An 18-µm microaggregate blood filter does not cause hemolysis during in vitro whole blood transfusions in sea turtles.

OBJECTIVE: Determine the hemolytic effect of an 18-µm microaggregate blood filter during in vitro sea turtle whole blood transfusions as well as describe the average diameter of leatherback (Dermochelys coriacea) and Kemp's ridley sea turtle (Lepidochelys kempii) RBCs. ANIMALS: 5 green (Chelonia mydas), 5 loggerhead (Caretta caretta), and 5 Kemp's ridley sea turtles (total n = 15). METHODS: Heparinized sea turtle blood was infused at 60 mL/h through a microbore extension set without and then with a postsyringe, inline 18-µm microaggregate blood filter. Pre- and postfiltration PCV, Hct, total solids, sodium, chloride, potassium, glucose, and free plasma hemoglobin concentrations were measured. With the use of light microscopy and archived blood smears, the maximum and minimum diameter of 20 RBCs from each of the 5 leatherback and 5 Kemp's ridley sea turtles were measured with a calibrated ocular micrometer using 400X magnification. RESULTS: There were no significant differences between pre- and postfiltration samples for Hct, total solids, sodium, chloride, potassium, glucose, and free plasma hemoglobin concentrations; however, there was a significant median postfiltration decrease in PCV of approximately 4%, representing a 13% decrease of the total RBCs transfused. Average maximum diameters for leatherback and Kemp's ridley sea turtle RBCs were 19.7 and 16.1 µm, respectively. CLINICAL RELEVANCE: Although the 18-µm microaggregate blood filter does not hemolyze transfused sea turtle RBCs and is likely safe for in vivo blood transfusions, the filter's pores may retain a small proportion of infused RBCs given their diameter.

Major crossmatch compatibility of rabbit blood with rabbit, canine, and feline blood.

OBJECTIVE: To evaluate the major crossmatch compatibility between rabbit recipients, rabbit donors, and the major canine and feline blood types. DESIGN: Prospective in vitro study in December 2021. SETTING: Academic veterinary teaching hospital. ANIMALS: Whole blood samples were collected from 11 healthy New Zealand White rabbits (Oryctolagus cuniculus) with no previous transfusion history. Three pigtail segments were acquired from dog erythrocyte antigen (DEA)-1-positive, DEA-1-negative, and feline type A blood units. Whole blood was collected from a healthy type B blood donor cat. INTERVENTIONS: Blood from each rabbit recipient underwent a major crossmatch using standard tube crossmatch methodology with itself and the following donor blood types: rabbit, DEA-1-positive, DEA-1-negative, feline type A, and feline type B. MEASUREMENTS AND MAIN RESULTS: Self-crossmatches and crossmatches between rabbit recipients and conspecific donors were negative for hemolysis and agglutination. Crossmatches between rabbit recipients and canine and feline donors yielded no hemolysis but produced varying degrees of macroscopic and microscopic agglutination. Rabbit recipients had 1.4 (95% confidence interval: 1.1-1.8) times the risk of macroscopic agglutination when major crossmatched with canine blood compared to feline blood. No significant difference in agglutination was found between DEA-1-positive and DEA-1-negative or feline type A and type B donors. CONCLUSIONS: These findings support allogeneic blood transfusions between rabbits being highly compatible and suggest rabbits have naturally occurring alloantibodies against both canine and feline red blood cells. However, feline red blood cells had a lower rate of in vitro incompatibility on major crossmatch, suggesting potentially higher in vivo compatibility if an emergency xenotransfusion is needed. Further prospective research is needed to determine if xenotransfusion is associated with a higher incidence of acute and delayed transfusion reactions in rabbits than allogeneic transfusions.

Traumatic telson avulsion and attempted external stabilization in Atlantic horseshoe crabs (Limulus polyphemus).

