Evaluation of circulating IGF-I and IGFBP-3 as biomarkers for tumors in dogs

Background@#Serum-based parameters are considered non-invasive biomarkers for cancer detection. In human studies, insulin-like growth factor-I and II (IGF-I and IGF-II) and insulin-like growth factor binding protein-3 (IGFBP-3) are useful as diagnostic or prognostic markers and potential therapeutic targets. @*Objectives@#This study examined the diagnostic utility of circulating IGF-I, IGF-II, and IGFBP-3 levels in healthy dogs and dogs with tumors. @*Methods@#The serum concentrations of these biomarkers in 86 dogs with tumors were compared with those in 30 healthy dogs using an enzyme-linked immunosorbent assay (ELISA). @*Results@#The ELISA results showed no difference between healthy dogs and dogs with tumors in the serum IGF-II concentrations. On the other hand, there was a significant difference in the circulating IGF-I and IGFBP-3 levels between healthy dogs and dogs with tumors. The concentrations of serum IGF-I (median [interquartile range], 103.4 [59.5–175] ng/mL) in dogs with epithelial tumors were higher than those (58.4 ng/mL [43.5–79.9]) in healthy dogs.Thus, the concentrations of serum IGFBP-3 (43.4 ng/mL [33.2–57.2]) in dogs with malignant mesenchymal tumors were lower than those (60.8 ng/mL [47.6–70.5]) in healthy dogs. @*Conclusions@#The serum IGF-I and IGFBP-3 levels can be used as diagnostic biomarkers in dogs with tumors.

Evaluation of circulating PD-1 and PD-L1 as diagnostic biomarkers in dogs with tumors

Background@#Programmed cell death protein-1 (PD-1) and programmed cell death ligand-1 (PD-L1) have important roles in tumor evasion of the immune system. @*Objectives@#This study aimed to assess the diagnostic utility of circulating PD-1 and PD-L1 levels in healthy dogs and dogs with tumors. @*Methods@#Circulating PD-1 and PD-L1 levels in the serum of 71 dogs with tumors were compared with those of 52 healthy dogs by performing enzyme-linked immunosorbent assay (ELISA). @*Results@#The ELISA results revealed higher circulating PD-1 and PD-L1 levels in dogs with tumors (2.9 [2.2–3.7] ng/mL; median [IQR] and 2.4 [1.4–4.4] ng/mL, respectively) than in healthy dogs (2.4 [1.9–3.0] ng/mL; p = 0.012 and 1.4 [0.9–2.1] ng/mL; p < 0.001, respectively). Especially, there was a significant difference in circulating PD-1 levels between healthy dogs and dogs with malignant epithelial tumors (2.4 [1.9–3.0] ng/mL and 3.1 [2.6–4.4] ng/mL, respectively; p < 0.01). In addition, there was a significant difference in circulating PD-L1 levels between healthy dogs and dogs with lymphomas (1.4 [0.9–2.1] ng/mL and 2.7 [1.6–5.8] ng/mL, respectively; p < 0.001). @*Conclusion@#This study indicates that circulating PD-1 and PD-L1 have potential as tumor diagnostic biomarkers in dogs with tumors.

Prevalence and antimicrobial resistance of Klebsiella species isolated from clinically ill companion animals

Background@#Klebsiella spp. is an important conditional pathogen in humans and animals.However, due to the indiscriminate use of antibiotics, the incidence of antimicrobial resistance has increased. @*Objectives@#The purpose of this study was to investigate antimicrobial resistance in strains of Klebsiella strains and the phylogenetic relatedness of extended-spectrum cephalosporin (ESC)-resistance among Klebsiella strains isolated from clinically ill companion animals. @*Methods@#A total of 336 clinical specimens were collected from animal hospitals.Identification of Klebsiella species, determination of minimum inhibitory concentrations, detection of ESC resistance genes, polymerase chain reaction-based replicon typing of plasmids by conjugation, and multilocus sequence typing were performed. @*Results@#Forty-three Klebsiella strains were isolated and, subsequently, 28 were identified as K. pneumoniae, 11 as K. oxytoca, and 4 as K. aerogenes. Eleven strains were isolated from feces, followed by 10 from ear, 7 from the nasal cavity, 6 from urine, 5 from genitals, and 4 from skin. Klebsiella isolates showed more than 40% resistance to penicillin, cephalosporin, fluoroquinolone, and aminoglycoside. ESCresistance genes, CTX-M groups (CTX-M-3, CTX-M-15, and CTX-M-65), and AmpC (CMY-2 and DHA-1) were most common in the K.pneumoniae strains. Some K. pneumoniae carrying CTX-M or AmpC were transferred via IncFII plasmids. Two sequence types, ST709 and ST307, from K. pneumoniae were most common. @*Conclusions@#In conclusion, this is the first report on the prevalence, ESCresistance genotypes, and sequence types of Klebsiella strains isolated from clinically ill companion animals. The combination of infectious diseases and antimicrobial resistance by Klebsiella in companion animals suggest that, in clinical veterinary, antibiotic selection should be made carefully and in conjunction with the disease diagnosis.

