pH-Dependent Solution Structure and Activity of a Reduced Form of the Host-Defense Peptide Myticin C (Myt C) from the Mussel Mytilus galloprovincialis

Myticin C (Myt C) is a highly variable host-defense peptide (HDP) associated to the immune response in the mediterranean mussel (Mytilus galloprovincialis), which has shown to be active across species due to its strong antiviral activity against a fish rhabdovirus found in fish cells overexpressing this HDP. However, the potential antimicrobial properties of any synthetic analogue of Myt C has not yet been analysed. Thus, in this work we have synthesised the sequence of the mature peptide of Myt C variant c and analysed the structure activity relationships of its reduced (non-oxidized) form (red-MytCc). In contrast to results previously reported for oxidized isoforms of mussel myticins, red-MytCc was not active against bacteria at physiological pH and showed a moderate antiviral activity against the viral haemorrhagic septicaemia (VHS) rhabdovirus. However, its chemotactic properties remained active. Structure/function studies in neutral and acid environments by means of infrared spectroscopy indicated that the structure of red-MytCc is pH dependent, with acid media increasing its alpha-helical content. Furthermore, red-MytCc was able to efficiently aggregate artificial phospholipid membranes at low pH, as well as to inhibit the Escherichia coli growth, suggesting that this activity is attributable to its more structured form in an acidic environment. All together, these results highlight the dynamic and environmentally sensitive behavior of red-Myt C in solution, and provide important insights into Myt C structure/activity relationships and the requirements to exert its antimicrobial/immunomodulatory activities. On the other hand, the pH-dependent direct antimicrobial activity of Myt C suggests that this HDP may be a suitable template for the development of antimicrobial agents that would function selectively in specific pH environments, which are sorely needed in this “antibiotic-resistance era”.     

The effect of erector spinae plane block on perioperative analgesic consumption and complications in dogs undergoing hemilaminectomy surgery: a retrospective cohort study

ObjectiveTo compare the perioperative use of analgesics and complication rates in dogs administered an erector spinae plane (ESP) block or a traditional opioid-based (OP) treatment as part of analgesic management during hemilaminectomy.Study designRetrospective cohort study.AnimalsMedical records of 114 client-owned dogs.MethodsGeneral data included demographics, duration of procedure, number of laminae fenestrated, perioperative use of steroid and non-steroidal anti-inflammatory drugs. Intra- and postoperative analgesics used in 48 hours and complications rates were compared between groups. Opioid use was expressed in morphine equivalents [ME (mg kg^?1)]. Continuous data were compared using the Mann–Whitney U test and incidence of events with a Fisher's exact tests. Multiple linear regression was used to evaluate association between perioperative ME consumption (dependent variable) with other independent variables. Data are presented as median (range). Differences were considered significant when p < 0.05.ResultsGroup ESP comprised 42 dogs and group OP 72 dogs. No differences were observed in the general data. Intraoperative ME was 0.65 (0.20–3.74) and 0.79 (0.19–5.60) mg kg^?1 in groups ESP and OP, respectively (p = 0.03). Intraoperative infusion of lidocaine was administered intravenously (IV) to 23.8% and 68% of groups ESP and OP, respectively (p < 0.0001). Intraoperative infusion of ketamine was administered IV to 21% and 40% of groups ESP and OP, respectively (p = 0.04). Regression analysis revealed the ESP block as the only independent variable affecting the perioperative ME consumption. Pharmacological intervention to treat cardiovascular complications was administered to 21.4% and 47.2% of dogs in groups ESP and OP, respectively (p = 0.008). There were no differences in postoperative complication rates.Conclusions and clinical relevanceESP block was associated with reduced perioperative opioid consumption, intraoperative adjuvant analgesic use and incidence of pharmacological interventions to treat cardiovascular complications in dogs undergoing hemilaminectomy.     

