Regenerative response in dogs naturally and experimentally infected with Babesia rossi.

BACKGROUND: The regenerative response following Babesia rossi infection in dogs is mild, despite severe hemolytic anemia. OBJECTIVE: We aimed to compare the admission absolute reticulocyte count (ARC) and reticulocyte indices in 103 dogs naturally infected with B. rossi with 10 dogs suffering from immune-mediated hemolytic anemia (IMHA) and 14 healthy control dogs. The regenerative response was also evaluated in five dogs experimentally infected with B. rossi. METHODS: This is a retrospective observational study of records generated on the ADVIA 2120 hematology analyzer. RESULTS: The median hematocrits (HCT) of the B. rossi and IMHA groups were significantly lower than the control group (p < .001 for both); however, no differences were seen between the B. rossi and IMHA groups. Compared with the control group, the median ARC was significantly higher in the B. rossi (p = .006) and IMHA (p = .019) groups but significantly lower in the B. rossi group than the IMHA group (p = .041). In the experimentally infected dogs, there was a sudden decrease in the ARC approximately 48 h after the detection of peripheral parasitemia, which was followed by an increase after treatment. Reticulocytes of naturally infected B. rossi dogs were larger, with more variation in cellular volume. The reticulocytes of the experimentally infected dogs decreased in size with decreasing hemoglobin concentrations as the study progressed. CONCLUSIONS: The regenerative response in dogs naturally infected with B. rossi is inadequate, given the severity of the anemia observed, and it might be a result of direct suppressive action by the parasite or host response on the bone marrow.

A prospective evaluation of the prevalence of thromboemboli and associated hemostatic dysfunction in dogs with carcinoma or sarcoma.

BACKGROUND: Knowledge of the prevalence of thromboemboli and the associated hemostatic status in dogs with carcinoma or sarcoma is unknown and might allow earlier intervention. OBJECTIVES: Estimate prevalence of thromboemboli and their association with hemostatic changes in dogs with carcinomas or sarcomas; estimate predictive values of hemostatic variables for thromboembolic disease in tumor-bearing dogs. ANIMALS: Thirty-two dogs with sarcoma, 30 with carcinoma, 20 healthy age-controlled dogs. METHODS: Prospective cross-sectional study. A hemostasis panel (platelet concentration, thromboelastography, fibrinogen and D-dimer concentration, factor X, VII and antithrombin activity) was performed in all dogs. Tumor-bearing dogs underwent complete post mortem and histopathological evaluation. Comparisons between healthy dogs and tumor-bearing dogs with and without intracavitary hemorrhage; and tumor-bearing dogs with and without microthrombi were analyzed. RESULTS: Thromboembolic disease was identified in 32/62 (52%, 95% CI: 39%-65%) tumor-bearing dogs. Microthrombi were identified in 31/62 (50%, 95% CI: 37%-63%) dogs, 21/31 (68%, 95% CI: 49%-83%) had exclusively intra-tumoral microthrombi, 10/31 (32%, 95% CI: 17%-51%) had distant microthrombi. Macrothrombi were identified in 3 tumor-bearing dogs. Hemostatic changes potentially consistent with overt and non-overt disseminated intravascular coagulation were identified in some tumor-bearing dogs. D-dimer concentrations were significantly higher (P = .02) and platelet concentration significantly lower (P = .03) in tumor-bearing dogs with microthrombi compared to tumor-bearing dogs without microthrombi. D-dimer concentration above 500 ng/mL was 80% sensitive and 41% specific for the prediction of microthrombi presence. CONCLUSION: The high microthrombi prevalence and concomitant hemostatic dysfunction in dogs with carcinomas or sarcomas has not previously been reported, though the clinical importance is unknown. Increased D-dimer concentration might increase suspicion of microthrombi.

COOLING BY DOUSING WITH COLD WATER DOES NOT ALTER THE PATHOPHYSIOLOGICAL BIOCHEMICAL CHANGES INDUCED BY CAPTURE IN BLESBOK (DAMALISCUS PYGARGUS PHILLIPSI).

