Galactose-Binding C-Type Lectin Promotes Cellular Aggregation of Coelomocytes in Sea Cucumber

Echinoderms have a large coelomic cavity containing coelomocytes. When the coelomic fluid is removed from the cavity, the cells aggregate immediately. We found that a fraction or an extract of the intestine of the sea cucumber, Apostichopus japonicus, markedly accelerated cellular movement and aggregation on a glass slide, and this effect was clearly inhibited by galactose. We successfully purified the aggregation-promoting factor, a 16 kDa protein, from the intestine. TOF-MS analysis followed by de novo sequencing revealed that the protein is a C-type lectin. RNA-seq data and cDNA cloning demonstrated the factor to be a novel lectin, named AjGBCL, consisting of 158 aa residues in the mature form. Microscopic observation revealed that most of the aggregating cells moved toward aggregates and not to an intestinal fragment, suggesting that AjGBCL is not a chemoattractant but a cellular aggregation-inducing factor that may induce aggregates to release chemoattractant. We report, for the first time, an endogenous molecule that promotes coelomocyte aggregation in echinoderms.

Prevalence of Anisakis Larvae in Different Fish Species in Southern Albania: Five-Year Monitoring (2016–2020)

Anisakidae are nematodes that commonly parasitize in the coelomic cavity and viscera of several fish species. They can be found in flesh, which is why they have an important economic and public health impact. The aim of the current work was to assess the presence and prevalence of Anisakis larvae in fish species caught in the coastal area of the Karaburun Peninsula in Vlora Bay (Albania). A total of 856 of wild teleosts and 219 specimens of farmed fish were collected over a 5-year period (from 2016 to 2020). The results showed that out of a total of 1075 analyzed samples, 361 (33.58%) were parasitized with L3 larvae. In particular, only Solea vulgaris returned negative results, while Sparus aurata, Dicentrarchus labrax, and Sardinella aurita showed the lowest prevalence (4.55%, 9.17%, and 10.53%, respectively) and mean abundance (0.84, 1.19, and 0.92, respectively). Conversely, Scomber japonicus and Scomber scombrus showed the highest prevalence (74.07% and 68.00%, respectively) and mean abundance (188.24 and 249.82, respectively). The data suggest that the coastal area of the Karaburun Peninsula (southern Albania) may be a high-risk area for zoonotic diseases, and the consumption of raw or undercooked fish caught in the Vlora district could result in the acquisition of human anisakiasis. For these reasons, it is necessary to improve the surveillance plan.

Detailed morphology of tentacular apparatus and central nervous system in Owenia borealis (Annelida, Oweniidae)

The Oweniidae are marine annelids with many unusual features of organ system, development, morphology, and ultrastructure. Together with magelonids, oweniids have been placed within the Palaeoannelida, a sister group to all remaining annelids. The study of this group may increase our understanding of the early evolution of annelids (including their radiation and diversification). In the current research, the morphology and ulta-anatomy of the head region of Owenia borealis is studied by scanning electron microscopy (SEM), 3D reconstructions, transmission electron microscopy (TEM), and whole-mount immunostaining with confocal laser scanning microscopy. According to SEM, the tentacle apparatus consists of 8–14 branched arms, which are covered by monociliary cells that form a ciliary groove extending along the oral side of the arm base. Each tentacle contains a coelomic cavity with a network of blood capillaries. Monociliary myoepithelial cells of the tentacle coelomic cavity form both the longitudinal and the transverse muscles. The structure of this myoepithelium is intermediate between a simple and pseudo-stratified myoepithelium. Overall, tentacles lack prominent zonality, i.e., co-localization of ciliary zones, neurite bundles, and muscles. This organization, which indicates a non-specialized tentacle crown in O. borealis and other oweniids with tentacles, may be ancestral for annelids. TEM, light, and confocal laser scanning microscopy revealed that the head region contains the anterior nerve center comprising of outer and inner (=circumoral) nerve rings. Both nerve rings are organized as concentrated nerve plexus, which contains perikarya and neurites extending between basal projections of epithelial cells (radial glia). The outer nerve ring gives rise to several thick neurite bundles, which branch and extend along aboral side of each tentacle. Accordingly to their immunoreactivity, both rings of the anterior nerve center could be homologized with the dorsal roots of circumesophageal connectives of the typical annelids. Accordingly to its ultrastructure, the outer nerve ring of O. borealis and so-called brain of other oweniids can not be regarded as a typical brain, i.e. the most anterior ganglion, because it lacks ganglionic structure.