Evaluation of circulating PD-1 and PD-L1 as diagnostic biomarkers in dogs with tumors

Background@#Programmed cell death protein-1 (PD-1) and programmed cell death ligand-1 (PD-L1) have important roles in tumor evasion of the immune system. @*Objectives@#This study aimed to assess the diagnostic utility of circulating PD-1 and PD-L1 levels in healthy dogs and dogs with tumors. @*Methods@#Circulating PD-1 and PD-L1 levels in the serum of 71 dogs with tumors were compared with those of 52 healthy dogs by performing enzyme-linked immunosorbent assay (ELISA). @*Results@#The ELISA results revealed higher circulating PD-1 and PD-L1 levels in dogs with tumors (2.9 [2.2–3.7] ng/mL; median [IQR] and 2.4 [1.4–4.4] ng/mL, respectively) than in healthy dogs (2.4 [1.9–3.0] ng/mL; p = 0.012 and 1.4 [0.9–2.1] ng/mL; p < 0.001, respectively). Especially, there was a significant difference in circulating PD-1 levels between healthy dogs and dogs with malignant epithelial tumors (2.4 [1.9–3.0] ng/mL and 3.1 [2.6–4.4] ng/mL, respectively; p < 0.01). In addition, there was a significant difference in circulating PD-L1 levels between healthy dogs and dogs with lymphomas (1.4 [0.9–2.1] ng/mL and 2.7 [1.6–5.8] ng/mL, respectively; p < 0.001). @*Conclusion@#This study indicates that circulating PD-1 and PD-L1 have potential as tumor diagnostic biomarkers in dogs with tumors.

Prevalence and antimicrobial resistance of Klebsiella species isolated from clinically ill companion animals

Background@#Klebsiella spp. is an important conditional pathogen in humans and animals.However, due to the indiscriminate use of antibiotics, the incidence of antimicrobial resistance has increased. @*Objectives@#The purpose of this study was to investigate antimicrobial resistance in strains of Klebsiella strains and the phylogenetic relatedness of extended-spectrum cephalosporin (ESC)-resistance among Klebsiella strains isolated from clinically ill companion animals. @*Methods@#A total of 336 clinical specimens were collected from animal hospitals.Identification of Klebsiella species, determination of minimum inhibitory concentrations, detection of ESC resistance genes, polymerase chain reaction-based replicon typing of plasmids by conjugation, and multilocus sequence typing were performed. @*Results@#Forty-three Klebsiella strains were isolated and, subsequently, 28 were identified as K. pneumoniae, 11 as K. oxytoca, and 4 as K. aerogenes. Eleven strains were isolated from feces, followed by 10 from ear, 7 from the nasal cavity, 6 from urine, 5 from genitals, and 4 from skin. Klebsiella isolates showed more than 40% resistance to penicillin, cephalosporin, fluoroquinolone, and aminoglycoside. ESCresistance genes, CTX-M groups (CTX-M-3, CTX-M-15, and CTX-M-65), and AmpC (CMY-2 and DHA-1) were most common in the K.pneumoniae strains. Some K. pneumoniae carrying CTX-M or AmpC were transferred via IncFII plasmids. Two sequence types, ST709 and ST307, from K. pneumoniae were most common. @*Conclusions@#In conclusion, this is the first report on the prevalence, ESCresistance genotypes, and sequence types of Klebsiella strains isolated from clinically ill companion animals. The combination of infectious diseases and antimicrobial resistance by Klebsiella in companion animals suggest that, in clinical veterinary, antibiotic selection should be made carefully and in conjunction with the disease diagnosis.

