RESUMO
X-linked hereditary nephropathy (XLHN) in Navasota dogs is a spontaneously occurring disease caused by a mutation resulting in defective production of type IV collagen and juvenile-onset renal failure. The study was aimed at examining the evolution of renal damage and the expression of selected molecules potentially involved in the pathogenesis of XLHN. Clinical data and renal samples were obtained in 10 XLHN male dogs and 5 controls at 4 (T0), 6 (T1), and 9 (T2) months of age. Glomerular and tubulointerstitial lesions were scored by light microscopy, and the expression of 21 molecules was investigated by quantitative real-time polymerase chain reaction with selected proteins evaluated by immunohistochemistry. No significant histologic lesions or clinicopathologic abnormalities were identified in controls at any time-point. XLHN dogs had progressive proteinuria starting at T0. At T1, XLHN dogs had a mesangioproliferative glomerulopathy with glomerular loss, tubular necrosis, and interstitial fibrosis. At T2, glomerular and tubulointerstitial lesions were more severe, particularly glomerular loss, interstitial fibrosis, and inflammation. At T0, transforming growth factor ß, connective tissue growth factor, and platelet-derived growth factor α mRNA were overexpressed in XLHN dogs compared with controls. Clusterin and TIMP1 transcripts were upregulated in later stages of the disease. Transforming growth factor ß, connective tissue growth factor, and platelet-derived growth factor α should be considered as key players in the initial events of XHLN. Clusterin and TIMP1 appear to be more associated with the progression rather than initiation of tubulointerstitial damage in chronic renal disease.
Assuntos
Doenças do Cão/genética , Doenças Genéticas Ligadas ao Cromossomo X/veterinária , Nefropatias/veterinária , Nefrite Hereditária/veterinária , Animais , Colágeno Tipo IV/genética , Progressão da Doença , Doenças do Cão/metabolismo , Doenças do Cão/patologia , Cães , Doenças Genéticas Ligadas ao Cromossomo X/genética , Doenças Genéticas Ligadas ao Cromossomo X/metabolismo , Doenças Genéticas Ligadas ao Cromossomo X/patologia , Imuno-Histoquímica/veterinária , Rim/metabolismo , Rim/patologia , Nefropatias/genética , Nefropatias/metabolismo , Nefropatias/patologia , Glomérulos Renais/metabolismo , Glomérulos Renais/patologia , Masculino , Nefrite Hereditária/genética , Nefrite Hereditária/metabolismo , Nefrite Hereditária/patologia , Fator de Crescimento Derivado de Plaquetas/metabolismo , Proteinúria/veterinária , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Fator de Crescimento Transformador beta/metabolismoRESUMO
OBJECTIVES: To describe how veterinarians utilise and perform urinalyses for dogs and cats. MATERIALS AND METHODS: A survey, developed and distributed through the Veterinary Information Network, enlisted veterinarians who perform urinalyses for dogs and cats. Participants were directed to question banks based on whether urinalyses were performed in-house, by an outside diagnostic laboratory, or using an in-house automated instrument. Participants using multiple methods were directed to questions that related to the chosen methods. RESULTS: A total of 1059 predominantly first-opinion clinicians from the USA and Canada completed the survey. Participants performed urinalyses much less frequently than blood work during a routine examination. The most common factors preventing participants from performing a urinalysis with blood work included clients' financial constraints, difficulty obtaining urine and lack of perceived diagnostic need. The most common reasons for submission to a diagnostic laboratory included efficiency, more trusted results and convenience. Speed of obtaining results was the most common reason for performing urinalyses in-house. Of the participants who performed in-house urinalyses, fewer always performed a manual sediment examination (79%) as compared with urine-specific gravity (99%) and manual dipstick (87%). CLINICAL SIGNIFICANCE: This survey documents that urinalysis is often not used in senior patients as recommended by recent clinical guidelines for dogs and cats which can result in decreased diagnosis and impaired management of subclinical disease. There is significant variability in urinalysis methods despite veterinary guidelines promoting standardisation, and this could lead to inaccurate results.
