Phylogenetic Relationships of Avian Cestodes from Brine Shrimp and Congruence with Larval Morphology.

Determining molecular markers for parasites provides a useful tool for their identification, particularly for larval stages with few distinguishable diagnostic characters. Avian cestodes play a key role in the food webs and biodiversity of hypersaline wetlands, yet they remain understudied. Using naturally infected Artemia, we identified cestode larvae (cysticercoids), assessed their genetic diversity, and explored phylogenetic relationships in relation to larval morphology and waterbird final hosts. We obtained partial 18S rDNA sequences for 60 cysticercoids of the family Hymenolepidae infecting Artemia spp. from seven localities and three countries (Spain, the USA, and Chile). We present the first DNA sequences for six taxa: Confluaria podicipina, Fimbriarioides sp., Flamingolepis liguloides, Flamingolepis sp. 1, Flamingolepis sp. 2, and Hymenolepis californicus. Intraspecific sequence variation (0.00-0.19% diversity) was lower than intergroup genetic distance (0.7-14.75%). Phylogenetic analysis revealed three main clades: 1-Flamingolepis, 2-Fimbriarioides, 3-Confluaria and Hymenolepis, all of which separated from hymenolepidids from mammals and terrestrial birds. This clear separation among taxa is congruent with previous morphological identification, validating the 18S gene as a useful marker to discriminate at generic/species level. Working with intermediate hosts allows the expansion of knowledge of taxonomic and genetic diversity of cestodes in wildlife, as well as elucidation of their life cycles.

Leakage of plastics and other debris from landfills to a highly protected lake by wintering gulls.

GENERAL CONTEXT: Gulls ingest plastic and other litter while foraging in open landfills, because organic matter is mixed with other debris. Therefore, gulls are potential biovectors of plastic pollution into natural habitats, especially when they concentrate in wetlands for roosting. NOVELTY: We quantified, for the first time, the flow of plastic and other anthropogenic debris from open landfills to a natural lake via the movement of gulls. We focused on Fuente de Piedra, an inland closed-basin lake in Spain that is internationally important for biodiversity. METHODOLOGY: In 2022, we sampled gull pellets regurgitated in the lake by lesser black-backed gulls Larus fuscus that feed on landfills, as well as their faeces, then characterized and quantified debris particles of ≥0.5 mm. By combining GPS and census data from 2010 to 2022, together with plastic quantification based on FTIR-ATR analysis, we estimated the average annual deposition of plastic and other debris by the wintering gull population into the lake. MAIN RESULTS: 86 % of pellets contained plastics, and 94 % contained other debris such as glass and textiles. Polyethylene (54 %), polypropylene (11.5 %) and polystyrene (11.5 %) were the main plastic polymers. An estimated annual mean of 400 kg of plastics were moved by gulls into the lake. Only 1 % of plastic mass was imported in faeces. DISCUSSION: Incorporating the biovectoring role of birds can provide a more holistic view of the plastic cycle and waste management. Biovectoring is predictable in sites worldwide where gulls and other waterbirds feed in landfills and roost in wetlands. We discuss bird deterrence and other ways of mitigating debris leakage into aquatic ecosystems.

Seed Size, Not Dispersal Syndrome, Determines Potential for Spread of Ricefield Weeds by Gulls.

Recent field data suggest that migratory gulls disperse many rice field weeds by gut passage (endozoochory), most of which are dry fruited and widely assumed to have no long-distance dispersal mechanisms, except via human activity. We investigated this mechanism with a feeding experiment, in which seeds of five common rice field weeds (in order of increasing seed size: Juncus bufonius, Cyperus difformis, Polypogon monspeliensis, Amaranthus retroflexus, and the fleshy-fruited Solanum nigrum) were fed to seven individuals of lesser black-backed gulls Larus fuscus held in captivity. We quantified seed survival after collecting faeces at intervals for 33 h after ingestion, then extracting intact seeds and running germination tests, which were also conducted for control seeds. All five species showed high seed survival after gut passage, of >70%. Gut retention times averaged 2-4 h, but maxima exceeded 23 h for all species. Germinability after gut passage was 16-54%, and gut passage accelerated germination in J. bufonius and S. nigrum, but slowed it down in the other species. All species had lower germinability after gut passage compared to control seeds (likely due to stratification prior to the experiment), but the loss of germinability was higher in smaller seeds. There was no evidence that the different dispersal syndromes assigned to the five species (endozoochory, epizoochory or barochory) had any influence on our results. In contrast, mean gut retention time was strongly and positively related to seed size, likely because small seeds pass more quickly from the gizzard into the intestines. Non-classical endozoochory of dry-fruited seeds by waterbirds is a major but overlooked mechanism for potential long-distance dispersal, and more research into this process is likely essential for effective weed management.

