Histology of dart tag insertion sites in the epaulette shark

Samples of dermal and epidermal tissues of epaulette sharks Hemiscyllium ocellatum were examined histologically to assess damage caused by tagging. Tissues from around tag sites were removed at time intervals ranging from 100 min to 284 days post-tagging. These samples showed acute and chronic responses to tagging. Acute responses consisted of localized tissue breakdown and haemorrhaging, and occurred within the first few hours after tag insertion. At 10 h post-tagging, an intermediate response was apparent. This phase was characterized by further haemorrhaging and red and white blood cell movement into the wound area. The chronic response observed in the 10–284-day post-tagging samples was characterized by fibrous tissue formation to sequester the tag. This tissue presumably protects the adjacent musculature from further trauma produced by movement of the tag and provides a continuous barrier between the internal and external milieu. Tissue repair appeared to progress consistently in all specimens and no secondary infections at the tag site were seen. Tagging produced only localized tissue disruption and did not appear to be detrimental to the long term health of individual sharks. Our findings show that spaghetti style dart tagging is an acceptable method for marking individuals (40–75+ cm total length) of this species.     

Observations on the process of wound repair in penaeid shrimp

The Petersen disk tag is a standard mark for penaeid shrimp, and attachment of the tag involves the insertion of a stainless steel pin through the shrimp's abdomen, resulting in a relatively large puncture wound. The wound healing process first observed at 24 hr post-tagging showed a pronounced hemocytic infiltration of the wound area. Hemocytes in contact with the pin became fusiform, began adhering to one another, and formed several concentric layers around the pin. Scattered foci of bacteria or nercrotic tissue in the vicinity of the would also became encapsulated by concentric layers of fusiform hemocytes, thereby forming nodules. Melanin appeared in association with the layers of hemocytes nearest the pin and in the nodules. Hemocytic infiltration was lollowed by the appearance of fibrocytes and the deposition of collagenlike fibers along the would channel 48 hr after wounding. Involution of epidermis and consequential cuticular involution into the would channel began at 96 hr after wounding. Complete epidermal and cuticular formation along the wound occurrd by 384 hr post-tagging.     

EFFECTIVENESS OF VARIOUS CATTLE EAR TAGS AS MARKERS FOR WEDDELL SEALS

Methods for estimating tag loss and sightability of tags are presented. Rates of tag loss were estimated for metal and several varieties of plastic cattle ear tags in a long-term population study of Weddell seals. Tags were lost at a higher rate in the first year after application than in subsequent years, and pups lost tags at a higher rate than older seals given the same tag types. Medium-sized plastic tags (Dalton, Allflex and Fearing) were retained at higher rates and were overlooked less frequently than metal tags on adult seals. Dalton Jumbo Rototags showed the lowest loss rates on pups in comparison to larger, flexible plastic tags, but were harder to read in the field.     

Molecular response of nasal mucosa to therapeutic exposure to broad-band ultraviolet radiation

Ultraviolet radiation (UVR) phototherapy is a promising new treatment for inflammatory airway diseases. However, the potential carcinogenic risks associated with this treatment are not well understood. UV-specific DNA photoproducts were used as biomarkers to address this issue. Radioimmunoassay was used to quantify cyclobutane pyrimidine dimers (CPDs) and (6–4) photoproducts in DNA purified from two milieus: nasal mucosa samples from subjects exposed to intranasal phototherapy and human airway (EpiAirway™) and human skin (EpiDerm™) tissue models. Immunohistochemistry was used to detect CPD formation and persistence in human nasal biopsies and human tissue models. In subjects exposed to broadband ultraviolet radiation, DNA damage frequencies were determined prior to as well as immediately after treatment and at increasing times post-treatment. We observed significant levels of DNA damage immediately after treatment and efficient removal of the damage within a few days. No residual damage was observed in human subjects exposed to multiple UVB treatments several weeks after the last treatment. To better understand the molecular response of the nasal epithelium to DNA damage, parallel experiments were conducted in EpiAirway and EpiDerm model systems. Repair rates in these two tissues were very similar and comparable to that observed in human skin. The data suggest that the UV-induced DNA damage response of respiratory epithelia is very similar to that of the human epidermis and that nasal mucosa is able to efficiently repair UVB induced DNA damage.     

