Avian influenza (AI) is a disease caused by influenza viruses type A that belong to the Orthomyxoviridae family. AI induces high economic losses in poultry production worldwide. Due to a possible outbreak, a national surveillance program was needed. From April to July 2016, 152 industrial poultry farms were randomly sampled. All samples were analyzed by competitive ELISA for Influenza type A viruses. Suspicious and positive sera were further analyzed by Hemagglutination Inhibition (HI) in order to serotype H5 or H7 low pathogenic avian influenza virus (LPAIV). The farms sampled showed 94.08%, 3.95% and 1.97% of negative, positive and suspicious results, respectively. However, serotyping revealed all positive and suspicious samples were negative to H5/H7 LPAIV. Our results show the absence of AI in the mainland Ecuadorian industrial poultry production.
Bovine anaplasmosis caused by Anaplasma marginale represents a serious threat to cattle farming worldwide, especially in the tropics and subtropics. In the present study, archived DNA samples from the blood of cattle (n=437) in the Nuwara Eliya, Galle, Ampara, Polonnaruwa, and Jaffna districts and buffalo (n=327) in the Galle, Polonnaruwa, Mannar, and Mullaitivu districts in Sri Lanka, were screened for A. marginale using a major surface protein 5 (msp5) gene-based PCR assay. The findings showed that 32.7 and 57.5% of cattle and buffalo, respectively, were A. marginale-positive. The rate of positivity differed significantly among geographical regions. In conclusion, the high rates of A. marginale infection in cattle and buffalo highlight the importance of effective control measures in Sri Lanka.
The aim of the present study was to evaluate the bacterial flora in the udder and intestinal environments in cows with and without protothecal mastitis. We used next-generation sequencing (NGS) analysis to identify 16S rRNA genes from bacterial flora present in 13 milk and 13 fecal samples from protothecal mastitic and healthy dairy cows in the Aichi region of Japan. Sequences associated with 5 species (Calothrix desertica, Corynebacterium simulans, Corynebacterium striatum, Empedobacter falsenii, and Rothia endophytica) showed the highest prevalence in samples of milk and feces from animals with protothecal mastitis. This range of species differed from those detected in the milk and feces from healthy cows.
In human medicine, computed tomography (CT) is the gold standard for visceral fat measurement. Research shows that the visceral fat area (VFA) of the umbilical slice is significantly correlated with the visceral fat volume (VFV). In veterinary medicine, however, few studies have evaluated visceral fat using CT. This study aimed to evaluate the visceral fat in dogs using CT images, and determine if the slice significantly correlated with VFV to simplify visceral fat measurements. This retrospective study includes data on 90 dogs that underwent whole-body CT scans for diagnostic purposes. VFV was calculated as the product of VFA and thickness in each CT slice; the correlation between VFV and VFA was analyzed at the level of each lumbar vertebra. Visceral fat percentage (VF%) was calculated as the ratio of the product of VFV and fat density to the body weight. Visceral fat area percentage (VFA%) was calculated as the ratio of VFA to the body area, and its correlation with the VF% and the body condition score (BCS) was analyzed. VFA was highly correlated with VFV at the level of each lumbar vertebra, with the highest correlation (r=0.964) at the L3 level. VFA% was significantly correlated with VF% (r=0.930) and weakly correlated with BCS (r=0.523). This study demonstrates that it is sufficient to use only the L3 slice for visceral fat evaluation and that the evaluation can be based on VFA% of the L3 level.
