Colocalization of glucagon-like peptide (GLP)-1 with GLP-2 in L-cells was investigated in the chicken ileum by using double immunofluorescent and immunocytochemical techniques. Ultrastructural features of L-cells were also clarified in this study. L-cells showing immunoreactivity for both GLP-1 and GLP-2 were distributed in the whole ileum. They showed comma-like or flask-like shape and were located in epithelium of crypts and lower part of intestinal villi. L-cells showing GLP-1-immunoreactivity only were found in epithelium of lower and middle parts of intestinal villi. Transmission electron microscopy indicated that L-cells identified by colloidal gold-labeled immunocytochemistry were covered apically with microvilli, open-type and contained many secretory granules in their perikarya. These secretory granules without halo were round to oval in shape and showed moderate electron density. The longest and shortest diameters of secretory granules were 355 ± 62 nm (mean ± SD) and 287 ± 48 nm, respectively. Double labeling immunocytochemistry using two different sizes of particles (6 and 12 nm in diameter) of colloidal gold revealed that GLP-1 colocalized with GLP-2 in the same secretory granules. This study advances new morphological data about the endocrine system of the chicken small intestine.
Internal hernia in avian species is very rare. A necropsy of a 2-week-old SPF White Leghorn chicken revealed that a loop of the duodenum and part of the pancreas (4 × 2 × 1 cm) was protruding through the abnormal foramen (2.5 cm in diameter) in the right posthepatic septum into the right ventral hepatic peritoneal cavity. The herniated loop was located underneath the ventral hepatic surface, leaving a groove on the right hepatic lobe (2 × 1.5 × 0.4 cm). The part of the pancreas involved in the hernia was grossly enlarged. Microscopically, a zone of pressure atrophy of hepatic tissue was characterized by crowdedness of hepatocytes with pyknotic nuclei and faint eosinophilic cytoplasm and indistinct narrow sinusoids. The pancreas revealed hypertrophy of the acinar cells with an increase in the secretory granules and basophilic cytoplasm. This is the first report of duodenum herniation into the right ventral hepatic peritoneal cavity resulting in groove formation on the ventral hepatic surface in a 2-week-old chicken
Porcine edema disease (ED) is a communicable disease of shoats caused by infection with Shiga toxin (Stx)-producing Escherichia coli. Stx2e is classified as a 1A5B-type toxin and is a decisive virulence determinant of ED. The single A subunit of Stx2e possesses enzymatic activity and is accompanied by a pentamer of B subunits, which binds to the host receptor and delivers the A subunit into the cell. In the present study, we used a mouse model to evaluate the immunogenicity of 3 ED vaccine candidates: a non-toxic mutant holotoxin mStx2e and 2 Stx2eB-based fusion proteins, Stx2eA2B-His and Stx2eB-His. Systemic inoculation of mice with mStx2e- and the Stx2eB-derived antigens induced anti-Stx2e IgG responses that were fully and partially capable of neutralizing Stx2e cellular cytotoxicity, respectively. Intranasal immunization with mStx2e protected the mice from subsequent intraperitoneal challenge with a lethal dose of Stx2e, whereas immunization with Stx2eA2B-His and Stx2eB-His afforded partial protection. Analysis of serum cytokines revealed that mStx2e, but not the Stx2eB-based antigens, was capable of inducing a Th2-type immune response. These results suggest that although the Stx2eB-based antigens elicited an immune response to Stx2e, they did so through a different mechanism to the Th2-type response induced by mStx2e.
Retinol-binding protein 4 (RBP4) is a specific transporter of retinol and was recently identified as an adipokine potentially involved in type 2 diabetes in humans and rodents. However, the function and structure of feline RBP4 have not been reported. In this study, we describe the molecular cloning and expression analysis of feline RBP4. The complete feline RBP4 cDNA encodes a precursor protein comprising an 18 amino acid signal peptide and a 183 amino acid mature protein. Feline RBP4 was mapped to chromosome D2. Mature feline RBP4 is 83–94% homologous to the RBPs of humans, cows and rodents. RT-PCR analysis revealed feline RBP4 expression in liver and adipose tissues.