Wild animals are commonly captured for conservation, research, and wildlife management purposes. However, capture is associated with a high risk of morbidity or mortality. Capture-induced hyperthermia is a commonly encountered complication believed to contribute significantly to morbidity and mortality. Active cooling of hyperthermic animals by dousing with water is believed to treat capture-induced pathophysiological effects, but remains untested. This study aimed to determine the pathophysiological effects of capture, and whether cooling by dousing with cold water effectively reduces these effects in blesbok (Damaliscus pygargus phillipsi). Thirty-eight blesbok were randomly allocated into three groups: a control group that was not chased (Ct, n=12), chased not cooled (CNC, n=14), and chased plus cooled group (C+C, n=12). The CNC and C+C groups were chased for 15 min prior to chemical immobilization on day 0. Animals in the C+C group were cooled with 10 L of cold water (4 C) for 10 min during immobilization. All animals were immobilized on days 0, 3, 16, and 30. During each immobilization, rectal and muscle temperatures were recorded, and arterial and venous blood samples collected. Blesbok in the CNC and C+C groups presented with capture-induced pathophysiological changes characterized by hyperthermia, hyperlactatemia, increased markers of liver, skeletal, and cardiac muscle damage, hypoxemia, and hypocapnia. Cooling effectively returned body temperatures to normothermic levels, but neither the magnitude nor the duration of the pathophysiological changes differed between the CNC and C+C groups. Therefore, at least in blesbok, capture-induced hyperthermia appears not to be the primary cause of the pathophysiological changes, but is more likely a clinical sign of the hypermetabolism resulting from capture-induced physical and psychological stress. Although cooling is still recommended to prevent the compounding cytotoxic effects of persistent hyperthermia, it is unlikely to prevent stress- and hypoxia-induced damage caused by the capture procedure.

Experimental infection with African Horse Sickness Virus in horses induces only mild temporal hematologic changes and acute phase reactant response.

OBJECTIVE: African Horse Sickness (AHS) is a vector-borne disease endemic to sub-Saharan Africa caused by African Horse Sickness Virus (AHVS). Infections in naïve horses have high morbidity and mortality rates. AHS pathogenesis is not well understood; neither the hematologic changes nor acute phase response occurring during infection has been fully evaluated. The study's objective was to characterize the hematologic changes and acute phase response during experimental infection with AHSV. ANIMALS: 4 horses negative for AHSV group-specific antibodies. PROCEDURES: In this prospective, longitudinal study conducted between November 23 and December 2, 2020, horses were experimentally infected with AHSV, and blood samples were obtained before inoculation and then every 12 hours until euthanasia. Hematologic changes and changes for serum amyloid A (SAA) and iron concentration were evaluated over time using a general linear model including natural logarithm of sampling time. RESULTS: All horses were humanely euthanized due to severe clinical signs typical of AHS. Median Hct increased significantly, and the median WBC count, monocyte count, eosinophil count, and myeloperoxidase index changed significantly in all horses over time. Horses developed marked thrombocytopenia (median, 48 X 103 cells/µL; range, 21 X 103 to 58 X 103 cells/µL) while markers of platelet activation also changed significantly. Median SAA increased and serum iron concentration decreased significantly over time. CLINICAL RELEVANCE: Results indicated severe thrombocytopenia with platelet activation occurs during infection with AHSV. Changes in acute phase reactants SAA and iron, while significant, were unexpectedly mild and might not be useful clinical markers.

The dataset for the inflammatory response during experimental infection and treatment of dogs with Babesia rossi.

Babesia rossi causes severe morbidity and mortality in dogs in sub-Saharan Africa. This was an experimental study designed to observe systemic changes caused by Babesia rossi infection within a canine disease model as well as investigate the influence of inoculum dose and treatment on the progression of inflammation and clinical disease. Six healthy male beagle dogs formed the study population, one dog was splenectomised and used to raise the infectious inoculum, three were administered a high B. rossi infectious dose and two a low infectious dose. Clinical examination, complete blood count (CBC) and C-reactive protein (CRP) were determined daily. Cytokines were quantified on stored plasma collected during the study, using a canine specific cytokine magnetic bead panel (Milliplex©). The experiment was terminated, and treatment administered once predetermined experimental or humane endpoints were reached. The data and information provided in the following article is the summary of all data points collected over the course of the eight-day experimental infection.

The prevalence of intra-tumoral and distant thrombi, as well as tumour-cell emboli in canine neoplasia.

