Surveillance for Toxoplasma gondii, Brucella spp., and Chlamydia spp. in Australian Fur Seal (Arctocephalus pusillus doriferus) Abortions.

The intracellular pathogens Toxoplasma gondii, Brucella spp., and Chlamydia spp. are all known causative agents of abortion in wildlife. Both T. gondii and Brucella spp. have been identified in marine mammal abortions and a limited number of studies have detected their potential presence in Australian fur seals (Arctocephalus pusillus doriferus), but data are sparse for these pathogens in Australian fur seal breeding colonies. Australian fur seals have been shown to have a high degree of third-trimester pregnancy loss in one of their largest breeding colonies. Additionally, pup production has declined at the largest breeding colony for the species. This study surveyed the presence of T. gondii, Brucella spp., and Chlamydia spp. as potential infectious causes of this reproductive loss. Aborted fetuses were collected from two of the largest breeding colonies for the species, Seal Rocks (n=19) and Kanowna Island (n=34). These were examined grossly and through histopathological evaluation, in conjunction with molecular testing for all three pathogens. Placentas were collected from full-term births during the pupping season from Kanowna Island (n=118). These were used to compare the molecular prevalence of the three pathogens in presumed successful pregnancies. Chlamydia spp. was not detected in aborted fetuses in this study. Brucella spp. was detected with PCR in both aborted fetuses (9.4%) and placentas from full-term births (3.4%), and T. gondii was detected using routine histopathology (n=2/53), immunohistochemistry (n=3/4), and PCR (n=4/53) in tissues from aborted fetuses. Toxoplasma gondii was present in 7.5% of third-trimester abortions and absent from all full-term placentas. Brucella spp. was detected in both aborted fetuses and full-term placentas. This is the first description of vertical transmission of T. gondii in a marine mammal from the southern hemisphere.

A Novel Marine Mammal Coxiella burnetii-Genome Sequencing Identifies a New Genotype with Potential Virulence.

The obligate intracellular bacterial pathogen Coxiella burnetii has been identified in a few species of marine mammals, some of which are showing population declines. It has been hypothesized that C. burnetii in marine mammals is a distinct genotype that varies significantly from the typical terrestrial genotypes. It appears to lack an IS1111. Isolates originating from Australian marine animals have a distinctly non-Australian profile of multiple-locus variable-number tandem-repeat analysis (MLVA). Extracted Coxiella DNA of Australian fur seal placental origin was sequenced using the Novaseq platform. Illumina 150 bp paired-end reads were filtered and trimmed with Trimgalore. The microbial community present in the sequenced genome was evaluated with Kraken and Bracken software using the NCBI database. A phylogenetic analysis was performed using 1131 core genes. Core genes were identified using Panaroo and inputted into Iqtree to determine the maximum-likelihood tree. A second phylogenetic tree was created using Rickettsiella grylii and using seven housekeeping genes. Results were compared with the C. burnetii Nine Mile RSA439 virulent genome. This new Australian marine mammal isolate of Coxiella (PG457) appears to be a novel genotype that lacks IS1111 and has a distinct MLVA signature (ms26, ms27, ms28, ms30, and ms31). The presence of genes for multiple virulence factors appears to give this genotype sufficient pathogenicity for it to be considered a possible causative agent of abortion in Australian fur seals as well as a potential zoonotic risk.

Pacific Gulls (Larus pacificus) as Potential Vectors of Coxiella burnetii in an Australian Fur Seal Breeding Colony.

