Klotho protein is recognized as having a renoprotective effect and is used as a biomarker for kidney injury. We investigated the level of Klotho protein in hyperoxaluria-induced kidney injury and the effects of vitamin E (Vit E) and vitamin C (Vit C) supplementation. Hyperoxaluria was induced by feeding 2% (w/v) Hydroxy-L-proline (HLP) in the drinking water for 21 days. Rats were divided into 5 groups; control (Group 1, n=7), HLP treated rats that received nothing else (Group 2, n=7), Vit E (Group 3, n=6), Vit C (Group 4, n=6) and both Vit E and Vit C (Group 5, n=7). Vit E (200 mg/kg) was injected on days 1, 6, 11 and 16, while Vit C (500 mg/kg) was given intravenously on days 1 and 11. The Klotho protein levels and oxidative status were measured. The expression level of kidney Klotho protein expression was significantly reduced by HLP-treatment, while the mRNA expression was higher (P<0.05), the plasma and kidney malondialdehyde and kidney superoxide dismutase activities were increased, and the kidney reduced glutathione and urinary total antioxidant status were decreased (P<0.05). All of these changes were ameliorated by administration of Vit E, Vit C or especially the co-administration of both. In conclusion, HLP-induced hyperoxaluria reduced the kidney Klotho protein level, which could be restored by Vit E and/or Vit C.
In South Korea, pigs were vaccinated once between 8 and 12 weeks of age because of the injection-site granulomas. Therefore this study was performed to determine the optimal age for single vaccination of growing pigs with the currently used type O FMD vaccine. With 498 pigs divided into four groups, seroprevalence of the antibody was analyzed with enzyme-linked immunosorbent assay. Although double vaccination is necessary to completely protect growing pigs from FMD virus infection with the current vaccine, the age of 8 weeks can be considered as the optimal age for piglet vaccination if the booster injection is unavailable.
We investigated the antidepressant-like effect of lactoferrin (Lf) in a repeated forced-swim test (FST) stress mouse model. FST was performed on days 1, 2, 7 and 14. Bovine Lf (bLf) or bovine serum albumin (BSA) was supplemented at 1% to the commercial diet after the first FST throughout the experimental period. The FST-control and FST+BSA group showed a marked increase in immobility time on day 2, which remained increased up to the 14th day, while the FST+bLf group showed a significant lower immobility time. Brain-derived neurotrophic factor (BDNF) content in the hippocampus significantly decreased in all of FST treated groups. These results suggest that bLf may improve the depressive-like symptoms induced by repeated FST.
Regulation of inflammation in intestinal mesothelial cells in the abdominal cavity is important for the pathogeny of clinical conditions, such as postoperative ileus, peritonitis and encapsulating peritoneal sclerosis. Here we have examined the inflammatory effect of lipopolysaccharide (LPS) and the anti-inflammatory effect of nicotinic acetylcholine receptor stimulation in rat intestinal mesothelial cells. LPS upregulated mRNA expression of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), monocyte chemotactic protein-1 (MCP-1) and inducible nitric oxide synthase (iNOS). The α7, α9 and α10 subunits of nicotinic acetylcholine receptor were detected in intestinal mesothelial cells. Nicotine (10 nM) significantly inhibited LPS-induced mRNA expression of IL-1β and iNOS, but not TNF-α and MCP-1. In addition, the α7 nicotinic acetylcholine receptor selective agonist, PNU-282987 (10 nM), significantly inhibited LPS-induced mRNA expression of IL-1β but not TNF-α, iNOS and MCP-1. Finally, we found that enteric nerves adhered to intestinal mesothelial cells located under the ileal serosa. In conclusion, intestinal mesothelial cells react to LPS to induce the production of nitric oxide from iNOS. The anti-inflammatory action of intestinal mesothelial cells expressing α7nAChR may be mediated via their connectivity with enteric nerves.
Macrophage polarization is a process by which macrophages acquire a distinct phenotypic and functional profile in response to microenvironmental signals. The classically and alternatively activated (M1 and M2, respectively) macrophage phenotypes are defined by the specific molecular characteristics induced in response to prototypic pro- and anti-inflammatory cues. In this study, we used LPS/IFN-γ and IL-4 to stimulate porcine alveolar macrophages (PAMs) in vitro and investigated the expression changes of several novel markers during macrophage polarization. Notably, we found that LPS/IFN-γ-stimulated PAMs express prototypical M1 molecules, whereas IL-4-stimulated PAMs express M2 molecules. We also demonstrated that replication of the highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV) strain HuN4 was effectively suppressed in LPS/IFN-γ-stimulated M1 PAMs (M1 type), but not IL-4 stimulated M2 PAMs. However, this was not observed with the classic, less pathogenic CH-1a strain. Moreover, we found that M2 marker expression gradually increased after PAM infection with PRRSV, whereas no significant changes were found with M1 marker expression, suggesting that PRRSV infection may skew macrophage polarization towards an M2 phenotype. Finally, we found that anti-viral cytokine expression was significantly higher in M1 macrophages than in M2 macrophages or nonpolarized controls. In summary, our results show that PRRSV replication was significantly impaired in M1 PAMs, which may serve as a foundation for further understanding of the dynamic phenotypic changes during macrophage polarization and their effects on viral infection.