Retention of loop,monel, and passive integrated transponder tags by wild,free‐ranging Lake Sturgeon (Acipenser fulvescens Rafinesque, 1817)

Tagging or marking of fish is instrumental to fisheries biologists and managers seeking to distinguish groups of fish, track movement or migration patterns, and monitor population characteristics. However, tag loss can inhibit the ability of biologists and managers to reach these objectives. The ability of Lake Sturgeon to live for long periods of time and reach large sizes, in combination with their dynamic spawning activity, requires tags to be retained under a variety of environmental and physically demanding conditions. This study evaluated tag retention of loop, monel, and passive integrated transponder (PIT) tags on wild, free‐ranging Lake Sturgeon in Lake Huron and the St. Clair – Detroit River System. Lake Sturgeon in this study were double‐tagged with both a PIT tag under one of the three anterior‐most dorsal scutes and an external tag (loop or monel) at the base of the dorsal fin. Fish were at large for up to 16 years. Overall, tag loss for PIT tags was 1% followed by monel tags at 12% and loop tags at 36%. Tag loss for loop tags was higher when the initial length of Lake Sturgeon tagged was smaller. Tag loss for monel tags increased with time at large but was not related to length at initial tagging. Monel tags left behind abrasion marks when attached to smaller Lake Sturgeon. PIT tag retention was higher than reported in previous studies that tagged other sturgeon species in different body locations. Monel tag retention was higher than other external tag types evaluated in previous studies while loop tags had similar retention rates to external tag types. Most previous studies on tag retention of sturgeon species were of shorter duration and conducted in laboratory settings, therefore loop tags may have performed more favorably during studies under short term laboratory settings. Results of this study suggest that PIT tags inserted below dorsal scutes represent a viable option for long‐term tracking of Lake Sturgeon. Monel tags attached at the base of the dorsal fin also seem to be a viable option relative to other external tag types, but should be limited to larger sturgeon as they can leave behind abrasion marks.     

Vanadate ingestion increases the gain in wound breaking strength and leads to better organized collagen fibers in rats during healing

Repair of incision wounds closed by suturing is evaluated by the progressive gain in wound breaking strength. Previously the closure of open wounds in rats ingesting vanadate, an inhibitor of tyrosine phosphate phosphatases, was shown to occur with deposition of more uniformly organized collagen fiber bundles. The hypothesis of this study was that deposition of more uniformly organized collagen fibers would enhance the gain in wound breaking strength of incisional wounds. Six adult rats received vanadate-supplemented saline drinking water for 1 week before placement of two 6-cm, parallel, suture-closed wounds on their backs. Six control rats received identical wounds and were given saline drinking water. The drinking water regimen was continued for 1 week after wounding, and then wound strength was tested with a tensiometer and tissue samples were obtained for histologic evaluation. Wound breaking strength doubled in vanadate-treated rats compared with controls. Bright-field and polarized light microscopy showed that the connective tissue matrix of granulation tissue from control rats was oriented perpendicular to the surface of the skin. In contrast, the connective tissue matrix of granulation tissue from vanadate-treated rats was oriented parallel to the skin surface. The gap in granulation tissue between the edges of the wounds in the vanadate-treated rats was greater than that in controls. Electron microscopy showed that wounds in the vanadate-treated contained uniform collagen fibers that were 20 percent greater in diameter and more evenly spaced than they were in controls. It is proposed that these changes in the organization of collagen fibers within incisional wounds were responsible for the increased wound breaking strength observed in rats ingesting vanadate.     

Capturing Tissue Repair in Zebrafish Larvae with Time-lapse Brightfield Stereomicroscopy

The zebrafish larval tail fin is ideal for studying tissue regeneration due to the simple architecture of the larval fin-fold, which comprises of two layers of skin that enclose undifferentiated mesenchyme, and because the larval tail fin regenerates rapidly within 2-3 days. Using this system, we demonstrate a method for capturing the repair dynamics of the amputated tail fin with time-lapse video brightfield stereomicroscopy. We demonstrate that fin amputation triggers a contraction of the amputation wound and extrusion of cells around the wound margin, leading to their subsequent clearance. Fin regeneration proceeds from proximal to distal direction after a short delay. In addition, developmental growth of the larva can be observed during all stages. The presented method provides an opportunity for observing and analyzing whole tissue-scale behaviors such as fin development and growth in a simple microscope setting, which is easily adaptable to any stereomicroscope with time-lapse capabilities.     

