Multi-Injection Pharmacokinetics of Meloxicam in Kemp’s Ridley (Lepidochelys kempii) and Green (Chelonia mydas) Sea Turtles after Subcutaneous Administration

The objective of this study was to determine the pharmacokinetics and safety of multiple injections of meloxicam (MLX) administered subcutaneously (SQ) in Kemp’s ridley (Lepidochelys kempii) and green (Chelonia mydas) sea turtles. Based on results from a previously published single-injection study, a multiple-injection regimen was derived for the Kemp’s ridleys, which consisted of administering MLX at a dose of 1 mg/kg SQ every 12 h for 5 days, and for green turtles at a dose of 1 mg/kg SQ every 48 h for three treatments. Six turtles of each species were used for the study, and blood samples were taken at multiple time intervals. The terminal half-life after the last dose for the Kemp’s ridley sea turtles was calculated at 7.18 h, and for the green sea turtles at 23.71 h. Throughout the multiple injections, MLX concentrations remained above 0.57 µg/mL, a concentration targeted in humans for the analgesic and anti-inflammatory effects. No negative side effects or changes to blood parameters evaluated were observed during the study in either species. The results of this study suggest MLX should be administered SQ to Kemp’s ridley sea turtles at a dosage of 1 mg/kg every 12 h and in green sea turtles at a dose of 1 mg/kg every 48 h. The novelty of this work is that it is a multiple-injection study. Multiple injections were administered and produced concentrations that were considered therapeutic in humans, and the turtles did not have any adverse side effects. Furthermore, there were large differences in the pharmacokinetic values between green and Kemp’s ridley sea turtles.

Evaluation of the Respiratory Microbiome and the Use of Tracheal Lavage as a Diagnostic Tool in Kemp’s Ridley Sea Turtles (Lepidochelys kempii)

Respiratory disease is a common cause of morbidity and mortality in sea turtles, including the Kemp’s ridley sea turtle (Lepidochelys kempii). Although culture-dependent methods are typically used to characterize microbes associated with pneumonia and to determine treatment, culture-independent methods can provide a deeper understanding of the respiratory microbial communities and lead to a more accurate diagnosis. In this study, we characterized the tracheal lavage microbiome from cold-stunned Kemp’s ridley sea turtles at three time points during rehabilitation (intake, rehabilitation, and convalescence) by analyzing the 16S rRNA gene collected from tracheal lavage samples. We retrospectively developed a radiographic scoring system to grade the severity of lung abnormalities in these turtles and found no differences in diversity or composition of microbial communities based on radiographic score. We also found that the culture isolates from tracheal lavage samples, as well as other previously reported sea turtle pathogens, were present in variable abundance across sequenced samples. In addition to the tracheal microbial community of live turtles, we characterized microbial communities from other segments of the respiratory tract (glottis, trachea, anterior lung, posterior lung) from deceased turtles. We found a high degree of variability within turtles and a high degree of dissimilarity between different segments of the respiratory tract and the tracheal lavage collected from the same turtle. In summary, we found that the pulmonary microbial community associated with pneumonia in sea turtles is complex and does not correlate well with the microbial community as identified by tracheal lavage. These results underscore the limitations of using tracheal lavage for identification of the causative agents of pneumonia in sea turtles.

Mitochondrial DNA Evaluation and Species Identification of Kemp’s Ridley Sea Turtle (Lepidochelys kempii) Bones After a 3-Year Exposure to Submerged Marine and Terrestrial Environments

Because Kemp’s ridley sea turtles (Lepidochelys kempii) are critically endangered and closely related to the vulnerable olive ridleys (L. olivacea), it is essential for forensic investigations and conservation efforts to distinguish these species when only skeletal elements remain. DNA extraction and analysis by DNA sequencing of genetic markers is the only method to determine species identity reliably, yet these methods are significantly compromised when DNA becomes degraded. To evaluate the role that time and environment play in obtaining high-quality DNA sequencing data, we placed skeletal elements of a terrestrial mammal (Bos taurus) and L. kempii in a supratidal and subtidal environment for 3 years. Bi-annual sampling revealed that after 3 years, mitochondrial DNA (mtDNA) consistently identified each species from each environment. Our results show that mtDNA recovery from bone and identification for Kemp’s ridley sea turtles was possible up to 3 years in both environments. All sequencing data obtained was accurate and robust, but DNA sequencing results were not consistent after 664-days of exposure. Our findings led us to conclude that if sufficient DNA is extracted from bone samples, then high-quality sequence data can be obtained, and the resulting sequence data accurately reflects the reference sequence for the given gene marker. This study provides evidence that DNA can be extracted and analyzed from challenging biological substrates, like bone, when these substrates are exposed to seasonally dynamic maritime environmental conditions for up to 3-years.

Characterization of oral and cloacal microbial communities in cold-stunned Kemp’s ridley sea turtles (Lepidochelys kempii) during the time course of rehabilitation

Microbial communities of animals play a role in health and disease, including immunocompromised conditions. In the northeastern United States, cold-stunning events often cause endangered Kemp’s ridley turtles (Lepidochelys kempii) to become stranded on beaches in autumn. These sea turtles are admitted to rehabilitation facilities when rescued alive and are presumed immunocompromised secondary to hypothermia. To better understand the role that microbes play in the health of cold-stunned sea turtles, we characterized the oral and cloacal microbiome from Kemp’s ridley turtles at multiple timepoints during rehabilitation, from admission to pre-release, by using Illumina sequencing to analyze the 16S rRNA gene. Microbial communities were distinct between body sites and among turtles that survived and those that died. We found that clinical parameters such as presence of pneumonia or values for various blood analytes did not correlate with oral or cloacal microbial community composition. We also investigated the effect of antibiotics on the microbiome during rehabilitation and prior to release and found that the type of antibiotic altered the microbial community composition, yet overall taxonomic diversity remained the same. The microbiome of cold-stunned Kemp’s ridley turtles gradually changed through the course of rehabilitation with environment, antibiotics, and disease status all playing a role in those changes and ultimately the release status of the turtles.