Morphological development and allometric growth of yellowtail kingfish Seriola lalandi V. larvae under culture conditions

Morphological development and allometric growth patterns of Seriola lalandi larvae were assessed to characterize normal growth patterns under culture conditions. Early ontogenetic stages of yellowtail kingfish exhibited an exponential growth in terms of standard length as a function of age. Five development stages were characterized from hatching to the juvenile stage: larval stage I (0–2 days post hatch, dph) with endogenous feeding, characterized by a small yolk sac, unpigmented eyes, primordial finfold surrounding the body and a closed mouth; larval stage II (2–15 dph) characterized by mouth opening, complete pigmentation of eyes and the beginning of the exogenous feeding; subsequently, in the larval stage III (15–25 dph) the posterior tip of notochord of the larvae bended upward and the first rays appeared in fins, concomitant with a change in swimming behaviour; thereafter, larval stage IV (post‐flexion stage; 25–30 dph) began when larvae resembled in morphology to a juvenile organism; however, caudal and dorsal fins were not completely development. Lastly, the juvenile stage was reached 30 dph characterized by a morphology and fin structures similar to those of the adults. Growth and development of structures and organs associated with vital functions such as feeding, sensorial and breathing systems seemed to be more critical previous to 23 dph, which was reflected with a positive allometric growth of head and eyes during this period. The results from this study can be used as a tool‐guide to assess normal development in larval research with S. lalandi to improve existing rearing protocols in hatchery production.     

Validation of a multiplex PCR assay to detect Babesia spp. and Anaplasma marginale in cattle in Uruguay in the absence of a gold standard test

Detection of bovine Babesia spp. and Anaplasma marginale is based on the reading of Giemsa-stained blood or organ smears, which can have low sensitivity. Our aim was to improve the detection of bovine Babesia spp. and A. marginale by validating a multiplex PCR (mPCR). We used 466 samples of blood and/or organs of animals with signs and presumptive autopsy findings of babesiosis or anaplasmosis. The primers in our mPCR amplified the rap-1a gene region of Babesia bovis and B. bigemina, and the msp-5 region of A. marginale. We used a Bayesian model with a non-informative priori distribution for the prevalence estimate and informative priori distribution for estimation of sensitivity and specificity. The sensitivity and specificity for smear detection of Babesia spp. were 68.6% and 99.1%, and for A. marginale 85.6% and 98.8%, respectively. Sensitivity and specificity for mPCR detection for Babesia spp. were 94.2% and 97.1%, and for A. marginale 95.2% and 92.7%, respectively. Our mPCR had good accuracy in detecting Babesia spp. and A. marginale, and would be a reliable test for veterinarians to choose the correct treatment for each agent.     

Epidemiological description of the sea lice (Caligus rogercresseyi) situation in southern Chile in August 2007

Salmon sea lice represent one of the most important threats to salmon farming throughout the world. Results of private monitoring efforts have shown an increase in the number of positive cages and cage-level abundance of sea lice in southern Chile since 2004. As a consequence, the Chilean Fisheries Service implemented an Official Surveillance Program in the main salmon production area of southern Chile to assess the situation of sea lice in fish farms. Results showed that the prevalence of sea lice in the fish farms was 53.4%, ranging from 3.5% in Puerto Aysén to 100% in the Seno de Reloncaví zone. The average sea lice abundance was 11.8 per fish (Geometrical mean (GM)=8.61, 95% CI (2.1-6.9)). The highest levels were found in Seno de Reloncaví (GM=24.99, 95% CI (15.9-39.2)), Hornopirén (GM=14.7, 95% CI (10.4-20.8)) and Chiloé norte (GM=9.75, 95% CI (1-1.9)), and the lowest loads were observed in Puerto Aysén (GM=1.35, 95%CI (1-1.9)) and Puerto Cisnes (GM=1.67, 95%CI (1.1-2.6)). Salmo salar and Oncorhynchus mykiss had the highest abundance levels (GM=6.93, 95% CI (5.7-8.5), and (GM=5.55, 95% CI (3.6-8.5), respectively). O. kisutch showed lower levels (GM=1.34, 95% CI (1-1.7)), apparently being more resistant to infestation. Sea lice in farmed salmon are widely distributed in different zones of southern Chile, and are becoming a serious threat to this industry. Prevalence and abundance levels were found to be generally high, decreasing in southern zones.     