Assuntos
Doenças do Gato , Doenças do Cão , Gatos , Cães , Animais , Doenças do Gato/diagnóstico , Doenças do Cão/diagnóstico , Urinálise/veterinária , Urinálise/métodosRESUMO
In 2006, Nabity et al. reported on an atypical presentation of Trypanosoma cruzi (T. cruzi) infection in an 8-month old English Mastiff from central Texas. Clinical signs and laboratory findings included lymphadenopathy, weight loss, amastigotes in lymph node aspirates, and initial serological results suggestive of either T. cruzi or Leishmania infection. Given the poor prognosis, the dog was euthanized and subsequent testing and culture of parasites from a lymph node revealed T. cruzi infection. Because different parasite discrete typing units (DTUs) are potentially associated with different disease outcomes in a variety of mammalian hosts, an understanding of these relationships in naturally infected dogs may be useful for informing canine prognosis and may also have human health implications. Here, we compared strains using culture versus culture-independent methods. We subjected archived cultured parasites harvested from the lymph node in the infected Mastiff to two independent approaches for determining parasite DTU, including sequencing of the TcSC5D gene and use of DTU-specific qPCR probes to hybridize the nuclear spliced leader intergenic region (SL-IR). Both approaches revealed T. cruzi discrete typing unit TcIV. Testing of multiple other tissues directly without culturing, including heart/tongue, intestine, trachea/lymph nodes, and uterus/ovary, provided further evidence of disseminated TcIV infection in this dog. We report T. cruzi DTU TcIV as the cause of a severe disseminated infection in a dog from an area with triatomine vectors in central Texas, adding to the limited body of clinicopathologic data that links specific parasite strains to disease outcomes in dogs in the US. Future studies to type parasites from asymptomatic dogs and those with diverse disease manifestations will be useful in informing the degree to which parasite genetics is associated with disease presentation and severity. If applied to antemortem samples, diagnostic typing of parasites from infected dogs may assist in determining prognosis.
Assuntos
Doença de Chagas/veterinária , Doenças do Cão/diagnóstico , Trypanosoma cruzi/classificação , Animais , Bancos de Espécimes Biológicos , Doença de Chagas/diagnóstico , Doenças do Cão/parasitologia , Cães , Feminino , Variação Genética , Genótipo , Insetos Vetores/parasitologia , Linfonodos/parasitologia , Reação em Cadeia da Polimerase , Proteínas de Protozoários/genética , Texas , Triatoma/parasitologia , Trypanosoma cruzi/isolamento & purificaçãoRESUMO
Serum canine α1-proteinase inhibitor (cα1-PI) concentrations were evaluated in dogs with pancreatitis (n=24), exocrine pancreatic insufficiency (EPI; n=29), chronic hepatitis (CH; n=11) or proteinuric chronic kidney disease (CKD-P; n=61) to determine whether systemic proteinase/proteinase-inhibitor balance is altered in these conditions. Dogs with CKD-P had significantly lower cα1-PI concentrations than dogs with pancreatitis, EPI or CH; 16% of dogs with CKD-P had serum cα1-PI concentrations below the reference interval. Serum and urine cα1-PI concentrations were inversely correlated in dogs with CKD-P, but not in dogs with CH. This suggests that renal loss of cα1-PI contributes to decreased serum concentrations in dogs with CKD-P, while hepatic cα1-PI synthesis with CH either is not compromised or is counterbalanced by extrahepatic production.
Assuntos
Doenças do Cão/sangue , Hepatite Crônica/veterinária , Pancreatopatias/veterinária , Inibidores de Proteases/sangue , Insuficiência Renal Crônica/veterinária , Animais , Cães , Feminino , Hepatite Crônica/sangue , Masculino , Pancreatopatias/sangue , Peptídeo Hidrolases , Insuficiência Renal Crônica/sangueRESUMO
A case of Trypanosoma cruzi infection in a young English Mastiff from Texas is presented. Clinical signs and laboratory findings included subcutaneous edema, lymphadenopathy, weight loss, and hypoalbuminemia. Cytology of a lymph node revealed numerous amastigotes. No trypomastigotes were observed in buffy coat preparation of peripheral blood, and on histologic evaluation, most organs contained numerous interstitial pseudocysts. Initial serology was positive for both T. cruzi and Leishmania, and immunohistochemistry supported a diagnosis of Leishmania. However, additional serology supported a T. cruzi infection, and cultivation of organisms isolated from a lymph node revealed morphology consistent with T. cruzi. In addition, PCR analysis resulted in a 504 bp fragment with 99% homology to a flagellar protein of T. cruzi. Although uncommon, autochthonous cases of both T. cruzi and Leishmania have been reported in the United States. Clinical signs observed with both diseases can show many similarities, cytology may be indistinguishable, as in this case, and serological cross-reactivity is common. This case demonstrates an unusual presentation of T. cruzi and the use of multiple testing strategies to support its diagnosis.