Macroscopic thromboembolic disease has been associated with canine neoplasia, whereas prevalence studies of concurrent microthrombi and tumour-cell emboli are lacking. This retrospective study investigated microthrombi and tumour cell emboli by reviewing pathology records of dogs diagnosed with lymphoma, sarcoma, carcinoma and mast cell tumours with a concurrent description of thrombi or emboli. Pathology reports and medical records of cases with either tumour biopsies and/or post mortems with a diagnosis of neoplasia were reviewed for the presence of microthrombi, macrothrombi and/or tumour-cell emboli and the association with tumour type. Of the 28 895 canine cases in the database, 21 252 (73.5%) were antemortem biopsy specimens and 7643 were post mortems (26.5%); 2274 solid tumours were identified, 2107 (92.7%) were antemortem biopsy diagnoses and 167 (7.3%) were post mortem diagnoses. The prevalence of solid tumour types in the database (28 895 cases) was 872 (3.0%) lymphoma, 722 (2.5%) sarcoma, 455 (1.6%) carcinoma and 225 (0.8%) mast cell tumour. The prevalence of microthrombi associated with these tumours was 58/2274 (2.6%). Intra-tumoral microthrombi were reported in 53/2274 (2.3%) cases, the majority in sarcomas (37/53, 69.8%). No macrothrombi were reported. Tumour-cell emboli were identified in 39/2274 (1.7%) cases, 31/39 (79.5%) were extra-tumoral or distant emboli, and carcinoma the most commonly associated tumour (29/39; 74.4%). Microthrombi were reported in 2.6% of cases, the majority in sarcomas and tumour-cell emboli were identified in 1.7% of cases, the majority carcinomas. Prospective investigations are necessary to explore the potential clinical and prognostic implications of microthrombi and tumour-cell emboli in canine neoplasia.

Mean Platelet Volume and Platelet Volume Distribution Width in Canine Parvoviral Enteritis.

Bacterial translocation from the damaged intestinal tract, reported in canine parvoviral (CPV) enteritis, is thought to be responsible for the systemic inflammatory response resulting from coliform septicemia, which could ultimately progress to septic shock and death. Alterations in platelet indices, specifically mean platelet volume (MPV), is a consistent finding in critically ill people and dogs with and without sepsis. Increased MPV has been reported to be an indirect indicator of platelet activation and of bone marrow response in people and dogs with sepsis. The study aim was to compare admission MPV and platelet volume distribution width (PVDW) in dogs with CPV enteritis to that of healthy aged-matched control dogs. Forty-eight dogs with CPV enteritis and 18 healthy age matched control dogs were included. CPV infection was confirmed with electron microscopy and concurrent blood-borne infections were excluded using PCR. EDTA whole blood samples were analyzed on an automated cell counter, ADVIA 2120, within 30-60 min from collection. There was no significant difference for platelet count between the groups. The MPV for CPV infected dogs (median: 14.0; IQR: 12.2-15.1) was significantly higher compared to controls (11.3; IQR: 10.3-13.1, P = 0.002). The PVDW for CPV infected dogs (66.9; IQR: 64.2-68.8) was significantly higher compared to controls (63.3; IQR: 60.2-65.1, P < 0.001). These findings suggest that significant platelet activation is present in dogs with CPV enteritis which may play a role in the disease outcome, similar to people with sepsis. Further studies are required to investigate the prognosticating ability of MPV in dogs with CPV enteritis.

Experimental Babesia rossi infection induces hemolytic, metabolic, and viral response pathways in the canine host.

