Accumulation of docosapentaenoic acid (n-3 DPA) in a novel isolate of the marine ichthyosporean Sphaeroforma arctica.

OBJECTIVE: Currently, there is lack of a consistent and highly enriched source for docosapentaenoic acid (n-3 DPA, C22:5), and this work report the isolation of microorganism that naturally produces n-3 DPA. RESULTS: In this work, we screened microorganisms in our culture collections with the goal to isolate a strain with high levels of n-3 DPA. We isolated a strain of Sphaeroforma arctica that produces up to 11% n-3 DPA in total fatty acid and has a high n-3 DPA to DHA/EPA ratio. The cell growth of the isolated strain was characterized using microscopy imaging and flow cytometer technologies to confirm the coenocytic pattern of cell divisions previously described in S. arctica. Our novel isolate of S. arctica grew more robustly and produced significantly more n-3 DPA compared to previously isolated and described strains indicating the uniqueness of the discovered strain. CONCLUSION: Overall, this work reports a first isolate n-3 DPA producing microorganism and establishes the foundation for future strain improvement and elucidation of the physiological function of this LC-PUFA for human nutrition and health.

Novel processing and staining methodology of bottlenose dolphin (Tursiops truncatus) teeth for age determination.

Age determination of bottlenose dolphins (Tursiops truncatus) is a critical tool in understanding both individual and population health. There are many methods of aging bottlenose dolphins including analysis of teeth, pectoral flipper radiographs, and epigenetics. The most common and oldest method for aging toothed cetaceans is the counting of growth layer groups (GLGs) in the teeth. Current techniques have technical and repeatability challenges. Therefore, a processing technique that results in better resolution of GLGs is needed. This study compares different decalcifications and different histochemical staining techniques. Decalcification was done using 10% EDTA, Kristensen's decalcification, and Rapid Decalcification Solution (RDO). Following decalcification and routine processing, GLGs were assessed using Hematoxylin and Eosin (H&E), hematoxylin, Giemsa, Wright-Giemsa, Toluidine Blue (T-Blue), Masson's Trichrome, and Congo Red staining techniques. Decalcification with Kristensen's and staining with Masson's Trichrome and Congo Red were determined to best highlight GLGs. This processing and staining was then applied to a sample population of 102 bottlenose dolphins that were evaluated independently and blindly by two observers. Of the 102 dolphin samples, 13 (12.7%) were unable to age due to no clear distinction or distortion between GLGs.

CLINICAL AND HISTOPATHOLOGIC OCULAR FINDINGS IN AQUARIUM-HOUSED COWNOSE RAYS (RHINOPTERA BONASUS).

Cownose rays (Rhinoptera bonasus) are susceptible to ocular disease with their prominent globes, but despite being popular animals housed in aquaria, there is little published information about their normal ocular anatomy and common pathologic ocular findings. A total of 63 live cownose rays (CNR) from three unrelated, separately housed groups had ocular examinations, and 5 adult rays were selected for ocular ultrasound. All examinations were performed out of the water, and most without anesthesia. Clinical findings were described, categorized, and scored by severity. Sixty-two of 63 rays (123 eyes) had clinical abnormalities, including 110 eyes with corneal pathology (mild = 76, moderate/severe = 34) and 74 eyes with intraocular pathology (mild = 44, moderate/severe = 30). Grey-to-white corneal opacities were the most common pathology (n = 58 rays/100 eyes) followed by cataracts (n = 41 rays/58 eyes), then persistent (or dysplastic) pupillary membranes (n = 14 rays/15 eyes). Most pathologic findings appeared inactive, but one aquarium had several CNR with active ocular pathology. There was a significant association between the diagnosis of moderate/severe corneal and intraocular pathology with age (P = 0.008 and P = 0.014, respectively) and weight (P = 0.001 and P = 0.039, respectively), as well as moderate/severe corneal pathology and group sampled (P = 0.03). There were no other significant variables identified. Additionally, histopathology of 14 eyes (11 rays) from two different facilities were examined, with keratitis (n = 8) and uveitis (n = 2) as the most common lesions. This study shows a high prevalence of pathologic ocular findings in cownose ray eyes with heavier adults more likely to be affected than lighter juveniles. Comprehensive ocular evaluation is important in this species and serial ocular exams and future studies should be pursued to monitor ocular disease progression and better understand possible etiologies.