Potential dispersal of aquatic snails by waterbird endozoochory in neotropical wetlands / Dispersão potencial de caramujos por endozoocoria de aves aquáticas em áreas úmidas neotropicais

Abstract Waterbird-mediated zoochory is one of the main ecological mechanisms by which non-flying freshwater invertebrates can disperse between isolated wetlands. Passive dispersal through gut passage inside waterbirds (endozoochory) may explain how many organisms spread in the landscape. Here, we evaluate the potential for dispersal of aquatic snails by three waterbird species in neotropical wetlands. A total of 77 faecal samples from Coscoroba coscoroba (n = 28), Dendrocygna viduata (n = 36) and Anas flavirostris (n = 13) were collected in the field and taken to the laboratory. There, the samples were examined under a stereomicroscope to check for the presence of gastropod shells. We found 496 intact gastropod shells, and Heleobia piscium was the most abundant species (n= 485). We also found two shells of Drepanotrema sp. and nine others distributed between two different morphotypes of Planorbidae. Snails were present in 20.8 % of all samples, and were more frequent in faeces of coscoroba swan (50%) than the other two bird species. Our data suggest that aquatic snails may disperse by avian endozoochory between neotropical wetlands, with vectors including migratory bird species.

Resumo A zoocoria mediada por aves aquáticas é um dos principais processos ecológicos que explicam como invertebrados não-voadores habitantes de água doce se dispersam entre áreas úmidas isoladas. A dispersão passiva que ocorre através no interior dos intestinos de aves aquáticas (endozoocoria) pode explicar como estes invertebrados se distribuem na paisagem. Neste trabalho, avaliamos o potencial de dispersão de caramujos aquáticos por endozoocoria promovida por três espécies de aves aquáticas em áreas úmidas neotropicais. No total, 77 amostras fecais de capororoca (Coscoroba coscoroba, n = 28), irerê (Dendrocygna viduata, n = 36) e marreca-pardinha (Anas flavirostris, n = 13) foram coletadas em campo e levadas ao laboratório. As amostras foram examinadas em estereomicroscópio para verificar a presença de conchas de gastrópodes. Encontramos 496 conchas intactas, sendo Heleobia piscium a espécie mais abundante (n = 485). Também encontramos duas conchas de Drepanotrema sp. e nove de outros dois morfotipos de Planorbidae. Os caramujos estiveram presentes em 20,8% de todas as amostras, sendo mais frequentes nas fezes do capororoca (50%). Nossos dados sugerem que caramujos aquáticos podem se dispersar por endozoocoria de aves entre áreas úmidas neotropicais, com vetores incluindo espécies de aves migratórias e residentes.

Attention by Selection: A Deep Selective Attention Approach to Breast Cancer Classification.

Deep learning approaches are widely applied to histopathological image analysis due to the impressive levels of performance achieved. However, when dealing with high-resolution histopathological images, utilizing the original image as input to the deep learning model is computationally expensive, while resizing the original image to achieve low resolution incurs information loss. Some hard-attention based approaches have emerged to select possible lesion regions from images to avoid processing the original image. However, these hard-attention based approaches usually take a long time to converge with weak guidance, and valueless patches may be trained by the classifier. To overcome this problem, we propose a deep selective attention approach that aims to select valuable regions in the original images for classification. In our approach, a decision network is developed to decide where to crop and whether the cropped patch is necessary for classification. These selected patches are then trained by the classification network, which then provides feedback to the decision network to update its selection policy. With such a co-evolution training strategy, we show that our approach can achieve a fast convergence rate and high classification accuracy. Our approach is evaluated on a public breast cancer histopathological image database, where it demonstrates superior performance compared to state-of-the-art deep learning approaches, achieving approximately 98% classification accuracy while only taking 50% of the training time of the previous hard-attention approach.