BACKGROUND: Babesia rossi is a leading cause of morbidity and mortality among the canine population of sub-Saharan Africa, but pathogenesis remains poorly understood. Previous studies of B. rossi infection were derived from clinical cases, in which neither the onset of infection nor the infectious inoculum was known. Here, we performed controlled B. rossi inoculations in canines and evaluated disease progression through clinical tests and whole blood transcriptomic profiling. RESULTS: Two subjects were administered a low inoculum (104 parasites) while three received a high (108 parasites). Subjects were monitored for 8 consecutive days; anti-parasite treatment with diminazene aceturate was administered on day 4. Blood was drawn prior to inoculation as well as every experimental day for assessment of clinical parameters and transcriptomic profiles. The model recapitulated natural disease manifestations including anemia, acidosis, inflammation and behavioral changes. Rate of disease onset and clinical severity were proportional to the inoculum. To analyze the temporal dynamics of the transcriptomic host response, we sequenced mRNA extracted from whole blood drawn on days 0, 1, 3, 4, 6, and 8. Differential gene expression, hierarchical clustering, and pathway enrichment analyses identified genes and pathways involved in response to hemolysis, metabolic changes, and several arms of the immune response including innate immunity, adaptive immunity, and response to viral infection. CONCLUSIONS: This work comprehensively characterizes the clinical and transcriptomic progression of B. rossi infection in canines, thus establishing a large mammalian model of severe hemoprotozoal disease to facilitate the study of host-parasite biology and in which to test novel anti-disease therapeutics. The knowledge gained from the study of B. rossi in canines will not only improve our understanding of this emerging infectious disease threat in domestic dogs, but also provide insight into the pathobiology of human diseases caused by Babesia and Plasmodium species.

Oxidative burst and phagocytic activity of phagocytes in canine parvoviral enteritis.

Canine parvoviral enteritis (CPE) is a severe disease characterized by systemic inflammation and immunosuppression. The function of circulating phagocytes (neutrophils and monocytes) in affected dogs has not been fully investigated. We characterized the functional capacity of canine phagocytes in CPE by determining their oxidative burst and phagocytic activities using flow cytometry. Blood was collected from 28 dogs with CPE and 11 healthy, age-matched, control dogs. Oxidative burst activity was assessed by stimulating phagocytes with opsonized Escherichia coli or phorbol 12-myristate 13-acetate (PMA) and measuring the percentage of phagocytes producing reactive oxygen species and the magnitude of this production. Phagocytosis was measured by incubating phagocytes with opsonized E. coli and measuring the percentage of phagocytes containing E. coli and the number of bacteria per cell. Complete blood counts and serum C-reactive protein (CRP) concentrations were also determined. Serum CRP concentration was negatively and positively correlated with segmented and band neutrophil concentrations, respectively. Overall, no differences in phagocyte function were found between dogs with CPE and healthy control dogs. However, infected dogs with neutropenia or circulating band neutrophils had decreased PMA-stimulated oxidative burst activity compared to healthy controls. Additionally, CPE dogs with neutropenia or circulating band neutrophils had decreased PMA- and E. coli-stimulated oxidative burst activity and decreased phagocytosis of E. coli compared to CPE dogs without neutropenia or band neutrophils. We conclude that phagocytes have decreased oxidative burst and phagocytic activity in neutropenic CPE dogs and in CPE dogs with circulating band neutrophils.

Effects of storage time and temperature on thromboelastographic analysis in dogs and horses.

BACKGROUND: The accessibility of thromboelastography (TEG) to general practitioners is limited by short sample storage times (30 minutes) and storage temperatures (20-23°C). OBJECTIVES: We aimed to evaluate the stability of canine and equine citrated blood samples when stored for extended periods of time, both at room temperature (RT) (20-23°C) and refrigerator temperature (FT) (2-7.5°C). METHODS: Citrated whole blood samples from healthy dogs and horses (n = 10 for each) were stored for 30 minutes (baseline) at RT before TEG analysis. Baseline values for TEG variables R, K, α, MA, LY30, and LY60 were compared with those from samples stored for 2, 8, and 22.5 h, at RT and FT. Results were compared using an ANOVA (P < .05). Total allowable analytical error (TEa ) based on biological variation data was used to evaluate stability. RESULTS: In dogs, statistically significant differences included shorter R, longer K, decreased MA, and increased LY60 at various time points and storage temperatures from 2 h onward. Only samples stored for 2 h at FT showed acceptable stability compared with TEa . In horses, statistically significant differences included shorter R and K, and decreased α, LY30, and LY60 at various time points and storage temperatures from 2 h onward. Samples were not stable at any time compared with TEa , regardless of the temperature. CONCLUSIONS: In this study, canine samples could be stored for up to 2 h at FT without affecting TEG results; equine samples should be stored for 30 minutes at RT.

Comparison of canine urine specific gravity measurements between various refractometers in a clinical setting.