Description of normal pulmonary radiographic findings in 55 apparently healthy juvenile Kemp's ridley sea turtles (Lepidochelys kempii).

A total of 55 digital radiographic studies from 53 individual juvenile Kemp's ridley sea turtles (Lepidochelys kempii) were retrospectively used to determine the normal radiographic anatomy of the lower respiratory tract in sea turtles that had been stranded due to hook-and-line injury and were otherwise clinically healthy. There were three or four projections available for each study: dorsoventral (DV), rostrocaudal (RoCd), and left and/or right lateral. The DV and RoCd were most conducive for assessing global lung volume and symmetry of lung volume. The DV and lateral views were most helpful for evaluating the main bronchus and its branching channels and for assessing lung margination. The RoCd view was most useful for assessing the symmetry of the lung opacity. The lateral views were most helpful for assessing the ventral margin of each lung lobe. On the lateral view, the main bronchus lay ventrally and coursed horizontally through the lung from cranial to caudal. On the DV view, the bronchus lay medially and was observed to be curvilinear coursing caudomedially. On the RoCd view, the main bronchus was located ventromedially. The RoCd view demonstrated the channels and niches end-on resulting in a reticulated or honeycomb appearance. The channels were seen as uniform striations coursing perpendicular to the main bronchus on the lateral views (vertical striations coursing dorsal to ventral) and DV views (horizontal striations coursing medially to laterally).

BARIUM SULFATE GELATIN AS A NOVEL APPROACH TO FACILITATE GASTROINTESTINAL TRACT POSITIVE CONTRAST STUDIES IN A CAPTIVE HARBOR SEAL (PHOCA VITULINA) AND CALIFORNIA SEA LIONS (ZALOPHUS CALIFORNIANUS) WITHOUT RESTRAINT.

Gastrointestinal tract contrast medium studies are a valuable diagnostic modality to evaluate gastrointestinal anatomy, motility, and pathology. Four positive contrast medium studies were performed twice on a harbor seal (Phoca vitulina) and once each on two California sea lions (Zalophus californianus) to evaluate for gastrointestinal abnormalities by using barium-impregnated gelatin. Because marine mammals in human care routinely receive plain gelatin as a component of their diet and as secondary reinforcement, feeding a barium gelatin mitigates the need for tube feeding or restraint. Operant conditioning was used to position the pinnipeds for voluntary radiographs. The barium gelatin permitted adequate evaluation of gastric emptying and intestinal transit times and good evaluation of the structure of the intestinal tract. Full evaluation of gastric anatomy was limited due to the gelatin blocks not conforming to the stomach in their solid form and the barium quickly exiting the stomach as barium gelatin blocks disintegrated and because orthogonal views could not always be acquired. Even with these limitations, barium gelatin resulted in diagnostically valuable contrast imaging in a stress-free patient setting with reduced risk of aspiration and eliminated the effects that anesthesia can have on gastric motility as well as other anesthesia-associated risks.

1H-filtered correlation experiments for assignment and determination of coupling constants in backbone labelled proteins.

The implementation of [13Calpha,13C',15N,2Halpha] labelled amino acids into proteins allows the acquisition of high resolution triple resonance experiments. We present for the first time resonance assignments facilitated by this new labelling strategy. The absence of 1JCalpha,Cbeta couplings enables us to measure 1JCalpha,C' scalar and 1DCalpha,C' residual dipolar coupling constants using modified HNCA experiments which do not suffer from sensitivity losses characteristic for 13C constant time experiments.