Atlantic horseshoe crabs (Limulus polyphemus) are prevalent in public aquarium touch pools. Despite their popularity, the literature concerning medical management under managed care is sparse. Noninfectious conditions include trauma to the exoskeleton and compound eyes; however, injury to the soft tissue connecting the telson and opisthosoma has not been previously reported. This report describes telson avulsion in three Atlantic horseshoe crabs at a public aquarium and attempted external stabilization with a vascular silicone tie in two of three affected animals. The horseshoe crab that received no veterinary intervention suffered a complete telson amputation 1 month after the initial injury. Although the two other horseshoe crabs did well postoperatively, and external stabilization prevented further avulsion or amputation, they still could not right themselves if flipped into dorsal recumbency. All three horseshoe crabs were ultimately euthanized due to animal welfare concerns. To avoid potentially serious telson ligament injury, horseshoe crabs should never be picked up by the telson. The authors recommend that telson ligament injuries be addressed promptly, any inciting cause be mitigated, and external stabilization be considered earlier.

Diagnostic agreement between three point-of-care glucose and ß-hydroxybutyrate meters and reference laboratory methods in stingrays.

Point-of-care (POC) glucose and ß-hydroxybutyrate (ß-HB) meters can potentially provide rapid insight into an elasmobranch's metabolic state in clinical and field research settings. This study evaluated the diagnostic agreement of three commercial POC meters against reference laboratory methods for glucose and ß-HB concentrations in stingrays. Blood was collected during anesthetized exams from 28 stingrays representing four species: cownose rays (Rhinoptera bonasus), Atlantic stingrays (Hypanus sabina), southern stingrays (Hypanus americanus), and yellow stingrays (Urobatis jamaicensis). Glucose and ß-HB concentrations were measured with each POC meter using whole blood and plasma; in parallel, plasma glucose and ß-HB concentrations were measured via reference laboratory methods. Agreement between POC meters and reference laboratory methods was assessed using Bland-Altman methods, Passing-Bablok regression, observed total error, percent relative error, and linear mixed effect models. Plasma glucose and ß-HB concentrations determined by reference laboratory methods ranged from <20-63 mg/dL to 0.05-5.38 mmol/L, respectively. One human POC meter-the Precision Xtra-showed the greatest agreement with reference laboratory methods when measuring glucose with whole blood [mean bias and 95% CI: 0 (-3-4) mg/dL] and ß-HB with plasma [mean bias and 95% CI: 0.1 (-0.04-0.2) mmol/L]. Stingray sex, weight, buffy coat, and packed cell volume did not significantly affect the agreement between POC meters and reference laboratory methods. Across all three POC meters, mean bias and imprecision for plasma ß-HB concentrations were relatively small (0-0.1 mmol/L and 0%, respectively). Utilizing POC meters to measure glucose and ß-HB in stingrays may be viable when reference methods are unavailable.

An integrative review of lateral line depigmentation in marine and freshwater fish.

OBJECTIVE: To qualitatively review reports on lateral line depigmentation (LLD) in marine and freshwater fish. SAMPLE: English-language publications concerning LLD published before March 1, 2020. PROCEDURES: Electronic searches of CAB abstracts, PubMed, and Web of Science databases and the proceedings of the International Association of Aquatic Animal Medicine were performed. Records were systematically screened and selected for inclusion in an integrative review. Bibliographies of records included in the review were examined to identify other records to be screened. Included records were qualitatively reviewed. Evidence level and quality were graded according to previously described criteria. Information pertinent to epidemiological factors, etiopathogenesis, clinical and histopathologic findings, treatment, and prevention of LLD was collected. RESULTS: 401 records were screened, and 24 unique publications (16 peer-reviewed articles, 1 textbook, and 7 abstracts) were included in the study; 12 (50%), 1 (4%), 6 (25%), and 5 (21%) were classified as evidence level I (experimental), II (quasi-experimental), III (nonexperimental), and V (clinical reports or clinician experience), respectively. Seventeen (71%) and 7 (29%) reports were classified as high quality and good quality, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: LLD should be considered a clinical observation indicative of a dermatologic response of fish to suboptimal conditions; LLD should continue to be adopted as the preferred term to describe the classic signs. Whereas gross findings are similar among species, histologic findings can vary. Evidence-based treatment of LLD for individual fish consists of source control (changing tanks or systems), topical treatment with 0.01% becaplermin gel, supportive care, and antimicrobial treatment when warranted. For schools of fish, treatment and prevention of LLD should be focused on improving suboptimal environmental and physiologic conditions.

