Mycobacterium kansasii Infection in a Farmed White-Tailed Deer (Odocoileus virginianus) in Florida, USA.

A 7-year-old farmed white-tailed deer doe was transported to a Levy County, Florida property and began to decline in health, exhibiting weight loss and pelvic limb weakness. The doe prematurely delivered live twin fawns, both of which later died. The doe was treated with corticosteroids, antibiotics, gastric cytoprotectants, and B vitamins but showed no improvement. The doe was euthanized, and a post mortem examination was performed under the University of Florida's Cervidae Health Research Initiative. We collected lung tissue after the animal was euthanized and performed histological evaluation, using H&E and Ziehl-Neelsen (ZN) staining, and molecular evaluation, using conventional PCR, followed by Sanger sequencing. The microscopic observations of the H&E-stained lung showed multifocal granuloma, while the ZN-stained tissue revealed low numbers of beaded, magenta-staining rod bacteria inside the granuloma formation. Molecular analysis identified the presence of Mycobacterium kansasii. This isolation of a non-tuberculous Mycobacterium in a white-tailed deer emphasizes the importance of specific pathogen identification in cases of tuberculosis-like disease in farmed and free-ranging cervids. We report the first case of M. kansasii infection in a farmed white-tailed deer (Odocoileus virginianus) in Florida. Although M. kansasii cases are sporadic in white-tailed deer, it is important to maintain farm biosecurity and prevent farmed cervids from contacting wildlife to prevent disease transmission.

EVALUATION OF A COMBINATION OF TILETAMINE-ZOLAZEPAM, MEDETOMIDINE, AND AZAPERONE WITH NASAL OXYGEN SUPPLEMENTATION FOR THE IMMOBILIZATION OF CAPTIVE CHACOAN PECCARIES (CATAGONUS WAGNERI) IN THE CHACO REGION OF PARAGUAY.

A combination of tiletamine-zolazepam, medetomidine, and azaperone was used to immobilize captive Chacoan peccaries (Catagonus wagneri) for health assessments and biological sample collection at the Centro Chaqueño para la Conservación e Investigación (CCCI) in the Paraguayan Chaco during July in 2017 and 2018. In total, 83 peccaries kept in 0.25-1.50 hectare enclosures were immobilized via dart-administered anesthetic. Mean animal weight was 33.89±3.74 kg (standard deviation; n=77). The mean intramuscular (IM) anesthetic drug and dosages were 0.03±0.00 mg/kg of medetomidine, 0.91±0.10 mg/kg of Zoletil 50 (tiletamine-zolazepam), and 0.30±0.03 mg/kg azaperone. The mean time to recumbency after darting was 6.07±2.65 min. The mean time to reach the anesthetic plane postdarting was 10.00±2.00 min. Muscle relaxation was adequate to allow minor veterinary procedures. A mean dosage of 0.15±0.02 mg/kg of atipamezole was given IM to reverse the medetomidine. Recoveries were smooth and animals were standing by 59.17±30.18 min postreversal. Full recovery and release back to enclosures occurred 90±30 min postreversal. A single dose of this drug combination provided adequate anesthesia for 88% of adult Chacoan peccaries; 12% needed a supplemental dose of tiletamine-zolazepam because of failure to receive the full dose from the anesthetic dart. Sex and age did not impact the dosage required to achieve immobilization. Confinement during recovery from anesthesia is required with this protocol. Aside from mild hypoxemia, no adverse effects from anesthesia were observed. However, oxygen supplementation as a part of this protocol is recommended to support circulatory and respiratory capacity.

A Mortality-Based Description of EHDV and BTV Prevalence in Farmed White-Tailed Deer (Odocoileus virginianus) in Florida, USA.