The response of the atrium to direct mechanical wounding in the adult heart of the newt,Notophthalmus viridescens

Summary Wound repair and proliferation were examined in the injured newt atrium with light- and electron-microscopic techniques including autoradiography. Hearts were injured by removing a piece approximately 0.5 mm² of the atrial wall. The five-day wound was an endothelial and mesothelial-lined blood clot bordered by a 150-m necrotic zone. Repair progressed from the periphery inward with areas of macrophage activity replaced by fibroblasts and connective tissue. The wound at 25 days consisted of a scar with few myocytes. There was no difference in the proliferative behavior between the right and left atria. Proliferative cells were localized to a 500-m reactive zone surrounding the wound. The maximum mesothelial cell thymidine-labeling index of 20.5% and mitotic index of 1.4% were seen 5 days after injury. The peak connective tissue cell thymidine-labeling index of 10.2% and mitotic index of 0.4% were seen 10 days after wounding. The peak thymidine-labeling index of 9.8% for myocardial cells was recorded 10 days after injury with a mitotic index of 0.2%. Proliferation returned to control levels by 25 days post-injury. Electron microscopy demonstrated that myocytes engaged in DNA synthesis were indistinguishable from control myocytes. Z-band material was not observed in mitotic myocytes, but myofilaments and junctions were present.     

Effects of Smart Position Only (SPOT) tag deployment on white sharks Carcharodon carcharias in South Africa

We present 15 individual cases of sub-adult white sharks that were SPOT tagged in South Africa from 2003-2004 and have been re-sighted as recently as 2011. Our observations suggest SPOT tags can cause permanent cosmetic and structural damage to white shark dorsal fins depending on the duration of tag attachment. SPOT tags that detached within 12-24 months did not cause long term damage to the dorsal fin other than pigmentation scarring. Within 12 months of deployment, tag fouling can occur. After 24 months of deployment permanent damage to the dorsal fin occurred. A shark survived this prolonged attachment and there seems little compromise on the animal's long term survival and resultant body growth. This is the first investigation detailing the long term effects of SPOT deployment on the dorsal fin of white sharks.     

Effects of passive integrated transponder tags on short-term feeding patterns in European perch (Perca fluviatilis)

We tested the feeding behaviour of small European perch (Perca fluviatilis) in a laboratory study during the first 24 h after handling and 23 mm passive integrated transponder (PIT) tag implantation. Feeding commenced almost immediately following tagging and overall feeding patterns were unaffected by tagging. However, untagged perch had more feeding events than PIT-tagged individuals. This discrepancy could be attributed to post-tagging effects or/and reduced room for food due to the presence of the tag in the body cavity.     

Predicting the factors influencing the inter- and intraspecific survival rates of riverine fishes implanted with acoustic transmitters

Biotelemetry is a central tool for fisheries management, with the implantation of transmitters into animals requiring refined surgical techniques that maximize retention rates and fish welfare. Even following successful surgery, long-term post-release survival rates can vary considerably, although knowledge is limited for many species. The aim here was to investigate the post-tagging survival rates in the wild of two lowland river fish species, common bream Abramis brama and northern pike Esox lucius, following their intra-peritoneal double-tagging with acoustic transmitters and passive integrated transponder (PIT) tags. Survival over a 2-year period was assessed using acoustic transmitter data in Cox proportional hazards models. Post-tagging survival rates were lowest in the reproductive periods of both species, but in bream, fish tagged just prior to spawning actually had the highest subsequent survival rates. Pike survival was influenced by sex, with males generally surviving longer than females. PIT tag detections at fixed stations identified bream that remained active, despite loss of an acoustic transmitter signal. In these instances, loss of the acoustic signal occurred up to 215 days post-tagging and only during late spring or summer, indicating a role of elevated temperature, while PIT detections occurred between 18 and 359 days after the final acoustic detections. Biotelemetry studies must thus always consider the date of tagging as a fundamental component of study designs to avoid tagged fish having premature end points within telemetry studies.     