The safety and biological effects of a long-term dose of D-allulose were evaluated in healthy dogs. For 12 weeks, the dogs were administered D-allulose (0.2 g/kg) or placebo daily. Plasma total cholesterol concentrations in the D-allulose group were significantly lower than those in the control group at and after week 2 (P<0.05). D-Allulose administration did not cause clinical signs or changes in hematological and biochemical levels, except for lipids. D-Allulose administration also did not influence body weight. Plasma glucose and insulin concentrations in the glucose tolerance test, performed one day after the termination of D-allulose administration, were not different between groups, suggesting no cumulative effects of D-allulose on glucose metabolism in healthy dogs. In conclusion, long-term administration of D-allulose caused no harmful effects in dogs.
Two cases of extralobar pulmonary sequestrations from a walrus (Odobenus rosmarus) and a Steller sea lion (Eumetopias jubatus) are described in the present study. Grossly, an independent, soft unilocular cystic mass was found within the abdominal cavities of both animals, adherent to the diaphragm in O. rosmarus and attached to the cardia of the stomach in E. jubatus. Histopathologically, the cysts were lined by pseudostratified ciliated columnar epithelium with abundant goblet cells, while the wall comprised of glands, hyaline cartilage, bronchiole- and alveolus-like structures, smooth muscles, and large, well-developed elastic and muscular arteries. The pinniped cases presented are exceptionally rare and to the best of the authors’ knowledge, marks the first descriptions of this congenital anomaly in wildlife.
An 8-year-old neutered female Yorkshire terrier with mediastinal neoplasm and subsequent cranial vena cava invasion developed multiple venous collaterals from the brachiocephalic venous trunks to the caudal vena cava. Collateral venous pathways have been described in dogs with obstruction or increased blood flow resistance of the caudal vena cava but cranial vena cava collaterals have not been reported until now in veterinary patients. In this report, the CTA characteristics of such peculiar vascular routes are described and compared to similar findings reported in human medical literature. The recognition of such ancillary CT finding could help radiologists to reach a more accurate diagnosis of superior vena cava syndrome.
In this study, two commercially available oral electrolyte solutions (OES) with high sodium (CF) or with high glucose and glycine (SL), and two prototype OES were evaluated in terms of rehydration and preventing catabolism. Prototype OES based on CF were prepared by doubling the glucose amount (CFG) or by doubling both glucose and glycine (CFGG). Thirty-two diarrheic calves were randomly assigned four groups with eight calves in each group. Blood volume increased with CF and CFGG compared with that of other OES. The catabolic preventive effect was excellent in CFGG and SL. Our results suggest that both the amount of sodium, glucose, and glycine, and ratio of these factors aid dehydration and provide energy.
Fernando NÁJERA, Andrew J. HEARN, Joanna ROSS, Diana A. Ramírez SALDIVAR, Meaghan N. EVANS, Sergio GUERRERO-SÁNCHEZ, Senthilvel K.S.S. NATHAN, Ignacio de Gaspar SIMÓN, David W. MACDONALD, Benoit GOOSSENS, Luis Revuelta RUEDA
[Advance Publication] Released: September 14, 2017
There is currently no available information regarding the veterinary management of Sunda clouded leopards (Neofelis diardi), either in captivity or in the wild. In this study, 12 Sunda clouded leopards were anesthetized between January 2008 and February 2014 for medical exams, and/or GPS-collaring. Seven wild-caught individuals were kept in captivity and 5 free-ranging animals were captured by cage traps. Two anesthesia combinations were used: medetomidine-ketamine (M-K) or tiletamine-zolazepam (T-Z). Atipamezole (0.2 mg/kg im) was used as an antagonist for medetomidine. Medetomidine (range: 0.039–0.054 mg/kg) and ketamine (range: 3–4.39 mg/kg) were administered during 5 immobilizations, resulting in median induction times of 7 min. After a median anesthesia time of 56 min, atipamezole was injected, observing effects of antagonism at a median time of 12 min. T-Z (range: 6.8–10.8 mg/kg) was administered on 7 occasions. Median induction times observed with this combination were shorter than with M-K (4 min vs. 7 min; P=0.04), and anesthesia and recovery times were significantly longer (244 min and 35 min vs. 56 min and 16 min, respectively; P=0.02). Lower heart rates were measured in the M-K group, while lower rectal temperatures were found in the T-Z group. Both combinations resulted in safe and reliable immobilizations, although given the favorable anesthesia and recovery times of M-K, we recommend this approach over T-Z for the veterinary handling of Sunda clouded leopards.
The study was aimed to investigate biofilm forming ability of Mycoplasma hyopneumoniae and to determine the minimum biofilm eradication concentrations of antibiotics. Biofilm forming ability of six strains of M. hyopneumoniae was examined using crystal violet staining on coverslips. The results demonstrated an apparent line of biofilm growth in 3 of the strains isolated from swine with confirmed cases of enzootic pneumonia. BacLight bacterial viability assay revealed that the majority of the cells were viable after 336 hr of incubation. Moreover, M. hyopneumoniae persists in the biofilm after being exposed to 10 fold higher concentration of antibiotics than the minimum inhibitory concentrations in planktonic cells. To the best of our knowledge, this is the first report of biofilm formation in M. hyopneumoniae. However, comprehensive studies on the mechanisms of biofilm formation are needed to combat swine enzootic pneumonia caused by resistant M. hyopneumoniae.
