Relationship between the number of Hepatozoon canis gamonts and hematobiochemical values in dogs

@#The occurrence of canine hepatozoonosis in Thailand is primarily caused by Hepatozoon canis. Recently, the relationship of hematology and biochemistry with this disease has been studied, but knowledge regarding the relationship between the quantity of H. canis intracellular gamonts and the hematological profile has not yet been reported. The objective of this study was to investigate the clinical, hematological and biochemical profile of H. canis-positive dogs and the relationship of the number of H. canis gamonts, animal signalment, and hematological and biochemical values. A total of 185 H. canis-positive blood samples were examined, including buffy coat smears and comprehensive data. The number of gamonts was randomly counted from buffy coat smears samples (75/185). The dogs infected with H. canis presented to the animal hospital mostly for health status checks, anorexia, or accidents. Observations from the physical examination on the first day of registration included systemic abnormalities such as digestive, integument, respiratory, urogenital, etc. Most of the dogs showed clinical signs of systemic abnormality in more than one system. Our study shows that plasma proteins are correlated with the number of H. canis gamonts, using Spearman’s rho correlation coefficient with significant difference (p <0.05). This finding could be applied to improve the diagnosis and treatment of canine hepatozoonosis.

Relationship between the number of Hepatozoon canis gamonts and hematobiochemical values in dogs.

The occurrence of canine hepatozoonosis in Thailand is primarily caused by Hepatozoon canis. Recently, the relationship of hematology and biochemistry with this disease has been studied, but knowledge regarding the relationship between the quantity of H. canis intracellular gamonts and the hematological profile has not yet been reported. The objective of this study was to investigate the clinical, hematological and biochemical profile of H. canis-positive dogs and the relationship of the number of H. canis gamonts, animal signalment, and hematological and biochemical values. A total of 185 H. canis-positive blood samples were examined, including buffy coat smears and comprehensive data. The number of gamonts was randomly counted from buffy coat smears samples (75/185). The dogs infected with H. canis presented to the animal hospital mostly for health status checks, anorexia, or accidents. Observations from the physical examination on the first day of registration included systemic abnormalities such as digestive, integument, respiratory, urogenital, etc. Most of the dogs showed clinical signs of systemic abnormality in more than one system. Our study shows that plasma proteins are correlated with the number of H. canis gamonts, using Spearman's rho correlation coefficient with significant difference (p <0.05). This finding could be applied to improve the diagnosis and treatment of canine hepatozoonosis.

The expressions in oxytocin and sex steroid receptors in the reproductive tissues of normal and unilateral cryptorchid dogs.

In males, oxytocin is involved with various physiological functions, such as reproductive tract contractility and testicular steroidogenesis. Due to the relationship between sex steroid hormones, oxytocin receptor (OTR) expression and cryptorchidism pathogenesis, this study aimed to investigate the mRNA expression and the localization of OTR in relation to sex steroid receptors in the male reproductive tract of both normal and unilateral abdominal cryptorchid dogs using quantitative PCR and immunohistochemistry. Male dogs were divided into two groups of normal and cryptorchid dogs. Samples from each cryptorchid dog were separated into two subgroups: scrotal and abdominal subgroups. The results showed that a lower percentage of positive OTR immunostaining in the testis and epididymis was observed in the cryptorchid group compared to the normal group. Within the cryptorchid group, the mRNA expression and the localization of OTR in the testis and epididymis of the abdominal subgroup was less than that of the scrotal subgroup. Moreover, the localization of OTR and estrogen receptor beta (ERß) in reproductive tissues was positively correlated only in the normal group and not in the cryptorchid group. In conclusion, this study proposed that OTR expression, as well as the correlation between the OTR and ERß in reproductive tissues of male dogs, can be disturbed by cryptorchidism. Furthermore, the OTR, ERß and their correlation may be involved with the pathogenesis of cryptorchidism. Therefore, the study of gene knockout models to confirm the effect of OTR and sex steroid receptors on canine cryptorchidism should be of interest for further investigation.

