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Wild adult females of a low trophic omnivore teleost species, the flathead grey mullet (Mugil cephalus), caught in the western Mediterranean were sampled. The lipid and fatty acid composition of ovaries, liver, muscle, and peritoneal fat were analysed at previtellogenesis, early-vitellogenesis —first observed at mid-summer (early August)—, late-vitellogenesis, and the post-spawning period —from mid-September to mid-October—. During ovarian development, the lipid content of muscle was low and constant (3.85%–4.92%), indicating that the muscle was not used to store lipids for gonadal growth. Although constant, lipid content in the liver was higher (18.46%–22.62%) than in the muscle, and HSI% increased during gonad development, suggesting a dynamism in the mobilization of the hepatic lipids. Total lipids in the gonads significantly increased with maturation (from 4.90% to 34.59%) in parallel with the GSI (from 0.8% to 15.5%) to decrease after spawning. Peritoneal fat was probably transitional fat that could be rapidly metabolized or transferred to other tissues but no specific function could be assigned because its presence in previtellogenic and early-vitellogenic females varied greatly. One of the main sources of lipids accumulated in the ovary was most likely diet. The total percentage of ΣMUFA, mainly 17:1 —previously not identified in high quantities in teleost vitellogenic ovaries and likely of bacterial origin— and 16:1, strongly increased in the ovaries with maturation. The 16:1 might be an important source of lipids for embryo development. High percentages of DHA, EPA, and ARA were found in the ovary during previtellogenesis available to be used during gonadal maturation. Understanding lipid and fatty acid changes in broodstock tissues can increase our knowledge of the nutritional requirements of the fish used in aquaculture breeding programs. info:eu-repo/semantics/acceptedVersion

Data de publicació electrònica: 01-01-2023 Includes supplementary materials for the online appendix. Sex determination (SD) shows huge variation among fish and a high evolutionary rate, as illustrated by the Pleuronectiformes (flatfishes). This order is characterized by its adaptation to demersal life, compact genomes and diversity of SD mechanisms. Here, we assembled the Solea senegalensis genome, a flatfish of great commercial value, into 82 contigs (614 Mb) combining long- and short-read sequencing, which were next scaffolded using a highly dense genetic map (28,838 markers, 21 linkage groups), representing 98.9% of the assembly. Further, we established the correspondence between the assembly and the 21 chromosomes by using BAC-FISH. Whole genome resequencing of six males and six females enabled the identification of 41 single nucleotide polymorphism variants in the follicle stimulating hormone receptor (fshr) consistent with an XX/XY SD system. The observed sex association was validated in a broader independent sample, providing a novel molecular sexing tool. The fshr gene displayed differential expression between male and female gonads from 86 days post-fertilization, when the gonad is still an undifferentiated primordium, concomitant with the activation of amh and cyp19a1a, testis and ovary marker genes, respectively, in males and females. The Y-linked fshr allele, which included 24 nonsynonymous variants and showed a highly divergent 3D protein structure, was overexpressed in males compared to the X-linked allele at all stages of gonadal differentiation. We hypothesize a mechanism hampering the action of the follicle stimulating hormone driving the undifferentiated gonad toward testis. This study was supported by the Spanish Ministry of Economy and Competitiveness, FEDER Grants (RTI2018-096847-B-C21, RTI2018-096847-B-C22 and RTI2018-097110-B-C21), Junta de Andalucía-FEDER Grant (P20-00938) and the European Union's Horizon 2020 research and innovation programme under grant agreement No. 817923 (AQUA-FAANG). We thank Geneaqua SL for their participation and financial support of sequencing. We acknowledge the bioinformatic support of the Centro de Supercomputación de Galicia (CESGA).

Der udvikles løbende nye redskaber til deliriumvurdering i engelsktalende lande. Vi ønskede at oversætte CAM-ICU-7 inspireret af WHOʼs anbefalinger og gøre den klar til implementering på danske intensivafdelinger. Selv om delirium er en tilstand, der kendes i alle lande, så kan det være vanskeligt at oversætte et vurderingsredskab uden at redskabets oprindelige betydning ændres. To dansk fødte intensivsygeplejersker oversatte fra engelsk til dansk, og en skotsk født sygeplejerske med mange års erfaring fra en dansk intensivafdeling tilbageoversatte til engelsk uden at have kendskab til den oprindelige udgave. Efterfølgende test og interview, hos erfarent personale viste ingen problemer, hvorimod mindre erfarne sygeplejersker havde meget relevante spørgsmål og kommentarer, der medførte ændringer i teksten. Selv om CAM-ICU-7 er et ret kort og simpelt redskab, kom der således flere ting frem som ville kunne influere på validiteten af oversættelsen, hvis ikke WHOʼs retningslinjer bliver anvendt.

