Total infectome investigation of diphtheritic stomatitis in yellow-eyed penguins (Megadyptes antipodes) reveals a novel and abundant megrivirus.

First identified in 2002, diphtheritic stomatitis (DS) is a devastating disease affecting yellow-eyed penguins (Megadyptes antipodes, or hoiho in te reo Maori). The disease is associated with oral lesions in chicks and has caused significant morbidity and mortality. DS is widespread among yellow-eyed penguin chicks on mainland New Zealand yet appears to be absent from the subantarctic population. Corynebacterium spp. have previously been suspected as causative agents yet, due to inconsistent cultures and inconclusive pathogenicity, their role in DS is unclear. Herein, we used a metatranscriptomic approach to identify potential causative agents of DS by revealing the presence and abundance of all viruses, bacteria, fungi and protozoa - together, the infectome. Oral and cloacal swab samples were collected from presymptomatic, symptomatic and recovered chicks along with a control group of healthy adults. Two novel viruses from the Picornaviridae were identified, one of which - yellow-eyed penguin megrivirus - was highly abundant in chicks irrespective of health status but not detected in healthy adults. Tissue from biopsied oral lesions also tested positive for the novel megrivirus upon PCR. We found no overall clustering among bacteria, protozoa and fungi communities at the genus level across samples, although Paraclostridium bifermentans was significantly more abundant in oral microbiota of symptomatic chicks compared to other groups. The detection of a novel and highly abundant megrivirus has sparked a new line of inquiry to investigate its potential association with DS.

Abundant dsRNA picobirnaviruses show little geographic or host association in terrestrial systems.

Picobirnaviruses are double-stranded RNA viruses known from a wide range of host species and locations but with unknown pathogenicity and host relationships. Here, we examined the diversity of picobirnaviruses from cattle and gorillas within and around Bwindi Impenetrable Forest National Park (BIFNP), Uganda, where wild and domesticated animals and humans live in relatively close contact. We use metagenomic sequencing with bioinformatic analyses to examine genetic diversity. We compared our findings to global Picobirnavirus diversity using clustering-based analyses. Picobirnavirus diversity at Bwindi was high, with 14 near-complete RdRp and 15 capsid protein sequences, and 497 new partial viral sequences recovered from 44 gorilla samples and 664 from 16 cattle samples. Sequences were distributed throughout a phylogenetic tree of globally derived picobirnaviruses. The relationship with Picobirnavirus diversity and host taxonomy follows a similar pattern to the global dataset, generally lacking pattern with either host or geography.

A novel gyrovirus is abundant in yellow-eyed penguin (Megadyptes antipodes) chicks with a fatal respiratory disease.

Yellow-eyed penguins (Megadyptes antipodes), or hoiho in te reo Maori, are predicted to become extinct on mainland Aotearoa New Zealand in the next few decades, with infectious disease a significant contributor to their decline. A recent disease phenomenon termed respiratory distress syndrome (RDS) causing lung pathology has been identified in very young chicks. To date, no causative pathogens for RDS have been identified. In 2020 and 2021, the number of chick deaths from suspected RDS increased four- and five-fold, respectively, causing mass mortality with an estimated mortality rate of >90%. We aimed to identify possible pathogens responsible for RDS disease impacting these critically endangered yellow-eyed penguins. Total RNA was extracted from tissue samples collected during post-mortem of 43 dead chicks and subject to metatranscriptomic sequencing and histological examination. From these data we identified a novel and highly abundant gyrovirus (Anelloviridae) in 80% of tissue samples. This virus was most closely related to Gyrovirus 8 discovered in a diseased seabird, while other members of the genus Gyrovirus include Chicken anaemia virus, which causes severe disease in juvenile chickens. No other exogenous viral transcripts were identified in these tissues. Due to the high relative abundance of viral reads and its high prevalence in diseased animals, it is likely that this novel gyrovirus is associated with RDS in yellow-eyed penguin chicks.

An alternative combination therapy with metronidazole, clindamycin and doxycycline for Babesia gibsoni (Asian genotype) in dogs in Hong Kong.

Babesia spp. are globally distributed hemoparasites that cause disease in many mammalian species. The species Babesia gibsoni (Asian genotype) is prevalent and endemic in many Asian countries but has also been reported in growing numbers in countries outside of Asia. Standard therapies for the treatment of B. gibsoni often fail to result in consistent and successful clearance of the organism. This study evaluated the use of a combination of three antibiotics: metronidazole, clindamycin and doxycycline after atovaquone and azithromycin failed to eliminate the infection on a polymerase chain reaction (PCR) test. The aim of this study was to determine whether the triple antibiotic combination was an appropriate alternative or additional treatment for the elimination of B. gibsoni. The medical records of 24 patients treated from December 2012 to July 2015 were retrospectively analyzed. The diagnosis of B. gibsoni was confirmed with a PCR test that was also used to assess treatment response. All patients were initially treated with the standard therapy, atovaquone and azithromycin with a 25% success rate clearing B. gibsoni. Dogs that remained positive on PCR using the standard therapy were then treated with the triple antibiotic protocol achieving an 87% success rate. The inclusion of an alternative and potentially effective protocol for the treatment of B. gibsoni would increase the options for the current therapeutic options, could aid in clearance of the organism and offer a more affordable option for clients.