Low-density Lipoprotein Improves Motility and Plasma Membrane Integrity of Cryopreserved Canine Epididymal Spermatozoa.

Cryopreservation of caudal epididymal spermatozoa is an effective technique to conserve genetic potentials of superior dogs when it is not possible to collect ejaculated spermatozoa. Although hen egg yolk is commonly supplemented into the semen extender, active substances within the egg yolk which protect sperm against cryoinjury remain to be discovered. Among its compositions, low-density lipoprotein (LDL) has been reported to have a cryoprotective property for sperm cryopreservation. However, the effects of LDL on dog epididymal spermatozoa during cryopreservation have not yet been investigated. This study aimed to investigate the effects of LDL on epididymal spermatozoa quality following cryopreservation and thawing. After routine castration of 12 dogs, caudal epididymides from individuals were separated from the testes and cut into a few pieces in a Tris-buffer. Spermatozoa recovered from each sample were examined at once for sperm quality and divided into six groups of extender: no LDL, 20% egg yolk, 4%, 8%, 16%, and 24% LDL, before cryopreservation. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, morphology, plasma membrane integrity, and acrosome integrity were evaluated. The results revealed that 4% LDL and 20% egg yolk yielded significantly higher sperm motility (57.69% and 52.69%, respectively, p<0.05) than other LDLs. In addition, 4% LDL yielded the significantly highest plasma membrane integrity (70.54%, p<0.05). In conclusion, the supplementation of 4% LDL in Tris-glucose extender could be applied for cryopreservation of canine epididymal spermatozoa.

Clinical severity score system in dogs with degenerative mitral valve disease.

BACKGROUND: Several risk factors already have been determined for dogs with degenerative mitral valve disease (DMVD). Risk factors often have been considered in isolation and have not always taken into account additional information provided by the history and physical examination (PE). HYPOTHESIS/OBJECTIVES: Data obtained from history and PE of dogs with DMVD provide prognostic information and can be used for risk stratification. ANIMALS: Client-owned dogs (n = 244) with DMVD recruited from first opinion practice. METHODS: Prospective longitudinal follow-up of dogs with DMVD. History and PE data were obtained at 6-month intervals and analyzed with time-dependent Cox models to derive relative risk of cardiac death. Independent hazard ratios were used to derive a clinical severity score (CSS), the prognostic value of which was evaluated by analyzing the median survival times for different risk groups and ROC analysis. Analysis of the progression of CSS over time also was undertaken. RESULTS: History of cough, exercise intolerance, decreased appetite, breathlessness (difficulty breathing) and syncope with PE findings of heart murmur intensity louder than III/VI and absence of respiratory sinus arrhythmia were independently associated with outcome and allowed development of the CSS. Clinical severity score distinguished groups of dogs with significantly different outcomes. CONCLUSIONS AND CLINICAL IMPORTANCE: Routinely obtained clinical findings allow risk stratification of dogs with DMVD. Results of ancillary diagnostic tests may be complementary to history and PE findings and always should be interpreted in conjunction with these findings.

Longitudinal electrocardiographic evaluation of dogs with degenerative mitral valve disease.

BACKGROUND: Increased heart rate (HR) and decreased heart rate variability (HRV) are evident in some dogs with degenerative mitral valve disease (DMVD). OBJECTIVES: Evaluation of the factors influencing HR and HRV (assessed by the vasovagal tonus index; VVTI) and their change over time in dogs with DMVD. ANIMALS: Client-owned dogs (n = 257) with DMVD recruited from first opinion practice. METHODS: Prospective longitudinal follow-up at six-monthly intervals of dogs with DMVD. Dogs followed up for at least 18 months (n = 102) were grouped according to their outcome as dogs dying/euthanized because of cardiac disease (n = 28; Group 1), noncardiac disease (n = 40; Group 2) and dogs alive (n = 34; Group 3). HR and VVTI were measured on 1-minute ECG recordings. Repeated measures linear models were constructed to investigate the factors that influence HR and VVTI and their changes over time. RESULTS: Heart rate and VVTI were affected by disease severity and were different in Cavaliers compared to other breeds. Group 1 and Group 2 dogs underwent an increase in HR and decrease in VVTI, evident at least 18 months before death. Group 1 had a further decrease in VVTI followed by an increase in HR approximately 1 year and 6 months before death, respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: Dogs with DMVD have an increase in HR and decrease in HRV over a year before death, with greater changes in those dogs dying/euthanized because of cardiac disease. Both HR and VVTI can potentially be regarded as biomarkers for all-cause mortality.