Burdock (BD) is a common vegetable with many pharmacological properties. However, few studies have examined the effect of BD on exercise performance and physical fatigue. We aimed to evaluate the potential beneficial effects of BD on fatigue and ergogenic functions following physical challenge in mice. Methods: Male ICR mice were divided into four groups to receive either vehicle, or BD at 348.5, 697 or 1,742.5 mg/kg/day, by daily oral gavage for 4 weeks. Exercise performance and fatigue were evaluated from forelimb grip strength, exhaustive swimming time, and post-exercise levels of physical fatigue-related biomarkers serum lactate, ammonia, glucose, and creatine kinase (CK). Results: BD supplementation elevated endurance and grip strength in a dose-dependent manner. It also significantly decreased lactate, ammonia, and CK levels after physical challenge. In addition, BD supplementation had few subchronic toxic effects. Conclusions: Supplementation with BD has a wide spectrum of bioactive effects, including health promotion, performance improvement, and fatigue reduction.
The objective of this study was to measure differences between arterial and venous blood gas parameters and to evaluate whether arterial blood gas values can be estimated from venous blood in Asiatic black bears (ABBs). Twelve healthy captive ABBs (8 males and 4 females; 8–16 years; 76.8–220 kg) were included in this study. The bears were immobilized with medetomidine and zolazepam-tiletamine using a dart gun. Arterial and venous samples were collected simultaneously at 5 and 35 min after recumbency (5- and 35-min points). Partial pressure of oxygen (PO2), partial pressure of carbon dioxide (PCO2), pH, bicarbonate (HCO3–), total carbon dioxide (TCO2), oxygen saturation of hemoglobin (SO2) and base excess (BEecf) were analyzed using a portable blood gas analyzer. There was no marked difference in measured and calculated variables over time in both venous and arterial blood except for PO2. However, arterial PO2, SO2 and pH were significantly higher and arterial PCO2, TCO2 and HCO3– were lower than those of venous samples at both 5- and 35-min points. In the regression analysis to estimate arterial values from venous values, PCO2, TCO2, HCO3–, BEecf and pH significantly showed over 0.45 in coefficient of determination value (R2), and there were little differences between actual and predicted arterial values. Although there were limits in venous gas values replaced those of arterial blood, if we could not get the arterial samples, the regression formulas for arterial values from venous blood in this study would be useful clinically, except for PO2 and SO2.
This study reports on two Hanwoo (a native Korean breed of cattle) calves, a 3- and 6-month-old presenting with diarrhea, anorexia, and blindness. Ophthalmoscopic examination revealed bilateral papilledema in both calves. Reverse-transcription polymerase chain reaction tests for bovine viral diarrhea virus, rotavirus, and coronavirus were all negative. The levels of serum vitamin A in the two affected calves were 0.317 µg/dl and 0.481 µg/dl, respectively. These values are much lower than the normal vitamin A levels; therefore, the calves were diagnosed with hypovitaminosis A.
Short chain fatty acids (SCFAs) play an important role in the maintenance of colonic homeostasis, and their depletion has been reported in various gastrointestinal disorders. Inflammatory colorectal polyps (ICRPs) are a recently recognized disease specific to miniature dachshunds (MDs), and fecal dysbiosis with a reduction of SCFA-producing bacteria has been reported with this disease. Therefore, this study was performed based on the hypothesis that a reduced SCFA concentration associates with the development of ICRPs. We recruited 11 ICRP-affected MDs and 25 control MDs. Their fecal SCFA concentrations and bacterial proportions were quantified using high performance liquid chromatography and quantitative real-time PCR, respectively. The feces of ICRP-affected MDs contained lower amounts of propionic acid and lower proportions of Bifidobacterium than the feces of control MDs. Furthermore, fecal proportions of Bifidobacterium, Firmicutes and Lactobacillus exhibited significant positive correlations with fecal concentrations of total SCFAs and/or propionic acid; fecal Escherichia coli proportions correlated negatively with fecal concentrations of total SCFAs, as well as acetic, propionic and butyric acid. This result indicates an association between fecal dysbiosis and fecal SCFA concentrations; these phenomena may contribute to ICRP pathogenesis in MDs. Potential therapeutic targeting of the reduced propionic acid concentration using probiotics, prebiotics or SCFA enemas merits further study.
Hydroxyethyl starches (HES) are commonly used synthetic colloidal solution in veterinary medicine. Despite of possible adverse effect to kidney injury in human, there is no report about nephrotoxic effects of HES in dogs. HES was administered to a Golden retriever (4-year-old, intact male) with ascites in order to increase plasma osmolality. Initially, the dog was mild azotemic, however, kidney function was rapidly deteriorated after several days of HES administration. Finally, histopathological examination revealed remarkable osmotic nephrosis. In the case reported herein, acute kidney injury was remarkably developed after HES administration. Clinical and histopathologic findings of acute kidney injury support nephrotoxic effects of HES to a dog.
Twenty-three isolates of Pasteurella multocida were tested for susceptibility to six aminoglycoside agents and screened by polymerase chain reaction for the presence of aminoglycoside resistance genes. In addition, mutations in the resistance-determining region of strains showing a high level of induced resistance to spectinomycin strains were examined. Susceptibility testing showed that all of the isolates were resistant to at least two types of aminoglycosides, and that the most effective antimicrobial was spectinomycin. The resistance genes aphA1, strB, and aacA4 were present in all 23 isolates. In the three induced spectinomycin-resistant strains, a 9-bp deletion in rpsE that encodes ribosomal protein S5 was detected.