This study describes a case of tetralogy of Fallot in a lamb showing failure to thrive and signs of respiratory distress. Physical examination, electrocardiography, thoracic radiographies, echocardiography and cardiac troponin I evaluation were performed. The value of cardiac troponin I was compared with the values of 10 healthy lambs of the same age and breed, and the affected animal demonstrated an increase in cardiac troponin I. Due to the poor prognosis, euthanasia was indicated, and necropsy confirmed the diagnosis. This is the first report of an increase in cardiac troponin I in a lamb with tetralogy of Fallot.
Canine aggression is one of the behavioral problems for which veterinary behaviorists are most frequently consulted. Despite this, the classification of canine aggression is controversial, and there are several classification methodologies. While the etiology of canine aggression differs among the types of aggression, the behavioral background underlying aggression is not well understood. Behavior trait-based evaluation of canine aggression would improve the effectiveness and efficiency of managing canine aggression problems. We developed a questionnaire addressing 14 behavioral items and items related to four types of canine aggression (owner-, child-, stranger- and dog-directed aggression) in order to examine the associations between behavioral traits and aggression in Shiba Inu. A total of 400 Shiba Inu owners recruited through dog events (n=134) and veterinary hospitals (n=266) completed the questionnaire. Factor analysis sorted the behavioral items from both the event and clinic samples into four factors: “sociability with humans,” “reactivity to stimuli,” “chase proneness” and “fear of sounds.” While “reactivity to stimuli” correlated significantly positively with all of the four types of aggression (P=0.007 to <0.001), “sociability with humans” correlated significantly negatively with child- and stranger-directed aggression (P<0.001). These results suggest that the behavioral traits involved in canine aggression differ among the types of aggression and that specific behavioral traits are frequently simultaneously involved in several types of aggression.
Antibiotics continue to be used as growth promoters in the poultry industry. Honeybee (Apis melifera) venom (HBV) possesses a number of beneficial biological activities, particularly for regulating the immune system. The aim of the present study was to evaluate the immunoprophylactic effects of HBV against Salmonella Gallinarum in broiler chicks as an initial step towards developing eco-friendly alternatives to reduce antibiotic use. HBV was administered using a spray technique. HBV improved body weight gain, particularly in the presence of infection. Moreover, HBV enhanced antibody production activity against formalin-killed S. Gallinarum. The CD4+:CD8+ T lymphocyte ratio, relative mRNA expression levels of interleukin-18 and interferon-γ, and serum lysozyme activity also increased following HBV administration before the infection period as well as during infection. HBV reinforced bacterial clearance and increased survivability against S. Gallinarum. Corresponding pathological analyses demonstrated that the HBV-sprayed group displayed mild and less severe abnormal changes compared with those in the control group. It was presumed that the prophylactic effects of HBV against S. Gallinarum were associated with its non-specific immune response stimulating activity. Thus, HBV may provide an alternative to reduce antibiotic use in the poultry industry.
The present study evaluated the effect of hemoplasmosis on cattle productivity. Prevalence of bovine hemoplasma was examined by polymerase chain reaction (PCR) using whole blood samples collected from 93 breeding cows and their 71 calves in Hokkaido, Japan. Monthly milk production records and other clinical data were compared between Mycoplasma wenyonii (Mw)-infected, “Candidatus Mycoplasma haemobos” (CMh)-infected, co-infected and PCR-negative groups. Blood chemical parameters were obtained from the 93 cows and 64 calves. PCR results showed that 89.2% (83/93) of cows and 14.1% (10/71) of calves were positive for bovine hemoplasma. Based on productivity data obtained from the 93 cows, Mw-infected, CMh-infected and co-infected cows had significantly lower monthly milk yield compared to PCR-negative cows. Furthermore, decline in milk yield was prolonged in CMh-infected and co-infected groups. No significant differences were found for other clinical findings among the four groups. Calf birth weight tended to be lower for Mw-infected, CMh-infected and co-infected groups compared to the PCR-negative group. There were no significant differences in all blood parameters of cows and calves among the four groups. In addition, no significant differences were found in any parameter between hemoplasma-infected and PCR-negative calves.