Feasibility of open-chest cardiopulmonary resuscitation through a transdiaphragmatic approach in dogs.

OBJECTIVE: To describe and evaluate the feasibility of a transdiaphragmatic (TD) approach for open-chest cardiopulmonary resuscitation (OCCPR) as an alternative to a traditional lateral thoracotomy (LT) in a canine cadaver model. STUDY DESIGN: Randomized noninferiority ex vivo study. ANIMALS: Fourteen canine cadavers weighing 17.4-30.2 kg. METHODS: An LT and a TD approach to the heart were performed in each cadaver. The order of procedures as well as an assignment to specific operators were randomized before starting the study. Data recorded included the time between incision and initiation of cardiac compressions; time between initiation of the first suture placement and closure of the intrapleural space; time between initiation of the first suture placement and final skin suture; trauma to pulmonary, cardiac, hepatic and neurovascular structures; distance between the caval foramen and diaphragmatic incision; the intercostal space entered during LT; and appropriate closure. RESULTS: The mean time between incision and initiation of cardiac compressions for the TD approach (85 ± 35 seconds) was noninferior to the LT (84 ± 28 seconds). The pleural space was closed faster after the TD approach (531 ± 276 seconds) than after the lateral approach (817 ± 294 seconds, P = .03). Total duration of closure did not differ between techniques (P = .11). There was no difference between the complication rates of each approach. CONCLUSION: The TD approach did not prolong the procedure or increase the complication rate compared with an LT. CLINICAL SIGNIFICANCE: This study provides evidence to support additional investigation of the TD approach for OCCPR to determine its efficacy and safety in live animals.

Acceptance, Benefits, and Challenges of Public Health-Oriented Pet Business Regulations in King County, Washington.

CONTEXT: New regulations were implemented in King County, Washington, in 2010 requiring pet businesses to obtain a permit from Public Health-Seattle & King County (Public Health) and undergo annual inspections to provide education and ensure compliance with regulatory standards. The regulations were developed as a tool for zoonotic disease control and prevention education for businesses and their customers, as well as for environmental protection. OBJECTIVE: To assess the acceptance, benefits, and challenges of the new regulations and identify ways for Public Health to improve educational efforts and assist businesses with compliance. DESIGN: Cross-sectional survey. SETTING: King County, Washington. PARTICIPANTS: Pet businesses with Public Health permits in 2013. MAIN OUTCOME MEASURE: Self-administered survey responses. RESULTS: The response rate was 40.5%. The majority of respondents provided grooming, pet day care, and kennel/boarding services from small, independent businesses. Sixty-one percent reported Public Health inspections as beneficial, especially concerning disinfection procedures and using an infection control plan. Almost three-fourths of respondents used the Public Health template to develop the infection control plan. Forty-four percent reported using the educational materials provided by Public Health, and 62% used educational materials from other sources. Most respondents reported that they gained benefits from the pet business permit, although fewer agreed that they obtained a good value from the permit and fee. The most common benefits reported were protection of animal and human health and establishing the credibility of the pet business. CONCLUSIONS: Major challenges with the implementation of the pet business regulations were not generally reported by respondents. Most respondents reported a collaborative relationship between Public Health and the pet businesses. Improvements in infection control practices and positive responses to the inspections were reported by pet businesses. Survey results were used to improve infection control plan templates, increase the use of educational materials, and improve the Web site and business portal performance.

The effect of Hetastarch (670/0.75) in vivo on platelet closure time in the dog.

OBJECTIVE: To evaluate the effect of 6% hydroxyethyl starch (HES) solution in vivo, with an average molecular weight of 670 kDa and degree of substitution of 0.75, on canine platelet function. DESIGN: Prospective, controlled-experimental study. SETTING: University of California, Davis, Veterinary Medical Teaching Hospital. ANIMALS: Seven healthy employee-owned dogs. INTERVENTIONS: Seven dogs were included in the treatment group. Four of these dogs also served as the control group. Platelet closure time (CT) was measured using a platelet function analyzer and collagen/ADP cartridges. Dogs were given 20 mL/kg of either sodium chloride 0.9% (control group, n=4) or HES (treatment group, n=7) IV over 1 hour. CT was measured before the infusion, and at 1, 3, 5, and 24 hours after the start of the infusion. MEASUREMENTS AND MAIN RESULTS: There was a significant change over time from 0 to 24 hours (P<0.001), a significant difference between groups across time (P<0.001), and a significant group-by-time interaction (P=0.007). At 3 hours, mean CT for the treatment group was 122.3+/-18.1 seconds, which was significantly different (P<0.001) from the control group (71.0+/-3.5 s). At 5 hours, mean CT for the treatment group was 142.7+/-33.9 seconds, which was significantly different (P=0.001) from the control group (75.0+/-8.6 s). Mean CT at 24 hours was within the reference interval for both the control and treatment group (66.0+/-2.9 and 81.8+/-11.9 s, respectively); however, CT in 3 individual dogs in the treatment group at this time point remained prolonged. CONCLUSIONS: A clinically relevant dose of HES 670/0.75 prolongs CT in dogs for up to 24 hours. This may be due to platelet dysfunction in addition to the effects of hemodilution, and therefore, may increase the risk of bleeding.

