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.

Fiber type and metabolic characteristics of skeletal muscle in 16 breeds of domestic dogs.

The domestic dog (Canis lupus familiaris) species comprises hundreds of breeds, each differing in physical characteristics, behavior, strength, and running capability. Very little is known about the skeletal muscle composition and metabolism between the different breeds, which may explain disease susceptibility. Muscle samples from the triceps brachii (TB) and vastus lateralis (VL) were collected post mortem from 35 adult dogs, encompassing 16 breeds of varying ages and sex. Samples were analyzed for fiber type composition, fiber size, oxidative, and glycolytic metabolic capacity (citrate synthase [CS], 3-hydroxyacetyl-coA dehydrogenase [3HAD], creatine kinase [CK], and lactate dehydrogenase [LDH] enzyme activities). There was no significant difference between the TB and VL in any of the measurements. However, there were large intra species variation, with some variables confirming the physical attributes of a specific breed. Collectively, type IIA was the predominant fiber type followed by type I and type IIX. The cross-sectional areas (CSA) of the fibers were all smaller when compared to humans and similar to other wild animals. There was no difference in the CSA between the fiber types and muscle groups. Metabolically, the muscle of the dog displayed high oxidative capacity with high activities for CS and 3HAD. Lower CK and higher LDH activities than humans indicate a lower and higher flux through the high energy phosphate and glycolytic pathways, respectively. The high variability found across the different breeds may be attributed to genetics, function or lifestyle which have largely been driven through human intervention. This data may provide a foundation for future research into the role of these parameters in disease susceptibility, such as insulin resistance and diabetes, across breeds.

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.

Deciphering Genetic Susceptibility to Tuberculous Meningitis.

Tuberculous meningitis (TBM) is the most severe form of extrapulmonary tuberculosis (TB) that arises when a caseating meningeal granuloma discharges its contents into the subarachnoid space. It accounts for ~1% of all disease caused by Mycobacterium tuberculosis and the age of peak incidence is from 2-4 years. The exact pathogenesis of TBM is still not fully understood and the mechanism(s) by which the bacilli initially invade the blood-brain-barrier are still to be elucidated. This study investigated the involvement of the host genome in TBM susceptibility, by considering common variants (minor allele frequency (MAF) >5%) using microarray genotyping and rare variants (MAF <1%) via exome sequencing. A total of 123 TBM cases, 400 pulmonary TB (pTB) cases and 477 healthy controls were genotyped on the MEGA array. A genome-wide association study (GWAS) comparing 114 TBM cases to 395 healthy controls showed no association with TBM susceptibility. A second analysis comparing 114 TBM cases to 382 pTB cases was conducted to investigate variants associated with different TB phenotypes. No significant associations were found with progression from pTB to TBM. Ten TBM cases and 10 healthy controls were exome sequenced. Gene set association tests SKAT-O and SKAT Common Rare were used to assess the association of rare SNPs and the cumulative effect of both common and rare SNPs with susceptibility to TBM, respectively. Ingenuity Pathway Analysis (IPA) of the top-hits of the SKAT-O analysis showed that NOD2 and CYP4F2 are both important in TBM pathogenesis and highlighted these as targets for future study. For the SKAT Common Rare analysis Centriolar Coiled-Coil Protein 110 (CCP110) was nominally associated (p = 5.89x10-6) with TBM susceptibility. In addition, several top-hit genes ascribed to the development of the central nervous system (CNS) and innate immune system regulation were identified. Exome sequencing and GWAS of our TBM cohort has identified a single previously undescribed association of CCP110 with TBM susceptibility. These results advance our understanding of TBM in terms of both variants and genes that influence susceptibility. In addition, several candidate genes involved in innate immunity have been identified for further genotypic and functional investigation.

Comparative antimicrobial efficacy of 4 surgical hand-preparation procedures prior to application of an alcohol-based hand rub in veterinary students.