Hemorrhagic disease (HD) caused by bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) is the most important viral disease of farmed and wild white-tailed deer (WTD; Odocoileus virginianus) and can cause substantial mortality in susceptible hosts. Captive cervid farming is an emerging industry in Florida, an HD-enzootic region. Morbidity and mortality due to HD are major concerns among deer farmers, but the impact of HD on Florida's cervid farming industry is unknown. Our primary objective was to determine the prevalence of epizootic hemorrhagic disease virus (EHDV) and bluetongue virus (BTV) among WTD submitted to the University of Florida Institute of Food and Agricultural Sciences Cervidae Health Research Initiative (CHeRI) for post-mortem diagnostics. Our secondary objectives were to identify the predominant circulating EHDV serotypes during each sampling year and to determine the age class with the greatest proportion of EHDV- and BTV-positive post-mortem specimens. From 2016 to 2020, spleen samples from 539 farmed WTD with unexplained mortality were tested for the presence of EHDV and BTV by RT-qPCR. Overall, the prevalence of EHDV, BTV, or EHDV/BTV coinfection was 26%, 16%, and 10%, respectively, and 44% of deer (237/539) were diagnosed with HD by RT-qPCR. The predominant circulating EHDV serotype varied by year. Overall, EHDV-2 was the most commonly identified serotype (55% of PCR-positive cases), and EHDV-1 was the least frequently identified serotype (16% of PCR-positive cases). The greatest proportion of EHDV/BTV positives among mortality cases was observed in young WTD aged 3-6 months (50%-82% positive). There was a significant difference in the prevalence of EHDV/BTV by age when comparing specimens from WTD over 1 year old (p = 0.029, n = 527). Among these samples, the number of reported mortalities and the prevalence of EHDV/BTV were highest in yearling animals (56%). These data provide the first estimate of EHDV and BTV prevalence and virus serotypes among farmed WTD in Florida, identify the WTD age groups with the greatest proportions of EHDV- and BTV-positive specimens, and suggest that HD caused by these two viruses may be a major source of mortality challenging the captive cervid farming industry in Florida.

Genome Sequence of a Yunnan Orbivirus Isolated from a Dead Florida White-Tailed Deer (Odocoileus virginianus).

We report the complete coding sequences of a Yunnan orbivirus isolated from a dead white-tailed deer (Odocoileus virginianus) in Florida in 2019. The prevalence of Yunnan orbivirus and its role in disease among farmed white-tailed deer remain to be determined.

Genome Sequence of a CHeRI Orbivirus 3 Strain Isolated from a Dead White-Tailed Deer (Odocoileus virginianus) in Florida, USA.

We report the genome sequence of an orbivirus isolated from a dead farmed white-tailed deer in Florida. The deer was coinfected with epizootic hemorrhagic disease virus type 2. Phylogenetic and genetic analyses supported the virus as the fourth strain of the CHeRI orbivirus 3 species.

Three New Orbivirus Species Isolated from Farmed White-Tailed Deer (Odocoileus virginianus) in the United States.

We report the detection and gene coding sequences of three novel Orbivirus species found in six dead farmed white-tailed deer in the United States. Phylogenetic analyses indicate that the new orbiviruses are genetically closely related to the Guangxi, Mobuck, Peruvian horse sickness, and Yunnan orbiviruses, which are thought to be solely borne by mosquitos. However, four of the six viruses analyzed in this work were found as co-infecting agents along with a known cervid pathogen, epizootic hemorrhagic disease virus-2 (EHDV-2), raising questions as to whether the new viruses are primary pathogens or secondary pathogens that exacerbate EHDV-2 infections. Moreover, EHDV-2 is known to be a Culicoides-borne virus, raising additional questions as to whether Culicoides species can also serve as vectors for the novel orbiviruses, if mosquitoes can vector EHDV-2, or whether the deer were infected through separate bites by the insects. Our findings expand knowledge of the possible viral pathogens of deer in the United States. Moreover, due to the close genetic relatedness of the three new orbiviruses to viruses that are primary pathogens of cattle and horses, our findings also underscore a crucial need for additional research on the potential role of the three new orbiviruses as pathogens of other animals.

Molecular characterization of a novel reassortment Mammalian orthoreovirus type 2 isolated from a Florida white-tailed deer fawn.