Coyotes are ubiquitous on the North American landscape as a result of their recent expansion across the continent. They have been documented in the heart of some of the most urbanized cities, such as Chicago, Los Angeles, and New York City. Here, we explored the genomic composition of coyotes in the New York metropolitan area to investigate if genomic demography and admixture differs from expected for urban-dwelling canids. We identified moderate-to-high estimates of relatedness among coyotes living in Queens and adjacent neighborhoods, suggestive of a relatively small population. Although we found low background levels of domestic dog ancestry across most coyotes in our sample, we identified a male suspected to be a first-generation coyote-dog hybrid, as well as his two putative backcrossed offspring that carried approximately 25% dog ancestry. The male coyote-dog hybrid and one backcrossed offspring each carried two mutations that are known to increase human-directed hypersociability in dogs and gray wolves. An additional, unrelated coyote with little dog ancestry also carries two of these mutations. These genetic patterns suggest that gene flow from domestic dogs may become an increasingly important consideration as coyotes continue to inhabit metropolitan regions.

Spotted wolffishAnarhichas minorreproduction in captivity is dependent on in vitro fertilization. However, it is often challenging to acquire sufficient fresh sperm to fertilize the eggs that are obtained. In this study, we evaluate the possibility to store spotted wolffish sperm by refrigeration. Spotted wolffish sperm has the particularity that is already motile on stripping, and currently it is not possible to immobilize and reactivate. Thus, sperm refrigeration protocols should focus in extending this motility period that usually lasts up to 2 days. In a first experiment, we evaluated the possibility that the motility period of the sperm was limited by contamination with urine. The urea concentration in the sperm obtained both by stripping (17.10 ± 1.98 mg/dL) and directly from the testis (12.59 ± 2.37 mg/dL) was similar (p > 0.05), which indicate that the sperm collection method used avoid contamination with urine. Afterwards, we tested the possibility that the sperm motility period was limited by energy stores. The ATP concentration (initial value 5.65 ± 0.86 nmol/109cells) remained stable (p = 0.099) during 30 h after sperm collection, and similar values (p = 0.329) were recorded at end of sperm storage in both diluted (3.88 ± 1.35 nmol/109cells) and undiluted samples (4.76 ± 1.08 nmol/109). This indicates that the low intracellular ATP consumption, derived from the slow sperm motility, can probably be compensated rapidly enough by mitochondrial synthesis of ATP in the spotted wolffish sperm. In both experiments, diluted sperm kept higher percentage of motile cells during the storage time.

Trachypithecus, which currently contains 20 species divided into four groups, is the most speciose and geographically dispersed genus among Asian colobines. Despite several morphological and molecular studies, however, its evolutionary history and phylogeography remain poorly understood. Phayre's langur ( Trachypithecus phayrei) is one of the most widespread members of the genus, but details on its actual distribution and intraspecific taxonomy are limited and controversial. Thus, to elucidate the evolutionary history of Trachypithecus and to clarify the intraspecific taxonomy and distribution of T. phayrei, we sequenced 41 mitochondrial genomes from georeferenced fecal samples and museum specimens, including two holotypes. Phylogenetic analyses revealed a robustly supported phylogeny of Trachypithecus, suggesting that the T. pileatus group branched first, followed by the T. francoisi group, and the T. cristatus and T. obscurus groups most recently. The four species groups diverged from each other 4.5-3.1 million years ago (Ma), while speciation events within these groups occurred much more recently (1.6-0.3 Ma). Within T. phayrei, we found three clades that diverged 1.0-0.9 Ma, indicating the existence of three rather than two taxa. Following the phylogenetic species concept and based on genetic, morphological, and ecological differences, we elevate the T. phayrei subspecies to species level, describe a new species from central Myanmar, and refine the distribution of the three taxa. Overall, our study highlights the importance of museum specimens and provides new insights not only into the evolutionary history of T. phayrei but the entire Trachypithecus genus as well.