We investigated various essential factors for optimum infrared thermography for cattle clinics. The effect of various factors on the detection of surface temperature was investigated in an experimental room with a fixed ambient temperature using a square positioned on a wall. Various factors of animal objects were examined using cattle to determine the relationships among presence of hair, body surface temperature, surface temperature of the eyeball, the highest temperature of the eye circle, rectum temperature and ambient temperature. Also, the surface temperature of the flank at different time points after eating was examined. The best conditions of thermography for cattle clinics were determined and were as follows: (1) The distance between a thermal camera and an object should be fixed, and the camera should be set within a 45-degree angle with respect to the objects using the optimum focal length. (2) Factors that affect the camera temperature, such as extreme cold or heat, direct sunshine, high humidity and wind, should be avoided. (3) For the comparison of thermographs, imaging should be performed under identical conditions. If this is not achievable, hairless parts should be used.
A locally bred, 12-year-old, intact female Satsuma dog presented with generalized alopecia. Erythema, crusts and desquamation were observed primarily on the truck. Papules and erosions were present in the pinnae, and there were multiple areas of skin necrosis on the right forelimb. The cutaneous lesions had not responded to treatment with systemic antibiotics and prednisolone. The dog also had progressive anemia. Babesia gibsoni was detected in the blood, and the dog was treated with antiprotozoal agents. The skin lesions and anemia improved, but relapsed after the treatment was discontinued. Histopathological examination of skin biopsies revealed findings suggestive of early leukocytoclastic vasculitis or ischemic vasculopathy.
A 12-week-old female Wire-haired miniature dachshund presented with non-progressive ataxia and hypermetria. Due to the animal’s clinical history and symptoms, cerebellar malformations were suspected. Computed tomography (CT) and magnetic resonance imaging (MRI) detected bilateral ventriculomegaly, dorsal displacement of the cerebellar tentorium, a defect in the cerebellar tentorium and a large fluid-filled cystic structure that occupied the regions where the cerebellar vermis and occipital lobes are normally located. The abovementioned cystic structure and the defect in the cerebellar tentorium were comparable to those seen in humans with Dandy-Walker syndrome. However, the presence of the cystic structure in the occipital lobe region was unique to the present case. During necropsy, the MRI findings were confirmed, but the etiology of the condition was not determined.
The intestinal helminth fauna of Tibetan sand foxes (Vulpes ferrilata) and red foxes (Vulpes vulpes) inhabiting in Qinghai, China, was evaluated by conducting necropsy of hunted foxes and fecal egg examination of field-collected feces. In northeast and south Qinghai, 36 foxes were necropsied, and the species of foxes and the parasites detected were identified by the DNA barcoding. In 27 red foxes and 9 Tibetan sand foxes examined, Mesocestoides litteratus (total prevalence: 64%), Toxascaris leonina (50%), Taenia pisiformis (8%) and Taenia crassiceps (8%) were found in both species of foxes. Echinococcus shiquicus (8%) and Taenia multiceps (6%) were found only in Tibetan sand foxes. Echinococcus multilocularis (3%) and Alaria alata (8%) were found only in red foxes. In the fecal egg examination of the rectal feces, 100% of taeniid cestodes, 73% of Toxascaris and 27% of Mesocestoides worm-positive samples showed egg-positive, indicating that coprological survey for parasite eggs could only provide partial information of intestinal parasite fauna. For field-collected feces, molecular identification of feces origins and fecal egg examination were performed. In 15 Tibetan sand fox and 30 red fox feces, we found E. multilocularis eggs in one feces of Tibetan sand fox. The present study indicated that the upper intestinal helminth fauna of the two fox species in Qinghai does not differ significantly and both species would play an important role in the maintenance of taeniid cestodes.