Left atrial enlargement (LAE) is a well-known negative prognostic factor in dogs with myxomatous mitral valve disease (MMVD). Left atrial-to-aortic root ratio (LA/Ao) is the most commonly used method to evaluate left atrial (LA) size in dogs, while the left atrial anteroposterior diameter (LAD) has been proposed as an additional measurement of LA size in different species. The aim of this study was to establish a normal reference range of LAD normalized to body weight (LADn) in dogs using allometric scales, and to evaluate the agreement between LADn and LA/Ao in the detection of LAE in dogs with MMVD. This was a retrospective, multicenter, observational study. We included 330 healthy dogs, 30 dogs with MMVD in ACVIM stage B1, 30 dogs in ACVIM stage B2, and 30 dogs in ACVIM stage C. The reference range for the LAD, depending on body weight, was between 16.91 mm and 49.68 mm. The reference range for the LADn in healthy dogs was between 10.49 and 15.72. LADn was significantly greater in dogs with MMVD compared to healthy dogs, and a significant difference in LADn was noted between different ACVIM stages (P<0.001). The most accurate cut-off value of LADn to differentiate between dogs in groups B2 and C was 20.3 (sensitivity, 83.3%; specificity, 83.3%). There was a misclassification rate of 37% between LADn and LA/Ao in the detection of LAE in group B1. This study provides a normal reference range for LAD in dogs, which can be used as an additional tool to assess LAE in dogs with MMVD.
The paracrine function of mesenchymal stem cells (MSCs) during transplantation has been recently studied due to its poor differentiation ratio. Dimethyloxalylglycine (DMOG) has been used to promote angiogenesis in experimental animal models, however, comparable approaches for canine MSCs are not sufficient. In the present study, we assessed whether DMOG improves angiogenesis in canine adipose tissue-derived mesenchymal stem cells (cAT-MSCs). cAT-MSCs were treated with DMOG and their effect on angiogenesis was investigated by cell proliferation assay, western blotting, and tube formation assay. Dimethyloxalylglycine preconditioning enhanced the expression of vascular endothelial growth factor (VEGF) among pro-angiogenic factors in cAT-MSCs via hypoxia-inducible factor-1α stabilization. Dimethyloxalylglycine primed-cAT-MSC-conditioned media increased angiogenesis in human umbilical vein endothelial cells. These results suggest that DMOG conditioning of cAT-MSCs augmented the secretion of VEGF, which acted as a prominent pro-angiogenic factor during angiogenesis. DMOG-primed cAT-MSCs may have the potential to induce beneficial effects in ischemic diseases in clinical trials.
Babesia rossi infection has been reported to be associated with the high prevalence of pancreatitis in dogs. In this study, we retrospectively investigated whether pancreatitis occurs in B. gibsoni-infected dogs. The clinical manifestations, and hematological and serum biochemical examination results, including canine pancreatic-specific lipase (cPL), in 20 B. gibsoni-infected dogs were analyzed. The cPL concentration exceeded 400 µg/l in only 2 dogs, and they were suspected of having pancreatitis. Although the cPL concentration did not correlate with the degree of anemia or the level of parasitemia, it correlated with the band neutrophil count, platelet count, and blood urea nitrogen (BUN) level. Our study suggested that the prevalence of pancreatitis is lower among B. gibsoni-infected dogs than B. rossi-infected dogs.
The accuracy of urine analyzers used for dogs and cats has remained uncertain. This study examines the agreement between results of urine analysis obtained using two devices marketed for animals and for humans and the results of quantitative biochemical analysis. The degrees of concordance for bilirubin and ketones in the same category were ~80%, but for pH these were only ~60% in dogs and cats. Degrees of concordance for protein and the UP/C ratio clearly differed between the devices for animals and humans. We found that values for bilirubin and ketones obtained using urine analyzers may be reliable, but pH is unlikely to be accurate enough to be clinically useful for dogs and cats.
An 18-month-old female Holstein Friesian heifer presented with a history of stunted growth and a recent onset of anorexia; she presented with cyanosis and eventually died. A postmortem examination revealed obstruction of the right ventricular outflow tract, ventricular septal defect, overriding aortic root, right ventricular hypertrophy, and an atrial septal defect, indicating a pentalogy of Fallot (POF). In addition to POF, the heifer also had pulmonary artery dilatation, although she did not present with patent ductus arteriosus. This heifer had the longest lifespan among the Holstein cattle reported to have POF, which may be secondary to delayed pulmonary obstruction due to deformation of one of the pulmonary valves.