With the expansion and diversification of global aquaculture, efforts continue to develop new bio-technologies for assisted reproduction in species that present reproductive dysfunctions. Flathead grey mullet (Mugil cephalus) held in intensive conditions in the Mediterranean region, display a severe reproductive dysfunction, where males do not produce fluent milt and females are arrested at previtellogenesis or early stages of vitellogenesis. In the present study, weekly injections of species-specific single-chain recombinant gonadotropins (rGths); follicle stimulating hormone (rFsh) (6 to 12 μg kg-1 doses) and luteinizing hormone (rLh) (2.5 to 24 μg kg-1 doses) were administered to induce vitellogenesis, from previtellogenesis / early vitellogenesis to the completion of vitellogenic growth in females and enhance spermatogenesis to produce adequate volumes of sperm from non-fluent males. During the experiment, all treated females (n = 21) developed oocytes in late vitellogenesis with 603 ± 8 μm diameter and all treated males produced fluent sperm. To induce oocyte maturation, ovulation and spawning, females were treated with either (i) a priming dose of 30 μg kg-1 of rLh and a resolving dose of 40 mg kg-1 of progesterone (P4), (ii) priming and resolving doses of 30 μg kg-1 of rLh, or (iii) priming and resolving doses of 40 mg kg-1 of P4 given 24:05 ± 0:40 h apart. Females were placed in spawning tanks with rGth treated males that had fluent sperm. Spontaneous spawns of fertilised eggs were obtained after inducing with rLh + P4 or rLh + rLh (priming and resolving injections) with a spawning success of the 85% (8 of 9 females) and 100% (n = 6), respectively. The eggs collected from the tanks presented 64 ± 22% fertilization with embryo development and 57 ± 24 % hatching. The treatment P4 + P4 had a lower ovulation success (50 % - 3 of 6 females) and spawning success (17 %) with no fertilised eggs. Success was independent of the initial gonadal stage of females. In comparison, control females did not show any advance in gonadal development from initial stages and control males did not produce fluent sperm. The present results confirm the possibility of controlling oogenesis from previtellogenesis to the completion of maturation and fertilised tank spawning using exclusively rFsh and rLh in a teleost species.

The expression of Interleukin-1ß (IL-1ß) and its antagonist and Interleukin-1 receptor antagonist (IL-1Ra) are correlated with greater human intervertebral disc (IVD) degeneration, suggesting that elevated IL-1β activity promotes disc degeneration. Many in vitro studies support such a mechanistic relationship, whereas few in vivo investigations have been reported. The present study tests the effect of increased IL-1β activity on intervertebral disc in mice with an IL-1Ra gene deletion. IL-1Ra-/- mice and wild-type (WT) C57Bl6J mice were examined at 3 and 12 months of age. Caudal IVD segments were evaluated for disc degeneration by histopathology, functional testing, and inflammatory gene expression relevant to IL-1β pathways. To test differences in injury response, pinprick annular puncture was performed on IL-1Ra-/- and WT mice and evaluated similarly. IL-1Ra-/- IVDs had significantly worse histopathology at 3 months compared to WT controls, but not at 12 months. IL-1Ra-/- IVDs exhibited significantly more viscous mechanical properties than WT IVDs. qPCR revealed downregulation of inflammatory genes at 3 and 12 months in IL-1Ra-/- IVDs, with concomitant downregulation of anabolic and catabolic genes. Annular puncture yielded no appreciable differences between 2-week and 6-week post-injured WT and IL1-Ra-/- IVDs in histopathology or biomechanics, but inflammatory gene expression was sharply downregulated in IL-1Ra-/- mice at 2 weeks, returning by 6 weeks post injury. In the present study, IL-1Ra deletion resulted in increased IVD histopathology, inferior biomechanics, and transiently decreased pro-inflammatory cytokine gene expression. The histopathology of IL-1Ra-/- IVDs on a C57BL/6J background is less severe than a previous report of IL1Ra-/- on a BALB/c background, yet both strains exhibit IVD degeneration, reinforcing a mechanistic role of IL-1β signaling in IVD pathobiology. Despite a pro-inflammatory environment, the annular puncture was no worse in IL-1Ra-/- mice, suggesting that response to injury involves pathways other than inflammation. Overall, this study supports the hypothesis that IL-1β-driven inflammation is important in IVD degeneration.

Throughout South Asia, cattle are regularly treated with non-steroidal anti-inflammatory drugs (NSAIDs) and their carcasses are left for scavengers to consume. Residues of the NSAID diclofenac in cattle carcasses caused widespread mortality and catastrophic population declines in three species of Gyps vulture during the 1990s and 2000s. Diclofenac is now banned, but other NSAIDs are used in its place. Different lines of evidence, including safety testing in Gyps vultures, have shown that some of these other NSAIDs are toxic, or probably toxic, to vultures. The NSAID nimesulide is widely available and commonly used, and has been found in dead vultures with signs of renal failure (i.e. visceral gout) and without the presence of diclofenac and/or other vulture-toxic NSAIDs. Nimesulide is therefore probably toxic to vultures. Here, we report safety testing of nimesulide in Gyps vultures. In a controlled toxicity experiment, we gave two vultures the maximum likely exposure of nimesulide calculated from initial pharmacokinetic and residue experiments in cattle. Two other control birds were given an oral dose of water. Both vultures dosed with nimesulide died within 30 h, after showing outward signs of toxicity and increases in biochemical indicators of renal failure. Post-mortem examinations found extensive visceral gout in both vultures. Both control vultures survived without biochemical indicators of renal failure. With this evidence, we call for an immediate and comprehensive ban of nimesulide throughout South Asia to ensure the survival of the region’s Critically Endangered vultures. More generally, testing the impacts of drugs on non-target species should be the responsibility of the pharmaceutical industry, before their veterinary use is licensed.

