Development of a Model of Natural Infection with Mycobacterium bovis in White-Tailed Deer

The objective of this study was to develop a suitable experimental model of natural Mycobacterium bovis infection in white-tailed deer (Odocoileus virginianus), describe the distribution and character of tuberculous lesions, and to examine possible routes of disease transmission. In October 1997, 10 mature female white-tailed deer were inoculated by intratonsilar instillation of 2 3 103 (low dose) or 2 3 105 (high dose) colony forming units (CFU) of M. bovis. In January 1998, deer were euthanatized, examined, and tissues were collected 84 to 87 days post inoculation. Possible routes of disease transmission were evaluated by culture of nasal, oral, tonsilar, and rectal swabs at various times during the study. Gross and microscopic lesions consistent with tuberculosis were most commonly seen in medial retropharyngeal lymph nodes and lung in both dosage groups. Other tissues containing tuberculous lesions included tonsil, trachea, liver, and kidney as well as lateral retropharyngeal, mandibular, parotid, tracheobronchial, mediastinal, hepatic, mesenteric, superficial cervical, and iliac lymph nodes. Mycobacterium bovis was isolated from tonsilar swabs from 8 of 9 deer from both dosage groups at least once 14 to 87 days after inoculation. Mycobacterium bovis was isolated from oral swabs 63 and 80 days after inoculation from one of three deer in the low dose group and none of four deer in the high dose group. Similarly, M. bovis was isolated from nasal swabs 80 and 85 days after inoculation in one of three deer from the low dose group and 63 and 80 days after inoculation from two of four deer in the high dose group. Intratonsilar inoculation with M. bovis results in lesions similar to those seen in naturally infected white-tailed deer; therefore, it represents a suitable model of natural infection. These results also indicate that M. bovis persists in tonsilar crypts for prolonged periods and can be shed in saliva and nasal secretions. These infected fluids represent a likely route of disease transmission to other animals or humans.

Susceptibility of Raccoons (Procyon lotor) to Infection with Mycobacterium bovis

Tuberculosis due to Mycobacterium bovis infection is endemic in white-tailed deer (Odocoileus virginianus) in the northeastern portion of the lower Michigan peninsula (USA). Various wild carnivores and omnivores, including raccoons (Procyon lotor), are infected with M. bovis within the endemic area. To investigate the pathogenesis of tuberculosis in raccoons and the likelihood of M. bovis transmission from infected raccoons to other susceptible hosts, we experimentally inoculated raccoons with single oral doses of M. bovis (ranging from 30 to 1.7 x 105 colony forming units [CFU]), five daily oral doses of M. bovis (ranging from 10 to 1 x 105 CFU), or a single intravenous (IV) dose of 1 x 105 CFU of M. bovis, from November 1998 through December 2000. Granulomatous lesions consistent with tuberculosis, or tissue colonization with M. bovis, were seen in one of five raccoons in the single low oral dose group, one of five raccoons in the multiple low oral dose group, two of five raccoons in the multiple medium oral dose group, five of five raccoons in the multiple high oral dose group, and five of five raccoons in the IV inoculated group. In orally inoculated raccoons, lesions were most common in the tracheobronchial and mesenteric lymph nodes and lung. Excretion of M. bovis in saliva or nasal secretions was noted in all IV inoculated raccoons and two of five multiple low oral dose raccoons. Mycobacterium bovis was not isolated from urine or feces from any experimentally inoculated raccoons. The need for multiple large oral doses to establish infection, and the low number of orally inoculated raccoons that excreted M. bovis in nasal secretions or saliva, suggest that widespread tuberculosis among raccoons is unlikely.

