Ferrets as a Mammalian Model to Study Influenza Virus-Bacteria Interactions.

Ferrets represent an invaluable model for the study of influenza virus pathogenicity and transmissibility. Ferrets are also employed for the study of bacterial pathogens that naturally infect humans at different anatomical sites. While viral and bacterial infection studies in isolation using animal models are important for furthering our understanding of pathogen biology and developing improved therapeutics, it is also critical to extend our knowledge to pathogen coinfections in vivo, to more closely examine interkingdom dynamics that may contribute to overall disease outcomes. We discuss how ferrets have been employed to study a diverse range of both influenza viruses and bacterial species and summarize key studies that have utilized the ferret model for primary influenza virus challenge followed by secondary bacterial infection. These copathogenesis studies have provided critical insight into the dynamic interplay between these pathogens, underscoring the utility of ferrets as a model system for investigating influenza virus-bacteria interactions.

Pharmacokinetic Study of Islatravir and Etonogestrel Implants in Macaques.

The prevention of HIV and unintended pregnancies is a public health priority. Multi-purpose prevention technologies capable of long-acting HIV and pregnancy prevention are desirable for women. Here, we utilized a preclinical macaque model to evaluate the pharmacokinetics of biodegradable ε-polycaprolactone implants delivering the antiretroviral islatravir (ISL) and the contraceptive etonogestrel (ENG). Three implants were tested: ISL-62 mg, ISL-98 mg, and ENG-33 mg. Animals received one or two ISL-eluting implants, with doses of 42, 66, or 108 µg of ISL/day with or without an additional ENG-33 mg implant (31 µg/day). Drug release increased linearly with dose with median [range] plasma ISL levels of 1.3 [1.0-2.5], 1.9 [1.2-6.3] and 2.8 [2.3-11.6], respectively. The ISL-62 and 98 mg implants demonstrated stable drug release over three months with ISL-triphosphate (ISL-TP) concentr54ations in PBMCs above levels predicted to be efficacious for PrEP. Similarly, ENG implants demonstrated sustained drug release with median [range] plasma ENG levels of 495 [229-1110] pg/mL, which suppressed progesterone within two weeks and showed no evidence of altering ISL pharmacokinetics. Two of the six ISL-98 mg implants broke during the study and induced implant-site reactions, whereas no reactions were observed with intact implants. We show that ISL and ENG biodegradable implants are safe and yield sufficient drug levels to achieve prevention targets. The evaluation of optimized implants with increased mechanical robustness is underway for improved durability and vaginal efficacy in a SHIV challenge model.

Ultra-long-acting in-situ forming implants with cabotegravir protect female macaques against rectal SHIV infection.

Ultra-long-acting delivery platforms for HIV pre-exposure prophylaxis (PrEP) may increase adherence and maximize public health benefit. We report on an injectable, biodegradable, and removable in-situ forming implant (ISFI) that is administered subcutaneously and can release the integrase inhibitor cabotegravir (CAB) above protective benchmarks for more than 6 months. CAB ISFIs are well-tolerated in female mice and female macaques showing no signs of toxicity or chronic inflammation. In macaques, median plasma CAB concentrations exceed established PrEP protection benchmarks within 3 weeks and confer complete protection against repeated rectal SHIV challenges. Implant removal via a small incision in 2 macaques at week 12 results in a 7- to 48-fold decrease in plasma CAB levels within 72 hours. Modeling to translate CAB ISFI dosing suggests that a 3 mL injection would exceed protective benchmarks in humans for over 5 months post administration. Our results support the clinical advancement of CAB ISFIs for ultra-long-acting PrEP in humans.

Effects of Buprenorphine Treatment on Influenza Pathogenesis in the Ferret (Mustela putorius furo).