Diclofenac caused the death of millions of vultures on the Asian subcontinent. Other non-steroidal anti-inflammatory drugs (NSAIDs) have since also been shown to be toxic to vultures with the exception of meloxicam. For this study, we evaluated the effect of diclofenac on renal uric acid transport and glomerulus filtration in an acute toxicity model. In a two-phase study with the same birds, healthy chickens (a validated model species) were treated intravenously with para-amino hippuric acid (PAH) and iohexol (IOH) in combination in phase 1. In phase 2, the same PAH and IOH combination was then combined with diclofenac (10 mg/kg). In both phases, blood and faeces were sequentially collected. In phase 1, the birds showed no signs of ill health. Moreover, PAH, IOH and uric acid clearance was rapid. In phase 2, two chickens showed early signs of hyperuricemia 8 hours after exposure and died approximately 24h later. Necropsy showed classic signs of renal damage and gout. Diclofenac had a rapid plasma half-life of elimination of less than 2 hours indicating that toxicity was likely due to an irreversible destruction of a physiological process. All the birds in phase 2 had decreased uric acid, PAH and IOH clearance in comparison to phase 1. The decrease in PAH clearance was variable between the birds (average of 71%) but was near 98% reduced in the two birds that died. It is concluded that diclofenac alters both renal perfusion and renal plasma flow, with death associated with tubular secretion being reduced to negligible functionality for a prolonged period. This would support previous in vitro findings of early cell death from ROS accumulation. However, further evaluation is needed to elucidate this final step.

Coyotes are ubiquitous on the North American landscape as a result of their recent expansion across the continent. They have been documented in the heart of some of the most urbanized cities, such as Chicago, Los Angeles, and New York City. Here, we explored the genomic composition of 16 coyotes in the New York metropolitan area to investigate genomic demography and admixture for urban-dwelling canids in Queens County, New York. We identified moderate-to-high estimates of relatedness among coyotes living in Queens (r = 0.0–0.5) and adjacent neighborhoods, suggestive of a relatively small population. Although we found low background levels of domestic-dog ancestry across most coyotes in our sample (5%), we identified a male suspected to be a first-generation coyote–dog hybrid with 46% dog ancestry, as well as his two putative backcrossed offspring that carried approximately 25% dog ancestry. The male coyote–dog hybrid and one backcrossed offspring each carried two transposable element insertions that are associated with human-directed hypersociability in dogs and gray wolves. An additional, unrelated coyote with little dog ancestry also carried two of these insertions. These genetic patterns suggest that gene flow from domestic dogs may become an increasingly important consideration as coyotes continue to inhabit metropolitan regions.

A recurring theme in right wing thinking is that older people are a burden on the young, absorbing an unfair share of national resources, including spending on state pensions, benefits, NHS and social care. This perspective is presented as a zero-sum game, in which generations must compete for scarce resources, their interests being in conflict. In contrast, others say that pensioners have by no means escaped the impact of austerity policies and that life expectancy has flatlined. In this chapter, we examine these views and the statistical evidence used to support them. We conclude that it is essential to distinguish both inter-cohort differences in the lifecourse, as well as intra-cohort inequalities of income, wealth and life chances that are associated with gender, class and ethnicity; that all generations need a secure income in retirement and sufficient health and social care; that older people’s past and current contributions to society are substantial but often invisible; and that intergenerational solidarity through families and social institutions promotes the welfare of all ages.

