Detection of anti-Leishmania infantum antibodies in sylvatic lagomorphs from an epidemic area of Madrid using the indirect immunofluorescence antibody test

An outbreak of human leishmaniosis was confirmed in the southwest of the province of Madrid, Spain, between July 2009 and December 2012. Incidence of Leishmania infection in dogs was unchanged in this period, prompting a search for alternative sylvatic infection reservoirs. We evaluated exposure to Leishmania in serum samples from animals in the area with an indirect immunofluorescence test (IFAT). Using promastigotes from six culture passages and a 1/25 threshold titer, we found anti-Leishmania infantum seroreactivity in 9.3% of cats (4 of 43), 45.7% of rabbits (16/35) and 74.1% of hares (63/85). Use of promastigotes from >10 in vitro passages resulted in a notably IFAT lower titer, suggesting antigenic changes during extended culture. Postmortem inspection of seropositive animals showed no clinical signs of infection. The results clearly suggest that asymptomatic hares were the main reservoir in the outbreak, and corroborate IFAT as a sensitive serological surveillance method to detect such cryptic Leishmania infections.     

Changes in amine concentrations during aging of red wine in oak barrels

This investigation studied the evolution of amines in red wines made with Merlot variety, during aging in American oak barrels (Quercus alba) and in French oak barrels (Quercus sessilis) from the Allier and Nevers regions. From the results obtained it was observed that the evolutions of the amines were similar in all three types of oak woods. Histamine and tyramine were produced at the beginning of the aging process, although they were not accumulated in the wines, probably due to their degradation. Putrescine was the most abundant amine in the wines; its concentration increased to an important extent during aging as it did not undergo degradation. The concentration of cadaverine increased slightly at the first stage of aging and, like putrescine, did not degrade at all. The volatile amines showed slight variations during aging, although in no cases were high accumulations observed in the wines. Dimethylamine and isobutylamine were degraded during storage in the barrels.     

Development of a method for the genetic identification of mussel species belonging to Mytilus, Perna, Aulacomya, and other genera

Legislation regarding the labeling of processed products is an important issue in the protection of consumer rights. This labeling is especially important in products that cannot be identified on the basis of their morphological characters, because these are removed from the animal in the transformation process. The goal of this study was the identification of mussel species using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and Forensically Informative Nucleotide Sequencing (FINS) methodologies. The molecular marker selected was 18S rDNA (nuclear small-subunit rDNA gene), which allows identification at the genus level and at the species level in some cases. The genera included in this study were Mytilus, Perna, Aulacomya, Semimytilus, Brachidontes, Choromytilus, and Perumytilus. Different markers were used for genetic identification at the species level. To identify the species included in the genus Perna and Choromytilus, a fragment of ITS 1 (Internal Transcribed Spacer 1) was amplified by multiplex PCR and digested with restrictases. The species of Mytilus were identified by length polymorphism and RFLP of the polyphenolic adhesive protein gene. This methodology was validated with products manufactured in the authors' pilot plant and applied to commercial samples. Therefore, this sequential method can be completely or partially used to determine the mussel genus or species present in any food product.     

Interannual differences in growth and hatch‐date distributions of early juvenile European anchovy in the Bay of Biscay: implications for recruitment

In order to understand better the recruitment variability in European anchovy in the Bay of Biscay, it is important to investigate the processes that affect survival during the early life stages. Anchovy juvenile growth trajectories and hatch‐date distributions were inferred over a 3‐year period based on otolith microstructure analysis. Otolith growth trajectories showed a characteristic shape depending on their hatch‐date timing. Earlier‐born juveniles had notably broader maximum increments than later born conspecifics, resulting in higher growth rates. This observation suggests that early hatching would be beneficial for larval and juvenile growth, and, therefore, survival. The estimated juvenile hatch‐date distributions were relatively narrow compared with the extended anchovy spawning season (March–August) in the Bay of Biscay and indicated that only individuals originated mainly from the summer months (June–August) survived until autumn. Hatch‐date distributions were markedly different among years and seemed to influence the interannual recruitment strength of anchovy. We conclude that years characterized by juvenile survivors originating from the peak spawning period (May and June) would lead to considerable recruitment success. Downwelling events during the peak spawning period seem to affect larval survival. Furthermore, size‐dependent overwinter mortality would be an additional process that regulates recruitment strength in the anchovy population in the Bay of Biscay.     