Vernalization,photoperiod and GA3 interact to affect flowering of Japanese radish (Raphanus sativus Chinese Radish Jumbo Scarlet)

Raphanus sativus L. Chinese Radish Jumbo Scarlet has characteristics that make it an excellent plant model for vernalization studies. This study further characterizes flower induction of R. sativus Chinese Radish Jumbo Scarlet. Seed were imbibed in distilled water containing 0, 10⁻⁵ M or 10⁻³ M GA₃ for 24 h and were then exposed to 6 ± 0.5°C (vernalized) for 0, 2, 4, 6, or 8 days. Seedlings were then grown under a short- (8 h) or long-day photoperiod (8 h with or without a 4-h night interruption; 2200–0200 h). Of unvernalized plants grown under long- and short-day conditions, 45 and 3% flowered, respectively. Saturation of the vernalization response occurred after a 4- or 8-day vernalization treatment when plants were placed under long- or short-days, respectively. Basal leaf number and days to anthesis decreased when seeds were cooled for 2 or 4 days and were imbibed with 10⁻³ M GA₃ compared to distilled water only. These data indicate that R. sativus Chinese Jumbo Scarlet has principally an obligate vernalization requirement when grown under short-days. GA₃ application only facilitated flowering when the length of the vernalization treatment was marginal. Taken together, these data support the use of this plant as a model plant for identifying vernalization responses under short-day conditions.     

Evaluation of passive integrated transponder tag retention from two tagging locations in juvenile pallid sturgeon

Population augmentation and propagation of pallid sturgeon (Scaphirhynchus albus) have been a focus of scientists since the mid‐1990s. Various tag types have been used to gain a better understanding of population characteristics, stocking success, and movement patterns. Passive integrated transponder (PIT) tags have been consistently used since the inception of recovery efforts to mark age‐1 and older pallid sturgeon. This tag has been successfully used with other sturgeon species, and tagging protocols for pallid sturgeon were developed in response to those successes. Tag retention rates in pallid sturgeon have been reported as variable or unknown and there has been no formal study to determine PIT tag retention in pallid sturgeon, particularly with age‐1 individuals that are propagated in hatcheries to be stocked into the wild. The objective was to evaluate retention rates of PIT tags inserted into the operculum and along the base of the dorsal fin of age‐1 pallid sturgeon (total fork length [FL] range = 214–358 mm). Tagged sturgeon (n = 80) were held in a large holding tank and inspected for tag loss twice a month for a total of 189 days. Final tag retention was 83% for tags inserted into the operculum and 85% for tags inserted near the dorsal fin. Tags shed from the operculum were from smaller fish (mean FL = 265 mm) and losses occurred during the first 60 days of the experiment. Tags shed from the dorsal site were predominantly from larger individuals (mean FL = 305 mm) and losses were continual throughout the experiment. Future research should determine size‐specific tag retention rates so that hatcheries can maximize retention in either tag placement location.     

Developmental structure of the vertebral column,fins, scutes and scales in bester sturgeon,a hybrid of beluga Huso huso and sterlet Acipenser ruthenus

In the larval bester, a hybrid sturgeon of beluga Huso huso and sterlet Acipenser ruthenus, development of cartilage around the notochord began 7 days post hatch (dph) (14·0 mm, total length, L_T). The vertebral cartilage develops in the following sequence: basidorsals and basiventrals, neural canals, neural spines and ribs. The development of ribs remained incomplete in the largest specimen (181 dph, 179 mm L_T) that was examined. Endoskeletal development of the fins began 4 dph for the dorsal and anal fins, 6 dph for the pectoral fin and 10 dph for the caudal and pelvic fins. Complete elements of all fins were observed by 91 dph and complete ossification of fin rays was observed by 122 dph in the double‐stained specimens. Observation of the histological sections, however, suggested that ossification occurred soon after the formation of the organic matrix in the fin rays. Dorsal scutes were first visible by 25 dph, followed by the lateral and ventral scutes, which were visible by 37 and 44 dph, respectively. The number of scutes was fixed at 44, 59 and 91 dph and ossification was complete by 59 (dorsal) and 91 dph (lateral and ventral scutes) in the double‐stained specimens. Ossification occurred soon after the formation of the scute organic matrix in the histological sections. Four types of scales were observed in the H. huso×A. ruthenus hybrid. Median predorsal, preanal and small scales on the anterior section of the head were visible by 59 dph. Scales on the caudal fin were visible by 91 dph and a variable assemblage of scales anterior to the anal fin was visible by 122 dph. Both the scutes and scales developed in a process that is similar to that of intramembranous ossification.     