The aquaculture of meagre (Argyrosomus regius) requires methods for the control of reproduction that enable the production of families from specific individuals for selective breeding programs. We experimentally determined the parameters required for an in vitro fertilisation protocol. A total of 14 females and 5 males (mean ± S.D. weights of 20.45 ± 6.22 and 15.94 ± 2.75 kg, respectively) were used. Selected females had vitellogenic oocytes >550 μm in diameter and males had fluid sperm upon application of abdominal pressure. Both sexes were treated with an injection of 15 μg kg−1 of gonadotropin-releasing hormone agonist (GnRHa) to induce oocyte maturation/ovulation and enhance sperm production. To determine the timing of ovulation and window of high egg viability, females were stripped serially every 2.5 h beginning 35 h after GnRHa treatment. Sperm was obtained 24 h after GnRHa treatment and was diluted 1/4 in modified Leibovitz for storage at 4 °C until use. Sperm quality parameters such as percentage initial spermatozoa motility, duration of motility, velocity and density were determined using computer assisted sperm analysis (CASA). In vitro inseminations were made in duplicate or triplicate batches of eggs from each spawn by mixing 0.5–1 mL of eggs, 20–40 μL diluted sperm (pooled from two males) and 100 mL of seawater. Fertilisation success was examined at spermatozoa (spz): egg ratios between ~2000 and 400,000 spz egg−1. The optimal time for stripping ovulated females was ≤3 h after ovulation, which was the window of optimal egg viability. Ovulation under the conditions of this study was close to 38 h after GnRHa treatment, with a range from 35 to 41 h. Beginning from 3 h after ovulation, egg viability declined probably due to overripening. Sperm diluted in Leibovitz maintained motility and velocity for as long as 7 h after collection. Spermatozoa motility (%) and average path velocity (VAP, μm/s) of sperm samples obtained from males before GnRHa injection declined rapidly after activation compared to the samples obtained 24 h post-injection, with significant decreases respectively after 75 and 45 s. A minimum ratio of 150,000 spermatozoa egg−1 was necessary to ensure high fertilisation success. The acquired knowledge of the present study will aid the aquaculture industry and future research on selective breeding programs for meagre. info:eu-repo/semantics/acceptedVersion

The present study investigated the relationship between reproductive success and stress‐coping styles in gilthead seabream Sparus aurata in captivity. To characterise stress‐coping styles, a total of 22 breeders were submitted to three different individual‐based tests, one group‐based test and post‐handling glucocorticoid quantification. To assess spawning participation, a microsatellite analysis was performed on a total of 2698 larvae, which allowed each offspring to be assigned unambiguously to a single parental couple. Overall, S. aurata showed defined proactive and reactive behavioural traits. Proactive breeders exhibited higher levels of activity and risk taking and lower glucocorticoid blood levels than reactive breeders. The stress‐coping style traits were consistent over time and context (different tests). Breeders that contributed to a higher number of progeny exhibited proactive behaviours, while those showing low progeny contribution exhibited reactive behaviour. Therefore, breeders with a high proportion of progeny (> 20%) had significantly higher activity and risk taking and lower cortisol than breeders with low progeny contribution (< 20%). In addition, males were more proactive than females and males exhibited significantly higher activity, risk taking and lower cortisol than females. This study is the first to establish in S. aurata breeders: (a) a relationship between stress‐coping styles and spawning success; (b) a relationship between stress‐coping styles and gender; and (c) the existence of proactive and reactive traits at the adult stage. info:eu-repo/semantics/acceptedVersion

6 pages, 4 figures, Consecutive treatments with recombinant follicle-stimulating and luteinizing hormones (rFsh and rLh, respectively) stimulate spermatogenesis and potentiate sperm production in pubescent specimens of the oligospermic Senegalese sole (Solea senegalensis). However, sperm production in response to the hormones is highly variable, and the steroidogenic potential of the testis may be diminished due to sustained hormone supply. Here, we compared the effectiveness of low (9 μg/kg) and high (18 μg/kg) doses of rFsh and rLh to improve sperm production in adult sole during late winter-early spring (onset of the natural spawning period), and in autumn under a controlled temperature of 12 °C (period of testicular recrudescence). Treatment with rFsh over six weeks during spring, followed by a single rLh injection, did not enhance sperm production, possibly because of an advanced stage of sexual maturation of the males, as reflected by high Lh plasma levels (~17 ng/ml) before rFsh treatment. In contrast, in autumn, when the Lh circulating levels were much lower (~3 ng/ml), the low doses of rFsh and rLh generated a four-times increase in sperm production, whereas the high doses of the hormones were ineffective. However, treatment with rLh, regardless of the effect of rFsh, improved the motility of spermatozoa during both spring and autumn. These data confirm that consecutive rFsh and rLh treatments increase sperm production and quality in adult sole males, although they seem to be highly sensitive to the rFsh dose. The efficiency of recombinant gonadotropins also appears to be season-dependent despite the asynchronous nature of the sole testis, This work was funded by the Spanish Ministry of Science and Innovation (MINECO) (AGL2017-84013-R to F.C. and AGL2013-41196-R to J.C.), and the Spanish National Institute for Agronomic Research (INIA)-European Fund for Economic and Regional Development (FEDER) (RTA2014-00048 to N.D.). Participation of FC, SR and WG was funded, respectively, by a “Ramon y Cajal” contract (MINECO), a predoctoral grant from AGAUR (Government of Catalonia), and a predoctoral grant from the National Board of Science and Technology (CONACYT, Mexico). This work was also supported by the CERCA Programme (Government of Catalonia).