BACKGROUND: Veterinary facilities might use multiple refractometers and individuals to measure urine specific gravity (USG). Previous comparison studies show conflicting results. Furthermore, the clinical significance of measurement differences and interobserver variabilities has not been assessed. OBJECTIVES: We aimed to determine statistically and clinically significant differences between four refractometers in measuring canine USG and subsequent categorization of urine concentrations and azotemia and determine the variability between different observers performing USG measurements. METHODS: Fifty-nine specimens were included for the USG measurements with four refractometers by different observers. Each refractometer pair was compared using Spearman's rank correlation, Bland-Altman difference plots, and Deming regression analyses. Calculated bias was compared to set performance goals. Interobserver agreement was evaluated, and intraclass correlation coefficients were used to determine differences in the categorization of urine concentrations and azotemia (prerenal or renal). RESULTS: There was excellent correlation (rs  = .99-1.00) between refractometers. All comparisons involving R4 showed significant constant and proportional biases. Mean bias met the clinical performance goals for all refractometers, except for comparisons with R4, where up to 17 results were outside the allowable bias. There was almost perfect agreement (rs  = .999) between observers and excellent agreement (ICC = .96-.99) for the classification of urine concentrations. In azotemic patients (22%), there was perfect agreement (ICC = 1.00) for the categorization of azotemia. CONCLUSIONS: In most cases, three of the refractometers evaluated in this study can be used interchangeably at all USG values, without affecting clinical decision-making. Multiple observers did not significantly affect decision-making.

Haemostatic changes associated with fluid resuscitation in canine parvoviral enteritis.

The haemostatic status of dogs with canine parvovirus (CPV) enteritis, within 24 h of admission after initial fluid administration, has been described previously, but the haemostatic status at admission and after standard fluid resuscitation, as well as after initial fluid redistribution, has not been investigated previously. The objective of this study was to characterise the haemostatic status at admission and describe the effect of crystalloid fluid resuscitation on haemostatic variables in dogs with CPV enteritis. Twenty-seven client-owned, hospitalised dogs with confirmed natural CPV infection and 15 healthy age-matched controls were included in a prospective, observational clinical study. The volume of resuscitation fluid, haematocrit (HCT), platelet count, thromboelastography (TEG) variables, antithrombin (AT) activity, fibrinogen- and C-reactive protein (CRP) concentrations were measured in all dogs at admission, after fluid resuscitation and, in 10 dogs, after receiving an additional 3 hours of maintenance-rate crystalloid fluids. For the CPV group at admission, the median TEG reaction time (R) and maximum amplitude (MA) or clot strength, as well as the median HCT, fibrinogen and CRP concentrations, were significantly increased compared to the controls. After fluid resuscitation, median R was significantly shorter, MA significantly increased and HCT and AT activity significantly decreased compared to admission values. The haemostatic variables remained unchanged after 3 h of maintenance-rate crystalloid therapy. The increased clot strength present in dogs with CPV enteritis at admission was exacerbated after fluid resuscitation and persisted for hours after large-volume crystalloid fluid administration.

Neutrophil Myeloperoxidase Index in Dogs With Babesiosis Caused by Babesia rossi.

Babesiosis caused by the virulent tick-borne hemoprotozoan, Babesia rossi, results in a marked systemic inflammatory host response in dogs. Neutrophils form part of the innate immune response and contains myeloperoxidase (MPO) as the predominant component of the neutrophil lysosomal protein in azurophilic granules. The neutrophil myeloperoxidase index (MPXI), determined on the ADVIA hematology analyzer, is a quantitative estimate of intracellular MPO content. Objectives of this study were to: (a) compare MPXI in dogs with babesiosis with healthy control dogs; (b) compare MPXI in dogs that died from babesiosis with dogs that survived and controls; and (c) correlate the MPXI with the previously determined segmented and band neutrophil count and cytokine concentrations in dogs with babesiosis. Data for 140 dogs naturally infected with B. rossi and 20 healthy control dogs were retrospectively evaluated. Neutrophil counts and MPXI were determined on an ADVIA 2120 analyzer. Cytokine concentrations [interleukin (IL)-2, IL-6, IL-8, IL-10, IL-18, granulocyte-macrophage colony stimulating factor (GM-CSF), and monocyte chemo-attractant protein-1 (MCP-1)] were determined using a canine-specific multiplex immunoassay. The mortality rate of the Babesia-infected dogs was 11% (15/140). MPXI was significantly higher in Babesia-infected dogs (P = 0.033), and in Babesia-infected non-survivors (P = 0.011), compared with healthy control dogs. In Babesia-infected dogs a significant positive correlation was found between MPXI and IL-10 (r = 0.211, P = 0.039) and a significant negative correlation was found between MPXI and IL-8 (r = -0.350, P < 0.001). In Babesia-infected non-survivors, significant positive correlations were found between MPXI and IL-2 (r = 0.616, P = 0.033), IL-6 (r = 0.615, P = 0.033), IL-18 (r = 0.613, P = 0.034), GM-CSF (r = 0.630, P = 0.028), and MCP-1 (r = 0.713, P = 0.009). In Babesia-infected survivors, a significant negative correlation was found between MPXI and IL-8 (r = -0.363, P = 0.001). MPXI was correlated with pro-inflammatory cytokines in Babesia-infected dogs that died. The potential of MPXI as a novel marker of inflammation and prognosis in dogs infected with B. rossi, thus warrants further investigation.