The molecular clock network in mast cells has been shown to be a factor responsible for circadian regulation of allergic inflammation. PF670462 is a selective inhibitor of casein kinase 1δ and ε (CK1δ/ε) that control the posttranslational modification of clock proteins. The aims of this study were to evaluate the effects of PF670462 on gene and protein expression of FcεRI, the high-affinity IgE receptor, in canine mast cells and on IgE-mediated immediate-type cutaneous reactions in dogs. PF670462 decreased mRNA expression of FcεRIα and β, but not γ, and protein expression of FcεRI in a canine mast cell line. Furthermore, PF670462 suppressed IgE-mediated immediate-type cutaneous erythema in dogs. These findings indicate that CK1δ/ε function as regulators for FcεRI expression and IgE-mediated cutaneous reactions in dogs.
In Taiwan, Corchorus capsularis L. has long been cultivated and the leaves are consumed as edible vegetable. This study is to investigate the protection effect of extract of C. capsularis leaves (ECC) on ethanol-induced acute gastric mucosal lesion (AGML) in rats. The results of phytochemical determination in ECC for total polyphenol, flavonoid and polysaccharide were 59.88 ± 0.61 mg/g, 86.39 ± 18.0 mg/g and 320.89 ± 6.99 mg/g, respectively. ECC showed significant activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging with IC50 of 0.25 mg/ml. In vivo studies, Sprague-Dawley (SD) rats were randomly divided into five groups: sham, vehicle (control) and low-, medium-, and high-dose ECC (LECC, MECC, HECC; 200, 400, and 1,000 mg/kg/day, respectively). ECC was able to decrease significantly the ulcer index (UI) caused by 80% ethanol in a dose dependent manner. There was no significant effect on growth trend and food intake rate after the administration of ECC in the experimental period. The serum lipid parameters in ECC groups revealed significant increase in glutathione peroxidase (GPx), superoxide dilmutase (SOD) and catalase (CAT), and a decrease in malondialdehyde (MDA). Significant amelioration on pathological lesion score was found in ECC groups compared with the control group (P<0.05). The overall results indicate that ECC has protective effects on ethanol-induced AGML in rats, which could be associated with its antioxidant activity.
Surveillance of Echinococcus multilocularis in 98 pet dogs kept in a rural area of Hokkaido, Japan, from March 2018 to March 2019 suggested infection in seven dogs (7.1%) by E. multilocularis-specific copro-DNA examination, and one of them excreted E. multilocularis eggs that were identified by sequence analyses. Among the infected dogs, three were not allowed to run free when outdoors. Based on detection of E. multilocularis eggs in fox feces collected from roadsides in the same area, dogs kept in rural areas may have a high probability of becoming infected after preying on infected voles along such roadsides, even in domesticated settings. Therefore, examination along with periodic deworming administration is considered necessary to prevent transmission from dogs to owners.
Pigmented viral plaque is most commonly seen in Pug dogs in association with canine papillomavirus (CPV). In the present study, nucleic acid sequence and localization of viral genes were examined in 4 cases of pigmented viral plaque in Pug dogs. The results of polymerase chain reaction and nucleic acid sequence analysis showed that the 3 cases with pigmented viral plaque were infected with CPV4, and 1 case with CPV18. In the case with CPV18-positive viral plaque, CPV18 gene was also detected in a lesion of cytokeratin-14- and P63-positive basal cell tumor that developed adjacent to a pigmented viral plaque. Moreover, CPV gene was detected in the squamous cells of pigmented viral plaques and the neoplastic cells of basal cell tumor by in situ hybridization. This is the first report of basal cell tumor associated with CPV18-infection in the dog. Infection of CPV18 may be associated with development of basal cell tumor.
A male dog developed multiple cutaneous masses at 15 different sites between the ages of 11.5 and 13-years. The masses were surgically removed and histopathologically examined. In gross appearance, the cut surfaces of the masses were white with partially red areas. Microscopy revealed that the tumors were located at the dermis and were composed of spindle cells and pleomorphic cells with abundant eosinophilic cytoplasm, accompanying giant cells. These cells were occasionally arranged in bundles with minimal stromal collagen. Immunohistochemical analysis revealed that the neoplastic cells were strongly positive for vimentin, partially positive for smooth muscle actin and desmin, and negative for cytokeratin. Based on these pathological findings, the tumor was diagnosed as multiple cutaneous pleomorphic leiomyosarcoma.