Assuntos
Doença de Chagas/veterinária , Doenças do Cão/diagnóstico , Trypanosoma cruzi/isolamento & purificação , Animais , Doença de Chagas/diagnóstico , Doença de Chagas/patologia , Reações Cruzadas , Diagnóstico Diferencial , Doenças do Cão/patologia , Cães , Evolução Fatal , Feminino , Imuno-Histoquímica/veterinária , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/veterinária , Trypanosoma cruzi/imunologiaRESUMO
BACKGROUND: Urine protein loss is common in dogs with chronic kidney disease (CKD). HYPOTHESIS/OBJECTIVES: To evaluate new biomarkers of glomerular and tubulointerstitial (TI) damage compared with histology and as survival indicators in dogs with naturally occurring, proteinuric CKD. ANIMALS: One hunderd and eighty dogs with naturally occurring kidney disease. METHODS: Retrospective study using urine, serum, and renal biopsies from dogs with kidney disease, 91% of which had proteinuric CKD. Biomarkers were evaluated and correlated with pathologic renal damage, and significant associations, sensitivities, and specificities of biomarkers for renal disease type were determined. RESULTS: Fractional excretions of immunogloblin M (IgM_FE) and immunoglobulin G (IgG_FE) correlated most strongly with glomerular damage based on light microscopy (r = 0.58 and 0.56, respectively; P < .01). Serum creatinine (SCr) correlated most strongly with TI damage (r = 0.70, P < .01). Urine IgM/creatinine and urine NAG/creatinine had the highest sensitivity (75%) and specificity (78%) for detection of immune complex-mediated glomerulonephritis. Although individually most biomarkers were significantly associated with decreased survival time (P < .05), in a multivariate analysis, SCr, IgM_FE, and glomerular damage based on transmission electron microscopy (TEM) were the only biomarkers significantly associated with survival time (SCr: P = .001; IgM_FE: P = .008; TEM: P = .017). CONCLUSIONS AND CLINICAL IMPORTANCE: Novel urine biomarkers and FEs are useful for detection of glomerular and TI damage in dogs with proteinuric CKD and might predict specific disease types and survival.
Assuntos
Doenças do Cão/patologia , Proteinúria/veterinária , Insuficiência Renal Crônica/veterinária , Animais , Biomarcadores/sangue , Biomarcadores/urina , Doenças do Cão/sangue , Doenças do Cão/urina , Cães , Feminino , Masculino , Proteinúria/sangue , Proteinúria/urina , Insuficiência Renal Crônica/sangue , Insuficiência Renal Crônica/urina , Estudos RetrospectivosRESUMO
BACKGROUND: Symmetric dimethylarginine (SDMA) is a small molecule formed by methylation of arginine, and released into blood during protein degradation. SDMA is primarily eliminated by renal excretion and is a promising endogenous marker of glomerular filtration rate (GFR). OBJECTIVES: To validate an assay for SDMA measurement, determine stability of SDMA in blood, and compare SDMA with serum creatinine concentration (sCr) and GFR for early detection of decreasing kidney function in dogs with chronic kidney disease (CKD). ANIMALS: Eight male dogs affected with X-linked hereditary nephropathy and 4 unaffected male littermates. METHODS: Prospective study validating SDMA measurement using liquid chromatography-mass spectrometry, assessing stability of SDMA in serum and plasma, and serially determining sCr, SDMA, and GFR (using iohexol clearance) in dogs during progression from preclinical disease to end-stage renal failure. Correlations were determined using linear regression. Timepoints at which sCr, SDMA, and GFR identified decreased renal function were compared using defined cutoffs, trending in an individual dog, and comparison with unaffected littermates. RESULTS: Symmetric dimethylarginine was highly stable in serum and plasma, and the assay demonstrated excellent analytical performance. In unaffected dogs, SDMA remained unchanged whereas in affected dogs, SDMA increased during disease progression, correlating strongly with an increase in sCr (r = 0.95) and decrease in GFR (r = -0.95). Although trending improved sCr's sensitivity, SDMA identified, on average, <20% decrease in GFR, which was earlier than sCr using any comparison method. CONCLUSIONS AND CLINICAL IMPORTANCE: Symmetric dimethylarginine is useful for both early identification and monitoring of decreased renal function in dogs with CKD.