Swine edema disease is caused by Shiga toxin (Stx) 2e–producing Escherichia coli (STEC). Addition of highly concentrated zinc formulations to feed has been used to treat and prevent the disease, but the mechanism of the beneficial effect is unknown. The purpose of the present study was to investigate the effects of highly concentrated zinc formulations on bacterial growth, hemolysin production, and an Stx2e release by STEC in vitro. STEC strain MVH269 isolated from a piglet with edema disease was cultured with zinc oxide (ZnO) or with zinc carbonate (ZnCO3), each at up to 3,000 ppm. There was no effect of zinc addition on bacterial growth. Nonetheless, the cytotoxic activity of Stx2e released into the supernatant was significantly attenuated in the zinc-supplemented media compared to that in the control, with the 50% cytotoxic dose values of 163.2 ± 12.7, 211.6 ± 33.1, and 659.9 ± 84.2 after 24 hr of growth in the presence of ZnO, ZnCO3, or no supplemental zinc, respectively. The hemolytic zones around colonies grown on sheep blood agar supplemented with zinc were significantly smaller than those of colonies grown on control agar. Similarly, hemoglobin absorbance after exposure to the supernatants of STEC cultures incubated in sheep blood broth supplemented with zinc was significantly lower than that resulting from exposure to the control supernatant. These in vitro findings indicated that zinc formulations directly impair the factors associated with the virulence of STEC, suggesting a mechanism by which zinc supplementation prevents swine edema disease.
Canine histiocytic sarcoma (HS) is a malignancy originating from the histiocytic cell lineage and characterized by poor response to chemotherapy and short survival time. Mutation of the TP53 gene and its association with poor prognosis has been reported in several canine tumors. However, the mutation of this gene has not been investigated in canine HS. The aim of this study was to examine a TP53 gene mutation in dogs with HS. Aberrations of the TP53 gene were examined by polymerase chain reaction-single strand conformational polymorphism analysis and DNA sequence analysis, revealing mutations of the TP53 gene in 12 (46%) of 26 dogs affected by HS. The incidence of the TP53 gene mutation was relatively high in canine HS compared with other canine tumors. Among these mutations, 10 of 12 dogs (83%) with a TP53 gene mutation harbored the same mutation: a 2-base (AT) insertion in exon 5, resulting in the introduction of a stop codon (c.446_447insAT, p.Tyr150SerfsX8). Further studies are needed to examine the functional change due to the mutation and its association with the pathogenesis of canine HS.
Gammaherpesviruses (GHVs) are members of an emerging subfamily of the family Herpesviridae. A recent study identified a novel GHV in domestic cats (Felis catus GHV1, FcaGHV1), and epidemiological surveys have found that FcaGHV1 is distributed worldwide. In this study, we investigated the prevalence of GHVs in domestic cats in Japan with a molecular epidemiological survey. Blood samples were collected from 1,738 domestic cats and GHV-derived DNA was detected with PCR in 1.3% (23/1,738) of the Japanese domestic cats. The FcaGHV1 detected in this study was very similar to FcaGHV1 detected in a domestic cat in North America. Older age (>5 years old) and Feline immunodeficiency virus infection were identified as risk factors for GHV infection.
The purpose of this research was to compare the accuracy of newly described P wave-related parameters (P wave area, Macrux index and mean electrical axis) with classical P wave-related parameters (voltage and duration of P wave) for the assessment of left atrial (LA) size in dogs with degenerative mitral valve disease. One hundred forty-six dogs (37 healthy control dogs and 109 dogs with degenerative mitral valve disease) were prospectively studied. Two-dimensional echocardiography examinations and a 6-lead ECG were performed prospectively in all dogs. Echocardiography parameters, including determination of the ratios LA diameter/aortic root diameter and LA area/aortic root area, were compared to P wave-related parameters: P wave area, Macrux index, mean electrical axis voltage and duration of P wave. The results showed that P wave-related parameters (classical and newly described) had low sensitivity (range=52.3% to 77%; median=60%) and low to moderate specificity (range=47.2% to 82.5%; median 56.3%) for the prediction of left atrial enlargement. The areas under the curve of P wave-related parameters were moderate to low due to poor sensitivity. In conclusion, newly P wave-related parameters do not increase the diagnostic capacity of ECG as a predictor of left atrial enlargement in dogs with degenerative mitral valve disease.
A 10-month-old male Welsh Corgi with a history of acute blindness underwent neuro-ophthalmological testing and magnetic resonance imaging (MRI). Vision testing revealed complete visual deficits but the electroretinograph and pupillary light reflex were normal in both eyes. The motor and sensory functions of the eyelids and eyes were also normal. The MRI revealed compression of the optic chiasm caused by severe ventriculomegaly in the lateral and third ventricles. Such lesions are associated with inflammatory stenotic lesions in the mesencephalic aqueduct. Moderate neutrophilic pleocytosis was observed during cerebrospinal fluid analysis and Acinetobacter lwoffii was isolated, leading to a diagnosis of Acinetobacter-positive obstructive hydrocephalus. This is the first reported case of culture-proven Acinetobacter-associated postencephalitic hydrocephalus with acute blindness in a dog.
Progressive rod-cone degeneration (PRCD) is an autosomal recessive disease caused by c.5G>A mutation of the PRCD exon 2. This mutation has been identified in various breeds, including Labrador Retriever. The present study aimed to examine the allelic frequency of PRCD in Labrador Retrievers in Japan. A domestic and a guide dog population were genotyped for PRCD using polymerase chain reaction-restriction fragment length polymorphism. The allelic frequency of c.5G>A in domestic and guide dog populations (0.114 and 0.026, respectively) differed significantly. The allele with c.5G>A mutation appeared to spread widely in the domestic population as compared to that in the guide dog population. This might be the result of mating control for PRCD in the guide dog population.