Aquaporin-2 (AQP2), a vasopressin-regulated water channel, plays an important role in renal water homeostasis. It has been reported that the level of AQP2 in human urine is altered during pregnancy. However, little is known about the level of urinary AQP2 in pregnant cattle. In this study, we examined the level of AQP2-bearing extracellular vesicles (uEV-AQP2), which account for most urinary AQP2, in both heifers and cows during the gestational and postpartum periods. The level of uEV-AQP2 was significantly decreased during gestation in comparison with the other cattle examined. Similarly, the levels of EV marker proteins in uEVs, including tumor susceptibility gene 101 (TSG101) protein and apoptosis-linked gene 2-interacting protein X (ALIX), were significantly decreased during gestation. There were significant correlations between the levels of uEV-AQP2 and uEV-TSG101, or uEV-ALIX. Immunohistochemistry data from pregnant and non-pregnant cattle supported the notion that the level of uEV-AQP2 was decreased during gestation. These data indicate that the level of uEV-AQP2 is decreased in pregnant cattle, possibly through a decrease in both the number of EVs released into the urine and renal AQP2 expression.
It is known that oxidative stress is related to disease in humans and dogs. Many traditional Chinese medicines have been reported to have anti-oxidative effects, but there are no reports that they have anti-oxidative effects in dogs. In this study, we examined the anti-oxidative effects of Juzen-taiho-to, a traditional Chinese medicine, in dogs. Five healthy female beagle dogs (38–41 months of age weighing 8.6–10.7 kg) were orally administered Juzen-taiho-to at 450 mg/kg with food for 28 days. Blood samples were taken from all five dogs on days 0, 7, 14, 21, and 28. Using the blood samples, improvement of the antioxidant level as assessed by the biological antioxidant potential (BAP), reduced oxidative stress level as assessed by derivatives of reactive oxygen metabolites (d-ROMs), and improvement of blood fluidity were examined. Regarding the antioxidant level and blood fluidity, no significant difference was observed, but the oxidative stress level on days 14, 21, and 28 was significantly lower than that on day 0. Thus, Juzen-taiho-to may have anti-oxidative effects in dogs by reducing oxidative stress and be useful for oxidative stress-related diseases in dogs.
Phenobarbital (PB) decreases the cyclosporine (CsA) blood level in humans. However, the interaction of PB with CsA has not been reported in cats. This study investigated the effects of multiple doses of PB on the pharmacokinetics of CsA in three healthy cats. The treatments included oral CsA 5 mg/kg alone and oral CsA 5 mg/kg plus PB 5 mg/kg for 4 weeks. Co-administration of PB with CsA resulted in significant decreases in the oral bioavailability of CsA though both the first pass and elimination phases. These preliminary results suggest that oral administration of multiple doses of PB increases the required CsA dosage in CsA-based immunosuppressive therapy in cats.
We have previously shown that secreted protein acidic and rich in cysteine (SPARC) promotes myogenic differentiation of rat skeletal muscle progenitor cells in vitro, and in vivo small interfering RNA (siRNA)-mediated transient suppression of SPARC expression in skeletal muscle of mice causes atrophic changes of myofibers, suggesting that SPARC plays a role in the maintenance of skeletal muscle function. In order to know the effect of long-term deficiency of SPARC on skeletal muscle, we performed phenotypic analyses of skeletal muscle of SPARC-null mice. Age-associated changes of myofiber diameters were comparable between wild type (WT) and SPARC-null mice at all ages examined, indicating that the growth of myofibers is unaffected by the absence of SPARC. On the other hand, accumulation of fibrillar collagen was significantly reduced in SPARC-null mice compared to WT mice after 5 months of age without significant changes of collagen I gene expression. The results obtained in the present study suggest that SPARC plays a role to maintain the stiffness of skeletal muscle by regulating collagen accumulation.