Assuntos
Arginina/análogos & derivados , Insuficiência Renal Crônica/veterinária , Animais , Arginina/sangue , Biomarcadores/sangue , Estudos de Casos e Controles , Doenças do Cão , Cães/sangue , Diagnóstico Precoce , Taxa de Filtração Glomerular/veterinária , Masculino , Insuficiência Renal Crônica/sangue , Insuficiência Renal Crônica/diagnóstico , Reprodutibilidade dos TestesRESUMO
BACKGROUND: Bacterial infection of the urinary tract is a common disorder in dogs and cats. Although microscopic examination of urine sediment is routinely used to screen for infection, this test can lack sensitivity or require expertise. A reliable in-clinic screening test would be a useful adjunct for the identification of dogs and cats with bacterial urinary tract infection (UTI). HYPOTHESIS: That a catalase-based urine test (Accutest Uriscreen™) is a more sensitive screening test for UTI in dogs and cats than urine microscopic sediment examination. ANIMALS: One hundred and sixty client-owned dogs and cats. METHODS: Surplus urine from animals presented to a veterinary teaching hospital was used in this prospective observational study. A routine urinalysis, aerobic bacterial culture, and the Uriscreen test were performed on cystocentesis samples. Sensitivity and specificity with 95% confidence intervals and positive and negative likelihood ratios were calculated for Uriscreen and microscopic sediment examination using culture results as the gold standard. RESULTS: Bacterial culture was positive in 27/165 (16.4%) samples. The sensitivity, specificity, and positive and negative likelihood ratios for the Uriscreen were 89%, 71%, 3.0, and 0.15, respectively. Sensitivity, specificity, and positive and negative likelihood ratios for urine sediment microscopic examination were 78%, 90%, 7.8, and 0.24, respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: The Uriscreen is a more sensitive screening test for UTI in dogs and cats than sediment examination; however, the urine sediment examination was more specific. A negative Uriscreen result helps exclude UTI; however, urine bacterial culture is still necessary to exclude or confirm UTI in all cases.
Assuntos
Doenças do Gato/microbiologia , Catalase/urina , Doenças do Cão/microbiologia , Infecções Urinárias/veterinária , Animais , Doenças do Gato/urina , Gatos , Doenças do Cão/urina , Cães , Valor Preditivo dos Testes , Estudos Prospectivos , Sensibilidade e Especificidade , Infecções Urinárias/microbiologia , Infecções Urinárias/urinaRESUMO
Tubulointerstitial fibrosis (TIF) plays a central role in the progression to end-stage renal disease. Tubular epithelial cells (TECs) undergo epithelial-mesenchymal transition (EMT) and may contribute to the progression of TIF. Using immunohistochemistry, the primary aim of this study was to assess the expression of ß-catenin, human leukocyte antigen-DR (HLA-DR) and vimentin in renal biopsies from dogs with spontaneous kidney diseases of varying severities. Morphological diagnosis, severity of inflammation, TIF, HLA-DR expression and clinicopathological variables were compared in dogs with renal injury to identify any potential relationship between the different factors; ß-catenin down-regulation was used as a marker of EMT. Fibrosis, HLA-DR expression, serum creatinine concentration (SCr), and urine protein-to-creatinine ratio (UPC) were all increased and ß-catenin expression decreased in dogs with primary glomerular disease compared with dogs with acute tubular necrosis. HLA-DR expression by TECs was positively correlated to fibrosis, inflammation, UPC, and SCr. ß-catenin expression was negatively correlated to fibrosis, inflammation and HLA-DR expression. The progression of renal failure correlated closely with tubulointerstitial damage. De novo HLA-DR expression associated with ß-catenin down-regulation by TECs may represent a possible step in the progression of TIF and EMT.