OBJECTIVE: To determine the influence of skin preparations before application of an alcohol-based hand rub (ABHR) on bacterial counts before and after elective surgery. STUDY DESIGN: Clinical prospective study. SAMPLE POPULATION: Veterinary students (n = 103) performing ovariohysterectomies on 140 dogs. METHODS: Participants were randomly assigned to 1 initial surgical preparation on the day of surgery: A - hand preparation with medicated solution (4% w/v chlorhexidine bigluconate followed by an ABHR; B - application of a medication solution (benzalkonium chloride 0.1%-1% and polymeric biguanide hydrochloride 0.01%-0.1%) followed by an ABHR; C - nonmedicated pH-neutral soap hand wash followed by ABHR, and D - direct application of an ABHR. Samples were taken by pressing the distal finger tips to an agar plate before the hand preparation, after the hand preparation (n = 3), after ABHR application, and 120 minutes later. Colony-forming units (CFUs) for samples were determined. Total log CFU and CFU log10 reduction were calculated and used for comparison with P < .05. RESULTS: Two hours after surgery commenced, the participants of groups that performed a hand preparation had lower total CFUs than those that did not perform a hand preparation (P = .001). In particular, the number of CFUs was lower when ABHR was performed after application of pHN compared to direct ABHR (P = .001). CONCLUSION: In this population, performing a hand preparation with pHN prior to applying an ABHR had better antimicrobial effect for the duration of surgery than not performing a hand preparation. CLINICAL SIGNIFICANCE: Surgeons should wash their hands prior to ABHR before starting their first surgery of the day, even when hands appear clean.

Influence of high-protein and high-carbohydrate diets on serum lipid and fructosamine concentrations in healthy cats.

OBJECTIVES: The aim of this study was to determine whether high-protein and high-carbohydrate diets exert differential effects on serum cholesterol, triglyceride and fructosamine concentrations in healthy cats. METHODS: A randomised, crossover diet trial was performed in 35 healthy shelter cats. Following baseline health assessments, cats were randomised into groups receiving either a high-protein or high-carbohydrate diet for 4 weeks. The cats were then fed a washout diet for 4 weeks before being transitioned to whichever of the two studied diets they had not yet received. Fasting serum cholesterol, triglyceride and fructosamine concentrations were determined at the end of each 4-week diet period. RESULTS: Cats on the high-carbohydrate diet had significantly lower serum cholesterol (P <0.001) concentrations compared with baseline measurements. Cats on the high-protein diet had significantly higher serum cholesterol (P <0.001) and triglyceride (P <0.001) concentrations, yet lower fructosamine (P <0.001) concentrations compared with baseline measurements. In contrast, overweight cats (body condition score [BCS] >5) had lower cholesterol (P = 0.007) and triglyceride (P = 0.032) concentrations on the high-protein diet than cats within other BCS groups. CONCLUSIONS AND RELEVANCE: Diets higher in protein and lower in carbohydrates appear beneficial for short-term glucose control in healthy cats. A high-protein diet was associated with significantly elevated cholesterol and triglyceride concentrations in healthy cats, even though the increase was significantly less pronounced in cats with a BCS >5. This finding suggests that overweight cats process high-protein diets, cholesterol and triglycerides differently than leaner cats.

The use of thalidomide to treat children with tuberculosis meningitis: A review.