Rehabilitation of a green sea turtle (Chelonia mydas) after collision with motorboat in the archipelago of Fernando de Noronha, Brazil

A juvenile green sea turtle (Chelonia mydas) of undetermined sex was rescued on October 16, 2019, in the archipelago of Fernando de Noronha, Pernambuco, Brazil. The animal was presence of active bleeding resulting from two transverse fractures in the carapace, with exposure of the coelomic cavity. Radiological and hematological tests were performed. The animal received therapeutic support and surgical reconstruction of the carapace. The animal was medically cleared and reintroduced into the natural environment 120 days after it was rescued. The protocol used here for both the rehabilitation and the surgical procedure was concluded to be effective.

The non-brain anterior nerve center and tentacle crown structure of Owenia borealis (Annelida, Oweniidae): the evolution of the nervous system and tentacles in Bilateria

The Oweniidae are marine annelids with many unusual features of organ system, development, morphology, and ultrastructure. Together with magelionds, oweniids have been placed within the Palaeoannelida, a sister group to all remaining annelids. The study of this group may increase our understanding of the early evolution of annelids (including their radiation and diversification) and of the morphology of the last common bilaterian ancestor. In the current research, scanning electron microscopy revealed that the tentacle apparatus consists of 10 branched arms. The tentacles are covered by monociliary cells that form a ciliar groove that extends along the oral side of the arm base. Light, confocal, and transmission electron microscopy revealed that head region contains two circular intraepidermal nerves (outer and inner) that give rise to the neurites of each tentacle, i.e., intertentacular neurites are absent. Each tentacle contains a coelomic cavity with a network of blood capillaries. Monociliar myoepithelial cells of the tentacle coelomic cavity form both the longitudinal and the circular muscles. The structure of this myoepithelium is intermediate between simple and pseudo-stratified myepithelium. Overall, tentacles lack prominent zonality, i.e., co-localization of ciliary zones, neurite bundles, and muscles. This organization, which indicates a non-specialized tentacle crown in O. borealis and other oweniids with tentacles, is probably ancestral for annelids and for all Bilateria. The outer circular nerve of O. borealis is a dorsal medullary commissure that apparently functions as an anterior nerve center and is organized at the ultrastructural level as a stratified neuroepithelium. Given the hypothesis that the anterior nerve center of the last bilateral ancestor might be a diffuse neural plexus network, these results suggest that the ultra anatomy of that plexus brain might be a stratified neuroepithelium. Alternatively, the results could reflect the simplification of structure of the anterior nerve center in some bilaterian lineages.

Eventration in Green Iguana (Iguana iguana)

Background: The reptile class could be considered one of the biggest vertebrate groups and are divided in orders and suborders according to their characteristics. These animals’ maintenance in captivity, either at home, captive bred or at zoos, can generate risk to their health, if the required cares are not given for each respective species. The lack of individual cares could lead to bone and muscular diseases and to traumatic lesions in soft tissues, mainly in the coelomic cavity. The report that is being presented aims to describe the case of a green iguana (Iguana iguana) that presented an increase of volume in the coelomic cavity. The animal belongs to the squad of the Zoo “Dr. Fábio de Sá Barreto”.Case: A green iguana arrived at the Zoo in February 2019 coming from another Zoo, with already an increase of volume in the coelomic cavity. The animal was put in quarantine and later on, it was put in display at a terrarium in the Zoo considered adequate to reptiles, with another seven green iguanas along with an argentine tegu (Salvator rufescens). Their feed was offered in the morning and was composed of fruits, vegetables and flowers like hibiscus. In the end of July 2019, it was reported by the attendant that the animal was presented with anorexia and prostration, and these symptoms progressed to neurologic signs, as it walked in circles. So, the animal was evaluated by the Zoo veterinarians and on exam they noticed lethargy, dehydration, absence of reflexes (pupillary, eyelid and painful), locomotion difficulty and when the iguana moves, it walks in circles. The increase in volume had the same size as reported in February and a soft consistency. After that, the animal was interned and treated according to the symptoms and the clinical evolution. Ten days after the hospitalization, the animal was still not eating, and locomotion stopped completely. It was performed in an ultrasonographic exam evaluating all the coelomic cavity, in which a great anechoic area was visualized, and a true hernia was diagnosed. However, the content of the hernia was not identified. In the next day, the animal died, and, in the necropsy, it was possible to verify that the increase in volume was actually a bladder eventration. The eventration occurred due to a laceration in the coelomic cavity musculature that allows the passage of the bladder to the subcutaneous space and its incarceration. So, the elimination of the urine and of nitrogen compounds was difficult and a large accumulation of uric acid from the bladder to the urodeo.Discussion: Iguana iguana is a uricotelic animal, which means that the main nitrogenous waste product is uric acid. Nevertheless, ammonia is also eliminated in less quantity, because of the excess of protein in the diet. These animals eliminate around 98 to 99% of the nitrogen compounds as uric acid and less than 1% as ammonia, which prove that it is possible for the accumulation of ammonia in reptiles, if any obstacle in its elimination exists. The excess of ammonia is extremely toxic to the organism, leading to emesis, irritability, lethargy, anorexia, ataxia, motor difficulties, behavioral and neurological changes, and could progress to coma or even death. The bladder incarceration reported in this case, made it impossible for the excretion of urine, uric acid and ammonia, and these compounds remained accumulated. So, the clinical signs, along with the necropsy findings, were suggestive of intoxication by ammonia accumulation which could be responsible for the signs presented by the animal and the evolution to neurologic symptoms, coma and death.