Effects of Deepwater Horizon oil on feather structure and thermoregulation in gulls: Does rehabilitation work?

Impacts of large-scale oil spills on avian species are far-reaching. While media attention often focuses on lethal impacts, sub-lethal effects and the impacts of rehabilitation receive less attention. The objective of our study was to characterize effects of moderate external oiling and subsequent rehabilitation on feather structure and thermoregulation in gulls. We captured 30 wild ring-billed gulls (Larus delawarensis) and randomly assigned each individual to an experimental group: 1) controls, 2) rehabilitated birds (externally oiled, rehabilitated by washing), or 3) oiled birds (externally oiled, not rehabilitated). We externally oiled birds with weathered MC252 Deepwater Horizon oil (water for controls) and collected feathers and thermography imagery (FLIR) approximately weekly for four weeks to investigate feather structure (quantified using a barbule clumping index) and thermoregulatory ability (characterized by internal body temperature and external surface temperature). Post-oiling feather clumping was significantly higher in oiled and rehabilitated birds compared to controls, but steadily declined over time in both groups. However, feather microstructure in rehabilitated birds was indistinguishable from controls within three weeks of washing whereas the feathers of oiled birds were still significantly clumped a month post oiling. Internal body temperatures didn't differ in any of the groups, suggesting birds maintain thermoregulatory homeostasis in spite of moderate external oiling. External temperatures for rehabilitated birds didn't differ from controls within a week of rehabilitation. Overall, rehabilitation procedures were effective and washed birds were in better condition compared to non-rehabilitated, oiled birds. This study provides evidence that the benefits of rehabilitation for moderately oiled birds likely outweigh the costs with regard to feather structure and thermoregulation. While feather preening and time were insufficient to reestablish baseline fine scale feather structure in moderately oiled birds, the significant clumping reduction over time may indicate that rehabilitation of lightly oiled birds may not be necessary and deserves further study.

Diagnostic Uncertainty and the Epidemiology of Feline Foamy Virus in Pumas (Puma concolor).

Feline foamy virus (FFV) is a contact-dependent retrovirus forming chronic, largely apathogenic, infections in domestic and wild felid populations worldwide. Given there is no current 'gold standard' diagnostic test for FFV, efforts to elucidate the ecology and epidemiology of the virus may be complicated by unknown sensitivity and specificity of diagnostic tests. Using Bayesian Latent Class Analysis, we estimated the sensitivity and specificity of the only two FFV diagnostic tests available-ELISA and qPCR-as well as the prevalence of FFV in a large cohort of pumas from Colorado. We evaluated the diagnostic agreement of ELISA and qPCR, and whether differences in their diagnostic accuracy impacted risk factor analyses for FFV infection. Our results suggest ELISA and qPCR did not have strong diagnostic agreement, despite FFV causing a persistent infection. While both tests had similar sensitivity, ELISA had higher specificity. ELISA, but not qPCR, identified age to be a significant risk factor, whereas neither qPCR nor ELISA identified sex to be a risk factor. This suggests FFV transmission in pumas may primarily be via non-antagonistic, social interactions between adult conspecifics. Our study highlights that combined use of qPCR and ELISA for FFV may enhance estimates of the true prevalence of FFV and epidemiological inferences.

Frequent cross-species transmissions of foamy virus between domestic and wild felids.