Ferrets are the gold-standard model for influenza A virus (IAV) research due to their natural susceptibility to human and zoonotic IAV, comparable respiratory anatomy and physiology to humans, and development of clinical signs similar to those seen in infected people. Because the presence and progression of clinical signs can be useful in infectious disease research, uncertainty in how analgesics alter research outcomes or compromise characteristics of disease progression have outweighed the concern regarding animal discomfort from these symptoms. Nonetheless, the principles of animal research require consideration of refinements for this important model for IAV research. Opioids offer a possible refinement option that would not directly affect the inflammatory cascade involved in IAV infection. Mirroring pathogenicity studies that use ferrets, 12 ferrets were inoculated intranasally with the A(H3N2) IAV A/Panama/2007/1999 and divided into 3 treatment groups ( n = 4 each), of which 2 groups received buprenorphine treatments on different schedules and the third received a saline control. The duration and location of viral replication, lymphohematopoietic changes, and clinical signs were comparable across all groups at all time points. High quantities of infectious virus in nasal wash specimens were detected in ferrets from all groups through day 5 after inoculation, and peak viral titers from the upper respiratory tract did not differ between ferrets receiving buprenorphine treatments on either schedule. Compared with the saline group, ferrets receiving buprenorphine exhibited transient weight loss and pyrexia, but all groups ultimately achieved similar peaks in both of these measurements. Collectively, these findings support the continued evaluation of buprenorphine as a refinement for IAV-challenged ferrets.

One Health Investigation of SARS-CoV-2 Infection and Seropositivity among Pets in Households with Confirmed Human COVID-19 Cases-Utah and Wisconsin, 2020.

Approximately 67% of U.S. households have pets. Limited data are available on SARS-CoV-2 in pets. We assessed SARS-CoV-2 infection in pets during a COVID-19 household transmission investigation. Pets from households with ≥1 person with laboratory-confirmed COVID-19 were eligible for inclusion from April-May 2020. We enrolled 37 dogs and 19 cats from 34 households. All oropharyngeal, nasal, and rectal swabs tested negative by rRT-PCR; one dog's fur swabs (2%) tested positive by rRT-PCR at the first sampling. Among 47 pets with serological results, eight (17%) pets (four dogs, four cats) from 6/30 (20%) households had detectable SARS-CoV-2 neutralizing antibodies. In households with a seropositive pet, the proportion of people with laboratory-confirmed COVID-19 was greater (median 79%; range: 40-100%) compared to households with no seropositive pet (median 37%; range: 13-100%) (p = 0.01). Thirty-three pets with serologic results had frequent daily contact (≥1 h) with the index patient before the person's COVID-19 diagnosis. Of these 33 pets, 14 (42%) had decreased contact with the index patient after diagnosis and none were seropositive; of the 19 (58%) pets with continued contact, four (21%) were seropositive. Seropositive pets likely acquired infection after contact with people with COVID-19. People with COVID-19 should restrict contact with pets and other animals.

Comparison of outcome in dogs undergoing single-incision laparoscopic-assisted intestinal surgery and open laparotomy for simple small intestinal foreign body removal.

OBJECTIVE: To evaluate and compare outcome in dogs that underwent single-incision laparoscopic-assisted intestinal surgery (SILAIS) and open laparotomy (OL) for simple foreign body removal. STUDY DESIGN: Retrospective study. ANIMALS: Twenty-eight client-owned dogs that underwent SILAIS (n = 13) or OL (n = 15). METHODS: Foreign body removal via SILAIS was performed with a commercially available single-port device. After laparoscopic evaluation, a wound retraction device (WRD) was inserted. The small intestine was extracorporeally explored, and foreign body removal was performed. The surgeon's hand was inserted through the WRD to palpate portions of the gastrointestinal tract not able to be extracorporeally evaluated. Open laparotomy with foreign body removal was performed via a ventral midline approach. Medical records were retrospectively reviewed, and perioperative data were collected. Follow-up data were collected with a standardized questionnaire with the referring veterinarian and/or owner via telephone interview. RESULTS: No postoperative complications were encountered in either the SILAIS or the OL group, and all dogs were successfully discharged from the hospital. Conversion from SILAIS to OL occurred in 3/13 cases. There was no significant difference in duration of hospitalization, duration of time to recovery, or surgical time between surgical approaches (SILAIS vs OL). CONCLUSION: Single-incision laparoscopic-assisted intestinal surgery for foreign body removal was not significantly different from OL in a variety of outcome measures in this cohort of dogs. Diagnostic imaging including ultrasonography or computed tomography may improve appropriate case selection for SILAIS for simple foreign body removal. CLINICAL SIGNIFICANCE: Single-incision laparoscopic-assisted intestinal surgery offers a minimally invasive technique for simple small intestinal foreign body removal. Additional study is required to compare SILAIS with OL.

