Genetic diversity of Trypanosoma cruzi infecting raccoons (Procyon lotor) in 2 metropolitan areas of southern Louisiana: implications for parasite transmission networks.

Trypanosoma cruzi, the aetiological agent of Chagas disease, exists as an anthropozoonosis in Louisiana. Raccoons are an important reservoir, as they demonstrate high prevalence and maintain high parasitaemia longer than other mammals. Given the complex nature of parasite transmission networks and importance of raccoons as reservoirs that move between sylvatic and domestic environments, detailing the genetic diversity of T. cruzi in raccoons is crucial to assess risk to human health. Using a next-generation sequencing approach targeting the mini-exon, parasite diversity was assessed in 2 metropolitan areas of Louisiana. Sequences were analysed along with those previously identified in other mammals and vectors to determine if any association exists between ecoregion and parasite diversity. Parasites were identified from discrete typing units (DTUs) TcI, TcII, TcIV, TcV and TcVI. DTUs TcII, TcV and TcVI are previously unreported in raccoons in the United States (US). TcI was the most abundant DTU, comprising nearly 80% of all sequences. All but 1 raccoon harboured multiple haplotypes, some demonstrating mixed infections of different DTUs. Furthermore, there is significant association between DTU distribution and level III ecoregion in Louisiana. Finally, while certain sequences were distributed across multiple tissues, others appeared to have tissue-specific tropism. Taken together, these findings indicate that ongoing surveillance of T. cruzi in the US should be undertaken across ecoregions to fully assess risk to human health. Given potential connections between parasite diversity and clinical outcomes, deep sequencing technologies are crucial and interventions targeting raccoons may prove useful in mitigating human health risk.

Toxicity of oil and dispersed oil on juvenile mud crabs, Rhithropanopeus harrisii.

In order to simulate an offshore oil spill event, we assessed the acute toxicity of the non-dispersed and the chemically dispersed water-accommodated fraction (WAF) of crude oil using Louisiana sweet crude and Corexit(®) 9500A with juvenile Harris mud crabs (Rhithropanopeus harrisii), an important Gulf of Mexico benthic crustacean. The chemical dispersion of crude oil significantly increased acute toxicity of the WAF in juvenile mud crabs compared to naturally dispersed oil. The majority of the mortality in the chemically dispersed treatments occurred within 24 h. While higher concentrations of chemically dispersed WAF had no survivors, at lower concentrations surviving juvenile crabs displayed no long-term effects. These results suggest that if the juvenile crabs survive initial exposure, acute exposure to dispersed or non-dispersed crude oil may not induce long-term effects.

Seroprevalence of Leptospira Among Selected Mammals on a Wildlife Management Area in Louisiana, USA.

From August to December 2018, we collected blood samples from 98 individuals of 11 mammal species to examine seroprevalence of leptospirosis at the Sherburne Wildlife Management Area in central Louisiana, US. Overall, 21.4% of individuals tested positive for antibodies of at least one Leptospira interrogans serovar and six individuals were reactive for two or more serovars. The most prevalent serovar we detected was serovar Bratislava (19.4%), followed by serovar Grippotyphosa (6.1%), serovar Icterohaemorrhagiae and serovar Pomona (2.0%), and serovar Canicola and serovar Hardjo (1.0%). We detected the highest prevalence in fox squirrels (Sciurus niger), hispid cotton rats (Sigmodon hispidus), and feral swine (Sus scrofa), with serovar Bratislava being the most reactive for these three species. Positive samples returned titer results of 100-400 for all species and serovars, with the exception of one feral swine that returned a titer of 1,600 to serovar Bratislava, indicating an active infection. Although the potential effects of leptospirosis on our study species remains unclear, our data contribute information necessary to understand and manage potential risks of Leptospira exposure to wildlife, domestic animals, and humans.

Raccoons As an Important Reservoir for Trypanosoma cruzi: A Prevalence Study from Two Metropolitan Areas in Louisiana.

Raccoons are an important reservoir for Trypanosoma cruzi infection, having been reported to maintain a high and lengthy parasitemia. Although raccoon populations have historically been abundant in Louisiana, the prevalence rate of T. cruzi infection in raccoons in this state is unknown. Here, we tested raccoon tissues from two urban areas in Louisiana, namely Orleans Parish (OP) and East Baton Rouge Parish (EBRP), to investigate prevalence in these areas using direct detection through polymerase chain reaction. Overall, 33.6% of raccoons tested were positive. The prevalence in OP (42.9%) was significantly higher than the prevalence in EBRP (23.2%). There was no significant difference in prevalence between sexes or based on age, but there was a significant difference in infection prevalence based on season of trapping. These results suggest the importance of raccoons as a reservoir host, maintaining T. cruzi infection and potentially posing a risk to human health.

Disseminated Mycobacterium kansasii infection in a white-tailed deer and implications for public and livestock health.

We document a case of Mycobacterium kansasii, a rare, zoonotic bacterium, in a white-tailed deer (WTD; Odocoileus virginianus) in East Baton Rouge Parish, Louisiana. Grossly, the deer had fibrinous pleuropneumonia with yellow, mineralized nodules scattered throughout the lungs and extending to the pleura. The kidneys were enlarged and had numerous pale foci in the cortex. Microscopically, the pulmonary architecture was replaced by variably sized, multifocal-to-coalescing granulomas with peripheral histiocytes and fewer multinucleate giant cells, and necrotic centers with mineralization and hemorrhage. The latter rarely contained one to a few acid-fast, slender, 7-µm long bacteria, for which beaded morphology was sometimes evident. Similar acid-fast bacteria were also within histiocytes in the kidney. PCR assay of fresh lung sample and subsequent sequencing revealed a non-tuberculosis mycobacterium, M. kansasii. These lesions were similar to those that result from infection with M. bovis in WTD. Both M. bovis and M. kansasii are zoonotic. WTD are a reservoir of M. bovis, which is a major concern in regions in which WTD and cattle can come into close contact.

Case Series: Virulent hemosporidiosis infections in juvenile great horned owls (Bubo virginianus) from Louisiana and California, USA.

A total of eight juvenile great horned owls (Bubo virginianus) were found lethargic and on the ground in spring 2015, 2016, and 2017, including one fledgling from Louisiana, USA and seven nestlings from California, USA. One bird survived to release after rehabilitation; seven birds died or were euthanized due to poor prognosis and were necropsied. Necropsy findings were similar and included general pallor of all tissues, particularly the subcutis and lungs, and enlarged liver and spleen. Histopathology revealed multi-organ necrosis, abundant meronts containing merozoites, and intracytoplasmic pigmented haemosporidian parasites in blood cells in one bird. Leucocytozoon lineages lSTOCC16 and BUVIR06 were identified by polymerase chain reaction and genetic sequencing. The systemic Leucocytozoon infections were likely associated with morbidity and mortality in these owls. A second parasite, Haemoproteus lineage hSTVAR01, was also identified in an owl from Louisiana. This is the first identification of Leucocytozoon lineages that have been associated with mortality in young great horned owls.