Fur seal feces-associated circular ssDNA virus (FSfaCV) was discovered in a pig for the first time in Japan using a next-generation sequencer with duplex-specific nuclease. Full genome of the virus showed approximately 92% similarity to FSfaCVs from New Zealand fur seals. Furthermore, we investigated the prevalence of the ssDNA virus in 85 piglets in Japan, and 65 piglets were positive (76%) for the virus.
A white, lobular mass was found in the right ovary of a pregnant Risso's dolphin (Grampus griseus) at necropsy. The mass was unilateral and occupied most of the pre-existing ovarian tissue. Histologically, the mass was composed of diffuse sheets of polyhedral cells with abundant eosinophilic cytoplasm and oval nuclei, separated by fibrous connective tissue. Only a few ovarian follicles were observed at the periphery of the mass. Immunohistochemically, the large eosinophilic cells were positive for vimentin and negative for pan-cytokeratins. Based on the histopathological features, the present case was diagnosed as luteoma. In human medicine, luteoma of pregnancy, a tumor-like proliferative lesion occurring in pregnant women, is well described. In veterinary medicine, luteoma associated with pregnancy has never been described. The present study would provide useful information for understanding the characteristics of luteoma in animals.
An evaluation of mouse red blood cell (RBC) and platelet (PLT) counting with an automated hematology analyzer was performed with three strains of mice, C57BL/6 (B6), BALB/c (BALB) and DBA/2 (D2). There were no significant differences in RBC and PLT counts between manual and automated optical methods in any of the samples, except for D2 mice. For D2, RBC counts obtained using the manual method were significantly lower than those obtained using the automated optical method (P<0.05), and PLT counts obtained using the manual method were higher than those obtained using the automated optical method (P<0.05). An automated hematology analyzer can be used for RBC and PLT counting; however, an appropriate method should be selected when D2 mice samples are used.
To investigate the genetic variation in Haemophilus parasuis (HPS) in Sichuan, China, 11 isolates were analyzed based on the outer membrane protein P2 (OMPP2) sequence. Sequence analysis showed that the 11 isolates shared 93.0% to 100% nucleotide homology with 15 reference strains, and the consistency between the 26 strains was 89.0%. The isolates of HPS-1, 2, 4, 5, 6, 7, 8, 10 and 11 had a 69-base deletion from 770 base pairs (bp) to 850 bp, which was infrequent in China. The phylogenetic tree showed that HPS-3 and HPS-8 had closer relationships with European and Japanese strains, but shared 98.7% nucleotide homology with the SW114 Japanese strain.
This study evaluated the accuracy of a newly developed veterinary portable blood glucose meter (PBGM) with hematocrit correction in dogs and cats. Sixty-one dogs and 31 cats were used for the current study. Blood samples were obtained from each dog and cat one to six times. Acceptable results were obtained in error grid analysis between PBGM and reference method values (glucose oxidation methods) in both dogs and cats. Bland–Altman plot analysis revealed a mean difference between the PBGM value and reference method value of −1.975 mg/dl (bias) in dogs and 1.339 mg/dl (bias) in cats. Hematocrit values did not affect the results of the veterinary PBGM. Therefore, this veterinary PBGM is clinically useful in dogs and cats.
This study was undertaken to investigate drug resistance in Escherichia coli (E. coli) strains isolated from bamboo rats in Zhejiang province of China. One hundred and fifty-four E. coli strains were isolated from dead bamboo rats. Polymerase chain reaction (PCR) was used to detect the representative genes encoding resistance to commonly used β-lactam antibiotics. Highest resistance was observed for cefradine (24.03%), followed by penicillin (20.78%) and ceftazidime (20.13%). The isolation rates of β-lactam resistance genes were 53.25, 48.7015.58 and 14.29% for bla TEM, bla CTX-M, bla OXA and bla SHV, respectively, while 62 (40.26%) E. coli isolates harbored multiple β-lactam resistance genes. These results also suggested that long term use of these antibiotics leads to antibimicrobial resistance. We believe that this study will provide a guideline for veterinarians and a research basis for examining resistance-encoding genes in other food animals like bamboo rats.
A 1-year-old neutered male Pekingese was presented for evaluation and further treatment of cluster seizures. The dog had behavioral abnormalities, and a prosencephalic lesion was suspected following neurological examination. The dog showed signs of learning difficulty. Magnetic resonance imaging of the brain revealed a remarkably smooth cerebral cortex with a reduced number of gyri, as well as a cystic lesion associated with the quadrigeminal cistern. A diagnosis of lissencephaly, concurrent with a quadrigeminal cisternal cyst, was made. High-dose and multiple anticonvulsants were necessary to control the seizures. This is the first report of lissencephaly in a Pekingese.
An approximately two-year-old, male 6.1 kg body weight, Korean wild raccoon dog (Nyctereutes procyonoides koreensis) was captured by the wildlife medical rescue center of Kangwon National University. Upon physical examination, the heart rate was 87 beats per min and there were no clinical signs. The hematological, and blood biochemical profiles revealed no remarkable findings; however, thoracic radiographs showed cardiac enlargement, especially in the right atrium. On electrocardiogram, sinus node dysfunction and bradyarrhythmia were revealed. Echocardiography showed a left-to-right shunting atrial septal defect. Based on these findings, this Korean wild raccoon dog was diagnosed with atrial septal defect. This is the rare case report of atrial septal defect in wildlife.