Bias between two methods of albumin measurement in the white rhinoceros, Ceratotherium simum.

BACKGROUND: The bromocresol green (BCG) method has been reported to overestimate serum albumin concentration in several species due to non-specific binding to globulins. As the white rhinoceros has high concentrations of serum globulins, significant differences in albumin measured by the BCG method, and the field method of agarose gel serum protein electrophoresis (SPE) are expected. OBJECTIVES: We aimed to compare the BCG and SPE methods for albumin determination in the serum of white rhinoceroses. METHODS: SPE and BCG albumin were measured in 82 white rhinoceros serum samples. Results were compared using Bland-Altman difference plots and Passing-Bablok regression analysis. RESULTS: BCG albumin showed a significant mean constant positive bias of 7 g/L, or 36%, which was more than the total allowable error of 15% and was clinically significant. Methods were not comparable within the inherent imprecision of each method. CONCLUSIONS: The BCG method overestimated albumin concentrations in this species compared with agarose gel SPE, and method-specific reference intervals should be used.

Conserving wildlife in a changing world: Understanding capture myopathy-a malignant outcome of stress during capture and translocation.

The number of species that merit conservation interventions is increasing daily with ongoing habitat destruction, increased fragmentation and loss of population connectivity. Desertification and climate change reduce suitable conservation areas. Physiological stress is an inevitable part of the capture and translocation process of wild animals. Globally, capture myopathy-a malignant outcome of stress during capture operations-accounts for the highest number of deaths associated with wildlife translocation. These deaths may not only have considerable impacts on conservation efforts but also have direct and indirect financial implications. Such deaths usually are indicative of how well animal welfare was considered and addressed during a translocation exercise. Importantly, devastating consequences on the continued existence of threatened and endangered species succumbing to this known risk during capture and movement may result. Since first recorded in 1964 in Kenya, many cases of capture myopathy have been described, but the exact causes, pathophysiological mechanisms and treatment for this condition remain to be adequately studied and fully elucidated. Capture myopathy is a condition with marked morbidity and mortality that occur predominantly in wild animals around the globe. It arises from inflicted stress and physical exertion that would typically occur with prolonged or short intense pursuit, capture, restraint or transportation of wild animals. The condition carries a grave prognosis, and despite intensive extended and largely non-specific supportive treatment, the success rate is poor. Although not as common as in wildlife, domestic animals and humans are also affected by conditions with similar pathophysiology. This review aims to highlight the current state of knowledge related to the clinical and pathophysiological presentation, potential treatments, preventative measures and, importantly, the hypothetical causes and proposed pathomechanisms by comparing conditions found in domestic animals and humans. Future comparative strategies and research directions are proposed to help better understand the pathophysiology of capture myopathy.

A clinical and pathological description of 320 cases of naturally acquired Babesia rossi infection in dogs.