Much of the morbidity and mortality caused by tuberculous meningitis (TBM) is mediated by a dysregulated immune response. Effective host-directed therapy is therefore critical to improve survival and clinical outcomes. Currently only one host-directed therapy (HDT), corticosteroids, is proven to improve mortality. However, there is no evidence that corticosteroids reduce morbidity and the mechanism of action for mortality reduction is uncertain. Further, it has no proven benefit in HIV co-infected individuals. One promising host-directed therapy approach is to restrict the immunopathology arising from tumour necrosis factor (TNF)-α excess is via TNF-α inhibitors. There are accumulating data on the role of thalidomide, anti-TNF-α monoclonal antibodies (infliximab, adalimumab) and the soluble TNF-α receptor (etanercept) in TBM treatment. Thalidomide was developed nearly seventy years ago and has been a highly controversial drug. Birth defects and toxic adverse effects have limited its use but an improved understanding of its immunological mechanism of action suggest that it may have a crucial role in regulating the destructive host response seen in inflammatory conditions such as TBM. Observational studies at our institution found low dosage adjunctive thalidomide safe in treating tuberculous mass lesions and blindness related to optochiasmatic arachnoiditis, with good clinical and radiological response. In this review, we discuss possible mechanisms of action for thalidomide, based on our clinico-radiologic experience and post-mortem histopathological work. We also propose a rationale for its use in the treatment of certain TBM-related complications.

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.

Thalidomide Use for Complicated Central Nervous System Tuberculosis in Children: Insights From an Observational Cohort.

BACKGROUND: Much of the neurological sequelae of central nervous system (CNS) tuberculosis (TB) is due to an excessive cytokine-driven host-inflammatory response. Adjunctive corticosteroids, which reduce cytokine production and thus dampen the inflammation, improve overall survival but do not prevent morbidity. This has prompted investigation of more targeted immunomodulatory agents, including thalidomide. METHODS: We describe a retrospective cohort of 38 children consecutively treated with adjunctive thalidomide for CNS TB-related complications over a 10-year period. RESULTS: The most common presenting symptom was focal motor deficit (n = 16), followed by cranial nerve palsies and cerebellar dysfunction. Three of the 38 children presented with large dural-based lesions, manifesting as epilepsia partialis continua (EPC), 4 presented with blindness secondary to optochiasmatic arachnoiditis, and 2 children developed paraplegia due to spinal cord TB mass lesions. Duration of adjunctive thalidomide therapy (3-5 mg/kg/day) varied according to complication type. In children compromised by TB mass lesions, the median treatment duration was 3.9 months (interquartile range [IQR], 2.0-5.0 months), whereas in children with optic neuritis it was 2.0 months (IQR, 1.3-7.3 months) and in EPC it was 1.0 months (IQR, 1-2.5 months). Satisfactory clinical and radiological response was observed in 37 of the children. None of the children experienced rashes, hepatitis, or hematologic derangements or complained of leg cramps. CONCLUSIONS: This study is the largest cohort of adult or pediatric patients treated with adjunctive thalidomide for CNS TB-related complications. The drug has proved to be safe and well tolerated and appears to be clinically efficacious. The potential role of thalidomide or analogues in the treatment of other tuberculous meningitis-related complications requires further exploration.

Host Directed Therapies for Tuberculous Meningitis.

A dysregulated host immune response significantly contributes to morbidity and mortality in tuberculous meningitis (TBM). Effective host directed therapies (HDTs) are critical to improve survival and clinical outcomes. Currently only one HDT, dexamethasone, is proven to improve mortality. However, there is no evidence dexamethasone reduces morbidity, how it reduces mortality is uncertain, and it has no proven benefit in HIV co-infected individuals. Further research on these aspects of its use, as well as alternative HDTs such as aspirin, thalidomide and other immunomodulatory drugs is needed. Based on new knowledge from pathogenesis studies, repurposed therapeutics which act upon small molecule drug targets may also have a role in TBM. Here we review existing literature investigating HDTs in TBM, and propose new rationale for the use of novel and repurposed drugs. We also discuss host variable responses and evidence to support a personalised approach to HDTs in TBM.

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.

Vitamin D status in dogs with babesiosis.