Emerging viral outbreaks resulting from host switching is an area of continued scientific interest. Such events can result in disease epidemics or in some cases, clinically silent outcomes. These occurrences are likely relatively common and can serve as tools to better understand disease dynamics, and may result in changes in behavior, fecundity, and, ultimately survival of the host. Feline foamy virus (FFV) is a common retrovirus infecting domestic cats globally, which has also been documented in the North American puma (Puma concolor). The prevalent nature of FFV in domestic cats and its ability to infect wild felids, including puma, provides an ideal system to study cross-species transmission across trophic levels (positions in the food chain), and evolution of pathogens transmitted between individuals following direct contact. Here we present findings from an extensive molecular analysis of FFV in pumas, focused on two locations in Colorado, and in relation to FFV recovered from domestic cats in this and previous studies. Prevalence of FFV in puma was high across the two regions, ∼77 per cent (urban interface site) and ∼48 per cent (rural site). Comparison of FFV from pumas living across three states; Colorado, Florida, and California, indicates FFV is widely distributed across North America. FFV isolated from domestic cats and pumas was not distinguishable at the host level, with FFV sequences sharing >93 per cent nucleotide similarity. Phylogenetic, Bayesian, and recombination analyses of FFV across the two species supports frequent cross-species spillover from domestic cat to puma during the last century, as well as frequent puma-to-puma intraspecific transmission in Colorado, USA. Two FFV variants, distinguished by significant difference in the surface unit of the envelope protein, were commonly found in both hosts. This trait is also shared by simian foamy virus and may represent variation in cell tropism or a unique immune evasion mechanism. This study elucidates evolutionary and cross-species transmission dynamics of a highly prevalent multi-host adapted virus, a system which can further be applied to model spillover and transmission of pathogenic viruses resulting in widespread infection in the new host.

In vitro evaluation and in vivo use of a novel surgical stent to minimize suture pressure necrosis.

OBJECTIVE: To evaluate the efficacy of stents to distribute pressure when incorporated into tension-relieving sutures. STUDY DESIGN: An in vitro study and case report. ANIMAL: One common bottlenose dolphin (Tursiops truncates). METHODS: Three novel silicone stents, a traditional stent, and a control were tested in vitro by using a suture simulator. Stent surface area was measured by using a pressure mapping sensor. Pressure was derived from the measured surface area and the downward force of the suture simulator. Novel silicone stents were also used in the closure of an incision in a bottlenose dolphin. RESULTS: No difference was found in surface area or pressure among the three silicone stents (adjusted P > .05). Silicone stents yielded an average, 2.69 cm2 more surface area and 842.37 kPa less pressure compared with the control as well as 1.67 cm2 more surface area and 110.67 kPa less pressure compared with the traditional stent. The traditional stent yielded 1.02 cm2 more surface area and 731.7 kPa less pressure compared with the control. Incision site and silicone stent assessment performed postoperatively revealed no obvious pressure necrosis. CONCLUSION: Stents improved distribution of suture pressure, and novel silicone stents were more effective compared with traditional stents. Novel silicone stents appear to have preliminary clinical success in vivo. CLINICAL SIGNIFICANCE: Our study provides evidence that stents effectively help distribute suture pressure, and their incorporation might minimize the risk of suture pressure necrosis. Novel silicone stents should be integrated into tension-relieving suture patterns when closing wounds and incisions, particularly in aquatic animals.

Teaching Wildlife Disease Outbreak Response Through a Collaborative One Health Workshop.

Issues in the fields of wildlife disease and One Health are often difficult to address by single research groups because of the many disciplines and areas of expertise required to effectively solve complex problems. Although collaborations are becoming increasingly prevalent in the professional realm, many undergraduate, graduate, and professional students are merely introduced to the idea of collaboration without fully understanding how team-based approaches function. In this report, we describe the framework for a one-day workshop hosted by the Colorado State University student chapter of the Wildlife Disease Association (CSU WDA), where we gathered students and professionals to collectively investigate a simulated wildlife disease outbreak. CSU WDA student members designed the workshop and recruited professionals who are experts in their respective fields to run an outbreak simulation during the event. Based on pre- and post-event evaluation responses, this workshop was effective in increasing participants' knowledge of disease ecology, pathology, genetics, and microbiology, as well as the importance of collaboration among disciplines as it pertains to wildlife disease outbreaks.

Effects of External Oiling and Rehabilitation on Hematological, Biochemical, and Blood Gas Analytes in Ring-Billed Gulls (Larus delawarensis).