Serum concentrations of non-esterified fatty acids (NEFA) and very low-density lipoprotein (VLDL) in close-up dairy cattle were compared in relation to parity. Data were obtained from 37 nulli/primiparous (NP) and 24 multiparous (MU, parity: 2–7) cows between 14 days and 1 day prepartum. A positive correlation (r=0.684, P<0.01) was found between serum NEFA and VLDL concentrations in NP cows. Among the VLDL constituents, the NEFA concentration was particularly correlated with the triglyceride (TG) concentration (r=0.658, P<0.01). However, no significant correlation was found between the concentrations of NEFA and VLDL or VLDL-TG in MU cows (r=–0.028 and 0.307). These results suggest the presence of higher hepatic secretion of NEFA-derived VLDL in NP cows.
The aim of the present study was to determine changes in body iron storage in adult dogs following phlebotomy. We performed repeated phlebotomies by removing 1% body weight (approximately 10% of the total blood volume) weekly for a total of 12 times using adult beagle dogs without an iron-restricted diet. After treatment, stored iron was decreased, as demonstrated by gradual reductions in serum ferritin levels and hepatic iron contents. Anemia and abnormalities in blood chemistry analysis were not observed; therefore, this method was considered safe and useful for control of stored iron levels in adult dogs.
A 7-year-old castrated male ferret developed unilateral cervical lymphadenomegaly over a 1-month period. Histological examination revealed proliferation of tumor cells in a diffuse and partially nodular pattern. The tumor cells were predominantly Hodgkin cells and binucleated Reed-Sternberg cells, characterized by abundant, clear, vacuolated cytoplasm, pleomorphic, ovoid nuclei with thick nuclear membranes and distinct nucleoli. Multinucleated cells, resembling lymphocytic and histiocytic (L&H) cells, were also observed. Immunohistochemically, the tumor cells expressed Pax-5, BLA-36 and vimentin. A small population of the tumor cells expressed CD20. This case showed proliferation of Hodgkin/Reed-Sternberg cells in conjunction with L&H cells that were histologically analogous to feline Hodgkin’s-like lymphoma. However, Pax-5 and BLA-36 expression along with rare CD20 expression were consistent with classical Hodgkin’s lymphoma in humans.
Acerodon jubatus (the Golden-Crowned flying fox) is an endemic species in the Philippines, which was suspected to be a host of the Reston strain of the Ebola virus. As nocturnal animals, the flying foxes spend daytime at the roosting site, which they use for self-maintenance and reproduction. To understand the variation in diurnal behavior and time allocation for various activities in the Golden-Crowned flying fox, we investigated their daytime behavior and activity budget using instantaneous scan sampling and all occurrence focal sampling. Data collection was performed from 0700 to 1800 hr during January 8–17, 2017. The most frequent activity was sleeping (76.3%). The remaining activities were wing flapping (5.0%), self-grooming (4.2%), hanging relaxation (3.4%), wing spread (2.9%), movement (2.4%), mating/courtship (2.4%), aggression (1.9%), hanging alert (1.2%), excretion (0.1%) and scent marks (0.05%). The frequency of sleeping, wing flapping, self-grooming, hanging relaxation, aggression, mating/courtship and movement behaviors changed with the time of the day. Females allocated more time for resting than males, while males spent more time on the activities that helped enhance their mating opportunities, for example, movement, sexual activity and territorial behavior.
A female Indian star tortoise (Geochelone elegans) was evaluated for anorexia and suspected coelomic masses. Clinical examination indicated follicular stasis. The tortoise was anesthetized and placed in dorsal left lateral recumbency for a right prefemoral approach. The ovaries and oviducts were exposed through the right prefemoral incision. The ovarian vasculature, the mesovarium, the oviduct, mesosalpinx and associated vasculature were ligated and transected. Bilateral oophorosalpingectomy was performed through the incision, without coelioscopy. Since then, the same procedure has been performed in five other tortoises, and all recovered well. These results suggest that bilateral oophorosalpingectomy, performed through a prefemoral incision, without coelioscopy, is a safe and practical approach for treating follicular stasis in tortoises.
Day-old chicks from 3 hatcheries were placed on bedding paper and brought to a commercial broiler farm between January and July 2016. Sixty-six samples of the paper, which were stained with meconium droppings of the chicks, were collected and examined for isolation of cephalosporin-resistant Enterobacteriaceae. Cefotaxime (CTX)-resistant Klebsiella pneumoniae (1 isolate) and Enterobacter cloacae (4 isolates) were isolated from 5 (7.58%) of the 66 samples. Conjugation experiments revealed that the blaCTX-M-25 gene conferring CTX resistance was transferred from the K. pneumoniae isolate and 2 of the 4 E. cloacae isolates to Escherichia coli DH5alpha via IncA/C plasmids carrying the gene. Our results suggested that the blaCTX-M-25 gene originating from chicks may be spread among commercial broiler farms.
