Mortality induced by drifting longline hooks and branchlines in loggerhead sea turtles,estimated through observation in captivity

• 1. Drifting longlines are considered a major threat to endangered sea turtle populations worldwide. However, for a number of reasons, the mortality rate of captured turtles is not known with any certainty.
• 2. Information on 409 loggerhead turtles (Caretta caretta), collected during the day‐to‐day activities of a turtle rescue centre in Lampedusa island, central Mediterranean, in the period 2001–2005 has been analysed.
• 3. Observations indicate that: (i) drifting longlines are a major cause of mortality for sea turtles in the area; (ii) in addition to the hook, the piece of line attached to it (branchline) can easily cause death if it is long enough and well‐anchored; (iii) hooks and branchlines cause death in the short and long term, respectively; (iv) a turtle with a hook in the lower oesophagus/stomach has a very low chance of surviving the combined effect of hook and branchline; (v) the mortality of turtles with a hook in the mouth or higher oesophagus is probably important, though less than that of turtles with a hook in the lower oesophagus/stomach; (vi) in the study fishery, the average mortality of a turtle caught by a drifting longline is probably much higher than 30%.
• 4. Without specific investigations on the mortality of turtles with hooks in the mouth or higher oeasophagus, which are usually removed, the mortality induced by drifting longlines will remain unknown, preventing a full understanding of the effect on population growth and the real effectiveness of conservation measures such as use of different hooks and fishing depths, and proposals for adequate fishery management measures.
• 5. The number of turtles captured by drifting longlines should be drastically reduced, and because of the above uncertainty and the socio‐economic importance of the fishery sector, an ecosystem‐based management scheme should be promoted that is not limited to addressing only the turtle issue.

Copyright © 2007 John Wiley & Sons, Ltd.     

Design and chemical synthesis of a homogeneous polymer-modified erythropoiesis protein

We report the design and total chemical synthesis of "synthetic erythropoiesis protein" (SEP), a 51-kilodalton protein-polymer construct consisting of a 166-amino-acid polypeptide chain and two covalently attached, branched, and monodisperse polymer moieties that are negatively charged. The ability to control the chemistry allowed us to synthesize a macromolecule of precisely defined covalent structure. SEP was homogeneous as shown by high-resolution analytical techniques, with a mass of 50,825 +/-10 daltons by electrospray mass spectrometry, and with a pI of 5.0. In cell and animal assays for erythropoiesis, SEP displayed potent biological activity and had significantly prolonged duration of action in vivo. These chemical methods are a powerful tool in the rational design of protein constructs with potential therapeutic applications.     

Residue level, persistence, and storage performance of citrus fruit treated with fludioxonil

The potential of postharvest dip treatments with fludioxonil (FLU) (a synthetic analogue of the bacterial metabolite of pyrrolnitrin), in controlling postharvest decay caused by Penicillium digitatum and Penicillium italicum of citrus fruit was investigated in comparison with the conventional fungicide imazalil (IMZ). The ultrastructural changes of fruit epicuticular wax was investigated as a function of water dip temperature, and the possible role of these changes was related to residue accumulation under FLU treatment. Residues retained by fruit were determined as a function of fungicide concentration, dip temperature, and fruit storage conditions. Scanning electron microscopy analysis revealed that fruit dipping in water at 30 or 40 degrees C did not cause differences in cuticular wax's ultrastructure in comparison to control fruit, while treatments at 50, 55, or 60 degrees C caused the disappearance of wax platelets, resulting in relatively homogeneous skin surface, due to partial "melting" of epicuticular wax. Residues of FLU in fruit treated at 20 or 50 degrees C were significantly correlated with the doses of fungicide applied. When equal amounts of fungicide were employed, the residue concentrations were notably higher (from 2.6- to 4-fold) in fruit treated at 50 degrees C than in fruit treated at 20 degrees C. The dissipation rate of FLU in "Salustiana" and "Tarocco" oranges was lower in fruit subjected to treatment at 50 degrees C. The minimal FLU concentration for almost complete decay control in artificially wounded fruit during 7-d storage at 20 degrees C was 400 mg/L active ingredient (ai) in fruit treated at 20 degrees C and 100 mg/L ai in fruit treated at 50 degrees C. Results on nonwounded Tarocco oranges subjected to 3 weeks of simulated quarantine conditions at 1 degrees C, plus 6 weeks of standard storage at 8 degrees C and an additional two weeks of simulated marketing period (SMP) at 20 degrees C revealed that almost complete decay control with FLU applications of 100 mg/L at 50 degrees C and 400 mg/L at 20 degrees C resulted in ca. 0.8 mg/kg FLU fruit residues, in agreement with results on wounded citrus fruit. When equal concentrations and temperatures were applied, FLU treatments were as effective as IMZ. In vitro trials showed a low sensitivity to FLU against P. digitatum and P. italicum isolates. MIC values for the complete inhibition of mycelium growth were >or=100 microg/mL, while ED(50) values ranged from 0.1 to 1 microg/mL for P. digitatum and from 1 to >100 microg/mL for P. italicum. The latter result suggests that care should be taken to avoid exclusive application of FLU in a sustainable program for management of fruit decay. However, integrating fungicide application and hot water dip may reduce the possibility of selecting fungicide-resistant populations of the pathogen, by increasing the effectiveness of the treatment.     