Babesia rossi causes the most severe clinical disease in dogs of all the babesia parasites. We included 320 naturally-infected dogs that presented for care at the Onderstepoort Veterinary Academic Hospital between 2006 and 2016. All dogs had mono-infections confirmed by multiplex PCR. The data allowed more accurate clinical classification of the disease and identified parameters that were associated with disease severity and death. Odds ratios for dying were significant (P < 0.05) for increased band neutrophil count, collapse at presentation; presence of cerebral signs; hypoglycaemia; hyperlactatemia; high urea, high creatinine; hyperbilirubinaemia; hypercortisolaemia; and hypothyroxinaemia. Joint component analysis confirmed that the variables with significant odds ratios grouped together with death. Yet, multivariate logistic regression was unable to identify a group of significant independent predictors of death. Receiver Operator Characteristic curves indicated that low total thyroid hormone, high bilirubin, high serum urea and high cortisol concentrations were the variables with the highest sensitivity and specificity for death. These data provide both the clinician and researcher with a set of easily-measured laboratory and clinical assessments to classify cases into those that are uncomplicated and those that are complicated. The disease is complex and multisystemic and probably involves mechanisms more proximal in the pathogenesis than those that have been evaluated.

Clinical findings, synovial fluid cytology and growth factor concentrations after intra-articular use of a platelet-rich product in horses with osteoarthritis.

Osteoarthritis is a common cause of lameness in horses, resulting in poor performance. Intra-articular platelet-rich plasma can deliver a collection of bioactive molecules, such as autologous growth factors and proteins involved in the quality of tissue repair. Horses (n=5) with osteoarthritis affecting antebrachiocarpal, middle carpal or metacarpophalangeal joints, and horses (n=5) without osteoarthritis of the corresponding joints (radiographically free of osteoarthritis), were used for the production of platelet-rich plasma which was subsequently injected into selected joints. Clinical and synovial fluid changes after intra-articular injection of platelet-rich plasma as well as synovial platelet-derived growth factor-BB and transforming growth factor-beta 1 concentration changes were evaluated in these joints and compared between normal joints and joints with osteoarthritis. A gravity filtration system produced a moderately concentrated platelet-rich plasma, representing a 4.7-fold increase in baseline platelet concentration. The synovial effusion score was significantly different between the control joints and joints with osteoarthritis on Day 0 with a higher score in the group with osteoarthritis. Within the control group, the synovial effusion score was significantly higher on Days 1 and 2 compared to Day 0. For both groups, the synovial fluid nucleated cell count, predominantly intact neutrophils, was significantly increased on Days 1 and 2, with no significant difference between groups. The mean synovial platelet-derived growth factor-BB and transforming growth factor-beta 1 concentrations were increased for both groups but significantly lowered in the group with osteoarthritis on Day 1 compared to normal joints. Concentrations for platelet-derived growth factor-BB remained unchanged on Day 5, compared to Day 1, with no significant difference between groups. In conclusion, intra-articular treatment with platelet-rich plasma resulted in increased synovial growth factor concentrations in joints but with lower concentrations in joints with osteoarthritis. A transient inflammatory reaction was seen both clinically as an increase in synovial effusion and cytologically in both normal joints and joints with osteoarthritis.

Disease severity and blood cytokine concentrations in dogs with natural Babesia rossi infection.

AIMS: Babesia rossi causes severe disease in dogs. Here, we describe the association between serum cytokine concentrations and disease severity. METHODS: Seventeen controls and 55 PCR confirmed B rossi-infected dogs were included. Diseased dogs were subdivided into 23 critically ill and 32 relatively well cases. Serum concentrations of 11 cytokines and biochemical markers of disease severity were determined. RESULTS: Significant differences were detected for IL-6, IL-8, IL-10, MCP-1 and TNF-α between the groups. Generally, the more complicated the disease, the more pro-inflammatory the cytokine milieu. IL-8 showed a reverse trend and was negatively correlated with disease severity. IL-6, MCP-1 and TNF-α were also significantly higher in the dogs that died (n = 9) compared to the dogs that survived (n = 46). IL-8 showed the opposite. MCP-1 and TNF-α were negatively correlated with biochemical markers of severity. Glucose was negatively correlated with IL-6. Cortisol, peripheral parasite density and band neutrophil count were positively correlated, whilst thyroid hormone was negatively correlated with IL-6, MCP-1 and TNF-α. CONCLUSIONS: As in malaria and sepsis, B rossi infection induces a pro-inflammatory cytokine storm that correlates with disease severity and adverse outcome. The multiplicity of cytokines involved argues for redundancy in the system once the disease is established.