Myeloid-derived growth factor regulates neutrophil motility in interstitial tissue damage

Neutrophil recruitment to tissue damage is essential for host defense but can also impede tissue repair. The cues that differentially regulate neutrophil responses to tissue damage and infection remain unclear. Here, we report that the paracrine factor myeloid-derived growth factor (MYDGF) is induced by tissue damage and regulates neutrophil motility to damaged, but not infected, tissues in zebrafish larvae. Depletion of MYDGF impairs wound healing, and this phenotype is rescued by depleting neutrophils. Live imaging and photoconversion reveal impaired neutrophil reverse migration and inflammation resolution in mydgf mutants. We found that persistent neutrophil inflammation in tissues of mydgf mutants was dependent on the HIF-1α pathway. Taken together, our data suggest that MYDGF is a damage signal that regulates neutrophil interstitial motility and inflammation through a HIF-1α pathway in response to tissue damage.     

Wound healing in tadpole tailfin pieces in vitro.

Explants of tail fins from R. catesbeiana tadpoles undergo reepithelialization of their cut surfaces (healing) when cultured in vitro in Hanks' balanced salt solution at 22 degrees C. Healing is initiated early and closure of the wound is complete by 12 to 24 hours. Morphogenesis continues for several days as further reorganization and migration of epidermal cells from the regions adjacent to the wound margins take place. The addition of serum to the culture media improves the general appearance of these tissues and promotes healing. The rate of healing is affected by temperature. Tail fins maintained at 10 degrees C do not heal while fins maintained at 30 degrees and 37 degrees, although healing more rapidly than at 22 degrees, undergo progressive degeneration in culture. Epidermal cell movements were also studied in explants consisting of a combination of intact tail fin plus tail fin deprived of its epithelium. Rapid and extensive migration of epidermal cells from the intact tail fin across the collagen lamella of the stripped fin is observed.     

A non‐invasive dolphin telemetry tag: Computer design and numerical flow simulation

The impact of devices attached to animals remains a challenge in telemetry studies of dolphins. It was hypothesized that the hydrodynamic design of a tag could provide stable attachment to the dorsal fin by means of resultant hydrodynamic force appearing when a dolphin is swimming. To verify this hypothesis the computer fluid dynamics (CFD) study of tag performance was carried out. A virtual model presenting authentic geometry of a dolphin with tag attached to the dorsal fin was constructed. The same model without tag was used as a reference object to calculate tag impact as regards drag, lift, and moments coefficients. Flow around the models was simulated for the range of velocities as well as the ranges of pitch and yaw angles. It was shown that in 33 of 35 CFD scenarios the streamlined shape of a tag generates the lift force that facilitates keeping a tag attached to the fin. Throughout the set of calculations the tag‐associated drag coefficient does not exceed 4%, which indicates low impact. Data obtained present a baseline for the further development of non‐invasive dolphin telemetry tags.     

An essential role for TH2-type responses in limiting acute tissue damage during experimental helminth infection

Helminths induce potent T helper 2 (TH2)-type immune responses that can mediate worm expulsion, but the role of this response in controlling the acute tissue damage caused by migrating multicellular parasites through vital tissues remains uncertain. We used a helminth infection model in which parasitic nematode larvae migrate transiently through the lung, resulting in hemorrhage and inflammation. We found that IL-17 initially contributed to inflammation and lung damage, whereas subsequent IL-4 receptor (IL-4R) signaling reduced elevations in IL-17 mRNA levels, enhanced the expression of insulin-like growth factor 1 (IGF-1) and IL-10 and stimulated the development of M2 macrophages, all of which contributed to the rapid resolution of tissue damage. These studies indicate an essential role for TH2-type immune responses in mediating acute wound healing during helminth infection.