NDUFS7 variant in dogs with Leigh syndrome and its functional validation in a Drosophila melanogaster model.

Two Jack-Russell Terrier × Chihuahua mixed-breed littermates with Leigh syndrome were investigated. The dogs presented with progressive ataxia, dystonia, and increased lactate levels. Brain MRI showed characteristic bilateral symmetrical T2 hyperintense lesions, histologically representing encephalomalacia. Muscle histopathology revealed accumulation of mitochondria. Whole genome sequencing identified a missense variant in a gene associated with human Leigh syndrome, NDUFS7:c.535G > A or p.(Val179Met). The genotypes at the variant co-segregated with the phenotype in the investigated litter as expected for a monogenic autosomal recessive mode of inheritance. We investigated the functional consequences of the missense variant in a Drosophila melanogaster model by expressing recombinant wildtype or mutant canine NDUFS7 in a ubiquitous knockdown model of the fly ortholog ND-20. Neither of the investigated overexpression lines completely rescued the lethality upon knockdown of the endogenous ND-20. However, a partial rescue was found upon overexpression of wildtype NDUFS7, where pupal lethality was moved to later developmental stages, which was not seen upon canine mutant overexpression, thus providing additional evidence for the pathogenicity of the identified variant. Our results show the potential of the fruit fly as a model for canine disease allele validation and establish NDUFS7:p.(Val179Met) as causative variant for the investigated canine Leigh syndrome.

Retrospective study and outcome of 307 cats with feline infectious peritonitis treated with legally sourced veterinary compounded preparations of remdesivir and GS-441524 (2020-2022).

OBJECTIVES: Feline infectious peritonitis (FIP) is a serious disease that arises due to feline coronavirus infection. The nucleoside analogues remdesivir and GS-441524 can be effective in its treatment, but most studies have used unregulated products of unknown composition. The aim of the present study was to describe the treatment of FIP using legally sourced veterinary-prescribed regulated veterinary compounded products containing known amounts of remdesivir (injectable) or GS-441524 (oral tablets). METHODS: Cats were recruited via email advice services, product sales contacts and study publicity. Cats were excluded if they were deemed unlikely to have FIP, were not treated exclusively with the veterinary compounded products, or if there was a lack of cat and/or treatment (including response) data. Extensive cat and treatment data were collected. RESULTS: Among the 307 cats recruited, the predominant type of FIP was most commonly abdominal effusive (49.5%) and then neurological (14.3%). Three treatment protocols were used; remdesivir alone (33.9%), remdesivir followed by GS-441524 (55.7%) and GS-441524 alone (10.4%). The median (range) initial treatment period duration and longest follow-up time point after starting treatment were 84 (1-330) days and 248 (1-814) days, respectively. The most common side effect was injection pain (in 47.8% of those given subcutaneous remdesivir). Of the 307 cats, 33 (10.8%) relapsed, 15 (45.5%) during and 18 (54.5%) after the initial treatment period. At the longest follow-up time point after completion of the initial treatment period, 84.4% of cats were alive. The cats achieving a complete response within 30 days of starting treatment were significantly more likely to be alive at the end of the initial treatment period than those cats that did not. CONCLUSIONS AND RELEVANCE: Legally sourced remdesivir and GS-441524 products, either alone or used sequentially, were very effective in the treatment of FIP in this group of cats. Variable protocols precluded statistical comparison of treatment regimens.

Clinical and MRI findings of a suspected cortical malformation presented as a giant cerebral pseudomass in a German Shepherd dog.

A 5-month-old German Shepherd dog was presented with cluster seizures. MR imaging showed a large irregular pseudomass in the central region of the cranial cavity, compatible with a malformation of cortical development. Despite the extensive changes, the patient was neurologically normal interictally 1 year following diagnosis.

ACADM Frameshift Variant in Cavalier King Charles Spaniels with Medium-Chain Acyl-CoA Dehydrogenase Deficiency.

A 3-year-old, male neutered Cavalier King Charles Spaniel (CKCS) presented with complex focal seizures and prolonged lethargy. The aim of the study was to investigate the clinical signs, metabolic changes and underlying genetic defect. Blood and urine organic acid analysis revealed increased medium-chain fatty acids and together with the clinical findings suggested a diagnosis of medium-chain acyl-CoA dehydrogenase (MCAD) deficiency. We sequenced the genome of the affected dog and compared the data to 923 control genomes of different dog breeds. The ACADM gene encoding MCAD was considered the top functional candidate gene. The genetic analysis revealed a single homozygous private protein-changing variant in ACADM in the affected dog. This variant, XM_038541645.1:c.444_445delinsGTTAATTCTCAATATTGTCTAAGAATTATG, introduces a premature stop codon and is predicted to result in truncation of ~63% of the wild type MCAD open reading frame, XP_038397573.1:p.(Thr150Ilefs*6). Targeted genotyping of the variant in 162 additional CKCS revealed a variant allele frequency of 23.5% and twelve additional homozygous mutant dogs. The acylcarnitine C8/C12 ratio was elevated ~43.3 fold in homozygous mutant dogs as compared to homozygous wild type dogs. Based on available clinical and biochemical data together with current knowledge in humans, we propose the ACADM frameshift variant as causative variant for the MCAD deficiency with likely contribution to the neurological phenotype in the index case. Testing the CKCS breeding population for the identified ACADM variant is recommended to prevent the unintentional breeding of dogs with MCAD deficiency. Further prospective studies are warranted to assess the clinical consequences of this enzyme defect.