Biomedical, psychological, environmental and behavioural factors associated with adult obesity in a nationally representative sample.

OBJECTIVE: To identify personality, biomedical and behavioural factors associated with adult obesity in a large longitudinal sample. METHOD: In total, 5360 participants with data on personality, neurological functioning, maternal smoking during pregnancy, education and occupation, physical exercise, adult self-reported BMI and obesity were included in the study. Obesity at 55 years was the outcome variable. RESULTS: The rates of obesity increased from 9.5 to 22.8% from age 33 to 55 years. Logistic regression analyses (adjusted estimates) showed that childhood neurological functioning (OR = 1.32: 1.07-1.63, P < 0.01), maternal smoking during pregnancy (OR = 1.42: 1.22-1.65, P < 0.001), educational qualifications (OR = 0.54: 0.37-0.79, P < 0.01), trait conscientiousness (OR = 0.80:0.74-0.86, P < 0.001) and physical exercise (OR = 0.87: 0.82-0.92, P < 0.001) were significant predictors of obesity at age 55 years for both men and women. Trait extraversion for men (OR = 1.16: 1.07-1.26, P < 0.001) and trait emotional stability for women (OR = 0.90: 0.82-0.99, P < 0.05) were also significant predictors of the outcome variable. CONCLUSION: Biomedical, psychological, environmental and behavioural factors were all associated with adult obesity.

Overexpression of Specific CD44 Isoforms Is Associated with Aggressive Cell Features in Acquired Endocrine Resistance.

While endocrine therapy is the mainstay of ER+ breast cancer, the clinical effectiveness of these agents is limited by the phenomenon of acquired resistance that is associated with disease relapse and poor prognosis. Our previous studies revealed that acquired resistance is accompanied by a gain in cellular invasion and migration and also that CD44 family proteins are overexpressed in the resistant phenotype. Given the association of CD44 with tumor progression, we hypothesized that its overexpression may act to promote the aggressive behavior of endocrine-resistant breast cancers. Here, we have investigated further the role of two specific CD44 isoforms, CD44v3 and CD44v6, in the endocrine-resistant phenotype. Our data revealed that overexpression of CD44v6, but not CD44v3, in endocrine-sensitive MCF-7 cells resulted in a gain in EGFR signaling, enhanced their endogenous invasive capacity, and attenuated their response to endocrine treatment. Suppression of CD44v6 in endocrine-resistant cell models was associated with a reduction in their invasive capacity. Our data suggest that upregulation of CD44v6 in acquired resistant breast cancer may contribute to a gain in the aggressive phenotype of these cells and loss of endocrine response through transactivation of the EGFR pathway. Future therapeutic targeting of CD44v6 may prove to be an effective strategy alongside EGFR-targeted agents in delaying/preventing acquired resistance in breast cancer.

May arsenic pollution contribute to limiting Artemia franciscana invasion in southern Spain?

