Identification and quantification of benzimidazole resistance polymorphisms in Haemonchus contortus isolated in Northeastern Brazil

Haemonchus contortus is the most prevalent nematode in Brazil. The objective of this study was to select 6 populations of H. contortus of known or suspected benzimidazole resistance status and characterize these using quantitative real-time polymerase chain reaction (qPCR) for single nucleotide polymorphisms (SNPs) F200Y, F167Y and E198A in the β-tubulin isotype 1 gene. qPCR was performed using DNA from a pool of 10 adult male H. contortus from a single animal per farm. Faecal egg count reduction test (FECRT) and egg hatch test (EHT) were used to determine the resistance status. Samples were obtained from 6 farms located in 5 counties in the Ceará State: Tauá, Boa Viagem, Quixadá, Santa Quitéria and Solonópole. The inbred-susceptible-Edinburgh (ISE) isolate was used as reference for comparative purposes in the qPCR. Benzimidazole resistance was detected by FECRT on all farms with efficacy values ranging from 0 to 51%. EC50 values as determined by EHT were all above 1.49 μg/ml. High frequencies of the resistant SNPs F200Y and F167Y alleles were detected but no resistance was detected at SNP E198A. Our results suggest that the SNPs F167Y and F200Y are both important for benzimidazole resistance in the studied populations.     

Subependymal Reaction Secondary to Choroid Plexus Papilloma in a Horse

Choroid plexus papilloma (CPP) is a rare, benign, intracranial neoplasm. The present report described a choroid plexus papilloma of the fourth ventricle with unusual proliferative reaction of ependymal/subependymal cells in a stallion. A single, soft neoformation, red-gray in color, with an irregular surface, resembling a cauliflower, expanding from the fourth ventricle to the cerebellar peduncles was observed. Microscopically, the neoformation was composed of a single layer of simple cuboidal to columnar epithelium with a well-vascularized papillary-arboriform pattern. Multifocal subependymal aggregates forming rosettes were observed near the mesencephalic aqueduct (MA). Glial-fibrillary-acidic-protein (GFAP) was markedly positive only in the neuropil adjacent to the proliferative areas of the MA. Ependymal/subependymal reactions secondary to CPP have never been described in equine or human neuropathology. The capabilities of ependymal cells to regenerate and eventually proliferate are controversial and unclear and, thus, require additional investigation to understand the adaptive mechanisms of this rare pathogenic condition.     

A new large‐scale and cost‐effective restoration method for cold‐water coral gardens

1. Gorgonians dwelling on the Mediterranean continental shelf are among the most frequent fishing bycatch taxa. These species display several traits, such as long lifespans and slow growth, which make them very vulnerable to the impacts caused by fishing activities with far‐reaching and long‐lasting effects.
2. Hence, restoration and mitigation actions are crucial to enhance and speed up the natural recovery of damaged cold‐water coral gardens. Given the growing concern to develop effective and affordable restoration actions, the present study aims to propose and technically validate a new large‐scale and cost‐effective restoration method.
3. This technique, named ‘badminton method', consists of attaching bycatch Eunicella cavolini colonies to cobble supports and returning them to the continental shelf by gently throwing the gorgonian transplants directly from a boat.
4. Two consecutive field experiments were conducted in order to find the best cobble type support and gorgonian size to be used: first, to evaluate the landing efficiency of gorgonian transplants at different depths (from 5 to 30 m) and second, to evaluate their capability to maintain a correct position over time.
5. Natural cobbles with large gorgonians attached were the best option. Field results and modelling approaches suggest that the transplants would correctly land on the continental shelf seabed in a predicted area of around 60 m². Moreover, they would successfully maintain an upright position ensuring gorgonian survival over time.
6. The success of this method highlights the feasibility of large‐scale and low‐cost restoration actions with promising results for the conservation and recovery of cold‐water coral gardens.

     

Growth, stress response and free amino acid levels in Senegalese sole (Solea senegalensis Kaup 1858) chronically exposed to exogenous ammonia

