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Abstract 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.

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.

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.

Abstract ‘EBVM Learning’ is a freely available resource created in 2015 by an international team with the support of RCVS Knowledge. The resource comprises a series of online modules teaching the fundamental concepts of evidence-based veterinary medicine (EBVM) (Ask, Acquire, Appraise, Apply & Assess) supported by case studies, exercises, worked examples and quizzes. The aim of the current study (undertaken in 2019) was to review ‘EBVM Learning’ to ensure its ongoing relevance and usefulness to the range of learners engaged in EBVM. Feedback was gathered from stakeholder groups using website statistics and feedback forms, a survey and semi-structured interviews to provide a combination of quantitative and qualitative data. Website statistics revealed an international audience and a steady increase in visitors exceeding 1,000 per month in August 2020. Feedback via the online form (n=35) and survey (n=71) indicated that the resource was well structured, with an appropriate level and amount of content, useful examples and quizzes and the majority of respondents would use it again. Semi-structured interviews of educators (n=5) and veterinarians (n=8) identified three themes: features of the ‘EBVM Learning’ resource (strengths, suggestions for improvement), embedding the resource in education (undergraduate, postgraduate) and promoting EBVM (challenges, motivation for engagement). At a project team workshop the results were used to plan updates to the existing content and to identify new ways to promote learning and engagement. An updated version of ‘EBVM Learning’ was developed. ‘EBVM Learning’ is helping to produce the next generation of evidence-based practitioners and enabling to engage in the concepts of EBVM as part of their clinical practice.

Babesia rossi infection causes a severe inflammatory response in the dog, which is the result of the balance between pro- and anti-inflammatory cytokine secretion. The aim of this study was to determine whether changes in cytokine concentrations were present in dogs with babesiosis and whether it was associated with disease outcome. Ninety-seven dogs naturally infected with B. rossi were studied and fifteen healthy dogs were included as controls. Diagnosis of babesiosis was confirmed by polymerase chain reaction and reverse line blot. Blood samples were collected from the jugular vein at admission, prior to any treatment. Cytokine concentrations were assessed using a canine-specific multiplex assay on an automated analyser. Serum concentrations of interleukin (IL)-2, IL-6, IL-8, IL-10, IL-18, granulocyte-macrophage colony stimulating factor (GM-CSF) and monocyte chemotactic protein-1 (MCP-1) were measured. Twelve of the Babesia-infected dogs died (12%) and 85 survived (88%). Babesia-infected dogs were also divided into those that presented within 48 hours from displaying clinical signs, and those that presented more than 48 hours after displaying clinical signs. Cytokine concentrations were compared between the different groups using the Mann-Whitney U test. IL-10 and MCP-1 concentrations were significantly elevated for the Babesia-infected dogs compared to the healthy controls. In contrast, the IL-8 concentration was significantly decreased in the Babesia-infected dogs compared to the controls. Concentrations of IL-6 and MCP-1 were significantly increased in the non-survivors compared to the survivors. Concentrations for IL-2, IL-6, IL-18 and GM-CSF were significantly higher in those cases that presented during the more acute stage of the disease. These findings suggest that a mixed cytokine response is present in dogs with babesiosis caused by B. rossi, and that an excessive pro-inflammatory response may result in a poor outcome.

Canine rabies can be effectively controlled by vaccination with readily available, high-quality vaccines. These vaccines should provide protection from challenge in healthy dogs, for the claimed period, for duration of immunity, which is often two or three years. It has been suggested that, in free-roaming dog populations where rabies is endemic, vaccine-induced protection may be compromised by immuno-suppression through malnutrition, infection and other stressors. This may reduce the proportion of dogs that seroconvert to the vaccine during vaccination campaigns and the duration of immunity of those dogs that seroconvert. Vaccination coverage may also be limited through insufficient vaccine delivery during vaccination campaigns and the loss of vaccinated individuals from populations through demographic processes. This is the first longitudinal study to evaluate temporal variations in rabies vaccine-induced serological responses, and factors associated with these variations, at the individual level in previously unvaccinated free-roaming dog populations. Individual-level serological and health-based data were collected from three cohorts of dogs in regions where rabies is endemic, one in South Africa and two in Indonesia. We found that the vast majority of dogs seroconverted to the vaccine; however, there was considerable variation in titres, partly attributable to illness and lactation at the time of vaccination. Furthermore, >70% of the dogs were vaccinated through community engagement and door-to-door vaccine delivery, even in Indonesia where the majority of the dogs needed to be caught by net on successive occasions for repeat blood sampling and vaccination. This demonstrates the feasibility of achieving population-level immunity in free-roaming dog populations in rabies-endemic regions. However, attrition of immune individuals through demographic processes and waning immunity necessitates repeat vaccination of populations within at least two years to ensure communities are protected from rabies. These findings support annual mass vaccination campaigns as the most effective means to control canine rabies.