Shiba dogs are predisposed to chronic enteropathy (CE) and have poorer prognosis than other dog breeds. The objective of this study was to investigate the significance of polymerase chain reaction for antigen receptor rearrangement (PARR) results on clinical findings and prognosis of Shiba dogs with CE. We retrospectively collected data on 22 Shiba dogs diagnosed as having CE. Fifty-nine percent of the dogs had clonality-positive results on PARR analysis. Furthermore, on histopathology, epitheliotropic behavior of small lymphocytes of the intestinal mucosa was observed significantly more frequently in dogs with clonal rearrangement of antigen receptor genes (P=0.027). The median overall survival time of clonality-positive dogs was 48 days (range, 4–239 days), compared to 271 days (range, 45–1,316+ days) in clonality-negative dogs. The median overall survival time of epitheliotropism-positive dogs was 76 days (range, 30–349 days) compared to 239 days (range, 4–1,316+ days) for epitheliotropism-negative dogs. Statistical analysis revealed that the clonality-positive result was associated with significantly shorter survival time (P=0.036). In contrast, presence or absence of epitheliotropism had no statistically significant effect on survival time (P=0.223). These cases might appropriately be diagnosed as small T-cell intestinal lymphoma; there are some common clinical and pathogenic features with human enteropathy-associated T-cell lymphoma type 2. The pathogenesis and poor prognosis for Shiba dogs with CE seem to be associated with this type of lymphoma, although further investigation is warranted.
Contrast-enhanced ultrasonography (CEUS) with microbubbles as a contrast agent allows the visualization and quantification of tissue perfusion. The assessment of canine intestinal perfusion by quantitative CEUS may provide valuable information for diagnosing and monitoring chronic intestinal disorders. This study aimed to assess the repeatability (intraday variability) and reproducibility (interday variability) of quantitative duodenal CEUS in healthy dogs. Six healthy beagles underwent CEUS three times within one day (4-hour intervals) and on two different days (1-week interval). All dogs were sedated with a combination of butorphanol (0.2 mg/kg) and midazolam (0.1 mg/kg) prior to CEUS. The contrast agent (Sonazoid®) was administered using the intravenous bolus method (0.01 ml/kg) for imaging of the duodenum. Time-intensity curves (TIC) were created by drawing multiple regions of interest (ROIs) in the duodenal mucosa, and perfusion parameters, including the time-to-peak (TTP), peak intensity (PI), area under the curve (AUC), and wash-in and wash-out rates (WiR and WoR, respectively), were generated. Intraday and interday coefficients of variation (CVs) for TTP, PI, AUC, WiR, and WoR were <25% (range, 2.27–23.41%), which indicated that CEUS was feasible for assessing duodenal perfusion in healthy sedated dogs. Further study of CEUS in dogs with chronic intestinal disorders is necessary to evaluate its clinical applicability.
To establish an accurate, rapid, and a quantifiable method for the detection of Riemerella anatipestifer infection, a widespread infectious disease in birds, we developed a TaqMan-based real-time PCR assay by using DtxR gene-specific primers and a TaqMan probe. The standard curve established with a linear correlation (R2) of 0.998 and efficiency of 99% between the Ct value and the logarithm of the plasmid copy number. The reproducibility and specificity of the real-time PCR assay were confirmed by using plasmids containing DtxR genes or DNAs extracted from well-known bacteria or viruses causing duck diseases. The real-time PCR assay was 100 times more sensitive than the conventional PCR. The results reveal that the established real-time PCR assay might be a useful method for diagnosis and quantitative detection of Riemerella anatipestifer in birds.
The olfactory organ of turtle consists of the upper chamber epithelium (UCE) and the lower chamber epithelium (LCE), detecting air-borne odorants and water-borne odorants, respectively. In this study, we investigated the number of olfactory receptor neurons (ORNs) in the UCE and LCE of soft-shelled turtle in order to find their possible differences among terrestrial, semi-aquatic and highly-aquatic turtles. The number of ORNs in the soft-shelled turtle was higher in the LCE than in the UCE, suggesting its close relationship to the environment the turtle lives. In addition, relative abundance of the ORNs in the LCE to the UCE varied in accordance with the size of individuals, although its functional significance remains elusive.
The aim of this study was to describe a ventral laparoscopic technique for bilateral medial iliac lymphadenectomy in dogs. Twelve intact male purpose-bred research dogs weighting less than 15 kg were positioned in dorsal recumbency, and a 3-portal technique was used. Bilateral dissection was performed with vessel-sealing devices while tilting the surgical table by up to 30° towards the contralateral side of the target medial iliac lymph node (MILN) without changing the surgeon’s position. Using a ventral laparoscopic approach, bilateral MILNs were identified and excised in all dogs. The mean time for unilateral and bilateral MILN dissections were 9.7 ± 3.8 min and 21.0 ± 6.0 min, respectively. The mean times for the right and left MILN dissections were 10.8 ± 4.3 min and 9.8 ± 2.5 min, respectively. The mean total surgery time was 43.7 ± 7.7 min. In total, 26 MILNs were dissected. Several complications, including, mild to moderate capillary hemorrhage from perinodal fat and vessels (controlled laparoscopically), mild spleen trauma caused by the first trocar insertion, and capsular damage of MILNs, were observed. However, there were no other major complications. All MILN samples were evaluated and deemed suitable for histopathologic diagnosis. Laparoscopic excision of MILNs is a useful method of excisional biopsy for histopathologic diagnosis. Using this ventral laparoscopic approach with the 3-portal technique, bilateral MILN dissection suitable for obtaining histopathologic samples could be achieved in a short time in dogs weighing less than 15 kg.
