence of E. cuniculi infection in cats in Japan.

Two Cheviot ewes homozygous for the A₁₃₆L₁₄₁R₁₅₄Q₁₇₁ (AL₁₄₁RQ) prion protein (PrP) genotype were exposed intracerebrally to brain pools prepared using four field cases of atypical scrapie from the United Kingdom. Animals were clinically normal until the end of the experiment, when they were culled 7 years post-inoculation. Limited accumulation of disease-associated PrP (PrP^Sc) was observed in the cerebellar molecular layer by immunohistochemistry, but not by western blot or enzyme-linked immunosorbent assay. In addition, PrP^Sc was partially localized in astrocytes and microglia, suggesting that these cells have a role in PrP^Sc processing, degradation or both. Our results indicate that atypical scrapie is transmissible to AL₁₄₁RQ sheep, but these animals act as clinically silent carriers with long incubation times.

Methylmercury (MeHg) is a persistent environmental contaminant that has been reported worldwide. MeHg exposure has been reported to lead to increased risk of cardiovascular diseases; however, the mechanisms underlying the toxic effects of MeHg on the cardiovascular system have not been well elucidated. We have previously reported that mice exposed to MeHg had increased blood pressure along with impaired endothelium-dependent vasodilation. In this study, we investigated the toxic effects of MeHg on a human endothelial cell line, EA.hy926. In addition, we have tried to elucidate the role of myristoylated alanine-rich C kinase substrate (MARCKS) in the MeHg toxicity mechanism in EA.hy926 cells. Cells exposed to MeHg (0.1–10 µM) for 24 hr showed decreased cell viability in a dose-dependent manner. Treatment with submaximal concentrations of MeHg decreased cell migration in the wound healing assay, tube formation on Matrigel and spontaneous nitric oxide (NO) production of EA.hy926 cells. MeHg exposure also elicited a decrease in MARCKS expression and an increase in MARCKS phosphorylation. MARCKS knockdown or MARCKS overexpression in EA.hy926 cells altered not only cell functions, such as migration, tube formation and NO production, but also MeHg-induced decrease in cell viability and NO production. These results suggest the broad role played by MARCKS in endothelial cell functions and the involvement of MARCKS in MeHg-induced toxicity.

We investigated hematologic characteristics of healthy Akita dogs. All were found to contain glucose transporters, GLUT1 and GLUT4, in erythrocyte membrane, whereas Beagle and any other Western dogs have only GLUT4. Of 47 Akitas, ten showed high K and low Na concentrations with elevated glutathione (GSH) in erythrocytes due to Na,K-ATPase activity in the membrane (HK). Akitas showed increased capacity for recycling vitamin C or ascorbic acid (AA) from oxidized ascorbic acid (DHA) compared to Beagle dogs. Particularly, HK Akitas performed even greater AA recycling and ferricyanide reduction than normal Akitas which have normal GSH, low K and high Na concentrations (LK). All HK Akitas also had stomatin in erythrocyte membrane, while half of LK Akitas had it at lower levels than HK Akitas. Stomatin did not have any influence on AA recycling. GLUT1, Na,K-ATPase and stomatin in erythrocytes are characteristics of Akita dogs, and the high prevalence of these proteins suggests their positive roles in biological efficiency.

The prevalence of Yersinia enterocolitica (Y. enterocolitica) and Yersinia pseudotuberculosis was examined in 151 pet animals including 108 rodents, 39 rabbits and four sugar gliders from 13 pet stores in the Yamaguchi Prefecture, Japan. Y. enterocolitica serogroup O:3 biotype 3 negative for the Voges-Proskauer reaction (O:3/3 variant VP-) was isolated from five Djungarian hamsters (Phodopus sungorus) raised at the same pet store. These pathogenic Y. enterocolitica isolates carried the virulence genes, yadA, ail and virF, and were shown to be clonal by pulsed-field gel electrophoresis with NotI digestion. This is a first report of pathogenic Y. enterocolitica O:3/3 variant VP- in pet Djungarian hamsters in Japan.

The aim of this prospective, randomized clinical trial was to compare the total number of anesthetic interventions (TNAI) performed by the anesthetist to treat cardiovascular depression or arousal/movement episodes in dogs receiving intrathecal and general anesthesia (GA), maintained using propofol-based TIVA (group P) or isoflurane (group I). Mean arterial pressure (MAP) before (T₀) and 12 min after intrathecal anesthesia (T₁) and intraoperative vasoactive consumption were also compared. The TNAI to deepen the anesthetic plane or to treat hemodynamic depression in the pre-surgical and intra-surgical period was calculated in forty-two client-owned dogs randomly assigned to group P or I. Ten dogs for each group complied with the inclusion criteria and were analyzed. In pre-surgical period, the TNAI was higher in Group I [2 (0–5)] than Group P [0 (0–2)] (P=0.022), and ephedrine consumption was also higher in Group I [75 (0–200) µg/kg)] than Group P [(0 (0–50)] (P=0.016). MAP (mmHg) in Group P was 79 (66–95) at T₀ and 65 (59–86) at T₁ and 67.5 (50–73) and 57 (53–66) in Group I, respectively. At T₀ and T₁, MAP was higher in Group P (P=0.005 and P=0.006, respectively). No differences were found between the two groups in the intrasurgical period (P>0.05). This study shows that the GA protocol can have a relevant impact on the TNAI performed by the anesthetist in the pre-surgical period of anesthesia, to treat cardiovascular depression or arousal/movement episodes in dogs receving intrathecal anesthesia.