Pasteurized tumoral autograft as a novel procedure for limb sparing in the dog: A clinical report

OBJECTIVE: To evaluate use of a pasteurized tumoral autograft prepared from the resected primary bone neoplasm for limb sparing in a dog with distal radial osteosarcoma (OSA). STUDY DESIGN: Clinical case report. ANIMALS: A 9-year-old male Maremma shepherd dog. METHODS: After right distal radial OSA removal, the tumoral autograft was pasteurized. The excised bone segment was placed in a sterile watertight box containing sterile saline solution preheated to 65 degrees C in a water bath. The box was kept immersed in the water bath at 65 degrees C for 40 minutes to kill the tumor cells. The autograft was then fixed in the host with a plate and screws based on standard AO/ASIF technique for carpal arthrodesis. Three doses of cisplatin (70 mg/m(2) intravenously) were administered, 3 weeks apart; the initial dose was administered the day after surgery. RESULTS: The autograft was incorporated in a manner comparable to an allograft, and after 708 days, the metallic implants were removed. A 1-month activity restriction as well as spoon splint to protect the leg from a full loading were used thereafter. Limb function was fair to good, and the dog remains disease free after 56 months. CONCLUSIONS: A pasteurized autograft consisting of the resected primary bone neoplasm is a valid alternative to a cortical bone allograft for limb sparing in dogs with appendicular OSA in terms of feasibility and pattern of healing. CLINICAL RELEVANCE: This procedure can be an alternative method of limb sparing when difficulties are encountered in establishing and maintaining a canine bone allograft bank.     

Cytokine expression, glucocorticoid and growth hormone changes after porcine reproductive and respiratory syndrome virus (PRRSV-1) infection in vaccinated and unvaccinated naturally exposed pigs

The objective of this paper was to study the changes of some cytokines and neuroendocrine hormones in vaccinated and unvaccinated pigs that were naturally infected by a PRRSV-1 (porcine reproductive and respiratory syndrome virus) heterologous field strain. We analyzed gene expression of pro-inflammatory (TNF-α, IL-1β, MCP-1, IL-6), pro-immune (IFN-γ) and anti-inflammatory cytokines (IL-10) in PBMC, as well as hormonal (GH and cortisol) levels in blood samples of pigs obtained in a field trial previously reported [Martelli P, Gozio S, Ferrari L, Rosina S, De Angelis E, Quintavalla C, et al. Efficacy of a modified-live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine in pigs naturally exposed to a heterologous European (Italian cluster) field strain: clinical protection and cell-mediated immunity. Vaccine 2009;27:3788-99]. All vaccinated pigs showed an increase in pro-inflammatory and pro-immune cytokine gene expression with respect to controls and a prompt increase in GH that could be consistently associated with pro-inflammatory cytokines in sustaining innate immunity; moreover, the higher levels of cortisol indicates the activation of the hypothalamus-pituitary-adrenal (HPA) axis response. In contrast, unvaccinated pigs showed down-regulation of the cortisol and GH responses, and the pro-inflammatory and pro-immune cytokines remained at a basal or low level, with an increase of TNF-α and IL-6 in association with a higher level of IL-10 in the late phase of natural infection. The associated trends of pro-inflammatory and anti-inflammatory cytokines together with the cortisol level demonstrate that a previous vaccination promotes an early immune responsiveness in pigs and a more efficient control of inflammation in the late phase of infection with a heterologous PRRSV isolate; both events could sustain clinical protection.     