Experimental Deer-To-Deer Transmission of Mycobacterium bovis

Objective—To determine whether Mycobacterium bovis can be transmitted from experimentally infected deer to uninfected in-contact deer. Animals—Twenty-three 6-month-old white-tailed deer. Procedure—On day 0, M bovis (2 X 108 colony-forming units) was administered by intratonsillar instillation to 8 deer; 3 control deer received saline (0.9% NaCl) solution. Eight in-contact deer were comingled with inoculated deer from day 21. On day 120, inoculated deer were euthanatized and necropsied. On day 180, 4 in-contact deer were euthanatized, and 4 new incontact deer were introduced. On day 360, all in-contact deer were euthanatized. Rectal, oral, and nasal swab specimens and samples of hay, pelleted feed, water, and feces were collected for bacteriologic culture. Tissue specimens were also collected at necropsy for bacteriologic culture and histologic analysis. Results—On day 90, inoculated and in-contact deer developed delayed-type hypersensitivity (DTH) reactions to purified protein derivative of M bovis. Similarly, new in-contact deer developed DTH reactions by 100 days of contact with original in-contact deer. Tuberculous lesions in in-contact deer were most commonly detected in lungs and tracheobronchial and medial retropharyngeal lymph nodes. Mycobacterium bovis was isolated from nasal secretions and saliva from inoculated and in-contact deer, urine and feces from in-contact deer, and hay and pelleted feed. Conclusions and Clinical Relevance—Mycobacterium bovis is efficiently transmitted from experimentally infected deer to uninfected in-contact deer through nasal secretions, saliva, or contaminated feed. Wildlife management practices that result in unnatural gatherings of deer may enhance both direct and indirect transmission of M bovis.

Rabies in Experimentally Infected Bears, Ursus spp., with Epizootiologic Notes

Ten black bears, Ursus americanus Pallas, and three brown bears, U. arctos Linnaeus, were inoculated with rabies virus from naturally infected foxes in Alaska. The bears were more resistant than canine species, requiring at least 1,000 MLD50 of virus for infection. Low titres or negative results were obtained in salivary glands titrated in mice. Clinical course of the disease, post mortem findings, and microscopic lesions are described. Microscopic lesions were more· severe in brown bears, in which the inflammatory response was distinguished by the presence of numerous eosinophils in the perivascular infiltrate and among cells diffusely infiltrating the parenchyma. In both species, inclusion bodies were found only in the Purkinje cells of the cerebellum. Rabies is discounted as a factor in unprovoked attacks by bears on man at high latitudes. The epizootiology of rabies in a region where bears are numerous is discussed, with the conclusion that rabid foxes usually do not excrete sufficient quantities of virus in the saliva to infect bears. German title: Tollwut bei experimentell infizierten Bären, Ursus spp., mit epizootiologischen Anmerkungen German abtract: Zehn Schwarzbären, Ursus americanus Pallas, und drei Braunbären, U. arctos Linnaeus, wurden mit einem von Füchsen in Alaska isolierten Feldstamm des Tollwutvirus infiziert. Die Untersuchungsergebnisse lassen erkennen, daß Bären eine größere Resistenz gegenüber Tollwutinfektion aufweisen als hundeartige Karnivoren, und zwar konnten sie nicht mit weniger als 1000 MLD50 des Tollwutvirus infiziert werden. Das Virus war selten nachweisbar in den Speicheldrüsen der tollwuterkrankten Bären. Klinik und Pathologie der Tollwut bei Bären wurden kurz beschrieben. Die im Gehirn vorkommenden entzündlichen Veränderungen waren bei Braunbären besonders schwer und unterschieden sich durch die Häufigkeit der eosinophilen Leukozyten in den perivasculären und Gewebs-Infiltraten. Bei beiden Arten wurden Einschlußkörperchen nur in den Purkinje-Zellen beobachtet. Die Epizootiologie der Tollwut auf der Alaska-Halbinsel, wo Bären häufig vorkommen, wurde besprochen. Die Ergebnisse deuten an, daß Füchse wenig Virus mit dem Speichel ausscheiden, und selten soviel, daß es für die Infektion von Bären ausreicht. French title: La rage expérimentale chez les ours, Ursus spp., avec observations épizootiologiques French abstract: Dix ours noirs, Ursus americanus Pallas, et trois ours bruns, U. arctos Linnaeus, ont été inoculés avec de virus rabique provenant des renards infectés naturellement dans l'Alaska. Les ours Ont été plus résistants au virus que des espèces canines, et pour produire l'infection chez les ours, au moins 1000 MLD50 ont été requis. La titration des glandes salivaires chez des souris a données des titres peu éléves ou des résultats négatifs. La course clinique de la maladie, les observations des autopsies, et les lésions microscopiques sont decrites. Les lésions microscopiques les plus sévères ont été observées chez les ours bruns, dans lesquels la réponse inflammatoire a été distinguée par la présence de nombreux éosinophiles dans l'infiltration périvasculaire et parmi les cellules infiltrées diffusément dans Ie parenchyme. Chez les deux espèces des ours, des corps d'inclusion ont été trouvés seulement dans les cellules de Purkinje du cervelet. On a discuté l'épizootiologie de la rage dans une région où des ours sont nombreux, avec la conclusion qu'il y a dans la salive des renards rabiques une quantité de virus insuffisante pour infecter les ours. é ó í á ú Spanish title: Rabia en osos, Ursus spp., infectados experimentalmente, con anotaciones epizootológicas Spanish abstract: Diez osos negros, Ursus americanus Pallas, y tres osos pardos, U. arctos Linea, se infectaron con una estirpe campal de virus rábico aislada de zorros en Alasca. Los resultados de la experiencia permiten reconocer que los osos presentan una resistencia mayor frente a la infección rábica que los carnívoros cánidos, pues no se pudieron infectar con menos de 1.000 DML50 de virus rábico. El virus era muy raras veces identificable en las glándulas salivales de los osos enfermos de rabia. Se describen sucintamente la clínica y patología de la rabia en los osos. Las modificaciones inflamatorias en el cerebro eran muy graves en el oso pardo y se distinguían por la frecuencia de los leucocitos eosinófilos en los infiltrados perivasculares e hísticos. En ambas especies solo se hallaron corpúsculos de inclusión en las células de Purkinje. Se discute la epizootología de la rabia en la península de Alasca, donde es frecuente Ia presencia de osos. Los resultados señalan que los zorros eliminan poco virus con la saliva y casi nunca en cantidad tal que fuese suficiente para infectar los osos.