Canine babesiosis is a virulent infection of dogs in South Africa caused principally by Babesia rossi. Hypovitaminosis D has been reported in a wide range of infectious diseases in humans and dogs, and low vitamin D status has been associated with poor clinical outcomes. However, the relationship between vitamin D status and canine babesiosis has not been investigated. The objective of this study was to examine the relationship between the presence and severity of B. rossi infection and vitamin D status of infected dogs. Owners with dogs with a confirmed diagnosis of B. rossi infection and of healthy control dogs were invited to enrol onto the study. Vitamin D status was assessed by measurement of serum concentrations of the major circulating vitamin D metabolite, 25-hydroxyvitamin D (25[OH]D). Dogs with babesiosis (n = 34) had significantly lower mean serum 25(OH)D concentrations than healthy dogs (n = 24) (37.76 ± 21.25 vs. 74.2 ± 20.28 nmol/L). The effect of babesiosis on serum 25(OH)D concentrations was still significant after adjusting for any effect of age, body weight and sex. There was a negative relationship between serum 25(OH)D concentrations and disease severity in dogs with babesiosis. Serum concentrations of creatinine and alanine aminotransferase and time to last meal were not associated with serum 25(OH)D concentrations in dogs with babesiosis. In conclusion, dogs with Babesia rossi infections had lower serum 25(OH)D concentrations than healthy dogs. The inverse correlation between 25(OH)D concentrations and the clinical severity score indicate that hypovitaminosis D might be a helpful additional indicator of disease severity.

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.

Identification of an iron-responsive subtype in two children diagnosed with relapsing-remitting multiple sclerosis using whole exome sequencing.

BACKGROUND: Multiple sclerosis is a disorder related to demyelination of axons. Iron is an essential cofactor in myelin synthesis. Previously, we described two children (males of mixed ancestry) with relapsing-remitting multiple sclerosis (RRMS) where long-term remission was achieved by regular iron supplementation. A genetic defect in iron metabolism was postulated, suggesting that more advanced genetic studies could shed new light on disease pathophysiology related to iron. METHODS: Whole exome sequencing (WES) was performed to identify causal pathways. Blood tests were performed over a 10 year period to monitor the long-term effect of a supplementation regimen. Clinical wellbeing was assessed quarterly by a pediatric neurologist and regular feedback was obtained from the schoolteachers. RESULTS: WES revealed gene variants involved in iron absorption and transport, in the transmembrane protease, serine 6 (TMPRSS6) and transferrin (TF) genes; multiple genetic variants in CUBN, which encodes cubilin (a receptor involved in the absorption of vitamin B12 as well as the reabsorption of transferrin-bound iron and vitamin D in the kidneys); SLC25A37 (involved in iron transport into mitochondria) and CD163 (a scavenger receptor involved in hemorrhage resolution). Variants were also found in COQ3, involved with synthesis of Coenzyme Q10 in mitochondria. Neither of the children had the HLA-DRB1*1501 allele associated with increased genetic risk for MS, suggesting that the genetic contribution of iron-related genetic variants may be instrumental in childhood MS. In both children the RRMS has remained stable without activity over the last 10 years since initiation of nutritional supplementation and maintenance of normal iron levels, confirming the role of iron deficiency in disease pathogenesis in these patients. CONCLUSION: Our findings highlight the potential value of WES to identify heritable risk factors that could affect the reabsorption of transferrin-bound iron in the kidneys causing sustained iron loss, together with inhibition of vitamin B12 absorption and vitamin D reabsorption (CUBN) and iron transport into mitochondria (SLC25A37) as the sole site of heme synthesis. This supports a model for RRMS in children with an apparent iron-deficient biochemical subtype of MS, with oligodendrocyte cell death and impaired myelination possibly caused by deficits of energy- and antioxidant capacity in mitochondria.

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.

Management of intracranial tuberculous mass lesions: how long should we treat for?