Thromboelastographic platelet mapping in dogs with complicated Babesia rossi infection.

BACKGROUND: Dogs with Babesia rossi infection display a normocoagulable thromboelastogram, despite being markedly thrombocytopenic, which is purportedly due to large-scale platelet activation. Thromboelastographic platelet mapping (TEG-PM) evaluates individual contributions of thrombin, fibrinogen, and platelets to clot formation, and may elucidate some of the pathomechanisms of thrombocytopenia-associated hemostatic alterations. OBJECTIVE: This study investigated potential differences in TEG-PM variables in dogs with complicated B rossi infection compared with healthy controls, and whether these variables correlated with platelet activation indices. METHODS: The maximum amplitude (MA) following thrombin generation (MAThrombin ) was determined using kaolin-activated TEG. The TEG-PM variables included MA following the addition of platelet agonists arachidonic acid (MAAA ) and adenosine diphosphate (MAADP ), and MA due to fibrin alone (MAFibrin ). In addition, platelet indices and fibrinogen concentrations were determined. RESULTS: Thirteen dogs with complicated B rossi infection and five healthy controls were included. The median MAFibrin and fibrinogen concentrations were significantly higher (P < 0.01 for both) and median platelet count was significantly lower (P < 0.01) in the babesiosis group vs the control group. No significant differences were found for MAThrombin and MAAA/ADP . maximum amplitude due to fibrin alone was positively correlated with fibrinogen concentration (r = 0.735), mean platelet volume (r = 0.517), and mean platelet mass (r = 0.498), and negatively correlated with hematocrit (r = -0.685), platelet count (r = -0.476), and plateletcrit (r = -0.479) (P < 0.05 for all). CONCLUSIONS: This study suggests that the presence of hyperfibrinogenemia offsets the severe thrombocytopenia associated with B rossi to result in normal thromboelastograms and lack of overt clinical bleeding.

In vitro Antimycobacterial, Apoptosis-Inducing Potential, and Immunomodulatory Activity of Some Rubiaceae Species.

Tuberculosis (TB), a disease caused by microorganisms of the Mycobacterium tuberculosis complex, infects almost one-third of the world's population. The TB epidemic has been further exacerbated by the emergence of multi, extensively, and totally-drug-resistant (MDR, XDR, and TDRTB) strains. An effective immune response plays a crucial role in determining the establishment of TB infection. Therefore, the modulation of the immune system has been considered as a vital approach for the treatment or control of various immune-related diseases such as TB. In this study, the antimycobacterial, immunomodulatory, and apoptosis-inducing effects of six Rubiaceae species were evaluated. A twofold serial dilution method was used to determine the minimum inhibitory concentration values of the plant extracts. The effect of the extracts on the activity of 15-lipoxygenase was investigated. The levels of six different cytokines, IL-2, IL-4, IL-5, IL-10, IFN-γ, and TNF-α, were measured in LPS-activated U937 cell line while the apoptosis-inducing effect of the extracts was evaluated using an annexin V/PI assay using a flow cytometer. The results obtained revealed that all the six extracts tested had antimycobacterial activity against M. tuberculosis H37Rv, M. tuberculosis ATCC 25177, and Mycobacterium bovis ATCC 27299 strains, with MIC values ranging from 39 to 312 µg/mL. The extracts of Cremaspora triflora and Cephalanthus natalensis were the most active against M. tuberculosis (MIC = 39 µg/mL), followed by Pavetta lanceolata and Psychotria zombamontana against M. bovis (MIC = 78 µg/mL). The extracts of P. zombamontana and Psychotria capensis had remarkable IC50 values of 4.32 and 5.8 µg/mL, respectively, better than that of quercetin. The selected extracts promoted Th1/Th2 balances in an in vitro model at the tested concentration which may suggest the therapeutic value of the plant in diseases where inflammation is a significant factor such as TB. The addition of the crude extracts of C. triflora, P. capensis, and P. zombamontana at the tested concentrations to the cell culture medium induced apoptosis in a time- and dose-dependent manner. This interesting preliminary result generated from this study encourages further investigations of these extracts owing to the LOX-inhibitory effect, immunomodulatory, and apoptotic-inducing properties in addition to their antimycobacterial properties.