Assuntos
Doenças do Cão/genética , Células Epiteliais/metabolismo , Fibrose/veterinária , Antígenos HLA-DR/genética , Nefropatias/veterinária , Túbulos Renais/metabolismo , beta Catenina/genética , Animais , Doenças do Cão/etiologia , Doenças do Cão/metabolismo , Doenças do Cão/patologia , Cães , Regulação para Baixo , Células Epiteliais/patologia , Transição Epitelial-Mesenquimal , Feminino , Fibrose/etiologia , Fibrose/genética , Fibrose/patologia , Antígenos HLA-DR/metabolismo , Humanos , Imuno-Histoquímica/veterinária , Inflamação/etiologia , Inflamação/genética , Inflamação/patologia , Inflamação/veterinária , Nefropatias/etiologia , Nefropatias/patologia , Túbulos Renais/patologia , Masculino , Vimentina/genética , Vimentina/metabolismo , beta Catenina/metabolismoRESUMO
BACKGROUND: Glomerulonephropathies are common causes of kidney disease in dogs. OBJECTIVE: To determine the prevalence of immune-complex glomerulonephritis (ICGN) in North American dogs biopsied for suspected glomerular disease. ANIMALS: Renal biopsies (n = 733) submitted to the Texas Veterinary Renal Pathology Service between January 1, 2007 and December 31, 2012 were reviewed. Dogs were included if the biopsy was performed for suspected glomerular disease. METHODS: Specimens were evaluated by light microscopy (LM), immunofluorescence (IF), and transmission electron microscopy (TEM). Findings were retrospectively evaluated to categorize the diagnosis for each case. For the diagnosis of ICGN, TEM findings were considered conclusive when LM and IF were equivocal. RESULTS: Of the 501 dogs included in the study, 241 (48.1%) had ICGN; 103 (20.6%) had primary glomerulosclerosis; 76 (15.2%) had amyloidosis; 45 (9.0%) had nonimmune complex (IC) glomerulopathy; 24 (4.8%) had non-IC nephropathy; and, 12 (2.4%) had primary tubulointerstitial disease. Many (66/241; 27.4%) ICGN cases required TEM for definitive diagnosis, including 14 cases (5.8%) that were not suspected on LM. Of cases not diagnosed as ICGN, a substantial proportion (60/260; 23.1%) required TEM to rule out immune complex deposits, including 14 of 189 cases (7.4%) presumptively diagnosed as ICGN on LM. CONCLUSIONS AND CLINICAL IMPORTANCE: Approximately half of all dogs biopsied for suspected glomerular disease had conditions other than ICGN. Renal biopsy is needed to accurately categorize the underlying disease and direct appropriate treatment. Additionally, TEM and IF evaluations by experienced nephropathologists are necessary to obtain an accurate diagnosis in many cases.