Recently, Coxiella burnetii has been described as a novel pathogen potentially contributing to decreased pup production in Australian fur seals (AusFS, Arctocephalus pusillus doriferus). Pacific gulls (PGs, Larus pacificus) are known to scavenge AusFS placental material during the fur seal breeding season. It is hypothesized that PGs may act as vectors for this pathogen. In the present study, cloacal swabs, oral swabs and serum were collected from PGs on Kanowna Island (KI, an AusFS breeding colony) and a nearby island, Seal Island (SI), not occupied by pinnipeds. All sample sets were evaluated with qPCR for the com1, htpAB and IS1111 markers. Most oral and cloacal swabs from KI tested positive on both the com1 (94.1%; 88.2%) and htpAB targets (76.5%; 76.5%). Amplification was very low from the SI oral swabs and cloacal swabs. Only the KI serum samples had amplification (17.7% for both com1 and htpAB). There was no IS1111 amplification in either colony. The results demonstrate that PGs can potentially act as vectors for the spread of C. burnetii. In some birds, C. burnetii was detectable in the serum, indicating that gulls can experience bacteraemia. It appears that different feeding strategies in the same species within the same ecosystem can have profound effects on the prevalence of pathogens. Further studies are required to better understand the epidemiology and potential risks of this organism.

MEDETOMIDINE-KETAMINE-BUPRENORPHINE ANESTHESIA OF THE SOLITARY SUBTERRANEAN CAPE DUNE MOLE-RAT (BATHYERGUS SUILLUS) AND THE CAPE MOLE-RAT (GEORYCHUS CAPENSIS).

A prospective, descriptive study was conducted to evaluate the safety and efficacy of a field-ready anesthetic drug combination of medetomidine-ketamine-buprenorphine for data logger implantation surgery or recheck in free-ranging Cape dune (Bathyergus suillus: n = 41) and Cape (Georychus capensis: n = 37) mole-rats. All anesthesia data were reported as mean (±standard deviation). Medetomidine-ketamine-buprenorphine doses were 0.1 (±0.03), 10.6 (±2.8), and 0.06 (±0.03) mg/kg, respectively, for Cape dune mole-rats; and 0.2 (±0.03), 19.4 (±4.0), and 0.14 (±0.03) mg/kg, respectively, for Cape mole-rats. Induction was calm and took 2.00 (range: 1.00-6.00) min for the Cape dune and 1.75 (range 1.25 to 8.16) min for Cape mole-rats. A surgical plane of anesthesia was achieved in most Cape dune mole-rats (92%) and Cape mole-rats (90%). The remainder required supplementation with a single intramuscular injection of ketamine (3-9 mg/kg) during surgery. Heart and respiratory rates were 149 (±37) beats and 24 (±8) breaths per minute, respectively, for Cape dune mole-rats and 179 (±40) beats and 25 (±10) and breaths per minute, respectively for Cape mole-rats. Surgical time for mole-rats ranged from 25 to 38 min. Recovery was calm and took 8.50 (range: 2.00-19.00) min for Cape dune mole-rats and 9.75 (range: 2.00-34.00) min for Cape mole-rats to recover. For recovery, atipamezole was administered intramuscularly at 0.5 (±0.15) mg/kg for Cape dune mole-rats and 1 (±0.15) mg/kg for Cape mole-rats. All mole-rats were returned to their original burrows within 48 h of recovery. The medetomidine-ketamine-buprenorphine combination induced a predictable, safe anesthesia in Cape dune and Cape mole-rats suitable for short intraabdominal surgery. This combination is suited to in situ studies where the use of a formal surgery or laboratory is not feasible.

Disentanglement of Cape fur seals (Arctocephalus pusillus pusillus) with reversible medetomidine-midazolam-butorphanol.

