Canine corneal fibroblast and myofibroblast transduction with AAV5

Objective The aims of this study were (1) to determine the efficacy of adeno‐associated vector serotype 5 (AAV5) for delivering gene therapy to canine corneal fibroblasts (CCFs) and myofibroblasts (CCMs) using enhanced green fluorescent protein (GFP) marker gene and (2) to evaluate the cytotoxicity of AAV5 to CCFs and CCMs using an in vitro model. Methods Healthy donor canine corneas were used to generate primary CCFs by growing cultures in minimal essential medium supplemented with 10% fetal bovine serum. Canine corneal myofibroblasts were produced by growing cultures in serum‐free medium containing transforming growth factor β1 (1 ng/mL). An AAV5 titer (6.5 × 10¹² μg/mL) expressing GFP under control of hybrid cytomegalovirus + chicken β‐actin promoters (AAV5‐gfp) was used to transduce CCF and CCM cultures. Delivered gene expression in CCFs and CCMs was quantified using immunocytochemistry, fluorescent microscopy, and real‐time PCR. Transduction efficacy of the AAV5 vector was determined by counting DAPI‐stained nuclei and EGFP‐positive cells in culture. Phase‐contrast microscopy, trypan blue, and dUTP nick end labeling (TUNEL) assays were used to determine the toxicity and safety of AAV5 in this canine corneal model. Results Topical AAV5 application successfully transduced a significant population of CCFs (42.8%; P < 0.01) and CCMs (28%; P < 0.01). Tested AAV5 did not affect CCF or CCM phenotype or cellular viability and did not cause significant cell death. Conclusions The tested AAV5 is an effective and safe vector for canine corneal gene therapy in this in vitro model. In vivo studies are warranted.     

Risk-based surveillance for avian influenza control along poultry market chains in South China: The value of social network analysis

Over the past two decades, the poultry sector in China went through a phase of tremendous growth as well as rapid intensification and concentration. Highly pathogenic avian influenza virus (HPAIV) subtype H5N1 was first detected in 1996 in Guangdong province, South China and started spreading throughout Asia in early 2004. Since then, control of the disease in China has relied heavily on wide-scale preventive vaccination combined with movement control, quarantine and stamping out. This strategy has been successful in drastically reducing the number of outbreaks during the past 5years. However, HPAIV H5N1 is still circulating and is regularly isolated in traditional live bird markets (LBMs) where viral infection can persist, which represent a public health hazard for people visiting them. The use of social network analysis in combination with epidemiological surveillance in South China has identified areas where the success of current strategies for HPAI control in the poultry production sector may benefit from better knowledge of poultry trading patterns and the LBM network configuration as well as their capacity for maintaining HPAIV H5N1 infection. We produced a set of LBM network maps and estimated the associated risk of HPAIV H5N1 within LBMs and along poultry market chains, providing new insights into how live poultry trade and infection are intertwined. More specifically, our study provides evidence that several biosecurity factors such as daily cage cleaning, daily cage disinfection or manure processing contribute to a reduction in HPAIV H5N1 presence in LBMs. Of significant importance is that the results of our study also show the association between social network indicators and the presence of HPAIV H5N1 in specific network configurations such as the one represented by the counties of origin of the birds traded in LBMs. This new information could be used to develop more targeted and effective control interventions.     

Surgical management of a canine intracranial abscess due to a bite wound

Objective – To describe the successful surgical management of a brain abscess in a dog secondary to bite wound. Case Summary – A 10‐year‐old neutered female Welsh Corgi/Chihuahua, weighing 5.3 kg, was presented for evaluation of seizures, ataxia, and falling to the left 8 days after a presumptive fight with another dog. On examination at presentation, the dog was alert, responsive, and ambulatory with tetra‐ataxia, falling to the left, left‐sided postural deficits, and absent left menace response. Within 24 hours, the dog progressed to nonambulatory tetraparesis with minimal motor, absent postural reactions of all limbs, left nasal hypalgesia, reduced gag reflex, and depressed mentation. Computed tomographic images of the brain were suggestive of a bite wound fracture of the right parietal bone with secondary meningoencephalitis, right parietal lobe abscessation, and white matter edema adjacent to the bone fracture. A modified right rostrotentorial craniectomy was performed, the abscess was identified, contents of the abscess were removed, and the surgical site was flushed extensively before closing. Corynebacterium spp. was cultured from within the abscess. Within hours of surgery, the dog was quiet but alert, responsive, and sitting up in her cage. In addition to surgical intervention, intensive care, broad‐spectrum IV antimicrobials, and supportive therapy led to significant neurologic improvement with only occasional seizures and mild postural reaction deficits of the left hindlimb remaining. New or Unique Information Provided – Abscess formation within the CNS is uncommon in dogs and cats and is associated with a high mortality rate. In veterinary medicine the management of brain abscesses is controversial with limited information available regarding treatment. This is the first case report that demonstrates surgical intervention in combination with antimicrobial therapy can be used successfully in the treatment of a canine brain abscess.     

A Genetic Study of Piglet Growth and Survival

The study analysed 954 Yorkshire sows from a research station in Sweden. The heritability for mean piglet weight at 3 weeks was 0.30 and the heritability for number dead up to 3 weeks was 0.09. Correlations showed that when the number of piglets born into a litter increased, it followed that each piglet weighed less at 3 weeks of age (genetic correlation -0.1). The higher the litter size at birth, the more piglets died, and if many piglets died in a litter, growth was low in the surviving piglets. The genetic correlation between mean birth weight and mean weight at 3 weeks was estimated to be 0.6.     

Multicenter, randomized controlled trial of pain-related behaviors following routine neutering in dogs

OBJECTIVE: To evaluate the degree of postoperative pain in dogs undergoing elective castration or ovariohysterectomy (OHE); determine whether an association exists between surgeon experience, incision length, or surgery duration and degree of postoperative pain; and determine whether analgesic treatment decreases expression of postoperative pain behaviors. DESIGN: Randomized controlled clinical trial. ANIMALS: 426 client-owned dogs undergoing OHE or castration. PROCEDURES: Dogs underwent OHE or castration performed by an experienced veterinarian or a fourth-year veterinary student. Dogs were randomly assigned to 1 of 4 treatment groups: no perioperative analgesic treatment (n = 44), preoperative administration of morphine (144), preoperative administration of nalbuphine (119), and postoperative administration of ketoprofen (119). Dogs were evaluated while in the hospital before anesthesia and for 4 hours after surgery and once a day at home for 3 days after surgery. RESULTS: Dogs in all 4 groups had significant increases in overall pain scores after surgery, compared with baseline scores. There were significant differences among groups, with control dogs having significantly higher increases in overall pain scores than dogs in the other groups. Factors that did not influence the frequency or severity of pain-related behaviors included breed, individual hospital, anesthetic induction protocol, surgeon experience, and duration of surgery. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that dogs expressed behaviors suggestive of pain following OHE and castration, that analgesic treatment mitigated the expression of pain-related behaviors, and that surgeon experience and surgery duration did not have any effect on expression of pain-related behaviors.