Oviductal prolapse associated with a leiomyoma in a Hermann’s tortoise (Testudo hermanni)

A twelve-year-old, female Hermann’s tortoise (Testudo hermanni) was presented with an oviductal prolapse after showing tenesmus for three days prior to presentation. Radiography revealed a soft tissue opacity in the caudal coelomic cavity that ultrasonographically presented as a congested, invaginated tubular structure and was deemed to comprise the left oviduct. After repositioning the oviductal prolapse, coelioscopy confirmed that the invaginated structure consisted of the anterior part of the left oviduct and incorporated a part of the ipsilateral mesovarium and mesosalpinx. A transplastron osteotomy was carried out to perform unilateral salpingectomy and bilateral ovariectomy. A definitive diagnosis of an oviductal leiomyoma was made based on histopathological and immunohistochemical examination of the invaginated part of the oviduct. In the present case, the authors highlight that neoplastic disorders should be considered as a cause of oviductal invagination and prolapse in chelonians. To the authors’ knowledge, this is the first report of the diagnosis and successful treatment of an oviductal leiomyoma in a Hermann’s tortoise.

Surgical management of oesophageal foreign body using a minimally invasive oesophagotomy technique (MIOT) in an Indian Flap-shell turtle (Lissemys punctata) - a case report

Accidental ingestion of fish hooks is common among turtles. Several techniques are available for retrieving fish hooks, ranging from simple non-invasive techniques to more complicated and invasive surgical procedures. The purpose of this case report is to document the successful surgical retrieval of an oesophageal foreign body (fish hook) using a minimally invasive oesophagotomy technique (MIOT) in an Indian Flap-shell turtle (Lissemys punctata). The adult male Indian Flap-shell turtle (Lissemys punctata) presented with a nylon thread protruding from its mouth. The dorsoventral radiographs confirmed the presence of a foreign body in the caudal oesophagus, just outside the coelomic cavity. After the induction of general anaesthesia, a sterile artificial insemination (AI) sheath was passed intraorally through the fishing line until it reached the hook. The fish hook was then retrieved by making a 5 mm incision at the point where the tip of the AI sheath was felt. The incision in the oesophagus and skin was closed in a routine manner. The turtle was kept in secure captivity for 20 days, following which it was released to its natural habitat. The oesophagotomy technique used in this case reduces the size of the incision considerably, thereby decreasing the possibility of postoperative complications.