Avian species experience extensive morbidity and mortality following large-scale oil spills, often resulting in oiled birds being rescued, and admitted to rehabilitation. Our objective was to experimentally establish time-specific, descriptive blood analyte data following sublethal oil exposure and subsequent rehabilitation. Thirty wild Ring-billed Gulls (Larus delawarensis) were randomly allocated to three treatment groups of 10 birds each. One treatment group served as controls and two treatment groups were externally oiled daily for 3 days with weathered MC252 oil collected from the Deepwater Horizon oil spill, mimicking the upper threshold of the US Fish and Wildlife Service's moderate oiling classification. Following external oiling, one oiled treatment group was cleaned via standard rehabilitation practices. Serial venous blood samples were collected for a month to measure packed cell volume, total solids, blood gas and select plasma biochemistry analytes, total white blood cell estimates and differentials, and reticulocyte estimates. We found that both sublethal oil exposure and aspects of captivity were associated with a mild non-regenerative anemia. No other differences in venous blood gas and biochemical analytes as well as white blood cell concentrations were observed among the three groups. These findings suggest that the mild anemia seen in oiled birds undergoing rehabilitation is possibly multifactorial and that moderately oiled gulls have subtle, but potentially not insignificant clinicopathological abnormalities following sublethal oil exposure. Oiled gulls did not develop any clinicopathological derangements post-rehabilitation, suggesting current standard practices for rehabilitation cause minimal morbidity in clinically stable, moderately oiled gulls.

Feline Foamy Virus Infection: Characterization of Experimental Infection and Prevalence of Natural Infection in Domestic Cats with and without Chronic Kidney Disease.

Foamy viruses (FVs) are globally prevalent retroviruses that establish apparently apathogenic lifelong infections. Feline FV (FFV) has been isolated from domestic cats with concurrent diseases, including urinary syndromes. We experimentally infected five cats with FFV to study viral kinetics and tropism, peripheral blood mononuclear cell (PBMC) phenotype, urinary parameters, and histopathology. A persistent infection of primarily lymphoid tropism was detected with no evidence of immunological or hematologic perturbations. One cat with a significant negative correlation between lymphocytes and PBMC proviral load displayed an expanded FFV tissue tropism. Significantly increased blood urea nitrogen and ultrastructural kidney changes were noted in all experimentally infected cats, though chemistry parameters were not outside of normal ranges. Histopathological changes were observed in the brain, large intestine, and other tissues. In order to determine if there is an association of FFV with Chronic Kidney Disease, we additionally screened 125 Australian pet cats with and without CKD for FFV infection and found that FFV is highly prevalent in older cats, particularly in males with CKD, though this difference was not statistically significant compared to controls. Acute FFV infection was clinically silent, and while some measures indicated mild changes, there was no overt association of FFV infection with renal disease.

Impact of body condition on influenza A virus infection dynamics in mallards following a secondary exposure.

Migratory waterfowl are often viewed as vehicles for the global spread of influenza A viruses (IAVs), with mallards (Anas platyrhynchos) implicated as particularly important reservoir hosts. The physical demands and energetic costs of migration have been shown to influence birds' body condition; poorer body condition may suppress immune function and affect the course of IAV infection. Our study evaluated the impact of body condition on immune function and viral shedding dynamics in mallards naturally exposed to an H9 IAV, and then secondarily exposed to an H4N6 IAV. Mallards were divided into three treatment groups of 10 birds per group, with each bird's body condition manipulated as a function of body weight by restricting food availability to achieve either a -10%, -20%, or control body weight class. We found that mallards exhibit moderate heterosubtypic immunity against an H4N6 IAV infection after an infection from an H9 IAV, and that body condition did not have an impact on shedding dynamics in response to a secondary exposure. Furthermore, body condition did not affect aspects of the innate and adaptive immune system, including the acute phase protein haptoglobin, heterophil/lymphocyte ratios, and antibody production. Contrary to recently proposed hypotheses and some experimental evidence, our data do not support relationships between body condition, infection and immunocompetence following a second exposure to IAV in mallards. Consequently, while annual migration may be a driver in the maintenance and spread of IAVs, the energetic demands of migration may not affect susceptibility in mallards.