Effects of no‐take area size and age of marine protected areas on fisheries yields: a meta‐analytical approach

Marine protected areas (MPAs) are often promoted as tools for biodiversity conservation as well as for fisheries management. Despite increasing evidence of their usefulness, questions remain regarding the optimal design of MPAs, in particular concerning their function as fisheries management tools, for which empirical studies are still lacking. Using 28 data sets from seven MPAs in Southern Europe, we developed a meta‐analytical approach to investigate the effects of protection on adjacent fisheries and asking how these effects are influenced by MPA size and age. Southern European MPAs showed clear effects on the surrounding fisheries, on the ‘catch per unit effort’ (CPUE) of target species, but especially on the CPUE of the marketable catch. These effects depended on the time of protection and on the size of the no‐take area. CPUE of both target species and the marketable catch increased gradually by 2–4% per year over a long time period (at least 30 years). The influence of the size of the no‐take area appeared to be more complex. The catch rates of the entire fishery in and around the MPA were higher when the no‐take areas were smaller. Conversely, catch rates of selected fisheries that were expected to benefit most from protection increased when the no‐take area was larger. Our results emphasize the importance of MPA size on its export functions and suggest that an adequate, often extended, time frame be used for the management and the evaluation of effectiveness of MPAs.     

Isolation and characterization of immortalized porcine aortic endothelial cell lines

Primary porcine endothelial cells have a limited life span in culture. After four to five passages, they tend to de-differentiate and eventually reach senescence. The aim of this work was to establish immortalized porcine aortic endothelial cell lines (AOCs) to facilitate in vitro studies of different pathological process involving the endothelium. Primary porcine aortic endothelial cells (PAECs) were transfected with a plasmid containing the SV40 genome and selected on the basis of morphological and phenotypical features. Flow cytometry analysis demonstrated uptake of acetylated low density lipoproteins (Ac-LDL) and constitutive expression of SLA class I, CD29, CD31, CD41/61, CD80/86, CD46, SWC3, and LAMP-1 antigens by all analyzed lines and showed little differences to primary cells. The functional similarity between primary and immortalized endothelial cells was demonstrated in a cytotoxicity assay using a human natural killer cell line (NKL) as effector. The AOCs cell lines should be valuable tools for in vitro study of the human immune response against pig endothelial cells. In addition, they would be very useful to gain insight in the pathogenesis of some viral haemorrhagic diseases of pig such as African swine fever (ASF) or classical swine fever (CSF).     

Crossability relationships between the common potato, Solanum tuberosum spp. tuberosum, and its wild diploid relatives S. kurtzianum and S. ruiz-lealii

The common potato, Solanum tuberosum spp. tuberosum (tbr), is a tetraploid species with a narrow genetic base, but with a large number of related species that harbor great genetic diversity for numerous characters of agronomic interest. S. kurtzianum (ktz) and S. ruiz-lealii (rzl) are wild diploid relatives with potential resistance to adverse biotic and abiotic factors. To evaluate if this wild germplasm can be incorporated by conventional crosses into the tbr gene pool, pollen-pistil compatibility relations and seed set in interspecific crosses with tbr were investigated. In 4x tbr × 2x ktz crosses and the reciprocals, 89% and 52.2% of genotypic combinations (respectively) were compatible at the pollen-pistil level. Seeds were obtained from some genotypic combinations in both directions of the cross, suggesting that functional 2n gametes might be produced by particular ktz genotypes. In 2x rzl × 4x tbr crosses and their reciprocals, 35% and 11.7% (respectively) of the genotypic combinations were compatible at the pollen-pistil level, but no seeds were obtained. These results indicate that the reproductive isolation between 4x tbr and 2x ktz is incomplete, and that gene exchange between them is feasible. Moreover, further studies will have to be carried out with rzl to ascertain its crossability with 4x tbr.     

Can place names be used as indicators of landscape changes? Application to the Doñana Natural Park (Spain)

This work broaches the possibility of using place names as indicators of original landscapes that have been much transformed. The reconstruction of landscape elements from place names is commonly disputed because such daring notion is impossible to demonstrate. The present case avoids this by making a preliminary study of changes in the landscape using conventional methods. With the knowledge gained from objective and reliable sources, the possibility is analyzed of whether place names are a reflection of landscape changes taking place over a considerable period of time (the last few centuries). It is concluded that, for the present case study, in natural areas with a high rate of change of land use (Doñana Natural Park), place names indicate not only changes in the landscape, but also how such changes are perceived. In the study area, this is especially clear regarding the fens.