This research described the common behaviour of grey mullet (Mugil cephalus) under rearing conditions. The different behaviours exhibited by mullets were videorecorded with submersible cameras installed inside of three tanks. A total of 690 minutes per day (07:30 - 18:30 hours) were recorded per tank during a week. Afterwards, an ethogram was elaborated to organize the different behaviours exhibited by juvenileM. cephalus, with two general categories: a) locomotion, including three different observed behaviours (resting, swimming and fast swimming) b) feeding, including three behaviours (surface feeding, bottom feeding and rubbing). The video recordings showed thatM. cephalusis a species with a constant locomotion associated to feeding, since they showed constantly movement during most of day light period the opposite to dark periods. Mullets were observed to be a non-aggressive fish species, due to the absence of dominance and aggression towards conspecifics, resulting in a high predisposition for adaptation to captivity. Finally, behavioural frequencies of grey mullet’s juveniles were not significantly different among the three tanks for most of the behavioural variables analysed (p>0.05) except for the variable bottom feeding (p=0.02). Results from this study could be of interest to the aquaculture industry to implement protocols and to optimize rearing techniques for the production of grey mullet.

Under intensive captive conditions, wild-caught flathead grey mullet (Mugil cephalus) females remained arrested in previtellogenic stages of gonadal development and no sperm could be obtained from males. With the aim to induce oogenesis from previtellogenesis to oocyte maturation, induce the release of sperm and obtain fertilized eggs, female and male flathead grey mullet were treated with Mugil cephalus single-chain recombinant gonadotropins (rGths), follicle-stimulating (rFsh) and luteinizing (rLh) hormones. In Experiment 1, fish were treated with a weekly dose of rFsh (15 μg kg−1), which in females significantly (P 75% motile spermatozoa) was obtained from males in both experiments, and in Exp. 2 the addition of rLh induced the production of higher quantities of sperm that were used to fertilise the eggs. Although fertilisation was low (0.4%), these fertilized eggs with embryo development produced viable larvae (71% hatching). In comparison, control females remained arrested at previtellogenesis and control males did not produce sperm. The study demonstrated that both rGths are effective to induce the process of oogenesis in female flathead grey mullet and to obtain flowing sperm from males, adding more data to confirm the roles of the Gths in teleost gametogenesis. This is the first report, in a teleost species, of the use of rGths (rFsh and rLh) to induce oogenesis from previtellogenesis through to maturation to obtain eggs and larvae. This advance provides the bases for the development of therapies for the use in the aquaculture of teleost of commercial interest or the conservation of endangered species. info:eu-repo/semantics/acceptedVersion

Treatments with homologous recombinant follicle-stimulating and luteinizing hormones (rFsh and rLh, respectively) are known to enhance spermatogenesis and sperm production in sole, but the response can be highly variable depending on the dose, duration and time of the year of the rFsh treatment. To further investigate the physiological effects of rFsh and rLh on sperm production in sole, here we examined the pattern of spermiation of F1 males, of approximately 450 g, treated with rFsh and rLh under controlled temperature. In an initial trial at 12 °C, males were weekly injected intramuscularly with 18 μg kg−1 rFsh over five weeks and subsequently treated with a single injection of 18 μg kg−1 rLh. Histological analysis indicated that the rFsh+rLh treatment increased gonad weight and stimulated spermatogenesis, and also enlarged the size of the seminiferous and efferent duct (ED) tubules, resulting in a doubling of sperm production with respect to the controls. Sperm counts in the ED and sequential stripping of males at 24, 48 and 72 h post rLh injection further revealed that only one batch of spermatids is recruited into spermatozoa (Spz) differentiation after a single rLh induction. A peak of sperm accumulation in the ED occurs at 48 h, coinciding with the upregulation of genes potentially involved in Spz maturation. In a second experiment, we tested the effect of two rFsh doses (10 or 18 μg kg−1) over five weeks as previously, followed by one rLh injection at 12 °C or 17 °C. The results confirmed that spermiation was the highest 48 h after rLh treatment at 12 °C, which was increased in a dose-dependent manner with the dose of rFsh previously supplied (from 0.36 to 0.95 × 109 Spz kg−1). However, sperm production elicited with the low rFsh dose was potentiated by ~3-fold (from 0.36 to 1.06 × 109 Spz kg−1) when the rLh treatment was given at 17 °C. These data suggest that in Senegalese sole sperm collection should be carried out at 48 h after rLh treatment, and that a low dose of rFsh at 12 °C is highly efficient for stimulating sperm production when rLh is administered at a temperature close to that occurring during maximum natural spermiation. info:eu-repo/semantics/acceptedVersion