This study aimed to investigate intestinal helminth infection in stray dogs in Osaka Prefecture by surveying coprological samples from dogs captured from 2006–2011. Of 212 fecal samples collected, overall prevalence of infection was 39.2%. The most common species was Toxocara canis (25.0%), followed by Trichuris vulpis (8.0%), Spirometra erinaceieuropaei (3.3%), Taeniidae (2.4%), Ancylostoma caninum (1.9%) and Toxascaris leonine (0.5%). In the molecular analysis, all of the taeniid eggs were negative for Echinococcus multilocularis and were identified as other taeniid species (e.g., Taenia pisiformis). Our results suggest that stray dogs remain important infection reservoirs of zoonotic parasites in Osaka Prefecture. Therefore, control of stray dogs is crucial for reducing the risk of public health problems due to parasitic infections.
Rodent models of diabetes develop a slowing of nerve conduction velocity and mild axonal atrophy, but generally lack overt degenerative neuropathy. Spontaneously diabetic Wistar Bonn Kobori (WBN/Kob) rats develop severe diabetic peripheral motor neuropathy with a slowing of nerve conduction velocity. We examined the effect of glycemic control, using insulin implant, on neuropathic changes in these rats. Animals were divided into 2 groups: WBN group (spontaneously occurring diabetes rats) and WBN + insulin group (spontaneously occurring diabetes rats treated with insulin implants until 90 weeks of age). Conduction velocity was measured in sciatic–tibial motor nerves. These nerves also underwent qualitative and quantitative histomorphologic analysis. Mild to severe hyperglycemia (>200 mg/dl) and glycosuria (>100 mg/dl) were observed in the WBN group. In contrast, the blood glucose level of the WBN + insulin group fluctuated between normoglycemia (<200 mg/dl) and hyperglycemia. Conduction velocity significantly decreased in WBN group compared with WBN + insulin group. Morphologic analysis of the sciatic and tibial nerves of WBN group showed severe changes, including axonal degeneration, myelin distention, endoneurial fibrosis and microangiopathy. Insulin treatment corrected these changes without microangiopathy. These results suggest that insulin could decrease axonal atrophy and myelin distension of peripheral nerve in diabetic WBN/Kob rats. Observation of WBN/Kob rats revealed changes of axon, myelin and capillary caused by diabetes, thus indicating that this animal is a suitable model for investigating diabetic peripheral neuropathy.
The aim of this study was to evaluate MHC class I expression and prognosis using tumor tissues surgically removed from 9 dogs with mammary gland carcinomas and from 13 dogs with complex carcinomas. We assessed MHC class I expression and its correlation with tumor size, B2M expression, infiltration of lymphocytes, histological grade and prognosis. Hematoxylin and eosin-stained sections were histologically graded using the Elston and Ellis grading method. MHC class I expression on tumor cells was evaluated using the avidin-biotin peroxidase complex method. Loss of MHC class I expression from canine mammary gland carcinomas was significantly correlated with poor prognosis (P<0.05). Loss of MHC class I expression showed no association with poor prognosis in canine mammary gland complex carcinomas, because the data were not balanced. Only 1 of 13 (7.6%) canine mammary gland complex carcinomas showed loss of MHC class I expression. All 13 of these dogs showed good prognosis. Thus, the low frequency of MHC class I expression loss from canine mammary gland complex carcinomas may be associated with good prognosis. Taken together, these results suggest that loss of MHC class I expression may be associated with poor prognosis in canine mammary gland carcinomas.
The purpose of the study was to describe the clinical and electroretinographic features of clinical cases of progressive retinal atrophy (PRA) in miniature schnauzer (MS) of South Korea. Sixty-six MS (14 normal and 52 affected) were included. All animals underwent routine ocular examinations. Electroretinogram (ERG) was recorded in the 14 normal and 15 affected dogs. For normal dogs, the mean age ± SD was 4.1 ± 2.4 years (1 to 9 years), and there were no ocular abnormalities on the basis of ocular examinations and ERG results. For the PRA-affected dogs, it was shown that the mean age ± SD was 4.3 ± 1.1 years (2 to 7 years), and 44 dogs (84.6%) were 3 to 5 years old. Most of the PRA-affected dogs had abnormal menace responses (98.1%) and pupillary light reflexes (PLRs, 88.5%); some dogs showed normal menace response (1.9%) and PLRs (11.5%). Ophthalmoscopic abnormalities in the affected group included one or more of the following changes: hyperreflectivity and discoloration of the tapetal area, attenuation of retinal vessels, depigmentation in non-tapetal area and optic disc atrophy. ERG in the affected dogs showed non-recordable responses in all cases tested with clinical signs of PRA. The present study showed that PRA in MS was mainly observed between the age of 3 to 5 years. ERG revealed abnormal rod and cone responses in affected dogs at the ages studied.