Ehlers-Danlos syndrome (EDS) is a group of disorders caused by abnormalities in the extracellular matrix (ECM). Transforming growth factor-β (TGF-β) plays a crucial role in formation of the ECM by the SMAD (Sma-and Mad-related protein, mothers against decapentaplegic homolog) pathway. It has been reported that loss of function of zinc transporter ZRT/IRT-like protein 13 (ZIP13) is the cause of the spondylocheiro dysplastic form of EDS (SCD-EDS: OMIM 612350). Our previous study suggested that TGF-β1 has a relationship with the skin pathological condition in the Zip13-Knockout (KO) mouse, which is a model of SCD-EDS. Thus far, effective treatment based on modern medicine for this syndrome has not yet been established. According to an approach of traditional Chinese medicine, the present study investigates the medicinal effects of Makomo (Zizania latifolia) on certain aspects of SCD-EDS, such as skin morphology and plasma TGF-β1, in Zip13-KO mice. Increases in densities of collagen fibers and fibrils without a significant change in thickness of the dermal layer were observed in the group of mice fed a Makomo-containing diet. No change in the amount of collagen suggests that Makomo feed does not elevate collagen synthesis, but changes the length of glycosaminoglycan chains and decreases the distance between collagen fibrils. In conclusion, the changes of the skin structure suggest that Makomo can increase the mechanical strength of skin.
To investigate the molecular pathways involved in successful embryo implantation in mammals, we developed a novel method for gene transduction into the murine endometrium using in vivo electroporation. Plasmid DNA with an enhanced green fluorescence protein (EGFP) gene was injected into the uterine cavity of non-pregnant female mice, and electrical pulses were subsequently applied to the uterine horn using plate electrodes. EGFP expression was found only in the uterine luminal epithelium (LE), but not in the stroma. EGFP fluorescence in the LE was limited to the site where the positive side of the electrodes was placed during electric stimulation. These results demonstrated that our novel method enabled us to transduce a gene into a desired location of the murine uterus.
During the autumn migration of many waterfowls, body mass is lowest upon arrival at the wintering area and gradually increases during the winter. Consequently, body mass is highest before the spring migration. We studied the pattern of body mass changes in the Black-Headed Gull (Larus ridibundus) from December 2010 to December 2016 in the Shinhama area of Chiba, Japan. Based on 327 captured animals, body mass increased during the wintering period, but tended to decrease before migration. In 2014-2016, a muscle mass reduction in females was observed, explaining the change in body mass. However, the observed weight loss may be caused by many factors, which may be related to the migratory ecology of the regional population.
The outcomes of cardiopulmonary resuscitation (CPR) were retrospectively evaluated in 146 exotic animals including 20 pet birds, 47 rabbits, 34 hamsters, 18 ferrets, 7 turtles, and 20 other small mammals in cardiopulmonary arrest (CPA) at presentation or during hospitalization at an animal clinic. The rates of return of spontaneous circulation, survival after CPR, and discharge were 9.3%, 2.3%, and 1.2%, respectively. The mean success rate of CPR in animals included in this study was lower than those previously reported in dogs and cats. This might have been because of the challenges in effective chest compression, airway management and monitoring as well as establishment of intravenous catheterization route in exotic animals.
10-Hydroxy-2-decenoic acid (10H2DA) is a fatty acid found in royal jelly (RJ). In healthy mice, it activates 5´-AMP-activated protein kinase (AMPK) and increases glucose transporter 4 (GLUT4) translocation. Therefore, we examined whether 10H2DA has a potential therapeutic effect against type 2 diabetes in obese/diabetic KK-Ay mice. 10H2DA (3 mg/kg body weight) was administered to female KK-Ay mice for 4 weeks by oral gavage. Phenotypes for body weight, plasma glucose by oral glucose tolerance test and insulin levels were measured. mRNA and protein levels were determined using qRT-PCR and Western blot analyses, respectively. Long-term administration of 10H2DA significantly improved hyperglycemia and insulin resistance in KK-Ay mice, but did not prevent obesity. 10H2DA increased the expression of phosphorylated AMPK (pAMPK) protein in skeletal muscles; however, this expression did not correlate with increased GLUT4 translocation. Furthermore, 10H2DA neither enhanced the expression of adiponectin receptor mRNA nor activated the insulin signaling cascade, such as GSK-3β phosphorylation, in the liver. We found that 10H2DA-treated mice had a significant increase in the expression of peroxisome proliferator-activated receptor gamma coactivator 1 alpha (Pgc-1α) mRNA in skeletal muscles compared with non-treated group (P=0.0024). These findings suggest that 10H2DA is involved in the improvement of type 2 diabetes, at least in part via activation of Pgc-1α expression, but does not prevent obesity.
Flying foxes have been considered to be involved in the transmission of serious infectious diseases to humans. Using questionnaires, we aimed to determine the direct and/or indirect contacts of flying foxes in an Indonesian nature conservation area with domestic animals and humans living in the surrounding area. We surveyed 150 residents of 10 villages in West Java. Villages were classified into 3 groups: inside and/or within 1 km from the outer border of the conservation area and 1–5 km or 5–10 km away from the reserve’s outer border. Data were collected by direct interview using a structured questionnaire consisting of the respondent characteristics (age, sex and occupation); histories of contacts between flying foxes and humans, dogs and other domestic animals; and knowledge about infectious diseases, mainly rabies, in flying foxes. We found that flying foxes from the nature conservation area often enter residential areas at night to look for food, especially during the fruit season. In these residential areas, flying foxes had direct contacts with humans and a few contacts with domestic animals, especially dogs. People who encounter flying foxes seldom used personal protective equipment, such as leather gloves, goggles and caps. The residents living around the conservation area mostly had poor knowledge about flying foxes and disease transmission. This situation shows that the population in this region is at a quite high risk for contracting infectious diseases from flying foxes.
This article released online on July 30, 2014 as advance publication has been retracted by the Editorial Board of Journal of Veterinary Medical Science due to a violation of the journal’s “Information for Authors”.
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