Life-long learning in laboratory animal science and ethics for veterinary and para-veterinary professionals in South Africa.

Veterinary and para-veterinary professionals working in the animal research sector are critical to ensure scientific quality and the humane care and use of animals. However, there are few focused education and training opportunities available for these professionals in South Africa. A survey of veterinarians working in animal research, undertaken by the South African Association for Laboratory Animal Science, identified the need for more advanced education and training opportunities beyond the routine Day 1 Skills currently provided for in undergraduate education. These could be broadly categorised into knowledge and skills relating to species-specific husbandry, procedures and clinical approaches, research-related biosecurity and biosafety, and study-specific ethical and animal welfare considerations. A subsequent workshop, attended by 85 veterinary and para-veterinary professionals in the animal research sector, identified 53 life-long learning needs, each with an associated learning outcome, for this professional community. These were grouped into five overarching themes: Personal development (9); Leadership and management skills (12); Education and training skills (5); Welfare, ethics and clinical skills (20); and Regulations and quality-assurance (7). Of the 53 learning outcomes, 14 were knowledge-based, ten were competencies, and 29 both knowledge and competence. These life-long learning opportunities, if available and implemented, will address important needs of veterinary and paraveterinary professionals in the animal research sector in South Africa. This would empower these professionals, assist in improving animal and human wellbeing, support high-quality ethical science, and maintain public confidence in the sector, thus enabling a more satisfactory career environment.

Learning outcomes for the education and training of laboratory animal caretakers in South Africa.

Education and training is essential for laboratory animal caretakers (LACs), but there are no courses available in South Africa. A national workshop was thus held to collaboratively establish the learning outcomes (LOs) for the education and training (E&T) of LACs. Eighty-five stakeholders from 30 institutions took part in small group discussions interspersed with plenary sessions to draw up the consensus LOs. One-hundred-and-twenty LOs were identified, grouped into the following three main themes and 15 topics: 1) Focus on animals (animal care and husbandry, animal ethics, animal welfare, basic biology, environment); 2) Focus on humans (administration, health and safety, lifelong learning, professionalism, psychological wellbeing); and 3) Focus on systems (biosecurity, equipment, jurisprudence, logistics, and quality management). This E&T framework provides a foundation for a career path in the laboratory animal science field. The psychological (i.e. mental and emotional) wellbeing of LACs forms a noteworthy component of the focus on humans, since working with research animals is stressful and coping mechanisms are needed in order to promote compassion satisfaction and prevent compassion fatigue and burnout. Approximately 75% of the LOs are knowledge-based, while 25% are competencies in practical skills. It is recommended that competencies should be assessed by direct observation of practical/procedural skills, where competence in a procedure or practical task is assessed against predetermined criteria. These LOs are published with the intent that they will promote animal and human wellbeing, support ethical science, maintain public confidence, and in so doing, contribute to a just and civilised society.

A comparative study of cytoplasmic granules imaged by the real-time microscope, Nile Red and Filipin in fibroblasts from patients with lipid storage diseases.

Cytoplasmic granules in fibroblasts, visualized without stains, or labelled with Nile red, Filipin, or anti-LAMP-1 (lysosome-associated membrane protein 1), were imaged using the real-time microscope (RTM). New advances in light microscope technology were applied to detect cytoplasmic granules (RTM-visible granules) and characterize them by imaging contrast, size, shape, cellular distribution, composition, motion dynamics and quantity. Appearing as solid spheroids or ring structures, the majority of the RTM-visible granules contained Nile-red labelled neutral lipids. A smaller subpopulation, appearing dimmer, with less imaging contrast, contained Filipin-labelled free cholesterol. Most lipid storage granules have a diameter ranging from 0.3 mum to 0.6 mum, with a small population measuring up to 1 mum. They typically clustered in the perinuclear region and displayed relatively small oscillatory motion. Immunofluorescence based on LAMP-1 labelling highlighted granular structures that were distinct and separate from RTM-visible granules and other structures in the light modality of the microscope. RTM-visible granules were associated with disease phenotypes that have increased cellular neutral lipid stores corresponding to the Nile red-labelled droplets (e.g. triacylglycerides, cholesterol esters). As predicted, the fibroblast strains with a defect resulting in Wolman disease, when compared to control samples, consistently had RTM-visible granules, higher in imaging contrast and with larger diameters, that were labelled with Nile red, and also an increased frequency of Filipin-cholesterol complexes. By comparison, in fibroblasts where the lipid storage is less evident (Gaucher and Farber diseases) or from GM(1) gangliosidosis, where the primary storage substances are oligosaccharides, fewer and smaller RTM-visible granules were observed. In some cases, changes in contrast and morphology in the unstained cytoplasmic compartments were more evident than in the labelled structures. In summary, applying the RTM imaging system to fibroblasts enables differences between the various disease types to be seen and, in specific examples, a unique phenotype can be readily discerned.

Arrhythmogenic right ventricular cardiomyopathy in a dog.