Stressful husbandry conditions are likely to affect growth and amino acid metabolism in fish. In this study, chronic ammonia exposure was used to test the effects of a stressor on growth and amino acid metabolism of Senegalese sole juveniles. The fish were exposed for 52 days to 11.6 mg L⁻¹ [low‐TAN (L‐TAN)] or 23.2 mg L⁻¹ [high‐TAN (H‐TAN)] of total ammonia nitrogen (TAN), or to 0 mg L⁻¹ (Control). Growth in L‐TAN groups was slightly but significantly different from the Control groups [relative growth rate (RGR=0.35±0.13 and 0.52±0.23% day⁻¹ respectively)]. In H‐TAN groups, growth was severely affected (RGR=0.01±0.13% day⁻¹). Stress parameters (plasma cortisol and glucose) showed slight or no significant differences between treatments. Plasma free amino acid (FAA) concentrations were affected after H‐TAN treatment. Increases in glutamine and aspartate concentrations in H‐TAN fish suggest alterations in amino acid metabolism related to nitrogen excretion processes. Some of the changes in FAA concentrations also suggest mobilization to energy supply and synthesis of metabolites related to stress response. Therefore, Senegalese sole seem to adapt to the L‐TAN concentration tested, but the H‐TAN concentration reduced growth and affected amino acid metabolism.     

Insights into population genetics,connectivity and demographic history of the longnosed skate Dipturus oxyrinchus (Linnaeus, 1758) in the western Mediterranean Sea

1. Genetic information is crucial for the conservation of Dipturus oxyrinchus (Linnaeus, 1758), a threatened large skate with declining populations over most of its geographical range. The main aim of the present study was to investigate the genetic structure, connectivity and demographic history of the longnosed skate in Sardinia (western Mediterranean Sea).
2. Patterns of population structure were assessed in 175 specimens from six sampling sites. Variation in two mitochondrial genes (cytochrome c oxidase subunit I (COI) and control region) highlighted high genetic diversity and low but significant genetic differentiation among sites, which clustered into three groups corresponding to the north‐west, north‐east and south Sardinian coasts.
3. The observed genetic structuring could presumably depend on a combination of past geological events, contemporary restrictions to dispersal and biological characteristics of the species (e.g. site‐fidelity, no pelagic larval stage, limited dispersal of juveniles and/or adults).
4. Demographic analyses showed signs of past population expansion, but substantial current stability of Sardinian populations. From a conservation perspective, these results are encouraging, and indicate that Sardinian populations are still large and stable, and seem not to have suffered negative side‐effects from the ever‐growing fishing pressure in the region.
5. The occurrence of genetic structuring strongly supported the close monitoring of populations to identify any erosion of their gene pool, and high genetic variability of the Sardinian D. oxyrinchus populations could thus represent priority populations for conservation purposes, providing potential sources for recolonization in cases of local extinctions in other areas of the distribution range of the species.
6. When the sequences from Sardinia were compared with those available from other areas, the data seem to exclude the possibility that the Atlantic and Mediterranean host totally isolated populations or even different species, as recently suggested. However, additional markers and a larger sampling sites are needed to confirm these findings.

     

Evaluation of different protein sources in fingerling grey mullet <Emphasis Type="Italic">Mugil cephalus</Emphasis> practical diets

An eight-week experiment was carried out to evaluate the effects of different protein sources (fish and haemoglobin meal, soybean meal and torula yeast), in practical diets, on growth, body composition and gut morphology of fingerling grey mullet (Mugil cephalus). Weight gain (%), SGR, FCR, N retention, PER, PGR, FDR and carcass composition of fish were not significantly affected by the dietary protein source. Fish fed the torula yeast based diet showed reduction in growth performance. Histological examinations performed on the alimentary tract of the fish showed a normal structural pattern in the experimental groups, as fundamental histological and histochemical aspects were similar if compared to the control group. The lower growth performance observed in fish fed a torula yeast based diet may be tentatively correlated with the presence of some detrimental morpho-functional aspects in the gut of these fish if compared to fish fed the other diets. Further studies are necessary to confirm this hypothesis.     

Effects of four egg desticking procedures on hatching rate and further survival and growth of larvae in the tench (Tinca tinca L.)

Four desticking procedures for tench eggs (A: tannic acid solution (1 g L⁻¹) for 15 s; B: alcalase enzyme solution (8 mL L⁻¹) for 60 s; C: alcalase enzyme solution (15 mL L⁻¹) for 120 s; D: Woynarovich and Woynarovich (1980) solution for 58 min followed by tannic acid solution (1 g L⁻¹) for 15 s) were tested to obtain data about influence on embryo survival to hatching stage and further survival and growth of the larvae. In the tannic acid and Woynarovich and Woynarovich (1980) treatment (A and D) few eggs stuck together and some were adhered to the incubator walls, whereas in the alcalase treatments (B and C) eggs neither stuck together nor adhered to the incubator walls. Percentages of hatched larvae did not show significant differences (mean values ranged between 47.4% in treatment A to 37.0% in treatment C). Larvae deformities observed were <0.5% in all cases. There were no significant differences among survival and growth rates of the larvae from different egg desticking origin, reaching, after 30 days, mean survival values around 90% and total length and weight of 12.5 mm and 19 mg respectively.     