In vitro comparison of the effects of two forms of hydroxyethyl starch solutions on platelet function in dogs.

OBJECTIVE: To evaluate the effect of 2 hydroxyethyl starch (HES) preparations (ie, HES solution with a molecular weight of 600 kd and a degree of substitution of 0.7 [HES 600/0.7] and a calcium-containing polyionic HES solution with a molecular weight of 670 kd and a degree of substitution of 0.75 [HES 670/0.75]) on canine platelet function. SAMPLE POPULATION: Blood samples from 10 healthy adult dogs. PROCEDURES: Dilution of citrated whole blood was performed with saline (0.9% NaCl) solution, HES 600/0.7, and HES 670/0.75 at ratios of 1:9 (ie, 1 part saline solution or colloid to 9 parts whole blood) and 1:3. Measurements of time to platelet plug formation in a capillary tube (ie, closure time) were made by use of a bench-top platelet function analyzer with collagen and ADP platelet agonists. RESULTS: Mean baseline closure time was 68.0 +/- 15.3 seconds. A 1:3 dilution of whole blood with saline solution, HES 600/0.7, and HES 670/0.75 resulted in mean closure times of 85.8 +/- 15.7 seconds, 100.6 +/- 18.6 seconds, and 101.6 +/- 16.2 seconds, respectively. Closure time following 1:3 dilution of whole blood with saline solution was significantly different from baseline and from 1:9 dilution with saline solution. Closure time following 1:3 dilution of whole blood with HES 670/0.75 was significantly different from baseline, 1:3 and 1:9 dilutions with saline solution, and 1:9 dilutions with HES 600/0.7 or HES 670/0.75. CONCLUSIONS AND CLINICAL RELEVANCE: Saline solution, HES 600/0.7, and HES 670/0.75 affect canine platelet function by prolonging closure times; HES solutions prolonged closure time to a greater extent than saline solution.

Ozone exposure enhances endotoxin-induced mucous cell metaplasia in rat pulmonary airways.

Coexposure to different airborne pollutants can be more toxic to airway epithelium than an inhalation exposure to a single pollutant. We have previously reported that coexposure to ozone, the primary oxidant gas in photochemical smog, and unique inflammatory biogenic substances such as allergens or bacterial endotoxin, results in augmented epithelial and inflammatory responses in rat nasal airways (M. V. Fanucchi et al., 1998, Toxicol. Appl. Pharmacol. 152, 1-9; J. G. Wagner et al., 2002a, Toxicol. Sci.67, 284-294). In the present study, we investigated the toxic interaction of ozone and endotoxin on the respiratory epithelium in the pulmonary airways of laboratory rodents. F344 rats were intranasally instilled with 0, 2, or 20 microg endotoxin dissolved in sterile saline (150 microl/nasal passage). Six h after instillation rats were exposed to air or 1 ppm ozone for 8 h. One day later, endotoxin and ozone exposures were repeated. Three days after the last exposure, rats were sacrificed, the lungs were lavaged with saline, and the collected bronchoalveolar lavage fluid (BALF) was analyzed for inflammatory cells and secreted mucosubstances (mucin 5AC). Lung tissues were processed for light microscopic examination and morphometric analysis of numeric density of epithelial cell populations and volume densities of intraepithelial mucosubstances (IM). Conducting airways were microdissected and analyzed by quantitative RT-PCR to determine steady-state mucin gene (rMuc5AC) mRNA levels in respiratory epithelium. Endotoxin instillation caused a dose-dependent increase in BALF neutrophils that was further increased twofold in ozone-exposed rats given 20 microg endotoxin. Mucin glycoprotein 5AC was elevated in BALF from rats exposed to 20 microg, but not 2 microg endotoxin. Exposure to ozone alone did not cause mucus hypersecretion, but ozone potentiated mucus secretion in rats given 2 or 20 microg endotoxin. Airways of rats exposed to air or ozone alone had scant amounts of IM. Endotoxin instillation induced a dose-dependent increase in IM in airway epithelium that was significantly increased (twofold) in rats that were also exposed to ozone. Expression of rMuc5AC was induced in axial pulmonary airways by 2 and 20 microg endotoxin, and was increased further by ozone-exposure in rats instilled with 20 microg endotoxin. These data demonstrate that ozone exposure potentiates neutrophilic inflammation and mucus production and secretion elicited by a biogenic substance in rat pulmonary airways.