Investigating laparoscopic psychomotor skills in veterinarians and veterinary technicians.

OBJECTIVES: To determine the influence of age, year of graduation, and video game experience on baseline laparoscopic psychomotor skills. STUDY DESIGN: Cross-sectional. SAMPLE POPULATION: Licensed veterinarians (n = 38) and registered veterinary technicians (VTs) (n = 49). METHODS: A laparoscopic box trainer was set up at the 2016 Ontario Veterinary Medical Association (OVMA) and the 2016 Ontario Association of Veterinary Technicians (OAVT) conferences held in Toronto, Ontario, Canada. Participants volunteered to perform a single repetition of a peg transfer (PT) exercise. Participants were given a short demonstration of the PT task prior to testing. A Spearman's rank correlation (rs ) was used to identify associations between baseline psychomotor skills and self-reported surgical and non-surgical experiences collected via survey. Mann-Whitney U tests were used to compare PT scores in veterinarians and VTs. A P-value of < .05 was considered significant. RESULTS: The mean age of participants was 36 years (range 21-67) and the majority were female (83%). In veterinarians, PT scores were highest in the most recent graduates (P = .01, rs = 0.42), and PT scores increased with self-reported VG experience (P = .02, rs = 0.38). PT scores correlated inversely with age (P = .02, rs = -0.37). No associations were observed in VTs (P > .05). Veterinary technicians that frequently used chopsticks scored higher than those without chopstick experience (P = .04). CONCLUSIONS: Age and year of graduation correlated inversely, while self-reported VG experience correlated positively with laparoscopic psychomotor skills of veterinarians, when assessed on a simulator. The use of chopsticks may contribute to the acquisition of psychomotor skills in VTs.

The Impact of Distraction on Laparoscopic Skills in Veterinary Medical Students.

OBJECTIVE: To investigate the impact of distraction on the performance of a simulator-based laparoscopic task in veterinary students. STUDY DESIGN: Prospective, randomized trial. SAMPLE POPULATION: Years 1-4 veterinary students (n=41). METHODS: Participants repeated a simulated laparoscopic peg transfer task to eliminate any learning effects and were subsequently randomized to receive either a cognitive (double-digit addition questions, n=21) or sensory distraction (dogs barking and anesthesia monitor alerts, n=20). The laparoscopic task scores were compared between baseline and in the presence and absence of each distraction. The number of addition questions attempted, and the number of questions answered correctly in 1 minute were compared between baseline and during a concurrent laparoscopic task. RESULTS: Baseline laparoscopic task scores were not significantly different between groups (P=.09). Laparoscopic task scores were significantly lower than baseline when performed with the cognitive distraction (P<.001) and significantly higher than baseline when performed with the sensory distraction (P=.005). Participants undergoing cognitive distraction attempted significantly fewer addition questions (P<.001) and answered significantly fewer addition questions correctly (P<.001) when a concurrent laparoscopic task was performed. CONCLUSION: Cognitive distraction had a negative impact on the performance of a laparoscopic task in this cohort of veterinary students, whereas sensory distraction had a positive effect.