Identification,partial characterization,and use of grey mullet ( Mugil cephalus ) vitellogenins for the development of ELISA and biosensor immunoassays

Vitellogenin (Vtg) has proven to be a sensitive and simple biomarker in determining sex, sexual maturity, and xenoestrogenic effects in fish. Thus, our investigation has been focused on identification, partial characterization, and quantification of grey mullet (Mugil cephalus) Vtg through the use of a variety of biochemical and immunological analytical techniques. Mullet is considered both a promising aquaculture candidate and an important species for improving sediment quality in polyculture systems. In the first part of this work, grey mullet Vtg was purified from plasma of 17β-estradiol (E2)-induced male fish by a one-step chromatographic protocol, and partially characterized. Specific polyclonal antibodies were then raised against the mullet Vtg, and both an indirect ELISA and an optical immunosensor were set up and validated to quantify plasma Vtg. The indirect ELISA and the optical immunosensor assay developed showed linear measuring in the range 56.8–1047.1 ng mL−1 and 70–739 ng mL−1 Vtg concentrations in standard solutions, respectively. The results obtained suggest that the indirect ELISA allows Vtg detection over a wide dynamic range, thus resulting more suitable for rapid and sensitive sample screening. Therefore, we suggest that the direct immunosensor is a promising tool which needs more investigation to improve the sensitivity.     

Enteroendocrine profile of α-transducin immunoreactive cells in the gastrointestinal tract of the European sea bass (Dicentrarchus labrax)

In vertebrates, chemosensitivity of nutrients occurs through the activation of taste receptors coupled with G-protein subunits, including α-transducin (G_αtran) and α-gustducin (G_αgust). This study was aimed at characterising the cells expressing G_αtran immunoreactivity throughout the mucosa of the sea bass gastrointestinal tract. G_αtran immunoreactive cells were mainly found in the stomach, and a lower number of immunopositive cells were detected in the intestine. Some G_αtran immunoreactive cells in the stomach contained G_αgust immunoreactivity. Gastric G_αtran immunoreactive cells co-expressed ghrelin, obestatin and 5-hydroxytryptamine immunoreactivity. In contrast, G_αtran immunopositive cells did not contain somatostatin, gastrin/cholecystokinin, glucagon-like peptide-1, substance P or calcitonin gene-related peptide immunoreactivity in any investigated segments of the sea bass gastrointestinal tract. Specificity of G_αtran and G_αgust antisera was determined by Western blot analysis, which identified two bands at the theoretical molecular weight of ~45 and ~40 kDa, respectively, in sea bass gut tissue as well as in positive tissue, and by immunoblocking with the respective peptide, which prevented immunostaining. The results of the present study provide a molecular and morphological basis for a role of taste-related molecules in chemosensing in the sea bass gastrointestinal tract.     

FOAM, a new simple benthic degradative module for the LAMP3D model: an application to a Mediterranean fish farm

The modelling framework already introduced by Doglioli, Magaldi, Vezzulli and Tucci to predict the potential impact of a marine fish farm is improved following different directions, namely (1) real historic current-metre data are used to force the simulations, (2) settling velocity values specifically targeting Mediterranean fish species are used, and (3) a new benthic degradative module, the Finite Organic Accumulation Module, is added to the modelling framework. The Finite Organic Accumulation Module uses the output of the other functional units of the modelling framework to calculate the organic load on the seabed. The Finite Organic Accumulation Module considers the natural capability of the seafloor in absorbing part of the organic load. Different remineralization rates reflect the sediment stress level according to the work of Findlay and Watling. Organic degradation for both uneaten feed and faeces is evaluated by changing the release modality (continuous and periodical) and by varying the settling velocities. It is found that the maximum impact on the benthic community is observed either for quickly sinking uneaten feed released twice a day, or for less intense near-bottom current conditions. If both the above-mentioned scenarios coexist, a high stress level is established in the sediment. The model also suggests that the use of self-feeders in cages can reduce farm impacts significantly. These results show how the new and more complete modelling framework presented here is able to improve the objectivity in the decision-making processes and how it may be successfully used for planning and monitoring purposes.