Challenges of stem cell therapies in companion animal practice

Regenerative medicine using stem cells from various sources are emerging treatment modality in several refractory diseases in veterinary medicine. It is well-known that stem cells can differentiate into specific cell types, self-renew, and regenerate. In addition, the unique immunomodulatory effects of stem cells have made stem cell transplantation a promising option for treating a wide range of disease and injuries. Recently, the medical demands for companion animals have been rapidly increasing, and certain disease conditions require alternative treatment options. In this review, we focused on stem cell application research in companion animals including experimental models, case reports and clinical trials in dogs and cats. The clinical studies and therapeutic protocols were categorized, evaluated and summarized according to the organ systems involved. The results indicate that evidence for the effectiveness of cell-based treatment in specific diseases or organ systems is not yet conclusive. Nonetheless, stem cell therapy may be a realistic treatment option in the near future, therefore, considerable efforts are needed to find optimized cell sources, cell numbers and delivery methods in order to standardize treatment methods and evaluation processes.

High-level mupirocin resistance in Gram-positive bacteria isolated from diseased companion animals

The purpose of this study was to investigate the high-level mupirocin resistance (HLMR) in Gram-positive bacteria isolated from companion animals. A total of 931 clinical specimens were collected from diseased pets. The detection of mupirocin-resistant bacteria and plasmid-mediated mupirocin resistance genes were evaluated by antimicrobial susceptibility tests, polymerase chain reactions, and sequencing analysis. Four-hundred and six (43.6%) bacteria were isolated and 17 (4.2%), including 14 staphylococci and 3 Corynebacterium were high-level mupirocin-resistant (MICs, ≥ 1,024 ug/mL) harboring mupA. Six staphylococci of HLMR strains had plasmid-mediated mupA-IS257 flanking regions. The results show that HLMR bacteria could spread in veterinary medicine in the near future.

Genome-wide association study of degenerative mitral valve disease in Maltese dogs

Genome-wide association study (GWAS) is a powerful tool for identifying the genetic causes of various diseases. This study was conducted to identify genomic variation in Maltese dog genomes associated with degenerative mitral valve disease (DMVD) development and to evaluate the association of each biological condition with DMVD in Maltese dogs. DNA was extracted from blood samples obtained from 48 Maltese dogs (32 with DMVD and 16 controls). Genome-wide single nucleotide polymorphism (SNP) genotyping was performed. The top 30 SNPs from each association of various conditions and genetic variations were mapped to their gene locations. A total of 173,662 loci were successfully genotyped, with an overall genotype completion rate of 99.41%. Quality control analysis excluded 46,610 of these SNPs. Manhattan plots were produced using allelic tests with various candidate clinical conditions. A significant peak of association was observed between mitral valve prolapse (MVP) and SNPs on chromosome 17. The present study revealed significant SNPs in several genes associated with cardiac function, including PDZ2, Armadillo repeat protein detected in velo-cardio-facial syndrome, catenin (cadherin-associated protein) alpha 3, low-density lipoprotein receptor class A domain containing protein 4, and sterile alpha motif domain containing protein 3. To our knowledge, this is the first study of a genetic predisposition to DMVD in Maltese dogs. Although only a limited number of cases were analyzed, these data could be the basis for further research on the genetic predisposition to MVP and DMVD in Maltese dogs.

Polymorphism in the serotonin transporter protein gene in Maltese dogs with degenerative mitral valve disease

Degenerative mitral valve disease (DMVD) is the most commonly acquired cardiac disease in dogs. This study evaluated the relationship between genetic variations in the serotonin transporter (SERT) gene of Maltese dogs and DMVD. Genomic DNA was extracted from blood samples collected from 20 client-owned DMVD Maltese dogs and 10 healthy control dogs, and each exon of the SERT gene was amplified via polymerase chain reaction. The resulting genetic sequences were aligned and analyzed for variations by comparing with reference sequences; the predicted secondary structures of these variations were modeled and cross-verified by applying computational methods. Genetic variations, including five nonsynonymous genetic variations, were detected in five exons. Protein structure and function of the five nonsynonymous genetic variations were predicted. Three of the five polymorphisms were predicted to be probable causes of damage to protein function and confirmed by protein structure model verification. This study identified six polymorphisms of the SERT gene in Maltese dogs with DMVD, suggesting an association between the SERT gene and canine DMVD. This is the first study of SERT mutation in Maltese dogs with DMVD and is considered a pilot study into clinical genetic examination for early DMVD diagnosis.

Prolonged reflex bradycardia after occlusion of patent ductus arteriosus with an Amplatz canine duct occluder in a dog

A 9-month-old intact male Maltese dog (1.52 kg) was diagnosed with a patent ductus arteriosus (PDA). Transcatheter occlusion of the PDA was performed by using the Amplatz canine duct occluder (ACDO). After occlusion, reflex bradycardia occurred and lasted for at least 15 h with normal systolic arterial pressure and slightly increased diastolic arterial pressure. The bradycardia slowly resolved, and the heart rate was normal in re-examinations after 7 and 30 days. This is the first case of reflex bradycardia after ACDO implantation, in which the bradycardia continued for a long time, even after recovery from anesthesia.