An 8-month-old Labrador retriever bitch was evaluated for sudden-onset, progressive abdominal distension. Physical examination revealed an exaggerated inspiratory effort, severe ascites, bilateral jugular vein distension, and hypokinetic femoral arterial pulses. Thoracic auscultation detected tachycardia with muffled heart sounds, without audible cardiac murmurs. Thoracic radiographs identified severe right ventricular enlargement and pleural effusion. The electrocardiogram was consistent with incomplete right bundle branch block or right ventricular enlargement. Echocardiography demonstrated severe right ventricular and atrial dilation, secondary tricuspid regurgitation, and thinning and hypocontractility of the right ventricular myocardium. Left heart chamber sizes were slightly decreased, with normal left ventricular contractility. A diagnosis of arrhythmogenic right ventricular cardiomyopathy was reached, based on the characteristic clinical, electrocardiographic, radiographic and echocardiographic findings, and the exclusion of other causes of isolated right ventricular failure. Treatment effected good control of clinical signs, until acutely decompensated congestive right heart failure led to euthanasia after 4 months. Arrhythmogenic right ventricular cardiomyopathy is a well-described clinical entity in humans, and has previously been documented in 3 male dogs. The condition is characterised by progressive fibro-adipose replacement of right ventricular myocardium, while the left ventricle usually remains unaffected. It should be considered a differential diagnosis in any young dog presented with isolated right heart failure, syncope, or unexplained ventricular tachyarrhythmias. This article reports the 1st case of arrhythmogenic right ventricular cardiomyopathy in a female dog, and highlights its echocardiographic features.

A preliminary study on the serum protein response in canine babesiosis.

Total serum protein, albumin, globulin, globulin fractions (alpha, beta and gamma globulins) and an acute-phase protein (alpha1-acid glycoprotein) were evaluated in dogs with naturally occurring mild (Group 1), severe (Group 2) or complicated babesiosis (Group 3). Results showed that the total serum protein, albumin, A/G ratio and alpha globulins were statistically different between Groups 1 and 2. There was no statistical difference between groups with total, beta and gamma globulins. The findings from this study suggest that dogs with mild and severe babesiosis had low total serum proteins, albumin, A/G ratio and alpha globulins; dogs with complicated babesiosis showed no typical serum protein changes or patterns; and that there was no evidence of an acute-phase response detectable on serum protein electrophoresis in any of the 3 groups. A marked acute-phase response was, however, present, as measured by the alpha1-acid glycoprotein, in all 3 groups. As this was a retrospective study, the possibility that the observed responses were due in part to concurrent disease could not be excluded.

Acute pancreatitis: a newly recognised potential complication of canine babesiosis.

This retrospective study describes 4 cases of canine babesiosis with histologically confirmed acute pancreatitis. In addition, 16 dogs with babesiosis are reported with serum amylase (>3500 U/l) and/or lipase (>650 U/l) activity elevations of a magnitude that would support a diagnosis of probable acute pancreatitis, although extra-pancreatic sources of the enzymes could not be excluded in these cases. Median time of pancreatitis diagnosis was 2.5 days post-admission, with primarily young (median age 3 years), sexually intact dogs affected. The development of pancreatitis was unrelated to the degree of anaemia at time of admission. In addition to pancreatitis, 80% of cases suffered from other babesial complications, namely icterus (13), acute respiratory distress syndrome (6), immune-mediated haemolytic anaemia (6), renal failure (3), haemoconcentration (2) and cerebral syndrome (2). Acute respiratory distress syndrome, renal failure and cerebral syndrome were associated with a poor prognosis, with 4 of the 5 dogs included in the overall 26% mortality rate having at least 1 of these complications. Haemolytic anaemia with ischaemia-reperfusion injury to the pancreas is proposed as a possible primary pathophysiological mechanism in babesial pancreatitis. Hypotensive shock, immune-mediated haemolytic anaemia, haemoconcentration and possibly altered lipid metabolism in babesiosis may also be involved. The previously postulated pro-inflammatory cytokine milieu of complicated babesiosis may underlie the progression, if not the primary initiation, of pancreatic pathology. Acute pancreatitis may represent the previously reported 'gut' form of babesiosis.

Estimating downwind concentrations of viable airborne microorganisms in dynamic atmospheric conditions.

A Gaussian plume model has been modified to include an airborne microbial survival term that is a best-fit function of laboratory experimental data of weather variables. The model has been included in an algorithm using microbial source strength and local hourly mean weather data to drive the model through a summer- and winter-day cycle. For illustrative purposes, a composite airborne "virus" (developed using actual characteristics from two viruses) was used to show how wind speed could have a major modulating effect on near-source viable concentrations. For example, at high wind speeds such as those occurring during the day, or with short travel times, near-source locations experience high viable concentrations because the microorganisms have not had time to become inactivated. As the travel time increases, because of slow wind speed or longer distances, die-off modulation by sunshine, relative humidity, temperature, etc., potentially becomes increasingly predominant.