A female Bernese Mountain Dog was diagnosed with a right middle lung lobe mass. The dog was positioned in a left lateral recumbency and one-lung ventilation was used under general anesthesia. Video-assisted thoracic surgery anatomical lobectomy was performed with 4 cm small thoracotomy and two 6-mm ports. Pulmonary vessels and bronchus were dissected and isolated individually at the hilum of the right middle lung lobe. Pulmonary vessels were ligated and were coagulated and transected using a vessel sealing device. The bronchus was ligated and transected. The mass in the right middle lung lobe was removed with a clean margin and without complications. Video-assisted thoracic surgery anatomical lobectomy was used to successfully remove a primary lung tumor in a dog.
Follicle development in post-weaning sows is influenced by various factors. To control ovulation time using hormone, factors that influence ovulation should be investigated. The present study was performed to evaluate the effect of GnRH (buserelin) administration in relation to season and sow parameters on ovulation time in weaned sows. Seventy-seven weaned sows were divided into the following groups: control (hot season, n=21; cool season, n=16) and treatment (hot season, n=22; cool season, n=18). Sows were kept in a close house equipped with an evaporative cooling system. Ovulation time was determined every 6 hr using transrectal ultrasonography. Administration of 10 µg buserelin at 72 hr after weaning affected estrus-to-ovulation interval (EOI) and weaning-to-ovulation interval (WOI) in sows (P<0.05). The percentage of sows that ovulated between 44–56 hr after injection was higher in the cool season than in hot season (P<0.05). Weaning-to-estrus interval (WEI) and injection-to-estrus interval (IEI) were affected by season (P<0.05). Body condition score (BCS) of sows influenced EOI (P<0.01). Sows with low backfat thickness, lactation length <20 days, or litter weight ≥67 kg, had delayed injection-to-ovulation interval (P<0.05). In conclusions, buserelin administration (10 µg, at 72 hr after weaning) advanced ovulation. Hot season prolonged ovulation time. Sows that were weaned with lactation length of at least 20 days, litter weight less than 67 kg, or BCS of at least 3, had better responses to buserelin injection. High backfat reserve after weaning is important for ovulation induction response by buserelin injection.
We examined the effect of human chorionic gonadotropin (hCG) treatment on double ovulation rate of first-wave follicles five days after estrus in lactating dairy cows. Cows were divided into two groups: 1) Ipsilateral group (IG; n=35), in which the first-wave dominant follicle (DF) was ipsilateral to the corpus luteum (CL), and 2) Contralateral group (CG; n=30), in which the first-wave DF was contralateral to the CL five days after estrus, then 1,500 IU of hCG was administrated. Double ovulation rate was significantly higher in the CG (26.7%) than in the IG (2.9%). This study demonstrate that the double ovulation rate of first-wave follicles was higher in the first-wave DF located in the contralateral ovary to the CL.
The objective of this study was to evaluate the breeding performance of Indonesian beef cattle (Ongole cross) as recipients for embryo transfer using Limousin embryos. As a result, the pregnancy rate was 35% (7 out of 20 cows). There was a significant difference (P<0.01) in the serum progesterone concentration between the nonpregnant and pregnant cows at the time of the embryo transfer (day 7 after the estrus). The pregnancy rate in the Indonesian beef cows was low, which may be due to their insufficient genetic quality and/or low physical conditions caused by the poor management, like a low-nutrition diet. The low progesterone concentration in the nonpregnant cows on day 7 may be associated with the failure of embryo implantation.
Dichlorodiphenyltrichloroethane (DDT) is an organochlorine insecticide that has been used for indoor residual spraying for the control of mosquito-borne diseases including malaria. However, due to its toxicity and environmental persistence, there are concerns about its potential deleterious effects in humans and wildlife. Therefore, the current study aimed to monitor and estimate the level of DDTs in human communities. The accumulation of DDT and its metabolites was evaluated in house rat (as sentinel) livers collected in an area where DDT was sprayed. DDTs were measured using a gas chromatography / Electron Capture Detector. The results revealed high concentrations of DDTs in the rat livers and the levels of DDTs were similar to findings reported from the same area in 2014.