Primary intestinal mast cell tumor in a Russian Blue cat: ultrasonographic and histopathological findings

A 11-year-old, female Russian Blue cat was presented with anorexia, vomiting, and diarrhea lasting for 3 days. Abdominal ultrasonography revealed a hypoechoic, non-circumferential, and eccentrically formed intestinal loop with altered wall layering and thickening of the tunica muscularis. After surgical resection, histopathologic examination confirmed an infiltrative, round-cell neoplasm composed of sheets and cords of neoplastic mast cells within a fibrotic, edematous stroma. The cat was alive and healthy 6 months after surgery. To the best of our knowledge, this is the first reported case of an intestinal mast cell tumor in a Russian Blue cat in South Korea.

Serum serotonin concentration in lean and obese dogs with myxomatous mitral valve disease

The aim of the present study is to investigate the potential influence of obesity as a factor in 5-hydroxytryptamine (5-HT) concentration in myxomatous mitral valve disease (MMVD) dogs. Fifty-five client-owned dogs were enrolled in a randomized trial. Dogs were classified by echocardiography into healthy (control), mild, and moderate to severe MMVD groups. Each group was subclassified by using a 9-point body condition score (BCS); lean (BCS 5–6/9) and obese groups (BCS 7.5–9/9). Dogs with moderate to severe MMVD had lower serotonin (5-HT) concentrations than the control group (p = 0.03). Dogs with moderate to severe MMVD (p = 0.017) had lower serum 5-HT concentrations than the control group in the obese group (BCS 7.5–9/9). Significant difference was found between the lean and obese groups (p = 0.015) which are not consider severe in the MMVD group. These results suggested that 5-HT concentration was decreased with the increasing severity of MMVD, and obesity might be taken into consideration when interpreting the serotonin concentration in MMVD dogs.

Balloon valvuloplasty for severe subaortic stenosis in a Pomeranian dog

A nine-month-old Pomeranian dog with exercise intolerance and syncope was presented. The dog was depressed with grade 4 systolic murmur on cardiac auscultation. Based on cardiac examination, the dog was diagnosed with severe subaortic stenosis with involvement of the anterior mitral valve. β-blocker administration was initiated and clinical signs were improved, but not fully resolved. Balloon valvuloplasty was performed and the dog survived for nearly one year without clinical sign and the cardiac troponin I level was normalized. This case describes successful management of severe subaortic stenosis in a small breed dog through balloon valvuloplasty.

Acquired myasthenia gravis in a domestic shorthair cat with cranial mediastinal mass

A 7-year-old castrated male domestic short-hair cat presented with anorexia, constipation, depression, and voice alteration. Physical and neurological examinations revealed hyperthermia (40.5℃), ventroflexion of the neck, reduced responses to external stimuli, generalized muscle weakness, and exercise intolerance. Thoracic radiographs revealed the presence of a cranial mediastinal mass. The history, clinical signs, and other examination results were compatible with acquired myasthenia gravis (MG). Acetylcholine receptor (AChR) antibody titers were determined to confirm MG and the serum AChR antibody concentration was 1.24 nmol/L (reference interval, < 0.3 nmol/L). This is the first diagnosis of acquired MG in a cat in Korea.

Combination therapy of cyclosporine and prednisolone in a dog with systemic lupus erythematosus

An 11-year-old, spayed female poodle presented with fever and shifting lameness. Physical examination revealed hyperthermia (40.6℃), and proteinuria was detected upon urinalysis. Increased neutrophils (83%) and decreased viscosity were revealed upon synovial fluid analysis. Serum antinuclear antibody was positive at 1 : 80. Based on these findings, the dog was diagnosed with systemic lupus erythematosus. Immunosuppressive therapy was initiated with prednisolone and cyclosporine, and the condition was markedly improved after the treatments. This case report describes the clinical and laboratory findings, imaging characteristics and successful outcomes after prednisolone plus cyclosporine therapy in a canine systemic lupus erythematosus case.