Limited information exists regarding the complex interactions between biological invasions, pollution, and climate change. Most studies indicate that pollution tends to favor invasive species. Here, we provide evidence that arsenic (As) pollution may have a role in limiting the invasion of the exotic brine shrimp Artemia franciscana. We tested As toxicity in natural populations of Artemia parthenogenetica (a native taxon) and A. franciscana from localities in southern Spain with differing degrees of As contamination. Tests were conducted both under current mean temperature conditions (25 °C), and as per a future climate scenario (i.e., an increase in mean temperature of 4 °C). Acute toxicity was estimated on the basis of the median lethal concentration (at 24 h), and chronic toxicity was evaluated by measuring Artemia survival and growth under sublethal exposures (after 26 days). At 25 °C, native A. parthenogenetica from the highly polluted Odiel and Tinto estuary was much more resistant to acute As stress (LC50-24 h, 24.67 mg L(-1)) than A. franciscana (15.78 mg L(-1)) and A. parthenogenetica from unpolluted sites (12.04 mg L(-1))-suggesting that local adaptation to polluted conditions may occur. At 29 °C, resistance of A. parthenogenetica from Odiel decreased significantly, and there were no statistical differences in sensitivity between the three species/populations, suggesting that climate change may enhance the probability of invasion. Resistance increased with developmental stage from nauplii to adults, and was extremely high in cysts which still hatched at As concentrations of up to 6400 mg L(-1). Under sublethal chronic exposure A. franciscana performed better (survival and growth) than A. parthenogenetica, and both species experienced a faster growth when exposed to As, compared with unexposed (control) individuals, probably due to the hormesis. We discuss the ecological implications of our results.

Comparing cestode infections and their consequences for host fitness in two sexual branchiopods: alien Artemia franciscana and native A. salina from syntopic-populations.

The American brine shrimp Artemia franciscana is invasive in the Mediterranean region where it has displaced native species (the sexual A. salina, and the clonal A. parthenogenetica) from many salt pond complexes. Artemia populations are parasitized by numerous avian cestodes whose effects have been studied in native species. We present a study from the Ebro Delta salterns (NE Spain), in a salt pond where both A. franciscana and native A. salina populations coexist, providing a unique opportunity to compare the parasite loads of the two sexual species in syntopy. The native species had consistently higher infection parameters, largely because the dominant cestode in A. salina adults and juveniles (Flamingolepis liguloides) was much rarer in A. franciscana. The most abundant cestodes in the alien species were Eurycestus avoceti (in adults) and Flamingolepis flamingo (in juveniles). The abundance of E. avoceti and F. liguloides was higher in the A. franciscana population syntopic with A. salina than in a population sampled at the same time in another pond where the native brine shrimp was absent, possibly because the native shrimp provides a better reservoir for parasite circulation. Infection by cestodes caused red colouration in adult and juvenile A. salina, and also led to castration in a high proportion of adult females. Both these effects were significantly stronger in the native host than in A. franciscana with the same parasite loads. However, for the first time, significant castration effects (for E. avoceti and F. liguloides) and colour change (for six cestode species) were observed in infected A. franciscana. Avian cestodes are likely to help A. franciscana outcompete native species. At the same time, they are likely to reduce the production of A. franciscana cysts in areas where they are harvested commercially.

Bird migratory flyways influence the phylogeography of the invasive brine shrimp Artemia franciscana in its native American range.

Since Darwin's time, waterbirds have been considered an important vector for the dispersal of continental aquatic invertebrates. Bird movements have facilitated the worldwide invasion of the American brine shrimp Artemia franciscana, transporting cysts (diapausing eggs), and favouring rapid range expansions from introduction sites. Here we address the impact of bird migratory flyways on the population genetic structure and phylogeography of A. franciscana in its native range in the Americas. We examined sequence variation for two mitochondrial gene fragments (COI and 16S for a subset of the data) in a large set of population samples representing the entire native range of A. franciscana. Furthermore, we performed Mantel tests and redundancy analyses (RDA) to test the role of flyways, geography and human introductions on the phylogeography and population genetic structure at a continental scale. A. franciscana mitochondrial DNA was very diverse, with two main clades, largely corresponding to Pacific and Atlantic populations, mirroring American bird flyways. There was a high degree of regional endemism, with populations subdivided into at least 12 divergent, geographically restricted and largely allopatric mitochondrial lineages, and high levels of population structure (Φ ST of 0.92), indicating low ongoing gene flow. We found evidence of human-mediated introductions in nine out of 39 populations analysed. Once these populations were removed, Mantel tests revealed a strong association between genetic variation and geographic distance (i.e., isolation-by-distance pattern). RDA showed that shared bird flyways explained around 20% of the variance in genetic distance between populations and this was highly significant, once geographic distance was controlled for. The variance explained increased to 30% when the factor human introduction was included in the model. Our findings suggest that bird-mediated transport of brine shrimp propagules does not result in substantial ongoing gene flow; instead, it had a significant historical role on the current species phylogeography, facilitating the colonisation of new aquatic environments as they become available along their main migratory flyways.