Case Report: Multiple Cavernous Pericardial Lymphangioma (Pericardial Lymphangiomatosis) in a Captive Peregrine Falcon (Falco peregrinus brookei)

A 12-year-old female peregrine falcon (Falco peregrinus brookei) from a private raptor breeding facility that presented a good body condition, died suddenly without showing previous symptoms. At necropsy, in the coelomic cavity, multiple cystic structures demarcated by a thin transparent wall and filled with a serous content were observed. They were firmly adhered to the cranial part of the epicardium and adjacent tissues and occupied the entire thoracic area of the coelomic cavity. Microscopically, emerging simultaneously from several areas the epicardium, multiple irregular channels and cystic spaces, lined by a single endothelial cell layer and separated by fibrovascular septa containing smooth muscle tissue, were observed. Immunohistochemical examination revealed that the neoplastic endothelial cells positively immunolabelled for the pan-endothelial marker factor VIII-related antigen but immunostained negative for cytokeratins (PCK26) while strong positivity for sarcomeric α-smooth muscle actin (α-SMA) was detected in the cystic walls. Based on the morphological and immunohistochemical findings, lesions were determined as consistent with a multiple cavernous pericardial lymphangioma, or pericardial lymphangiomatosis, a rare vascular neoplasm. The animal also showed a diffuse chronic perihepatitis, a necrotic area in the liver and foci of cartilaginous metaplasia and calcification in the aorta and vena cava. Literature review, particularly on the epidemiology of lymphangioma, demonstrated the rarity of this tumor in the different animal species and in this location, particularly in birds, being the first report of this type of tumor in a peregrine falcon.

Use of Cost-effective Vacuum-assisted Closure Technique for Shell Fracture Repair and Coelomic Cavity Rupture Healing in a Chelonia carbonaria

Background: The vacuum-assisted closure (VAC) therapy system has been used as a noninvasive wound management technique for shell damage in Chelonians. These animals are excellent candidates for VAC therapy because of their unique shell anatomyconsists of dermal bones, which make bandage placement easier. Beyond that, they are suited for this technique behaviorally, because they are not inclined to remove the vacuum system intentionally. Considering the possibility of Testudines shell repair with the use of less invasive techniques that result in additional dermal bone lesions, the objective of the present study is to describe the adaptation of a vacuum dressing protocol using low-cost and easily accessible materials for post-traumatic shell healing of a specimen of Chelonoidis carbonaria.Case: A specimen of tortoise (Chelonoidis carbonaria), a male, weighing 630 g, approximately 3-year-old, was received at the Veterinary Hospital - Wild Animals Sector in the Federal University of Pará (UFPA) because of being run over by a vehicle. On physical examination, fractures of the dermal plaques and underlying bone structures were found, with rupture of the coelomic cavity. In addition, there was exposure and incarceration of an intestinal loop, with the presence of bleeding. According to the findings of the physical screening examination, the patient's prognosis was defined as good, as described in the literature that specifically focused on chelonian shell injuries. The animal was sent to the diagnostic imaging sector. Then, drug therapy was provided for pain control, vitamins were administered for nutritional support, and antibiotics and anti-inflammatory drugs were instituted. For the closure of the shell and coelomic cavity, a bandage was used with the VAC therapy system adapted as described for Chelonians in previous reports. After a complete osteosynthesis and closure of the coelomic cavity, repair of the integumentary component of the shell was possible. For this procedure, the animal did not need to be anesthetized. The patient was physically restrained by being placed in suspension on a support with a diameter smaller than the plastron. Complete asepsis of the shell was necessary. On top of the lesion, a polypropylene mesh and color less dental resin were applied. The animal continued to be evaluated after the repair to check for potential complications. This procedure ensured that the repaired plates remained stable. Furthermore, the animal did not seem to have any discomfort with the resin when moving, so the animal was discharged after 25 days of hospitalization.Discussion: Radiography was important to determine the condition of the animal and clinical prognosis, and thus, to perform the proper treatment. The VAC therapy system was successful in assisting the patient's recovery. It enabled the reduction of the healing time since shell injuries usually require four to eighteen months to heal. In this report, the healing process only required 17 days, demonstrating that the VAC therapy system is a beneficial treatment to treat traumatic injuries in Testudines. The restoration protocol of the integumentary component using dental resin is less invasive, and this type of material has been used previously by other authors. Drug treatment with aminoglycosides and sulphonamides administered prophylactically has proven to be effective and has been used successfully in reptiles. These drugs may be combined with maintenance fluid therapy to prevent adverse reactions from aminoglycosides, such as nephrotoxicity. It was concluded that the use of the VAC therapy system reduced the time of post-traumatic healing of the carapace and proved to be an innovative approach to treat traumatic injuries in Testudines in a less invasive way.