Primary renal fibrosarcoma with local invasion into the mesenteric membrane of a mongrel dog / 대한수의학회지

A 14-year-old, 7.4 kg, neutered male mongrel dog presented with vomiting, anorexia, and hematuria starting 3 days prior to admission. Serum biochemical profiles indicated severe azotemia. Computed tomography revealed loss of normal left kidney structure. The organ was 1.5 to 2 times larger than the right kidney with mixed attenuation. Histopathologic examination was performed after nephrectomy. The renal mass and mesenteric membrane were positive for vimentin and stained blue with Masson's trichrome. In conclusion, this was a rare occurrence of primary renal fibrosarcoma, most likely originated from the renal capsule, with local invasion into the mesenteric membrane.

Characteristic Clinical Features and Survival in Cats with Symptomatic Hypertrophic Cardiomyopathy / 동물의과학연구지

Hypertrophic cardiomyopathy (HCM) is recognized as the most common feline cardiac disease. Several studies have evaluated the population characteristics and survival time of cats with HCM; however, these reports included large numbers of asymptomatic HCM. The purpose of this study was to evaluate the characteristics and survival time of cats with symptomatic HCM admitted to emergency service. Medical records were examined to verify clinical diagnosis of HCM. Asymptomatic cats diagnosed with HCM were also excluded from the study. From a total of 13 cats, eight were classified in the arterial thromboembolism (ATE) group while the other five were in the congestive heart failure (CHF) group. Middle-aged (6 years, range 1.4~7 years) male cats (53.8%) were included in this study. Pelvic limb paralysis, depression, and respiratory distress were common clinical signs found in symptomatic HCM cats. Hematologic evaluation found that enzymes related to muscle damage and tissue necrosis were elevated in both groups and highly elevated in the ATE group. Left atrium was remarkably enlarged in ATE group cats. The median survival time of cats in the ATE group was significantly shorter than that of cats in the CHF group (P=0.002). Prospective investigation based on a large population would be required to clarify the effects of various factors on prognosis of HCM cats.

Drug-induced blood cell dyscrasia associated with phenobarbital administration in a dog / 대한수의학회지

A 13-year-old, spayed, female Chihuahua dog was referred for evaluation of fever, lethargy, and dyspnea. Hematologic evaluation revealed severe neutropenia, thrombocytopenia, and mild anemia. The dog had been undergoing phenobarbital therapy for the past 7 weeks because of generalized seizures due to meningoencephalomyelitis of unknown etiology. After ruling out other possible causes of cytopenias, a tentative diagnosis was made of drug-induced blood cell dyscrasia. The neutropenia and thrombocytopenia resolved after discontinuation of phenobarbital (8 days and 15 days after discontinuation, respectively). This is the first case report in Korea to demonstrate blood dyscrasia associated with idiosyncratic adverse effects of phenobarbital.

Alopecia areata in a spayed pug: clinical and immunohistochemical findings / 대한수의학회지

A two-year-old spayed female pug presented with symmetrical hyperpigmented alopecic lesions on her axillary and inguinal regions. There were no remarkable findings in dermatologic examinations and hormonal assays. Histological examination of biopsied tissues revealed prominent lymphocytic perifolliculitis along with shrunk hair follicles. Immunohistochemistry for CD3, CD79a, CD4, and CD8 showed a positive stain for CD4 antigen around hair bulbs, suggesting CD4 positive T lymphocyte infiltration. This case suggests the possibility that CD4 T lymphocyte-mediated inflammatory reaction could be a main mechanism in canine alopecia areata. Additional studies are warranted to investigate the immunological mechanism in canine species.

Identification of Staphylococcus pseudintermedius in canine infective endocarditis using PCR-RFLP analysis / 동물의과학연구지

A 5-year-old, 8.95 kg, female Schnauzer presented anorexia with a 3-day history and increased heart sound intensity. Based on the clinical and echocardiographic findings along with the positive blood culture result, the dog was diagnosed with infective endocarditis (IE). Using proper antibiotics treatment, clinical signs were improved within 3 days and resolved within 1 week. For exact identification of the causative agent, multiplex polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) methods were performed. The etiological agent was confirmed as Staphylococcus pseudintermedius with antibiotics resistance genes such as beta-lactamase (blaZ) and methicilline resistance (mecA). The bacterial virulence factors included pyogenic toxin genes such as staphylococcal enterotoxins A, B, C, D, and E and toxic shock syndrome toxin 1. Diagnosis of IE is challenging due to a variety of non-specific clinical presentations, rapid disease progression, and lack of a confirmative diagnostic technique. This report demonstrated that such molecular diagnostics could be very useful for diagnosing and identifying characteristics of the causative organism for prediction of prognosis and proper treatment. To our knowledge, this is the first report on the isolation of S. pseudintermedius using molecular diagnostics from a clinical case of canine IE.