A 2-year-old male mixed dog was referred to us for further evaluation and treatment of a 4-week-history of oliguria and abdominal distension after a surgical repair of urethral injury. To relieve the urethral stricture, we placed a self-expanding aortic stent graft extension with a partial coverage with an expanded polytetrafluoroethylene (ePTFE). After the placement of the stent, the dog presented with a normal urinary voiding, despite the presence of urinary incontinence. The current case indicates that the ePTFE-covered, self-expanding ASGE is an effective intervention for the treatment of severe urethral stricture in the dog.
We report here the clinical presentation, magnetic resonance imaging (MRI) findings and successful surgical management associated with triceps tendon avulsion in a dog. A definitive diagnosis of triceps tendon avulsion was made based on MRI with evidence of displacement of the triceps tendon. Surgical correction of triceps tendon avulsion was performed with two horizontal mattress sutures using polyester and two tunnels drilled in the olecranon to reattach the tendon to the proximal olecranon. At 9 months, there was no evidence of lameness on the left thoracic limb. This is the first case report to describe MRI evaluation for the diagnosis of the triceps tendon avulsion.
Trans-ε-viniferin is a naturally occurring polyphenol belonging to the stilbenoid family that has been isolated from Vitis amurensis, one of the most common wild grapes in Asia. We investigated the effects of trans-ε-viniferin on in vitro maturation (IVM) and developmental competence after in vitro fertilization (IVF) or parthenogenesis (PA). We observed that trans-ε-viniferin treatment during IVM did not improve nuclear maturation rates of oocytes in any group, but significantly increased (P<0.05) intracellular glutathione (GSH) levels and reduced reactive oxygen species (ROS) levels in the 0.5 μM treatment group. Trans-ε-viniferin treatment during IVM of recipient oocytes promoted higher (P<0.05) expression of DNA methyltransferase-1 (DNMT1) mRNA in the 0.5 μM treatment group as compared with the control group. However, the expression of essential transcriptional and apoptosis-related genes did not significantly differ from that of the control. In cumulus cells, pro-apoptosis gene expressions were changed as apoptosis decreased. Oocytes treated with trans-ε-viniferin during IVM did not have significantly different cleavage rates or blastocyst formation rates after PA, but total cell numbers were significantly higher (P<0.05) in the 0.5 and 5.0 μM treatment groups compared with those in the control group. IVF embryos showed similar results. In conclusion, these results indicate that trans-ε-viniferin treatment during porcine IVM increased the total cell number of blastocysts, possibly by increasing intracellular GSH synthesis, reducing ROS levels, increasing DNMT1 gene expression of oocytes and decreasing pro-apoptosis gene expressions of cumulus cells.
In this study, we attempted to express twelve glycoproteins of equine herpesvirus-1 (EHV-1) in 293T cells and to characterize these using monoclonal antibodies (MAbs) and horse sera against EHV-1. Expression of glycoprotein B (gB), gC, gD, gG, gI and gp2 was recognized by immunoblot analysis using horse sera, but that of gE, gH, gK, gL, gM and gN was not. Four MAbs recognized gB, four recognized gC and one recognized gp2. Two MAbs against gB cross-reacted with EHV-4. Interestingly, coexpression of gE and gI and gM and gN enhanced their antigenicity. Furthermore, immunoblot analysis of gp2 showed that different molecular masses of gp2 were recognized by the MAb against gp2 and horse sera against EHV-1. In this study, it was demonstrated that at least six glycoproteins were immunogenic to horses, and coexpression of gE and gI and gM and gN was important for enhancement of antigenicity.