Trachypithecus, which currently contains 20 species divided into four groups, is the most speciose and geographically dispersed genus among Asian colobines. Despite several morphological and molecular studies, however, its evolutionary history and phylogeography remain poorly understood. Phayre’s langur (Trachypithecus phayrei) is one of the most widespread members of the genus, but details on its actual distribution and intraspecific taxonomy are limited and controversial. Thus, to elucidate the evolutionary history of Trachypithecus and to clarify the intraspecific taxonomy and distribution of T. phayrei, we sequenced 41 mitochondrial genomes from georeferenced fecal samples and museum specimens, including two holotypes. Phylogenetic analyses revealed a robustly supported phylogeny of Trachypithecus, suggesting that the T. pileatus group branched first, followed by the T. francoisi group, and the T. cristatus and T. obscurus groups most recently. The four species groups diverged from each other 4.5–3.1 million years ago (Ma), while speciation events within these groups occurred much more recently (1.6–0.3 Ma). Within T. phayrei, we found three clades that diverged 1.0–0.9 Ma, indicating the existence of three rather than two taxa. Following the phylogenetic species concept and based on genetic, morphological, and ecological differences, we elevate the T. phayrei subspecies to species level, describe a new species from central Myanmar, and refine the distribution of the three taxa. Overall, our study highlights the importance of museum specimens and provides new insights not only into the evolutionary history of T. phayrei but the entire Trachypithecus genus as well.

Trachypithecus, which currently contains 20 species divided into four groups, is the most speciose and geographically dispersed genus among Asian colobines. Despite several morphological and molecular studies, however, its evolutionary history and phylogeography remain poorly understood. Phayre’s langur (Trachypithecus phayrei) is one of the most widespread members of the genus, but details on its actual distribution and intraspecific taxonomy are limited and controversial. Thus, to elucidate the evolutionary history of Trachypithecus and to clarify the intraspecific taxonomy and distribution of T. phayrei, we sequenced 41 mitochondrial genomes from georeferenced fecal samples and museum specimens, including two holotypes. Phylogenetic analyses revealed a robustly supported phylogeny of Trachypithecus, suggesting that the T. pileatus group branched first, followed by the T. francoisi group, and the T. cristatus and T. obscurus groups most recently. The four species groups diverged from each other 4.5–3.1 million years ago (Ma), while speciation events within these groups occurred much more recently (1.6–0.3 Ma). Within T. phayrei, we found three clades that diverged 1.0–0.9 Ma, indicating the existence of three rather than two taxa. Following the phylogenetic species concept and based on genetic, morphological, and ecological differences, we elevate the T. phayrei subspecies to species level, describe a new species from central Myanmar, and refine the distribution of the three taxa. Overall, our study highlights the importance of museum specimens and provides new insights not only into the evolutionary history of T. phayrei but the entire Trachypithecus genus as well.

Throughout South Asia, cattle are regularly treated with non-steroidal anti-inflammatory drugs (NSAIDs) and their carcasses are left for scavengers to consume. Residues of the NSAID diclofenac in cattle carcasses caused widespread mortality and catastrophic population declines in three species of Gyps vulture during the 1990s and 2000s. Diclofenac is now banned, but other NSAIDs are used in its place. Different lines of evidence, including safety testing in Gyps vultures, have shown that some of these other NSAIDs are toxic, or probably toxic, to vultures. The NSAID nimesulide is widely available and commonly used, and has been found in dead vultures with signs of renal failure (i.e. visceral gout) and without the presence of diclofenac and/or other vulture-toxic NSAIDs. Nimesulide is therefore probably toxic to vultures. Here, we report safety testing of nimesulide in Gyps vultures. In a controlled toxicity experiment, we gave two vultures the maximum likely exposure (MLE) of nimesulide calculated from initial pharmacokinetic and residue experiments in cattle. Two other control birds were given an oral dose of water. Both vultures dosed with nimesulide died within 30 h, after showing outward signs of toxicity and increases in biochemical indicators of renal failure. Post-mortem examinations found extensive visceral gout in both vultures. Both control vultures survived without biochemical indicators of renal failure. With this evidence, we call for an immediate and comprehensive ban of nimesulide throughout South Asia to ensure the survival of the region's Critically Endangered vultures. More generally, testing the impacts of drugs on non-target species should be the responsibility of the pharmaceutical industry, before their veterinary use is licensed.