Anaesthesia in pinnipeds is considered a much higher risk than in most terrestrial mammals because of their frequent proximity to water and physiological and anatomical adaptations related to diving, which also influence their anaesthesia management. Anaesthetising and immobilising entangled seals does not allow for selection of animals that are at a safe distance from the water's edge. Medetomidine-midazolam-butorphanol (MMB) sedation was trialled on eight entangled Cape fur seals (CFS) (Arctocephalus pusillus pusillus) to determine if it was safe to use on animals that entered the water post-darting. The MMB was given at an estimated dose of 0.03 mg/kg, 0.2 mg/kg and 0.2 mg/kg, respectively, via remote darting. Sedation was reversed with intramuscular atipamezole (0.15 mg/kg) and naltrexone (0.4 mg/kg) to antagonise the effects of medetomidine and butorphanol, respectively. Moderate sedation was achieved in six animals. Six of the animals entered the water after being darted. There was a single mortality and a single animal that was too lightly sedated for capture. The preliminary results indicate that MMB produces suitable sedation for disentanglement of CFS. Additionally, MMB might be suitable for application to field-based biological research.

Pharmacokinetics and pharmacodynamic effects in koi carp (Cyprinus carpio) following immersion in propofol.

OBJECTIVE: To test the hypothesis that plasma propofol concentration (PPC) is associated with anesthetic effect in koi carp administered propofol by immersion. STUDY DESIGN: Prospective study. ANIMALS: Twenty mature koi carp (mean ± standard deviation, 409.4 ± 83.7 g). METHODS: Fish were immersed in propofol (5 mg L-1). Physiological variables and induction and recovery times were recorded. In phase I, blood was sampled for PPC immediately following induction and at recovery. In phase II, following induction, fish were maintained with propofol (4 mg L-1) via a recirculating system for 20 minutes. Following established induction, blood was sampled at 1, 10 and 20 minutes. In phase III (n = 19), fish were anesthetized as in phase II with blood sampled nine times in a sparse sampling strategy. Simultaneously, a pharmacodynamics rubric was used to evaluate anesthetic depth. PPC was determined using high performance liquid chromatography with fluorescence detection. Following evaluation of normality, data were analyzed using paired t test or Spearman correlation test (significance was set at p < 0.05). RESULTS: In phase I, mean PPCs at induction (20.12 µg mL-1) and recovery (11.62 µg mL-1) were different (p < 0.001). In phase II, only mean PPCs at induction (17.92 µg mL-1) and 10 minutes (21.50 µg mL-1) were different (p = 0.013). In phase III, a correlation between PPCs and the pharmacodynamic rubric scores was found (p < 0.001, r = -0.93). There was no correlation between PPCs and recovery time (p = 0.057, r = 0.433). A two-compartment open model was chosen for the pharmacokinetic model. Absorption rate constant, elimination rate constant and intercompartmental rate constant were 0.48, 0.006 and 0.02 minute-1, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Measurable PPCs were achieved in koi carp anesthetized with propofol by immersion. Anesthetic depth of fish was negatively correlated with PPCs, but recovery time was not.

Fatal disseminated toxoplasmosis in a zoological collection of meerkats (Suricata suricatta).

Two confirmed cases of fatal disseminated toxoplasmosis occurred in an urban zoological collection of meerkats (Suricata suricatta). Both cases are suspected to be the result of feral cats gaining access to the enclosure. Toxoplasmosis has rarely been documented in meerkats. Subsequent to prophylactic treatment of all the animals and structural changes being implemented within the enclosure, no new cases have been recorded to date. Very little information is available on the disease in viverrids.

Subcutaneous xanthomatosis in a great white pelican (Pelecanus onocrotalus).

A great white pelican (Pelecanus onocrotalus) was referred for assessment of a subacute-onset, nonpainful swelling located in the pectoral region. Physical examination revealed a firm, round, well-circumscribed subcutaneous mass approximately 10 cm in diameter. Cytological evaluation of a fine needle aspirate of the mass was consistent with a mesenchymal tumor. The mass was excised, and a diagnosis of xanthomatosis was made based on histopathologic results. Avian xanthomatosis is a nonneoplastic condition of unknown etiology. Possible causes of this condition include trauma, metabolic or nutritional disorders. Similar lesions were not observed in the nine conspecifics that were fed the same diet and housed in the same enclosure. To our knowledge, this is the first report of xanthomatosis in the family Pelecanidae.