PENYULUHAN MENGENAI OPTIMALISASI UPAYA PELESTARIAN PENYU DI PANTAI MAPAK INDAH, KELURAHAN JEMPONG BARU, KECAMATAN SEKARBELA

Penyu merupakan salah satu biota laut yang dilindungi karena keberadaannya sudah terancam punah, terutama untuk beberapa jenis penyu, yaitu penyu belimbing (Dermochelys coriacea), penyu kemp’s ridley (Lepidochelys kempii) dan penyu sisik (Eretmochelys imbricata). Terbatasnya informasi yang diterima masyarakat menyebabkan upaya pelestarian penyu dilakukan secara tradisional sehingga tidak berjalan optimal, salah satunya yang terjadi di Pantai Mapak Indah, Kelurahan Jempong Baru, Kecamatan Sekarbela, Kota Mataram. Kegiatan pengabdian kepada masyarakat ini bertujuan untuk memberikan penyuluhan dan pendampingan kepada masyarakat, khususnya Kelompok Pelestari Penyu Pantai Mapak Indah mengenai bentuk-bentuk upaya pelestarian penyu yang baik dan benar. Berdasarkan pengamatan dan hasil wawancara selama kegiatan pengabdian ditemukan beberapa kegiatan dalam upaya pelestarian penyu di Pantai Mapak Indah yang dirasa kurang sesuai sehingga perlu untuk diluruskan dan diperbaiki. Rekomendasi yang diberikan dalam kegiatan pengabdian kepada masyarakat ini antara lain terkait dengan lokasi pengambilan telur, desain sarang peneluran, waktu pelepasan tukik, pemberian pakan tukik, dan keramaian akibat tingginya aktivitas warga di lokasi pelestarian penyu. Masyarakat menyambut baik rekomendasi yang diberikan dan berharap dapat mendapatkan pendampingan secara kontinu sehingga kegiatan pelestarian penyu dapat semakin dioptimalkan. Dengan demikian, kegiatan penyuluhan ini sangat bermanfaat bagi masyarakat khususnya pengelola wisata penyu Pantai Mapak Indah karena menambah wawasan mereka mengenai praktek konservasi penyu yang baik dan benar

Steatitis in Cold-Stunned Kemp’s Ridley Sea Turtles (Lepidochelys kempii)

The pathogenesis of steatitis that infrequently occurs in cold-stunned Kemp’s ridley sea turtles (KRT; Lepidochelys kempii) has been undetermined. The objectives of this study were to investigate the clinical (n = 23) and histologic findings (n = 11) in cold-stunned KRT, and to compare plasma concentrations of α-tocopherol (vitamin E), thiobarbituric acid reactive substances (TBARS), and the TBARS to vitamin E (T/E) ratio (an assessment of oxidative stress) between cold-stunned KRT with clinically and/or histologically confirmed steatitis (n = 10) and free-ranging KRT (n = 9). None of the cold-stunned turtles had clinically detectable steatitis at admission, and the median number of days to diagnosis of steatitis was 71 (range 33–469). Histologic findings of affected adipose tissue included heterophilic (n = 9) and/or histiocytic (n = 5) steatitis, fat necrosis (n = 7), myonecrosis (n = 2), and intralesional bacteria (n = 6). Cold-stunned KRT had significantly lower plasma vitamin E concentrations (median = 3.5 nmol/g), lower plasma TBARS concentrations (median = 1.6 nmol/g), and higher T/E ratios (median = 0.37), than controls (208.8 nmol/g; 2.1 nmol/g; 0.01, respectively). These results suggest a multifactorial etiology for the development of steatitis in KRT during rehabilitation, including tissue injury, septicemia, and various factors resulting in imbalances of anti-/oxidative status. By highlighting the need to provide more effective vitamin E supplementation, and the need to re-assess specific components of the diet, this study may lead to reduced incidence and improved medical management of steatitis in cold-stunned sea turtles.

Comparison of 2 glucose analytical methodologies in immature Kemp’s ridley sea turtles: dry chemistry of plasma versus point-of-care glucometer analysis of whole blood

Blood glucose measurements provide important diagnostic information regarding stress, disease, and nutritional status. Glucose analytical methodologies include dry chemistry analysis (DCA) of plasma and point-of-care (POC) glucometer analysis of whole blood; however, these 2 methods differ in cost, required sample volume, and processing time. Because POC glucometers use built-in equations based on features of mammalian blood to convert whole blood measurements to plasma equivalent units, obtained glucose data must be compared and validated using gold-standard chemistry analytical methodology in reptiles. For in-water, trawl-captured, immature Kemp’s ridley sea turtles ( Lepidochelys kempii) from Georgia, USA, we observed significant, positive agreement between the 2 glucose determination methods; however, the glucometer overestimated glucose concentrations by 1.4 mmol/L on average in comparison to DCA and produced a wider range of results. The discordance of these results suggests that POC glucometer glucose data should be interpreted in the context of methodology- and brand-specific reference intervals along with concurrent packed cell volume data.