The lung cytokine gene expression profiles of mice infected with 2 strains of H4N8 viruses isolated from shorebirds and reference H4 viruses from ducks are compared. Major differences between the two H4N8 strains of shorebirds, one of which causes a severe respiratory disease in mice, are in the PB1 and NS1 genes. In mice with H4N8 virus induced pneumonia, overall expression of TNF-α, IL-6 and IL-12 genes was markedly higher than in mice infected with other H4 viruses tested, although gene expression of type I interferon was not increased until day 4 post viral infection. In contrast, in mice infected with a comparison H4N8 strain, gene expression of type I interferon peaked on day 1 post viral infection. Overall, the cytokine response corresponds with the severity of disease caused by shorebird H4N8 virus. The results obtained in this study provide valuable information to understand the immunopathology induced by a low pathogenic avian influenza virus, which may be useful in preparation for outbreaks of novel influenza A virus.
To investigate the potential of adeno-associated viruses serotype 2 (AAV2)-mediated RNA interference (RNAi) as an antiviral agent against rabies, recombinant AAV2 vectors expressing siRNA targeting the nucleoprotein (N) gene of rabies virus (RABV) (rAAV-N796) were constructed and evaluated. When NA cells pretreated with rAAV-N796 were challenged with RABV, there was a 37.8 ± 3.4% to 55.1 ± 5.3% reduction in RABV virus titer. When cells pre-challenged with RABV were treated with rAAV-N796, there was a 4.4 ± 1.4 to 28.8 ± 3.2% reduction in RABV virus titer. Relative quantification of RABV transcripts using real-time PCR and Western blot revealed that the knockdown of RABV-N gene transcripts was based on the rAAV-N796 inoculation titer. When any NA cells were treated with rAAV-N796 before or after challenged with RABV, significant reduction in virus titer was observed in both administrations. Mice treated intracerebrally with rAAV-N796 exhibited 50 ± 5.3 and 62.5 ± 4.7% protection when challenged intracerebrally or intramuscally, respectively, with lethal RABV. When mice treated intramuscularly with rAAV-N796 were challenged intramuscularly with lethal RABV, they exhibited 37.5 ± 3.7% protection. When mice were intracerebrally and intramuscularly with rAAV-N796 24 hr after exposure to RABV infection, they exhibited 25 ± 4.1% protection The N gene mRNA levels in the brains of challenged mice with three different administrations were reduced (55, 68, 32 and 25%, respectively). These results indicated that AAV2 vector-mediated siRNA delivery in vitro in NA cells inhibited RABV multiplication, inhibited RABV multiplication in vivo in the mice brain and imparted partial protection against lethal rabies. So, it may have a potential to be used as an alternative antiviral approach against rabies.
Ascitic feline coronavirus (FCoV) RNA was examined in 854 cats with suspected feline infectious peritonitis (FIP) by RT-PCR. The positivity was significantly higher in purebreds (62.2%) than in crossbreds (34.8%) (P<0.0001). Among purebreds, the positivities in the Norwegian forest cat (92.3%) and Scottish fold (77.6%) were significantly higher than the average of purebreds (P=0.0274 and 0.0251, respectively). The positivity was significantly higher in males (51.5%) than in females (35.7%) (P<0.0001), whereas no gender difference has generally been noted in FCoV antibody prevalence, indicating that FIP more frequently develops in males among FCoV-infected cats. Genotyping was performed for 377 gene-positive specimens. Type I (83.3%) was far more predominantly detected than type II (10.6%) (P<0.0001), similar to previous serological and genetic surveys.
Molecular analysis of parapoxvirus envelope genes was performed. Parapoxvirus DNA was detected in eight calves from eight farms in Iwate Prefecture, Japan, between April and September 2010. Seven of the detected viruses were identified as bovine papular stomatitis virus (BPSV) by sequencing, because their nucleotide identity was more than 96.8% similar compared with BPSV strain V660. Among them, two formed a subgroup, because their amplicons were digested with Xmn I (a marker for BPSV) and Hinc II and exhibited a T61C nucleotide substitution in the sequenced region. The remaining virus was pseudocowpox virus that had not been reported previously in Japan. Our results demonstrate the presence of a new BPSV variant in Japan with genetic variability in the envelope gene.