Scraping Behavior in Male White-tailed Deer as a Potential Means of Transmitting Chronic Wasting Disease

Chronic wasting disease (CWD) has become a concern for wildlife managers and hunters across the United States. High prevalence of chronic wasting disease (CWD) in older male white-tailed deer (Odocoileus virginianus) suggests that sex-specific social behavior may contribute to the spread of the disease among males. Scraping is a marking behavior performed by male white-tailed deer during the rut in which a pawed depression and associated over-hanging branch are marked with saliva, glandular secretions, urine, and feces. We placed 71 and 35 motion-activated cameras on scrapes in DeSoto National Wildlife Refuge in western Nebraska and eastern Iowa from Oct. – Nov. 2005 and Sept. – Nov. 2006, respectively. We recorded 5009 encounters and 1830 direct interactions. We developed an ethogram of behaviors of interest at scrapes. We found that males interacted with scrapes more frequently than females (P < 0.001). Male interactions were more complex, with 69% consisting of ≥2 observed behaviors versus 25% and 13% for females and fawns. We identified individual male deer ≥2.5 years old and determined the minimum number of different scrapes individuals visited and the number of individuals that visit a single scrape. Individuals that appeared on camera ≥5 times visited a mean of 3.9 scrapes (range = 1-15) and traveled a mean minimum distance of 978 m between consecutive scrapes. A mean of 5.1 individuals visited a single scrape, and up to 43% of individuals returned to a scrape previously visited at least once. We modeled Risk Values based on frequency of occurrence, duration, and Threat Values of each behavior, for contacting and transmitting CWD prions at scrapes. Adult males had the highest total Risk Values for contacting CWD prions (114.1) and shedding prions (59.4). The “grasp-lick branch” behavior had the highest Risk Value for adult males for both contacting and transmitting prions. Our study reveals a sex specific social behavior in male white-tailed deer that has the potential to spread chronic wasting disease between adult males in the population.