HIGH PREVALENCE OF CYTAUXZOON FELIS IN BOBCATS (LYNX RUFUS) ACROSS OKLAHOMA AND OCCURRENCE IN WEST TEXAS, USA.

Cytauxzoonosis is a fatal tick-borne disease in domestic cats caused by infection with the apicomplexan Cytauxzoon felis. Bobcats are the natural wild-vertebrate reservoirs for C. felis, and infections are typically subclinical and chronic in this species. The present study was done to determine the prevalence and geographic distribution of C. felis infection in wild bobcats from Oklahoma and the occurrence in northwestern Texas. Tongue samples from 360 bobcats were collected from 53 counties in Oklahoma and 13 samples from three counties in Texas. For DNA extracted from each tongue sample, a probe-based droplet digital PCR assay was performed targeting the C. felis mitochondrial gene cytochrome c oxidase subunit III (cox3). Prevalence of C. felis infection was calculated for each county sampled, and data from individual counties were combined according to geographic regions and compared using chi-square tests. Overall prevalence of C. felis in bobcats from Oklahoma was 80.0% (95% confidence interval [CI], 75.6-83.8). The prevalence of infection was >90% for bobcats from central, northeastern, south-central, and southeastern regions of Oklahoma, but <68% for bobcats from northwestern and southwestern regions. Bobcats from central counties in Oklahoma were 25.693 times more likely to be infected with C. felis compared to all other bobcats sampled from the state. Higher prevalence estimates of C. felis in bobcats appeared to be in counties where known tick vectors are most common. Occurrence of C. felis in bobcats from northwestern Texas was 30.8% (95% CI, 12.4%-58.0%) based on 13 samples. Results of this study support the utilization of bobcats as sentinel animals to identify geographic areas with risk of C. felis infection to domestic cats.

A Serodiagnostic IgM ELISA to Detect Acute Cytauxzoonosis.

Cytauxzoonosis is a tick-borne infectious disease affecting domestic cats with high mortality and limited treatment modalities. Because early diagnosis and therapeutic intervention are crucial to survival of infected cats, the objective of this study was to develop an ELISA capable of detecting cytauxzoonosis and differentiating acute vs. chronic infection in clinical feline blood samples. A microsphere immunoassay (MIA) was developed to evaluate the production of Cytauxzoon felis-specific IgM and IgG antibodies in serial plasma samples from cats with experimental C. felis infection by targeting a C. felis-specific transmembrane protein (c88). Recombinant c88 protein was utilized to develop indirect ELISAs to detect IgM and IgG antibodies in clinical plasma samples from: PCR-positive cats with acute C. felis infection (n = 36), C. felis-negative cats with pyrexia (n = 10), healthy C. felis-negative cats (n = 22), and chronic C. felis carriers (n = 4). Anti-c88 IgM antibodies were detectable at day 12 post-tick infestation in cats with experimental C. felis infection (within 24 hours of developing clinical signs), while anti-c88 IgG was detectable at day 15 post-tick infestation - indicating IgM could be used to detect early infection. Using a cut-off value of 19.85 percent positive, the C. felis IgM ELISA detected acute cytauxzoonosis in 94.44% (34/36) of cats presented with clinical signs of acute cytauxzoonosis with 100% specificity (indicating a "Strong Positive" result). When a lower cutoff of 8.60 percent positive was used, cytauxzoonosis was detected in the 2 remaining PCR-positive cats with 87.88% specificity (indicating of a "Weak Positive" result). One C. felis-negative, febrile cat had high IgG, and chronic carriers had variable IgM and IgG results. Combined interpretation of IgM and IgG ELISAs did not reliably differentiate acute vs. chronic infection. While further validation on assay performance is needed, the C. felis IgM ELISA is a promising test to detect acute cytauxzoonosis and can be utilized to develop a point-of-care test for clinical use.

Clinical and Histopathologic Features of a Feline SARS-CoV-2 Infection Model Are Analogous to Acute COVID-19 in Humans.

The emergence and ensuing dominance of COVID-19 on the world stage has emphasized the urgency of efficient animal models for the development of therapeutics for and assessment of immune responses to SARS-CoV-2 infection. Shortcomings of current animal models for SARS-CoV-2 include limited lower respiratory disease, divergence from clinical COVID-19 disease, and requirements for host genetic modifications to permit infection. In this study, n = 12 specific-pathogen-free domestic cats were infected intratracheally with SARS-CoV-2 to evaluate clinical disease, histopathologic lesions, and viral infection kinetics at 4 and 8 days post-inoculation; n = 6 sham-inoculated cats served as controls. Intratracheal inoculation of SARS-CoV-2 produced a significant degree of clinical disease (lethargy, fever, dyspnea, and dry cough) consistent with that observed in the early exudative phase of COVID-19. Pulmonary lesions such as diffuse alveolar damage, hyaline membrane formation, fibrin deposition, and proteinaceous exudates were also observed with SARS-CoV-2 infection, replicating lesions identified in people hospitalized with ARDS from COVID-19. A significant correlation was observed between the degree of clinical disease identified in infected cats and pulmonary lesions. Viral loads and ACE2 expression were also quantified in nasal turbinates, distal trachea, lungs, and other organs. Results of this study validate a feline model for SARS-CoV-2 infection that results in clinical disease and histopathologic lesions consistent with acute COVID-19 in humans, thus encouraging its use for future translational studies.

