Limb-girdle muscular dystrophy type 2A (LGMD2A or LGMDR1) is a neuromuscular disorder caused by mutations in the calpain 3 gene (CAPN3). Previous experiments using adeno-associated viral (AAV) vector–mediated calpain 3 gene transfer in mice indicated cardiac toxicity associated with the ectopic expression of the calpain 3 transgene. Here, we performed a preliminary dose study in a severe double-knockout mouse model deficient in calpain 3 and dysferlin. We evaluated safety and biodistribution of AAV9-desmin-hCAPN3 vector administration to nonhuman primates (NHPs) with a dose of 3 × 10¹³ viral genomes/kg. Vector administration did not lead to observable adverse effects or to detectable toxicity in NHP. Of note, the transgene expression did not produce any abnormal changes in cardiac morphology or function of injected animals while reaching therapeutic expression in skeletal muscle. Additional investigation on the underlying causes of cardiac toxicity observed after gene transfer in mice and the role of titin in this phenomenon suggest species-specific titin splicing. Mice have a reduced capacity for buffering calpain 3 activity compared to NHPs and humans. Our studies highlight a complex interplay between calpain 3 and titin binding sites and demonstrate an effective and safe profile for systemic calpain 3 vector delivery in NHP, providing critical support for the clinical potential of calpain 3 gene therapy in humans.

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Blood transfusion has been described in ferrets as a treatment for oestrus‐associated anaemia and as a life‐saving therapy following trauma, iatrogenic (usually surgery‐induced) anaemia, autoimmune haemolytic anaemia and pure red cell aplasia. Although blood banking is a common method for storage of feline and canine blood it is not currently done with ferret blood. The aim of this study was to determine the shelf‐life of ferret blood using the anticoagulant citrate‐phosphate‐dextrose‐solution with adenine (CPDA). Two male ferrets were used as blood donors. From each ferret, 6 ml of blood was taken from the cranial vena cava and stored in 10 ml polyethylene terephthalate (PET) blood tubes containing 1 ml of CPDA solution. Blood was taken from each ferret once per month for five months. These 10 blood samples were stored in a laboratory refrigerator at 4°C for four weeks. Biochemical (glucose, pH, lactate, potassium, sodium) and haematological (haematocrit, light microscopic blood smear examination) analyses were performed on the stored blood at days 0, 7, 14, 21 and 28. Biochemical analyses revealed a progressive decrease from day seven in the stored blood pH, glucose and sodium, with a concomitant increase in lactate and potassium. These results are attributable to the ongoing metabolism and deterioration of the red blood cells (RBC) while in storage, and are more rapid than described for human or canine stored blood. Haematological analyses revealed a progressive elevation of the haematocrit due to the appearance of hypochromic red blood cells and echinocytes beginning at day 7. Haemolysis was observed in the microhaematocrit capillary tube sample by day 21, and microscopic clots were visible on the blood smear by day 28. The low blood pH and the appearance of many hypochromic RBCs and some echinocytes from day 7 in CPDA‐stored ferret blood, suggest stored ferret blood has a short shelf‐life when compared with stored human or canine blood. We recommend that ferret blood stored in CPDA should not be used for transfusion after seven days of storage at 4°C.

Case series summary: In October 2011, an abnormally large morbidity and mortality event was noted in the intensive care unit (ICU) of a veterinary school hospital in Nantes, France. Cats, and cats only, transferred from the emergency room presented with fever, ulcers on the tongue and cutaneous lesions around venepuncture or surgical incision sites, leading to suspicion of a feline calicivirus-associated virulent systemic disease confirmed with reverse transcriptase-polymerase chain reaction. A total of 14 cats were suspected. The clinical features and the origin of the contamination were described for each cat. The median length of incubation was 4.5 days. Fifty-seven percent of the cats were euthanased (8/14) and 21% died (3/14), with a combined mortality of 79% (11/14) – the highest ever reported. Median survival was 12 days. The recovery rate was 21% (3/14). Relevance and novel information: Eight outbreaks have been reported, in veterinary clinics or in group-housed cats. The main unusual aspects of the present outbreak were: (1) the extreme flare-up of lesions at sites of skin breach, precluding any puncture/incision; (2) the suggested better survival rate at home than in hospital; and (3) the immediate control of the outbreak after recognition of the disease. Other striking but less unusual features of this outbreak were: (4) the increasing of the virulence of the calicivirus with the passage of time; and (5) the primary role that the caregivers’ hands played in the spread of the outbreak.

Upper urinary tract urolithiasis is an emerging disease in cats, with 98% of kidney stones composed of calcium oxalate. In humans, disturbances in the intestinal and urinary microbiota are suspected to contribute to the formation of calcium oxalate stones. We hypothesized that similar mechanisms may be at play in cats. This study examines the intestinal and urinary microbiota of nine cats with kidney stones compared to nine healthy cats before, during, and after treatment with the antibiotic cefovecin, a cephalosporin. Initially, cats with kidney stones displayed a less diverse intestinal microbiota. Antibiotic treatment reduced microbiota diversity in both groups. The absence of specific intestinal bacteria could lead to a loss of the functions these bacteria perform, such as oxalate degradation, which may contribute to the formation of calcium oxalate stones. This study confirms the presence of a distinct urobiome in cats with kidney stones, characterized by greater richness and diversity compared to healthy cats. These findings highlight the potential of microbiota modulation as a strategy to prevent renal lithiasis in cats.