Tuberculous intracranial mass lesions are common in settings with high tuberculosis (TB) incidence and HIV prevalence. The diagnosis of such lesions, which include tuberculoma and tuberculous abscesses, is often presumptive and based on radiological features, supportive evidence of TB elsewhere and response to TB treatment. However, the treatment response is unpredictable, with lesions frequently enlarging paradoxically or persisting for many years despite appropriate TB treatment and corticosteroid therapy. Most international guidelines recommend a 9-12 month course of TB treatment for central nervous system TB when the infecting Mycobacterium tuberculosis ( M.tb) strain is sensitive to first-line drugs. However, there is variation in opinion and practice with respect to the duration of TB treatment in patients with tuberculomas or tuberculous abscesses. A major reason for this is the lack of prospective clinical trial evidence. Some experts suggest continuing treatment until radiological resolution of enhancing lesions has been achieved, but this may unnecessarily expose patients to prolonged periods of potentially toxic drugs. It is currently unknown whether persistent radiological enhancement of intracranial tuberculomas after 9-12 months of treatment represents active disease, inflammatory response in a sterilized lesion or merely revascularization. The consequences of stopping TB treatment prior to resolution of lesional enhancement have rarely been explored. These important issues were discussed at the 3 rd International Tuberculous Meningitis Consortium meeting. Most clinicians were of the opinion that continued enhancement does not necessarily represent treatment failure and that prolonged TB therapy was not warranted in patients presumably infected with M.tb strains susceptible to first-line drugs. In this manuscript we highlight current medical treatment practices, benefits and disadvantages of different TB treatment durations and the need for evidence-based guidelines regarding the treatment duration of patients with intracranial tuberculous mass lesions.

Prevalence of Salmonella in juvenile dogs affected with parvoviral enteritis.

Salmonellosis is a disease of major zoonotic importance and canine parvovirus is a potentially fatal cause of canine enteritis with a world-wide distribution. Persistent isolation of Salmonella during routine environmental sampling surveys of a hospital ward, reserved for the treatment of dogs with canine parvovirus infection, prompted investigation into a possible source. We hypothesised that dogs affected by canine parvovirus would have a higher prevalence of faecal salmonellae compared to an apparently healthy cohort. Seventy-four client-owned dogs naturally infected with canine parvovirus and 42 apparently healthy client-owned dogs were included in the study. This prospective, longitudinal, observational study was conducted over an 18-month period. Fresh faecal samples were collected from dogs aged 6 weeks to 9 months diagnosed with canine parvovirus infection and admitted for treatment, and from apparently healthy dogs presented for vaccination or routine hospital procedures. Faeces were submitted for the isolation, antimicrobial susceptibility testing and serotyping of salmonellae. The prevalence of faecal Salmonella shedding was 22% and 31% for the affected and apparently healthy dogs, respectively, which was not statistically different. No significant associations between Salmonella status and possible risk factors or continuous variables such as age, body weight and duration of hospitalisation were identified. All the Salmonella isolates (n = 32) were resistant to penicillin G, lincomycin and tylosin. Salmonellae from nine different serotypes were identified. The prevalence of Salmonella shedding in both groups was higher than that commonly reported, yet similar to those in previous reports on young dogs, shelter dogs or dogs fed a raw meat diet.

Prevalence of canine Babesia and Ehrlichia co-infection and the predictive value of haematology.

Canine babesiosis and ehrlichiosis are important tick-borne infections in South Africa. Many South African general veterinary practitioners perceive co-infection with Ehrlichia spp. as a common occurrence in dogs with babesiosis. Studies about the prevalence of co-infection in South African dogs are lacking. This retrospective study aimed to determine the prevalence of Ehrlichia co-infection in dogs with babesiosis. Additionally, the predicative value of specific haematological variables for co-infection was evaluated. The study population consisted of 205 dogs diagnosed with canine babesiosis presented to the Onderstepoort Veterinary Academic Hospital (OVAH) in 2006 and between 2011 and 2013. The Babesia-infected dogs were grouped based on presence or absence of an Ehrlichia spp. co-infection. Ehrlichia spp. co-infection was confirmed using polymerase chain reaction. Positive and negative predictive values (PPVs and NPVs) of leukopenia or thrombocytopenia for co-infection were also calculated. The prevalence of Babesiaspp. and Ehrlichia spp. co-infection in this cohort of dogs was 2%. In the babesiosis dogs, the PPV of leukopenia for co-infection with Ehrlichia spp. was 1.3%, and the NPV 97.4%. Similarly, the PPV and NPVs of thrombocytopenia for co-infection were 2.1% and 100%, respectively. Co-infection with Ehrlichia spp. was a rare occurrence in dogs with babesiosis presented to the OVAH. Normal leukocyte or platelet counts confidently ruled out the presence of concurrent ehrlichiosis in this cohort of dogs. However, the diagnosis of Ehrlichia co-infection based on the presence of thrombocytopenia or leukopenia would have been associated with false positive results in more than 97.4% of cases.