Capillary gas chromatography with atomic emission detection for pesticide analysis in soil samples

A method for the simultaneous determination of 10 pesticides (organochlorines, organophosphorus compounds, and pyrethrins) in soils using capillary gas chromatography with atomic emission detection (GC-AED) is reported. Soil samples are first "cleaned-up" with 25 mL of an ascorbic acid solution (pH 2.15). The aqueous phase is extracted with ethyl acetate, and the solid residue is then extracted twice with 10 mL of ethyl acetate. The three resultant organic extracts are combined, concentrated to dryness, and reconstituted in 1 mL of acetone. The pesticides are selectively detected by monitoring chlorine and bromine in the first run and sulfur emission line wavelength in the second run. Each chromatographic run takes 19 min. Detection limits are between 25 and 75 pg, depending on the compound, which corresponds to 1.7 and 5.0 ng/g in the soil samples, respectively. Recoveries of the pesticides from spiked preparations result in an overall mean recovery of 95.3% (n = 120) at fortification levels ranging from 10 to 60 ng/g, depending on the compound. The method is reliable and can be useful for routine monitoring in soils.     

Authentication of anglerfish species (Lophius spp) by means of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and forensically informative nucleotide sequencing (FINS) methodologies

Lophius represents the most important genus of the family Lophiidae from a commercial point of view. The main marketing formats of the species included in this genus are tails and cheeks, making impossible the species identification on the basis of their morphological characters. In the present study, two methods based on the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and phylogenetic analysis of DNA sequences [forensically informative nucleotide sequencing (FINS)] were developed to differentiate the seven species contained in the genus Lophius. In both cases, the molecular marker studied was the cytochrome oxidase subunit I gene (COI). The RFLP analysis of the PCR products digested with the endonuclease Mbo I generated species-specific restriction profiles, and the phylogenetic analysis showing a neighbor-joining tree with independent nodes was strongly supported for all of the studied species. These methods were applied to 40 commercial samples, allowing us to detect the samples incorrectly labeled. The fraudulent labeling ratio was higher in processed products (68.75%) than whole fish (31.25%). The species subjected to mislabeling were L. budegassa (68.75%), L. vomerinus (18.75%), and L. piscatorius (12.5%). Therefore, both methodologies can be independently used to authenticate the species belonging to the genus Lophius, being useful to check the fulfillment of labeling regulations of seafood products and to verify the correct traceability of commercial trade and the control of fisheries.     

Potential Polyunsaturated Aldehydes in the Strait of Gibraltar under Two Tidal Regimes

Diatoms, a major component of the large-sized phytoplankton, are able to produce and release polyunsaturated aldehydes after cell disruption (potential PUAs or pPUA). These organisms are dominant in the large phytoplankton fraction (>10 µm) in the Strait of Gibraltar, the only connection between the Mediterranean Sea and the Atlantic Ocean. In this area, the hydrodynamics exerts a strong control on the composition and physiological state of the phytoplankton. This environment offers a great opportunity to analyze and compare the little known distribution of larger sized PUA producers in nature and, moreover, to study how environmental variables could affect the ranges and potential distribution of these compounds. Our results showed that, at both tidal regimes studied (Spring and Neap tides), diatoms in the Strait of Gibraltar are able to produce three aldehydes: Heptadienal, Octadienal and Decadienal, with a significant dominance of Decadienal production. The PUA released by mechanical cell disruption of large-sized collected cells (pPUA) ranged from 0.01 to 12.3 pmol from cells in 1 L, and from 0.1 to 9.8 fmol cell⁻¹. Tidal regime affected the abundance, distribution and the level of physiological stress of diatoms in the Strait. During Spring tides, diatoms were more abundant, usually grouped nearer the coastal basin and showed less physiological stress than during Neap tides. Our results suggest a significant general increase in the pPUA productivity with increasing physiological stress for the cell also significantly associated to low nitrate availability.     

