A Tissue Digestion Protocol for Measuring Sarcoptes scabiei (Astigmata: Sarcoptidae) Density in Skin Biopsies.

Sarcoptic mange is a parasitic skin disease caused by the burrowing mite Sarcoptes scabiei that affects a diversity of mammals, including humans, worldwide. In North America, the most commonly affected wildlife includes wild canids, such as coyotes and red foxes, and more recently American black bears in the Mid-Atlantic and Northeast United States. Currently, surveillance for sarcoptic mange in wildlife is syndromic, relying on detection of clinical signs and lesions, such as alopecia and crusting of skin. When possible, skin scrapes are used to identify the causative mite. While skin scrapes are a valuable diagnostic tool to identify mites, this approach has significant limitations when used for quantification of mite burden. To further investigate mite burden in cases of sarcoptic mange, 6-mm punch biopsies were collected from affected skin of red foxes (Vulpes vulpes Linnaeus [Carnivora: Canidae]), a species historically affected by sarcoptic mange, frequently with high mite burdens and severe skin disease, and validated on skin tissue from mange-affected American black bears (Ursus americanus Pallas [Carnivora: Ursidae]) and coyotes (Canis latrans Say [Carnivora: Canidae]). Biopsies were digested by incubating the tissue in potassium hydroxide (KOH) at 55°C. The greatest tissue clearance and lowest mite degradation resulted after 12 h of tissue digestion. The purpose of this manuscript is to describe a methodology for host tissue digestion and mite quantification in cases of sarcoptic mange. This method will provide a valuable surveillance and research tool to better understand sarcoptic mange in wild and domestic animals, with applications to a diversity of other ectoparasitic diseases.

Bayesian Phylogenetic Analysis of Avipoxviruses from North American Wild Birds Demonstrates New Insights into Host Specificity and Interspecies Transmission.

Avian pox is commonly diagnosed in a variety of North American wild and domestic birds, yet little is known about the evolutionary relationships among the causative poxviruses. This study aimed to determine the phylogenetic relationships among isolates identified in different avian host species to better characterize the host range of specific viral strains and compare the genetic variability within and between viral clades. Skin lesions grossly and microscopically consistent with poxvirus infection from 82 birds collected in Canada, the United States, and the U.S. Virgin Islands were included in this study. A total of 12 avian species were represented; the most common species sampled were wild turkeys (Meleagris gallopavo), mourning doves (Zenaida macroura), and American crows (Corvus brachyrhynchos). Poxvirus samples from these birds were genotyped using PCR that targeted the 4b core protein gene followed by amplicon sequencing. Bayesian phylogenetic analyses of these viruses, in conjunction with publicly available sequences, representing avipoxvirus strains from six continents revealed statistically significant monophyletic clades based on genetic distances of sequences within and between observed clades. Genetic variation within the fowlpox clade was low compared to the canarypox clade. Host and geographic origins of viral isolates revealed overall clustering of viral strains within avian species, with a few exceptions. No genetic differences were observed between viruses from Canada and the United States within individual species. These results are novel in their characterization and comparison of the phylogenetic relationships of poxvirus isolates in wild bird species from North America. Further, we provide new data on the level of host specificity and specific strains circulating in North America.

El análisis filogenético bayesiano de los avipoxvirus de las aves silvestres de América del Norte demuestra nuevos conocimientos sobre la especificidad del huésped y la transmisión interespecífica. La viruela aviar se diagnostica comúnmente en una variedad de aves silvestres y domésticas de América del Norte, pero se sabe poco sobre las relaciones evolutivas entre los poxvirus. Este estudio tuvo como objetivo determinar las relaciones filogenéticas entre aislamientos identificados en diferentes especies de hospedadores aviares para caracterizar mejor el rango de hospedadores de cepas virales específicas y comparar la variabilidad genética dentro y entre los clados virales. Se incluyeron en este estudio lesiones cutáneas que eran consistentes macro y microscópicamente con la infección por poxvirus de 82 aves recolectadas en Canadá, Estados Unidos y las Islas Vírgenes de los Estados Unidos. Un total de 12 especies de aves fueron representadas; las especies más comunes en la muestra fueron los pavos silvestres (Meleagris gallopavo), huilota común (Zenaida macroura) y cuervos americanos (Corvus brachyrhynchos). Las muestras de poxvirus de estas aves fueron genotipadas mediante PCR que se enfocó en el gene de la proteína central 4b seguido de secuenciación de amplicón. Los análisis filogenéticos bayesianos de estos virus, junto con las secuencias disponibles públicamente, que representan cepas de avipoxvirus de seis continentes revelaron clados monofiléticos estadísticamente significativos basados en distancias genéticas de las secuencias dentro y entre los clados observados. La variación genética dentro del clado de la viruela del pollo fue baja en comparación con el clado de virus de canario. El huésped y los orígenes geográficos de los aislamientos virales revelaron un agrupamiento general de cepas virales dentro de las especies aviares, con algunas excepciones. No se observaron diferencias genéticas entre los virus de Canadá y los Estados Unidos dentro de las especies individuales. Estos resultados son novedosos en la caracterización y comparación de las relaciones filogenéticas de los aislados de poxvirus en especies de aves silvestres de América del Norte. Además, se proporcionan nuevos datos sobre el nivel de especificidad del huésped y las cepas específicas que circulan en América del Norte. Key words: Bayesian analysis, mourning dove, phylogenetic, poxvirus, sequencing, wild turkey, 4b gene.

Antibodies to type A influenza virus in wild waterbirds from Argentina.

Limited information exists on avian influenza (AI) virus infection in South American wild birds. As part of a national surveillance program in Argentina, indigenous waterbirds were screened for antibodies to AI virus. From November 2006 to July 2007, serum samples from 540 waterbirds of 12 species were tested for type-specific antibodies to AI virus with the use of a commercially available blocking enzyme-linked immunosorbent assay (bELISA) and the agar-gel immunodiffusion (AGID) test. Thirty-three percent (176/540) of serum samples were positive with the bELISA and 12% (64/540) were positive with the AGID test. The bELISA detected antibodies to AI virus in eight of the 12 species, and the AGID detected positives in only five species. These results provide insight into AI virus circulation in Argentinean waterbirds and preliminary data to guide further surveillance efforts.