DIAGNOSING AND CATEGORIZING LEPROSY IN LIVE EURASIAN RED SQUIRRELS (SCIURUS VULGARIS) FOR MANAGEMENT, SURVEILLANCE, AND TRANSLOCATION PURPOSES.

The presence of Mycobacterium lepromatosis and Mycobacterium leprae in Eurasian red squirrel (Sciurus vulgaris, ERS) carcasses throughout the British Isles, and leprosy as a disease, have recently been reported using histological and molecular diagnostic methods. In 2016, the first longitudinal study of ERS affected by leprosy was initiated. One of the main challenges was the reliable diagnosis of leprosy in live ERS, which is important for (a) welfare and case management and (b) surveillance or pretranslocation screening efforts. We explored diagnostic methods ranging from detailed clinical assessment and informative categorization of observed lesions, thermal imaging, serology (antiphenolic glycolipid-I antibody [αPGL-I] detection) to molecular methods (polymerase chain reaction [PCR]). For PCR the ear was established as the optimal sampling site. Based on the experiences from this 2-yr study we propose an objective categorization system for clinical lesions and a diagnostic framework for the combination of the diagnostic tools we found to be effective in live ERS: clinical assessment, αPGL-I serology, and PCR. Thermal imaging did not offer additional information for leprosy diagnostics in ERS. We propose an amended definition of leprosy lesions in ERS as "skin areas of local hair loss, in which a firm-rubbery, glossy swelling develops, that may ulcerate" and standardized terminology for describing ERS leprosy status. The information presented forms the basis of a consistent, reliable diagnostic and reporting system for leprosy cases in ERS.

CLINICAL PROGRESSION OF LEPROSY IN EURASIAN RED SQUIRRELS (SCIURUS VULGARIS) IN A NATURALLY INFECTED WILD POPULATION.

Leprosy has been described in Eurasian red squirrel (Sciurus vulgaris; ERS) carcasses since 2014. Studies of ERS carcasses have not provided information about incubation or disease progression in this host but have provided important insights into pathogen presence and distribution throughout the United Kingdom. Here we present field study data on 31 live ERS from an island population naturally infected with Mycobacterium leprae that were assessed longitudinally over a 2-yr time period. Clinical assessment, serologic (anti-phenolic glycolipid-I antibody [αPGL-I] detection) and molecular methods (polymerase chain reaction) were used to diagnose and categorize ERS at each assessment as a leprosy case, a leprosy suspect, colonized by M. leprae, or a contact ERS. Eight ERS (25.8%) were identified as leprosy cases: four at initial assessment, two at 6 mon and two at 24 mon after initial assessment. One ERS was categorized a leprosy suspect when it developed typical lesions 12 mon after initial assessment, despite negative serologic and molecular test results at this time, though M. leprae DNA had been isolated during the initial assessment. Seven ERS (22.6%) were categorized as colonized and of these, six were reassessed but did not develop clinical signs of leprosy within 6 (n = 2), 12 (n = 3), and 18 (n = 1) mon. Most (48.4%, n = 15) were categorized as contact ERS. Progression of leprosy lesions varied between ERS, but always increased in severity over time and was paralleled with increased antibody response. Based on our dataset, we propose the hypotheses: 1) leprosy in ERS is a chronic, slowly progressing disease in this species, similar to that described for other hosts; 2) lesions can undergo repeated ulceration-healing cycles; and 3) in some instances M. leprae DNA and αPGL-I antibodies are detectable before the onset of clinical signs of disease. Future studies addressing the progression of leprosy in ERS should follow affected animals over a longer time period and include tissue samples to pair molecular diagnostics with serologic results.

British Red Squirrels Remain the Only Known Wild Rodent Host for Leprosy Bacilli.

Eurasian red squirrels (Sciurus vulgaris) in the British Isles are the most recently discovered animal reservoir for the leprosy bacteria Mycobacterium leprae and Mycobacterium lepromatosis. Initial data suggest that prevalence of leprosy infection is variable and often low in different squirrel populations. Nothing is known about the presence of leprosy bacilli in other wild squirrel species despite two others (Siberian chipmunk [Tamias sibiricus], and Thirteen-lined ground squirrel [Ictidomys tridecemlineatus]) having been reported to be susceptible to experimental infection with M. leprae. Rats, a food-source in some countries where human leprosy occurs, have been suggested as potential reservoirs for leprosy bacilli, but no evidence supporting this hypothesis is currently available. We screened 301 squirrel samples covering four species [96 Eurasian red squirrels, 67 Eastern gray squirrels (Sciurus carolinensis), 35 Siberian chipmunks, and 103 Pallas's squirrels (Callosciurus erythraeus)] from Europe and 72 Mexican white-throated woodrats (Neotoma albigula) for the presence of M. leprae and M. lepromatosis using validated PCR protocols. No DNA from leprosy bacilli was detected in any of the samples tested. Given our sample-size, the pathogen should have been detected if the prevalence and/or bacillary load in the populations investigated were similar to those found for British red squirrels.

Red squirrels in the British Isles are infected with leprosy bacilli.

Leprosy, caused by infection with Mycobacterium leprae or the recently discovered Mycobacterium lepromatosis, was once endemic in humans in the British Isles. Red squirrels in Great Britain (Sciurus vulgaris) have increasingly been observed with leprosy-like lesions on the head and limbs. Using genomics, histopathology, and serology, we found M. lepromatosis in squirrels from England, Ireland, and Scotland, and M. leprae in squirrels from Brownsea Island, England. Infection was detected in overtly diseased and seemingly healthy animals. Phylogenetic comparisons of British and Irish M. lepromatosis with two Mexican strains from humans show that they diverged from a common ancestor around 27,000 years ago, whereas the M. leprae strain is closest to one that circulated in Medieval England. Red squirrels are thus a reservoir for leprosy in the British Isles.