Mammalian orthoreovirus (MRV) is the type species of the genus Orthoreovirus and causes a range of significant respiratory, nervous or enteric diseases in humans and animals. In 2016 a farmed white-tailed deer (Odocoileus virginianus) fawn became ill, displaying clinical signs of lethargy, dehydration, and profuse foul-smelling diarrhea. A necropsy was performed after the three-week-old fawn died and various tissue samples were submitted to the University of Florida's Cervidae Health Research Initiative for diagnostic evaluation. Aliquots of homogenized heart, liver, and spleen tissues were inoculated onto Vero E6 cells. After virus-specific cytopathic effects (CPE) were detected in Vero cells inoculated with spleen homogenate, infected cells were fixed in glutaraldehyde and analyzed by transmission electron microscopy (TEM), which revealed icosahedral virus particles approximately 75 nm in diameter with morphologies consistent with those of reoviruses within the cytoplasm of the infected cells. RNA extracted from virions in the spent media of infected cells with advanced CPE was used to prepare a cDNA library, which was sequenced using an Illumina MiSeq sequencer. Complete coding sequences for ten separate reovirus segments were attained, and these indicated the isolated agent was a MRV. Genetic and phylogenetic analyses based on the outer capsid sigma-1 (σ1) protein gene sequences supported the Florida white-tailed fawn isolate as a type 2 MRV that branched as the sister group to a MRV-2 strain previously characterized from the urine of a moribund lion (Panthera leo) in Japan. However, analyses based on 7/10 genes (L1-L2, M2-M3, S2-S4) supported the white-tailed deer MRV as the closest relative to a type 3 MRV strain isolated from a dead mink in China. These data suggest the white-tailed deer MRV may have resulted from the natural reassortment of MRVs originating from multiple wildlife species. To our knowledge, this is the first detection of MRV-2 infection in a white-tailed deer. Continued surveillance efforts are needed to determine whether this MRV-2 strain poses a health threat to farmed white-tailed deer populations.

Complete Genome Sequence of Mobuck Virus Isolated from a Florida White-Tailed Deer (Odocoileus virginianus).

Here, we report the complete genome sequence of mobuck virus isolated from a Florida white-tailed deer (Odocoileus virginianus) in 2017. This is the second report of mobuck virus in the United States and expands the known geographic range of this novel orbivirus into Florida.

Semen analysis parameters from a captive population of the endangered Chacoan peccary (Catagonus wagneri) in Paraguay.

Baseline information about the reproductive physiology of an endangered species is vital to captive breeding programs. This study analyzed reproductive parameters from eleven captive Chacoan peccaries (Catagonus wagneri) (mean age: 10.5 ±â€¯1.2 years old) in Paraguay. After immobilization, testis length, width and firmness (hard to flaccid, 1-3) were assessed, followed by electroejaculation and analysis of semen. Samples were examined for percentage and progressive motility, total cell count, percentage live spermatozoa and morphology. Mean (±SEM) testis volume and firmness were 24.7 ±â€¯1.8 cm3 and 2.1 ±â€¯0.1, respectively. Mean ejaculate volume was 2.9 ±â€¯0.7 ml with a pH of 7.7 ±â€¯0.3. Few male peccaries had motile spermatozoa (n = 5/9) with a mean percentage and progressive motility of 18.3 ±â€¯8.5% and 0.6 ±â€¯0.3, respectively. The mean percentage of live spermatozoa was 25.1 ±â€¯5.6%. Male peccaries had a low percentage of normal spermatozoa (12.4 ±â€¯2.5%). The mean total count of spermatozoa per ejaculate was also quite low at 1.58 ±â€¯1.01 million total spermatozoa per ejaculate. Spermatozoa defects were predominantly primary (77.7%) with the most common spermatozoa defects being tapered head (19.0 ±â€¯7.4%), diadem/crater (17.7 ±â€¯2.8%), and excess residual cytoplasm (9.6 ±â€¯2.5%). Male age was not correlated with semen parameters (percent live: r=-0.19; motility percentage: r = 0.01; percent normal spermatozoa: r = 0.38; total count: r = 0.29; p>0.05). Evaluation of additional males from this population as well as other captive populations at various time points during the year is warranted.