Vitamin A comprises vitamin A1 and vitamin A2; vitamin A1 is retinol and its fatty-acid esters and vitamin A2 is 3,4-didehydroretinol and its fatty-acid esters. Although vitamin A1 is generally recognized as the major vitamin A, vitamin A2 is found in some birds and mammals that eat fish containing vitamin A2. Plasma concentration of retinyl esters, but not retinol, is known to increase postprandially in humans. The objectives of this study were to confirm the presence of vitamin A2 in fish fed to penguins, and in penguin plasma, and the postprandial changes in vitamin A concentration in penguin plasma. Blood was collected from six male African penguins (Spheniscus demersus) before and after feeding on jack mackerels (Trachurus japonicus) along with a vitamin premix containing vitamin A1. Vitamin A1 concentration in fish was much higher than the requirement, and was 5-fold higher than the vitamin A2 concentration. Vitamin A2 was present in plasma but its concentration was at least 100-fold below that of plasma retinol, suggesting that vitamin A2 is much less bioavailable than vitamin A1 in penguins. Plasma retinol and retinyl palmitate concentrations were found to be stable after the meal. Plasma retinol concentration is suggested to be homeostatically controlled in penguins against the rapid flow of vitamin A1 after meal. The absorbed vitamin A1 is thought to be transported to the liver via the portal vein for storage in penguins, resulting in stable retinyl palmitate concentration in plasma after meal.
Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy (TSE) that affects members of the Cervidae family, including deer (Odocoileus spp.), elk (Cervus Canadensis spp.), and moose (Alces alces spp.). While CWD is a neurodegenerative disease, lymphoid accumulation of the abnormal isoform of the prion protein (PrPSc) is detectable early in the course of infection. It has been shown that a large portion of the PrPSc lymphoid accumulation in infected mule deer takes place on the surface of follicular dendritic cells (FDCs). In mice, FDC expression of PrPC has been shown to be essential for PrPSc accumulation. FDCs have been shown to normally express high levels of PrPC in mice and humans but this has not been examined in natural hosts for CWD. We used double immunofluorescent labeling and confocal microscopy to determine the PrPC expression characteristics of B and T lymphocytes as well as FDCs in palatine tonsils of CWD-negative mule deer and elk. We detected substantial PrPC colocalization with all cellular phenotypic markers used in this study, not just with FDC phenotypic markers.
Pulmonary mycosis is a fungal disease that commonly affects bottlenose dolphins (Tursiops truncatus) and is generally treated by the oral administration of azoles, such as itraconazole (ITZ) and voriconazole (VRZ). However, antifungal susceptibility testing of clinical isolates has not been well performed as a routine clinical examination in aquaria. In this study, we collected fungal species from the blowholes of 14 bottlenose dolphins, of which 12 were treated with ITZ or VRZ. All dolphins were housed in the Port of Nagoya Public Aquarium. The fungal species Candida albicans, C. tropicalis, C. glabrata, Aspergillus fumigatus, and A. niger were isolated. E-tests were performed to determine the minimum inhibitory concentrations (MICs) of ITZ and VRZ on these isolates. VRZ-resistant C. tropicalis (MIC: >32 µg/ml) and A. niger (MIC: >32 µg/ml) were isolated from three dolphins treated with ITZ or VRZ. Additionally, azole-resistant isolates of C. albicans and C. glabrata were collected from two dolphins that had never received azole therapy. To the best of our knowledge, our study is the first to report the isolation of VRZ-resistant C. albicans, C. tropicalis, and A. niger from the blowholes of bottlenose dolphins. Thus, antifungal susceptibility testing is a crucial strategy for selecting antifungal agents to treat respiratory fungal infections in bottlenose dolphins in aquaria.
An African penguin (Spheniscus demersus) presented with a large peri-cloacal mass. The mass was diagnosed as a pyogranuloma histologically, with multifocal to coalescing inflammatory responses in the subcutis and keratinized simple stratified squamous epithelium overlying the surface. The patient was prescribed 125 mg/kg oral clavulanic acid/amoxycillin twice a day and 20 mg/kg oral terbinafine once a day for 14 days, but there was no change in the size of the mass 4 days after therapy, so a decision was made to administer 1 mg/kg of intralesional methylprednisolone. Fourteen days later, the mass resolved with no further recurrence. This case demonstrated that intralesional methylprednisolone used with antibiotic and antifungals was effective in resolving a peri-cloacal pyogranuloma and without any side effects.