Clinicopathologic features of a feline SARS-CoV-2 infection model parallel acute COVID-19 in humans.

The emergence and ensuing dominance of COVID-19 on the world stage has emphasized the urgency of efficient animal models for the development of therapeutics and assessment of immune responses to SARS-CoV-2 infection. Shortcomings of current animal models for SARS-CoV-2 include limited lower respiratory disease, divergence from clinical COVID-19 disease, and requirements for host genetic modifications to permit infection. This study validates a feline model for SARS-CoV-2 infection that results in clinical disease and histopathologic lesions consistent with severe COVID-19 in humans. Intra-tracheal inoculation of concentrated SARS-CoV-2 caused infected cats to develop clinical disease consistent with that observed in the early exudative phase of COVID-19. A novel clinical scoring system for feline respiratory disease was developed and utilized, documenting a significant degree of lethargy, fever, dyspnea, and dry cough in infected cats. In addition, histopathologic pulmonary lesions such as diffuse alveolar damage, hyaline membrane formation, fibrin deposition, and proteinaceous exudates were observed due to SARS-CoV-2 infection, imitating lesions identified in people hospitalized with ARDS from COVID-19. A significant correlation exists between the degree of clinical disease identified in infected cats and pulmonary lesions. Viral loads and ACE2 expression were quantified in nasal turbinates, distal trachea, lung, and various other organs. Natural ACE2 expression, paired with clinicopathologic correlates between this feline model and human COVID-19, encourage use of this model for future translational studies.

A probe-based droplet digital polymerase chain reaction assay for early detection of feline acute cytauxzoonosis.

Cytauxzoonosis is a tick-borne disease of domestic cats with high mortality and narrow therapeutic window, particularly in the southcentral and southeastern United States. The causative agent is the apicomplexan protozoal parasite Cytauxzoon felis and is primarily transmitted by Amblyomma americanum, the lone star tick. Currently there is no vaccine available to prevent cytauxzoonosis and treatment is often ineffective if not initiated early enough in the course of disease. Early diagnosis and therapeutic intervention are therefore crucial for the survival of infected cats. Several methods are available for diagnosis of cytauxzoonosis, with PCR being the most sensitive. However, current PCR assays, which employ double-stranded DNA intercalating dyes to detect C. felis infection, have inherent limitations such as the potential for false positive detection of non-specific amplification products and inability to provide absolute quantification of parasite load. The objective of this study was to develop a probe-based droplet digital PCR (ddPCR) assay capable of detection and quantification of C. felis load over time and during treatment. The C. felis ddPCR assay was able to (i) reliably detect and quantify C. felis DNA in clinical blood samples from cats with acute cytauxzoonosis and (ii) monitor clinical parasite load in response to anti-protozoal treatment through absolute quantification of C. felis DNA over time. When tested on blood samples from cats with experimental C. felis infection, the assay was able to detect infection in cats as early as 24 h prior to the development of clinical signs. In addition, we demonstrate that this probe-based design can be utilized in traditional real-time PCR systems, with similar detection capabilities as compared to ddPCR. The C. felis probe-based ddPCR was also able to detect infection in samples with lower parasite loads when compared to existing nested PCR assays, although these results were not significant due to small sample size. To the author's knowledge, this is the first reported probe-based ddPCR assay to detect Cytauxzoon felis infection in domestic cats.

Younger, more senior, and most vulnerable? Interaction effects of age and job seniority on psychological distress and quality of life among firefighters.

OBJECTIVES: Firefighters are exposed to repeated traumatic events while the robustness of cumulative effects of repeated exposure to trauma on psychological distress among them are inconsistent. Considering the length of service and seniority are risk factors, the purpose of this study was to examine the effects of the interaction between age and seniority on psychological distress and quality of life among firefighters. METHODS: Participants were 229 firefighters of the Hualien County Fire Bureau in Taiwan who worked full time in response to emergency services and disaster rescues activities. Probable posttraumatic stress disorder (PTSD), depression, anxiety, agoraphobia, and quality of life were assessed among firefighters using psychodiagnostics questionnaires. Firefighters were stratified based on age into a young and a mature group, and based on length of service into a junior and a senior group, yielding four groups: "young-senior," "young-junior," "mature-senior," and "mature-junior." RESULTS: A majority of the firefighters were dissatisfied with their health condition. All the firefighters scored relatively lower at the social domain of their quality of life compared with physical, emotional, and environment domains. In the post hoc multiple comparison analysis, the young-senior firefighters were found to be most vulnerable to psychological distress, manifested in having a higher prevalence of, and suffering from more, severe probable PTSD, depression, anxiety, and agoraphobia symptoms than the other groups of firefighters. CONCLUSIONS: Age and job seniority have significant effects on psychological distress among firefighters. Being young and starting young to serve as a firefighter for a relatively long time is a significant risk factor for psychological distress. (PsycInfo Database Record (c) 2020 APA, all rights reserved).