A 40-year chronology of the vulnerability of spinetail devil ray (Mobula mobular) to eastern Pacific tuna fisheries and options for future conservation and management

1. Tuna fisheries are among the largest and most valuable fisheries in the world, but most interact with many non-target species, including several of high conservation importance. The spinetail devil ray (Mobula mobular) – listed as ‘Endangered’ on the IUCN Red List of Threatened Species – is a commonly discarded bycatch species, particularly in the eastern Pacific Ocean, yet insufficient data exist to undertake a traditional population assessment.
2. A new ecological risk assessment approach designed for data-limited settings – Ecological Assessment of the Sustainable Impacts of Fisheries (EASI-Fish) – was used to reconstruct the historical vulnerability status of M. mobular and to simulate potential changes in its status under 45 hypothetical conservation and management measures. These involved various temporal closures of the eastern Pacific Ocean tuna fishery, decreasing post-capture mortality by improved handling and release practices, and combinations of the two.
3. The species was classified as ‘Least Vulnerable’ between 1979 and 1993, but became ‘Most Vulnerable’ from 1994, which coincided with a rapid spatial expansion of the industrial purse-seine fishery, and especially from 2011 following the rapid increase in the number of sets made on floating objects. Simulating the conservation and management measures in place in 2018 revealed that 31 of the 45 scenarios resulted in a change in classification of the species to ‘Least Vulnerable’, which primarily involved a reduction of post-capture mortality by as little as 20%.
4. It is fortuitous in that education of fishers to implement appropriate best handling and release practices is simpler, more rapid and more cost-effective than the implementation of fishery closures or gear modifications, which can be expensive and complex to implement and monitor and will probably result in substantial reduction in the catches of target species.

     

Vertical and seasonal variation in the δ¹³C of leaf-respired CO₂ in a mixed conifer forest

The C-isotopic composition (δ13C) of leaf respiration (δ(LR)) has previously been shown to vary among functional groups, plant organs and times of day. We here investigated vertical and seasonal variation in δ(LR) through deep (~35 m) forest canopies. We measured δ(LR), δ13C of leaf bulk organic matter (δ(LB)), specific leaf area, net photosynthesis (A) and dark respiration in shade, middle and sun foliage in four conifer species from May to August. We used Keeling plots to estimate δ(LR); we developed a novel technique for ensuring that the respiratory substrate was not changing over the course of the measurement. Variables δ(LR) and δ(LB) displayed a vertical pattern in Abies grandis, Pseudotsuga menziesii and Thuja plicata, but were independent of canopy position in Larix occidentalis. Vertical gradients in δ(LB) (3.6‰) and δ(LR) (2.8‰) were similar. The respiratory enrichment (δ(LR)-δ(LB)) was smaller in expanding (3‰) than mature (4-8‰) foliage. There was a linear relationship between the respiratory enrichment and A. Our data support the hypothesis that δ(LR) values are related to patterns of C allocation among metabolic pathways. We demonstrated that considerable variation in δ(LR) occurs vertically through the canopy (3‰ gradient) and seasonally (3-7‰). Understanding sources of variation in respiratory signals is fundamental to comprehending C dynamics and for global model applications.     

Development of a method for the genetic identification of flatfish species on the basis of mitochondrial DNA sequences

In the present study a method for genetic identification of flatfish species was developed. The technique is based on DNA sequencing of amplified DNA by PCR and subsequent phylogenetic analysis ( FINS). A phylogenetic tree using the cytochrome oxidase subunit I (COI) was constructed and the bootstrap values calculated. The mentioned technique allows the genetic identification of more than 50 flatfish species in fresh, frozen, and precooked products. This analytical system was validated and subsequently applied to 30 commercial samples, obtaining 13 that were incorrectly labeled (43%). Four of the mislabeled samples were whole fish (31%), and nine were fillets (69%). The species with the higher rate of incorrect labeling were Pleuronectes platessa (17%) and Solea solea (10%). Other species incorrectly labeled were Hipoglossus hipoglossus (7%), Reinharditus hippoglossoides, Limanda ferruginea, and Microstomus kitt (3% each species). Therefore, this molecular tool is appropriate to clarify questions related with the correct labeling of commercial products, the traceability of raw materials, and the control of imported flatfish, and also can be applied to questions linked to the control of fisheries.