Aortic Arch Baroreceptor Stimulation in an Experimental Goat Model: A Novel Method to Lower Blood Pressure.

The effect of aortic baroreceptor stimulation on blood pressure manipulation was assessed using the goat species Capra aegagrus hircus. The aim of this study was to manipulate blood pressure with future intention to treat high blood pressure in humans. The ages of the animals ranged from 6 months to 2 years. A standard anesthesia protocol was used. A lateral thoracotomy was performed to gain access to the aortic arch. Data was collected with the Vigileo system. Pre stimulation blood pressure was compared with maximum post stimulation blood pressure values. Results were analyzed with the Wilcoxon signed rank test. In the study 38 animals were enrolled. Baroreceptor stimulation was performed for each animal using 3 different electrodes each of which emits an electrical impulse. In the pilot phase of the study, the median baseline blood pressure prior to stimulation of the baroreceptors was 110.8 mmHg. After stimulation the median blood pressure decreased to 88 mmHg. The average decrease in blood pressure was 22.8 mmHg. This decrease of blood pressure after stimulation of the baroreceptors is statistically significant (p < 0.0001) and the proof of concept was shown. During the extended phase all three probes had a significant effect on blood pressure lowering (p < 0.0001). The study confirmed that aortic baroreceptor stimulation has an effect on blood pressure lowering. This is a novel field of blood pressure manipulation. The hemodynamic effects of long-term aortic baroreceptor stimulation are unknown. Further investigations need to be done to determine whether a similar effect can be induced in different species such as primates and humans.

Childhood tuberculosis is associated with decreased abundance of T cell gene transcripts and impaired T cell function.

The WHO estimates around a million children contract tuberculosis (TB) annually with over 80 000 deaths from dissemination of infection outside of the lungs. The insidious onset and association with skin test anergy suggests failure of the immune system to both recognise and respond to infection. To understand the immune mechanisms, we studied genome-wide whole blood RNA expression in children with TB meningitis (TBM). Findings were validated in a second cohort of children with TBM and pulmonary TB (PTB), and functional T-cell responses studied in a third cohort of children with TBM, other extrapulmonary TB (EPTB) and PTB. The predominant RNA transcriptional response in children with TBM was decreased abundance of multiple genes, with 140/204 (68%) of all differentially regulated genes showing reduced abundance compared to healthy controls. Findings were validated in a second cohort with concordance of the direction of differential expression in both TBM (r2 = 0.78 p = 2x10-16) and PTB patients (r2 = 0.71 p = 2x10-16) when compared to a second group of healthy controls. Although the direction of expression of these significant genes was similar in the PTB patients, the magnitude of differential transcript abundance was less in PTB than in TBM. The majority of genes were involved in activation of leucocytes (p = 2.67E-11) and T-cell receptor signalling (p = 6.56E-07). Less abundant gene expression in immune cells was associated with a functional defect in T-cell proliferation that recovered after full TB treatment (p<0.0003). Multiple genes involved in T-cell activation show decreased abundance in children with acute TB, who also have impaired functional T-cell responses. Our data suggest that childhood TB is associated with an acquired immune defect, potentially resulting in failure to contain the pathogen. Elucidation of the mechanism causing the immune paresis may identify new treatment and prevention strategies.