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Efforts to breed Attwater's prairie chickens (APC; Tympanuchus cupido attwateri) in captivity to supplement wild populations of this endangered bird have been negatively affected by infections with Avipoxvirus and reticuloendotheliosis virus (REV). Because REV can be integrated into the genome of fowlpox virus (FPV) and may be transmitted in that manner, identifying the source of avipox disease in APC is important to mitigate the impact of this virus. Tissue samples from APC were collected from breeding programs in Texas from 2016 to 2020. These samples consisted of 11 skin lesions and three internal organs from a total of 14 different birds that died of unknown causes or were euthanized. Avipoxvirus was detected by PCR and isolation in embryonating chicken eggs in all skin lesion samples but was not detected in internal organs. Using sequence analysis of FPV polymerase and 4b genes, we determined that 10 out of 11 Avipoxvirus detections resided within the fowlpox clade and a single sample resided within the canarypox clade. REV sequences were detected in all FPV positive samples and in all internal organ tissues but were not detected in the sample matching the canarypox clade. Analysis of REV sequences and PCR detection showed the REV infecting APC was consistent with REV-A and had little variability on analysis of the U3 region of the long terminal repeat. The results of this study indicate control of REV in APC breeding colonies may benefit by a vaccination program targeting FPV and REV. However, a commercially available vaccine for REV is not available at this time.

Secuenciación genética de un virus de la viruela aviar de un gallo grande de las praderas Attwater y evaluación de su papel potencial en los brotes del virus de la reticuloendoteliosis. Los esfuerzos para criar gallos de las praderas grandes de Attwater (APC; Tympanuchus cupido attwateri) en cautiverio para complementar las poblaciones silvestres de esta ave en peligro de extinción se han visto afectados negativamente por infecciones con Avipoxvirus y con el virus de la reticuloendoteliosis (REV). Debido a que el virus de la reticuloendoteliosis puede integrarse en el genoma del virus de la viruela del pollo (FPV) y puede transmitirse de esa manera, identificar la fuente del virus pox en gallos de las praderas grandes es importante para mitigar el impacto de este virus. Se recolectaron muestras de tejido de gallos de las praderas grandes de programas de reproducción en Texas entre los años 2016 a 2020. Estas muestras consistieron en 11 lesiones cutáneas y tres órganos internos de un total de 14 aves diferentes que murieron por causas desconocidas o fueron sacrificadas. El Avipoxvirus se detectó mediante PCR y por aislamiento en huevos embrionados de pollo en todas las muestras de lesiones cutáneas, pero no se detectó en los órganos internos. Utilizando el análisis de secuencia de la polimerasa del virus de la viruela del pollo y de los genes 4b, se determinó que diez de las once detecciones de Avipoxvirus residían dentro del clado de la viruela aviar del pollo y una sola muestra residía dentro del clado de la viruela del canario. Se detectaron secuencias del virus de la reticuloendoteliosis en todas las muestras positivas para virus de la viruela de pollo y en todos los tejidos de órganos internos, pero no se detectaron en la muestra que coincidía con el clado de la viruela del canario. El análisis de las secuencias del virus de la reticuloendoteliosis y la detección por PCR mostró que los virus de reticuloendoteliosis que infectan a gallos de las praderas grandes eran compatible con virus de la reticuloendoteliosis A y tenía poca variabilidad en el análisis de la región U3 de la región repetida terminal larga. Los resultados de este estudio indican que el control del virus de la reticuloendoteliosis en colonias reproductoras de gallos de las praderas grandes puede beneficiarse de un programa de vacunación dirigido los virus de la viruela del pollo y de la reticuloendoteliosis. Sin embargo, una vacuna disponible comercialmente contra el virus de la reticuloendoteliosis no está disponible en este momento.

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The processing of blood samples can be delayed during health assessments of wildlife populations in distant locations. The use of whole blood preservatives may be useful in these situations. However, there is scant information regarding their use in nonmammalian species. This study tested the efficacy of two cell preservatives on whole blood collected from 12 Attwater's prairie chickens (Tympanuchus cupido attwateri). The preservatives used were Streck Cell Preservative© (SCP), a proprietary proteinaceous stabilizer developed for human flow cytology and validated in other mammalian species, and formalin, which is commonplace in histopathology, but its use in whole blood has been limited to fish. Grouped blood samples were treated with heparin, SCP, or formalin and analyzed at 0, 1, 4, and 7 days after collection for packed cell volume (PCV), complete blood count (CBC), and cellular morphology. SCP effectively preserved most cell types in Attwater's prairie chicken blood samples over a period of 7 days, with the exception of monocyte cell counts, which were significantly reduced from day 0. Formalin maintained total white blood cell counts at baseline levels measured by hemocytometer, but irregular staining characteristics prevented accurate analysis of differential counts or cellular morphology. Both preservatives altered PCV compared with the heparin control, but these values remained constant over time, highlighting the need for method-specific reference intervals. The validation of SCP in Attwater's prairie chickens supports its potential for use in other avian species for the collection of accurate hematologic data when the processing of blood samples may be delayed.

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A retrospective study of mortality and neonatal treatment for 975 Attwater's prairie chickens ( Tympanuchus cupido attwateri) was conducted from 2009-2015. Gross necropsy, histopathology, and medical records for chicks less than 8 wk of age at the Houston Zoo were reviewed and summarized. The total mortality rate was 36% ( n = 352). Yolk sac infection was the most common cause of death (21%; n = 73) followed by maladaptation (19%; n = 68), musculoskeletal abnormalities (19%; n = 66), necrotic enteritis (13%; n = 44), and mucoid enteritis (11%; n = 39). Gavage feeding of a highly digestible formula was associated with a higher proportion of necrotic enteritis and a reduced occurrence of both mucoid enteritis and maladaptation. Meloxicam administration had a significantly reduced incidence of mucoid enteritis. Survival past 10 days of age, prevention of neonatal bacterial infection, and supportive treatment with meloxicam are important factors to increase chick survival and improve captive breeding success for Attwater's prairie chickens. Gavage feeding of a diet low in digestible carbohydrates has the potential for reducing disease due to necrotic enteritis in this species.

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The invasive red imported fire ant (Solenopsis invicta) has negatively affected a host of taxonomic groups throughout its acquired North American range. Many studies have hypothesized indirect trophic impacts, but few documented those impacts. We evaluated invertebrate abundance as a factor limiting juvenile survival of the endangered Attwater's prairie-chicken (Tympanuchus cupido attwateri), and whether fire ants reduce invertebrate numbers and biomass. From 2009-2013, we monitored survival of Attwater's prairie-chicken broods (n = 63) with radio telemetry during the first 2 weeks post-hatch and collected daily invertebrate samples at brood sites. Broods located in areas with the highest median invertebrate count (338 invertebrates/25 sweeps) had a survival probability of 0.83 at 2 weeks post-hatch compared to 0.07 for broods located in areas with the lowest median invertebrate count (18 invertebrates/25 sweeps). During 2011-2012, we evaluated the reduction of fire ants on invertebrate numbers and biomass by aerially treating areas with Extinguish Plus™ in an impact-reference study design. Treated fields had 27% more individual invertebrates and 26% higher invertebrate biomass than reference fields. Our results clearly document that invertebrate abundance affects Attwater's prairie-chicken brood survival and that fire ants may indirectly contribute to low brood survival by suppressing invertebrate abundance. We posit that within the fire ant's acquired North American range, fire ants are likely contributing to declines of other insectivorous species. Published 2015. This article is a U.S. Government work and is in the public domain in the USA.