Increase in milt production by hormonal treatment in the pejerrey fish Odontesthes bonariensis (Valenciennes 1835)

In spite of interest in the cultivation of the pejerrey fish Odontesthes bonariensis (Cuvier & Valenciennes 1835), there are few studies on subjects required to advance this activity. One of the problems is the synchronization of female and male maturation to provide eggs and sperm for larval production. The low volume of expressible milt, either in wild or culture fish, is a major problem. The aim of this work was to study the effectiveness of the administration of different hormones on sperm production in pejerrey. Milt production was enhanced by the injection of human chorionic gonadotropin (hCG) (16.7‐fold increase, 625 IU kg^?1), carp pituitary extracts (13.5‐fold increase, 30 mg kg^?1), salmon pituitary extracts (12.8‐fold increase, 30 mg kg^?1), salmon‐type gonadotropin‐releasing hormone analogue (GnRH) (16.7‐fold increase, 10 μg kg^?1) and mammalian‐type GnRH analogue (10.8‐fold increase, 20 μg kg^?1). Sperm concentration, motility and the fertilization rate were not statistically different compared with control groups. It was also demonstrated that sperm could be obtained off‐season. Taken together, hCG is recommended to stimulate pejerrey spermiation because it is effective in low doses is inexpensive and is widely available.     

A modelling approach to infer the effects of wind farms on landscape connectivity for bats

Little is known about the potentially disrupting effects of wind farms on the habitat connectivity of flying vertebrates at the landscape scale. We developed a regional-scale model to assess the wind farm impact on bat migration and commuting routes. The model was implemented for the bat Nyctalus leisleri in a region of central Italy currently undergoing considerable wind farm development. A Species Distribution Model (SDM) for N. leisleri was generated using the MaxEnt algorithm based on 47 presence records (reduced to 19 after the autocorrelation procedure) and 10 environmental variables derived from topographic and land cover maps. We used the SDM to create a map of connectivity using the software UNICOR to identify potential commuting corridors (PCCs). The incidence of each wind farm on bat flight corridors was assessed by overlaying the existing (380) and planned (195) turbine locations onto the PCCs. The SDM was statistically robust (AUC > 0.8). Most of the corridors were concentrated in the western part of the region, which hosts the largest suitable areas for the species; most of the existing (54 %) and planned (72 %) wind farms interfered with important corridors connecting the western and the eastern parts of the region. Our results provide key information on the impact of the wind farm industry on biodiversity on a regional scale. The novel approach adopted, based on SDM and connectivity analysis, could be easily extended to other flying vertebrates and landscapes and constitutes a promising planning tool necessary for harmonizing the development of renewable energy infrastructures with issues of biodiversity conservation.     

Expert knowledge for translating land cover/use maps to General Habitat Categories (GHC)

Monitoring biodiversity at the level of habitats and landscape is becoming widespread in Europe and elsewhere as countries establish international and national habitat conservation policies and monitoring systems. Earth Observation (EO) data offers a potential solution to long-term biodiversity monitoring through direct mapping of habitats or by integrating Land Cover/Use (LC/LU) maps with contextual spatial information and in situ data. Therefore, it appears necessary to develop an automatic/semi-automatic translation framework of LC/LU classes to habitat classes, but also challenging due to discrepancies in domain definitions. In the context of the FP7 BIO_SOS (www.biosos.eu) project, the authors demonstrated the feasibility of the Food and Agricultural Organization Land Cover Classification System (LCCS) taxonomy to habitat class translation. They also developed a framework to automatically translate LCCS classes into the recently proposed General Habitat Categories classification system, able to provide an exhaustive typology of habitat types, ranging from natural ecosystems to urban areas around the globe. However discrepancies in terminology, plant height criteria and basic principles between the two mapping domains inducing a number of one-to-many and many-to-many relations were identified, revealing the need of additional ecological expert knowledge to resolve the ambiguities. This paper illustrates how class phenology, class topological arrangement in the landscape, class spectral signature from multi-temporal Very High spatial Resolution (VHR) satellite imagery and plant height measurements can be used to resolve such ambiguities. Concerning plant height, this paper also compares the mapping results obtained by using accurate values extracted from LIght Detection And Ranging (LIDAR) data and by exploiting EO data texture features (i.e. entropy) as a proxy of plant height information, when LIDAR data are not available. An application for two Natura 2000 coastal sites in Southern Italy is discussed.