The combined prognostic potential of serum high-sensitivity cardiac troponin I and N-terminal pro-B-type natriuretic peptide concentrations in dogs with degenerative mitral valve disease.

BACKGROUND: Identification of factors associated with decreased survival in dogs with degenerative mitral valve disease (DMVD) will allow more accurate prognosis. N-terminal pro-B-type natriuretic peptide (NT-proBNP) is negatively associated with survival in dogs with DMVD. In human patients, multimarker strategies provide superior risk stratification compared with single markers. HYPOTHESIS: High-sensitivity cardiac troponin I (hscTnI) and other clinical variables will be associated with survival time in dogs with DMVD. Measuring hscTnI and NT-proBNP in combination will be prognostically superior to measurement of either marker alone. The rate of change of these markers will vary according to cause of death. ANIMALS: Client-owned dogs (n = 202) with DMVD of varying severity and age-matched healthy control dogs (n = 30) recruited from first opinion private practice. METHODS: Prospective cohort study relating clinical variables at enrollment in dogs with DMVD to survival time (all-cause, cardiac, and noncardiac mortality). Multivariable Cox regression analysis was used to identify factors associated with survival. Measurements were obtained approximately every 6 months. Repeated measures models were constructed to assess changes over time. RESULTS: hscTnI, LVEDDN, heart rate, and age were independently associated with decreased survival time (all-cause mortality). Survival times were shortest in dogs in which both serum hscTnI and NT-proBNP were increased. hscTnI and NT-proBNP increased more rapidly in dogs that died of cardiac disease. CONCLUSIONS AND CLINICAL IMPORTANCE: Serum hscTnI has prognostic value in dogs with DMVD. Measurement of NT-proBNP and hscTnI is prognostically superior to measuring either alone. Serial measurement strategies provide additional prognostic information.

N-terminal pro B-type natriuretic peptide and left ventricular diameter independently predict mortality in dogs with mitral valve disease.

OBJECTIVES: To determine whether natriuretic peptide concentrations would predict all cause mortality in dogs with degenerative mitral valve disease. METHODS: One hundred dogs with naturally occurring degenerative mitral valve disease were prospectively recruited for this longitudinal study. Analysis of outcome was undertaken for 73 dogs for which the outcome was known. Dogs underwent physical examination, electrocardiography and echocardiography. Natriuretic peptide concentrations were measured by Enzyme-linked immunosorbent assay. The ability of natriuretic peptide concentrations, clinical, electrocardiographic and echocardiographic data, to predict all cause mortality was determined using univariable and multivariable Cox proportional hazards analyses. RESULTS: Thirty dogs died during the period of follow-up. Two variables were independently predictive of all cause mortality; these were the normalised left ventricular end-diastolic diameter and the N-terminal pro B-type natriuretic peptide concentration. An increase of the left ventricular end-diastolic diameter by 0.1 increased the hazard of all cause mortality by 20% (95% confidence interval: 4 to 37%, P=0.01) and a 100 pmol/l increase in N-terminal pro B-type natriuretic peptide increased the hazard by 7% (95 confidence interval: 2 to 11%, P=0.003). CLINICAL SIGNIFICANCE: N-terminal pro B-type natriuretic peptide concentration and left ventricular end-diastolic diameter are significantly and independently predictive of all cause mortality in dogs with degenerative mitral valve disease.