Accuracy of Lumbosacral Pedicle Screw Placement in Dogs: A Novel 3D Printed Patient-Specific Drill Guide versus Freehand Technique in Novice and Expert Surgeons.

OBJECTIVE: The aim of this study was to compare the accuracy of pedicle screw placement at the canine lumbosacral junction using a novel unilateral three-dimensional printed patient-specific guide (3D-PSG) versus a freehand drilling technique. Additionally, accuracy of screw placement between a novice and an experienced surgeon was determined. STUDY DESIGN: Preoperative computed tomography images from 20 lumbosacral cadaveric specimens were used to design a novel unilateral 3D-PSG for the L7 and sacral vertebrae which was printed in acryl-nitrile butadiene styrene plastic. A novice and an expert surgeon each placed 3.5mm cortical screws in 10 cadavers; on the left using the unilateral 3D-PSG and by the freehand (anatomic landmark) technique on the right. RESULTS: Sixty screws were placed using the unilateral 3D-PSG and 60 using the freehand technique. There was no statistical difference in accuracy for the comparison between methods performed by the expert (p = 0.679) and novice (p = 0.761) surgeon, nor between an expert and novice surgeon overall (p = 0.923). Unexpectedly, the use of a unilateral 3D-PSG increased variability for the expert surgeon in our study (p = 0.0314). CONCLUSION: Using a novel unilateral 3D-PSG did not improve the accuracy of screw placement for lumbosacral stabilization by a novice surgeon compared with an expert surgeon in lumbar spine surgery. This may reflect a suboptimal PSG design.

The Prospects of Non-EEG Seizure Detection Devices in Dogs.

The unpredictable nature of seizures is challenging for caregivers of epileptic dogs, which calls the need for other management strategies such as seizure detection devices. Seizure detection devices are systems that rely on non-electroencephalographic (non-EEG) ictal changes, designed to detect seizures. The aim for its use in dogs would be to provide owners with a more complete history of their dog's seizures and to help install prompt (and potentially life-saving) intervention. Although seizure detection via wearable intracranial EEG recordings is associated with a higher sensitivity in humans, there is robust evidence for reliable detection of generalized tonic-clonic seizures (GTCS) using non-EEG devices. Promising non-EEG changes described in epileptic humans, include heart rate variability (HRV), accelerometry (ACM), electrodermal activity (EDA), and electromyography (EMG). Their sensitivity and false detection rate to detect seizures vary, however direct comparison of studies is nearly impossible, as there are many differences in study design and standards for testing. A way to improve sensitivity and decrease false-positive alarms is to combine the different parameters thereby profiting from the strengths of each one. Given the challenges of using EEG in veterinary clinical practice, non-EEG ictal changes could be a promising alternative to monitor seizures more objectively. This review summarizes various seizure detection devices described in the human literature, discusses their potential use and limitations in veterinary medicine and describes what is currently known in the veterinary literature.

Owner's Perception of Seizure Detection Devices in Idiopathic Epileptic Dogs.

Accurate knowledge of seizure frequency is key to optimising treatment. New methods for detecting epileptic seizures are currently investigated in humans, which rely on changes in biomarkers, also called seizure detection devices. Critical to device development, is understanding user needs and requirements. No information on this subject has been published in veterinary medicine. Many dog health collars are currently on the market, but none has proved to be a promising seizure detector. An online survey was created and consisted of 27 open, closed, and scaled questions divided over two parts: part one focused on general questions related to signalment and seizure semiology, the second part focused specifically on the use of seizure detection devices. Two hundred and thirty-one participants caring for a dog with idiopathic epilepsy, were included in the study. Open questions were coded using descriptive coding by two of the authors independently. Data was analysed using descriptive statistics and binary logistic regression. Our results showed that the unpredictability of seizures plays a major part in the management of canine epilepsy and dog owners have a strong desire to know when a seizure occurs. Nearly all dog owners made changes in their daily life, mainly focusing on intensifying supervision. Owners believed seizure detection devices would improve their dog's seizure management, including a better accuracy of seizure frequency and the ability to administer emergency drugs more readily. Owners that were already keeping track of their dog's seizures were 4.2 times more likely to show confidence in using seizure detection devices to manage their pet's seizures, highlighting the need for better monitoring systems. Our results show that there is a receptive market for wearable technology as a new management strategy in canine epilepsy and this topic should be further explored.