Background Dysphagia related to intubation affects almost one in five patients in the intensive care unit; it may vary due to fluctuations in cerebral status and fatigue and lead to pneumonia, re-intubation and increased mortality. Taking care of patients' nutritional needs is an integral part of nursing and nurses must therefore be able to assess risk of dysphagia at all hours. However, Danish validated methods for use in the critically ill patient population are lacking. Aim To translate and adapt the Yale Swallow Protocol method for implementation in a Danish intensive care unit setting. Methods Translation and adaption followed WHO's recommendations for translation and adaption of instruments. Face validity of the Danish version was evaluated in four group interviews. These were digitally recorded, transcribed and analysed using deductive content analysis. All interviewees gave informed consent. Findings The translated version of the Yale Swallow Protocol was easy to understand, although nurses needed to have the method explained and needed training in using the method. In addition, the Yale Swallow Protocol must be adapted to local practice in terms of identifying patients at risk of dysphagia and deciding where to document results of the screening. Conclusion The translated version of the Yale Swallow Protocol proved conceptually and culturally sound and acceptable to intensive care unit nurses. Implementation should be followed by theoretical and practical training to build a nursing vocabulary relating to dysphagia and strengthen nurses' sensitivity to clinical manifestations consistent with dysphagia.

Diclofenac has been responsible for the deaths of millions of vultures on the Asian subcontinent. While the pathology of toxicity is well described, the mechanism of toxicity remains elusive. However, it was postulated that toxicity could be related to the unique avian renal vascular structure known as the renal portal valve and that that diclofenac altered valve functionality with subsequent renal ischaemia. While plausible, the valva renalis portalis has only been described in a small number of other bird species such as the chicken (Gallus domesticus), the domestic duck (Anas platyrhynchos domesticus) and ostrich (Struthio camelus) but not a raptor. The aim of this study was to evaluate the renal anatomy and related vasculature of the Cape griffon vulture (Gyps coprotheres) (CGV), a species sensitive to the toxic effects of diclofenac, using gross anatomy, histology and vascular casting. The vasculature of the vulture was found to be almost identical to that of the domestic chicken with the valva renalis portalis present in the v. iliaca externa between the v. renalis renalis cranialis and the v. renalis caudalus. The valve was ring-shaped with finger-like processes and histologically was composed of smooth muscle. The valve was also well vascularized and was associated with a nerve plexus. Based on the findings of this study, the proposed mechanism of toxicity is anatomically possible.

Under intensive captive conditions, wild-caught flathead grey mullet (Mugil cephalus) females remained arrested at early gonad development and no sperm could be obtained from males. With the aim to induce and complete oogenesis, induce the release of sperm and obtain fertilized eggs, adult female and male grey mullet were treated with M. cephalus single-chain recombinant gonadotropins (rGths), follicle-stimulating (rFsh) and luteinizing (rLh) hormones. In Experiment 1, fish were treated with a weekly dose of rFsh (15 μg kg−1), which in females significantly (P < 0.001) increased plasma concentration of 17β-estradiol and induced vitellogenic oocyte growth up to a maximum mean diameter of 425 ± 19 μm after 9 weeks of treatment. In Experiment 2, fish were treated with weekly injections of both rFsh and rLh at different doses (from 2.5 to 12 μg kg−1). Oocyte diameter reached 609 ± 5 μm, from which final oocyte maturation and ovulation was induced with 30 μg kg−1 of rLh and 40 mg kg−1 of progesterone. Good quality sperm (> 75% motile spermatozoa) was obtained from males in both experiments, and in Exp. 2 the addition of rLh induced the production of higher quantities of sperm that were used to fertilise the eggs. Although fertilisation was low (0.4 %), these fertilized eggs with embryo development produced viable larvae (71% hatching rate). This is the first report in a teleost species, to obtain larvae from eggs that were from immature females induced through to maturation with rGths. In comparison, control females remained arrested as immature fish and control males did not produce sperm. The study demonstrated that both rGths are effective to induce the entire process of oogenesis in sexually immature female grey mullet and to obtain flowing sperm from males, adding more data to confirm the roles of the Gths in teleost gametogenesis. This advance provides the bases of a therapy for the use in the aquaculture of teleost of commercial interest or the conservation of endangered species.