Assuntos
Amiloidose/veterinária , Complexo Antígeno-Anticorpo/imunologia , Doenças do Cão/epidemiologia , Doenças do Cão/imunologia , Glomerulonefrite/veterinária , Amiloidose/epidemiologia , Amiloidose/imunologia , Animais , Complexo Antígeno-Anticorpo/ultraestrutura , Biópsia/veterinária , Distribuição de Qui-Quadrado , Intervalos de Confiança , Cães , Feminino , Glomerulonefrite/epidemiologia , Glomerulonefrite/imunologia , Masculino , Microscopia Eletrônica de Transmissão/veterinária , Microscopia de Fluorescência , América do Norte/epidemiologia , Prevalência , Estudos Retrospectivos , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Sensitive and specific biomarkers for early tubulointerstitial injury are lacking. HYPOTHESIS: The excretion of certain urinary proteins will correlate with the state of renal injury in dogs with chronic kidney disease. ANIMALS: Twenty-five male colony dogs affected with X-linked hereditary nephropathy (XLHN) and 19 unaffected male littermates were evaluated. METHODS: Retrospective analysis of urine samples collected every 2-4 weeks was performed. Urine proteins evaluated were retinol binding protein (uRBP/c), ß2-microglobulin (uB2M), N-acetyl-ß-D-glucosaminidase (uNAG/c), neutrophil gelatinase-associated lipocalin (uNGAL/c), and immunoglobulin G (uIgG/c). Results were correlated with serum creatinine concentration (sCr), glomerular filtration rate (GFR), urine protein : creatinine ratio, and histopathologic analysis of serial renal biopsies. Analytical validation was performed for all assays; uNAG stability was evaluated. RESULTS: All urinary biomarkers distinguished affected dogs from unaffected dogs early in their disease process, increasing during early and midstages of disease. uRBP/c correlated most strongly with conventional measures of disease severity, including increasing sCr (r = 0.89), decreasing GFR (r = -0.77), and interstitial fibrosis (r = 0.80), P < .001. However, multivariate analysis revealed age, sCr, uIgG/c, and uB2M, but not uRBP/c, as significant independent predictors of GFR (P < .05). CONCLUSIONS AND CLINICAL IMPORTANCE: All urinary biomarkers were elevated before sCr increased, but typically after proteinuria developed in dogs with progressive glomerular disease because of XLHN. uRBP/c measurement might be promising as a noninvasive tool for diagnosis and monitoring of tubular injury and dysfunction in dogs.
Assuntos
Doenças do Cão/urina , Doenças Genéticas Ligadas ao Cromossomo X/veterinária , Nefrite Hereditária/veterinária , Acetilglucosaminidase/urina , Animais , Biomarcadores/urina , Biópsia/veterinária , Creatinina/urina , Doenças do Cão/genética , Doenças do Cão/patologia , Cães , Doenças Genéticas Ligadas ao Cromossomo X/genética , Doenças Genéticas Ligadas ao Cromossomo X/patologia , Doenças Genéticas Ligadas ao Cromossomo X/urina , Taxa de Filtração Glomerular/veterinária , Histocitoquímica/veterinária , Modelos Lineares , Lipocalinas/urina , Masculino , Nefrite Hereditária/genética , Nefrite Hereditária/patologia , Nefrite Hereditária/urina , Proteínas de Ligação ao Retinol/urina , Estudos Retrospectivos , Microglobulina beta-2/urinaRESUMO
BACKGROUND: Autosomal recessive hereditary nephropathy (ARHN) was diagnosed in 2 English Springer Spaniels (ESS), a breed not previously reported to be affected by hereditary nephropathy (HN). OBJECTIVE: To identify and characterize the genetic cause of ARHN in ESS. ANIMALS: Sixty-three ESS (2 with ARHN, 2 obligate carriers, and 59 others), 2 mixed-breed dogs with X-linked HN, and 2 English Cocker Spaniels (ECS) with ARHN were included. METHODS: ARHN was diagnosed based on transmission electron microscopy and immunostaining of kidney. DNA from affected dogs was screened for the mutation known to cause ARHN in ECS. Quantities of COL4A3, COL4A4, and COL4A5 mRNA transcripts in renal cortex were determined using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) for ARHN-affected dogs and 7 other dogs. The coding regions of COL4A3 and COL4A4 were sequenced for the 2 ARHN-affected ESS and an unaffected dog. Exon 30 of COL4A4 was sequenced for all 63 ESS. RESULTS: qRT-PCR indicated a significant reduction in transcript levels of both COL4A3 and COL4A4 mRNA in the kidney of ARHN-affected ESS. Sequencing identified a single nucleotide substitution in COL4A4 at base 2806 resulting in a premature stop codon. Thirteen of 25 related dogs were identified as carriers. CONCLUSIONS AND CLINICAL IMPORTANCE: A mutation highly likely to cause ARHN in ESS has been identified.