Cestodes change the isotopic signature of brine shrimp, Artemia, hosts: implications for aquatic food webs.

To reach the final host (greater flamingos), the cestode Flamingolepis liguloides alters the behaviour of its intermediate host, the brine shrimp, Artemia parthenogenetica, causing it to spend more time close to the water surface. During summer 2010, we showed that the prevalence of this cestode was consistently higher at the top of the water column in the Odiel salt pans in south-western Spain. We used stable nitrogen (N) and carbon (C) isotopic analysis to test the hypothesis that cestodes also alter resource use by Artemia. In early summer, we compared stable isotopes in infected hosts at the surface with those from uninfected hosts at the bottom of the water column. In late summer, we compared infected and uninfected Artemia from the bottom. δ(15)N was consistently enriched in infected individuals compared with uninfected hosts, especially in Artemia with multiple infections of F. liguloides (family Hymenolepididae) and those with mixed infections of F. liguloides and cestodes of the family Dilepididae. Infected individuals from the surface were enriched in δ(13)C compared with uninfected ones from the bottom, but the opposite was found when comparing uninfected and infected Artemia from the same depth. This may be caused by the increase in lipid concentration in infected Artemia. Isolated cysticercoids of F. liguloides were significantly enriched in δ(13)C compared with cysticercoids in infected hosts, but surprisingly were not enriched in N. Our findings illustrate the way cestodes can alter food webs and highlight the importance of considering the parasitic status of prey in studies of trophic ecology in saline wetlands.

Effects of lead exposure on oxidative stress biomarkers and plasma biochemistry in waterbirds in the field.

Medina lagoon in Andalusia has one of the highest densities of spent lead (Pb) shot in Europe. Blood samples from waterbirds were collected in 2006-2008 to measure Pb concentration (PbB), δ-aminolevulinic acid dehydratase (ALAD), oxidative stress biomarkers and plasma biochemistry. PbB above background levels (>20 µg/dl) was observed in 19% (n=59) of mallards (Anas platyrhynchos) and in all common pochards (Aythya ferina) (n=4), but common coots (Fulica atra) (n=37) and moorhens (Gallinula chloropus) (n=12) were all <20 µg/dl. ALAD ratio in mallards and coots decreased with PbB levels >6 µg/dl. In mallards, an inhibition of glutathione peroxidase (GPx) and an increased level of oxidized glutathione (oxGSH) in red blood cells (RBC) were associated with PbB levels >20 µg/dl. In coots, PbB levels were negatively related to vitamin A and carotenoid levels in plasma, and total glutathione in RBCs; and positively related with higher superoxide dismutase and GPx activities and % oxGSH in RBCs. Overall, the results indicate that previously assumed background levels of PbB for birds need to be revised.

Documenting dermatology practice: ratio of cutaneous tumors biopsied that are malignant.

BACKGROUND: Skin biopsy is an important part of dermatology practice. Little is known regarding dermatologist biopsying practices. OBJECTIVE: The objective was to assess the ratio of malignant lesions biopsied in practice and factors that influence this ratio. METHODS: Using a dermatopathology database, we analyzed tumors biopsied by dermatologists to determine the ratio of malignant to all tumors (malignancy ratio). Additional analyses were performed for factors influencing this ratio. RESULTS: Over 6 months, 11,072 tumors were submitted. Overall, the malignancy ratio was 41.7%, 3.8% for pigmented lesions. Practice type, location, or use of dermatoscopy did not influence this ratio; provider age was positively correlated with a higher malignancy ratio (r=0.51; p=0.02) and was a predictor of malignancy ratio (b=0.02; p=0.04). CONCLUSION: Approximately 40% of biopsied tumors were malignant. Dermatologists exhibit a wide range in their individual practice patterns. Increasing provider age predicted higher malignancy ratios with a 2% increase for each incremental year.