Both sexually selected traits and mate preferences for these traits can be context dependent, yet how variation in preferred traits could select for context dependent preferences has rarely been examined. The signal reliability hypothesis predicts that mate preferences vary across contexts (e.g., environments) in relation to the reliability of the information preferred traits provide in those contexts. Extensive variation in copy number of mc4r B alleles on the Y-chromosome that associates with male size in Xiphophorus multilineatus allowed us to use a split-sibling design to determine if male size is more likely to provide information about male genotype (i.e., dam) when males were reared in a warm as compared to a cold environment. We then examined strength of preference for male size by females reared in the same two environments. We found that males were larger in the cold environment, but male size was more variable across dams in the warm environment, and therefore male size would be a more reliable indicator of dam (i.e., genetics) in the warm environment. Females reared in the warm environment had stronger mate preferences based on male size than cold reared females, with a significant influence of dam on strength of preference. Therefore, strength of female preference for male size was influenced by the temperature in which they were reared, with the direction of the difference across treatments supporting the signal reliability hypothesis. Understanding how the reliability of male traits can select for contextual variation in the strength of the female mate preferences will further our discovery of adaptive mate preferences. For example, a relationship between the strength of a female\'s mate preference and their growth rates was detected in the context where females had a preference based on male size, supporting a hypothesis from previous work with this species of disassortative mating in relation to growth rates to mitigate a documented growth-mortality tradeoff.

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Kryptolebias marmoratus (Kmar), a teleost fish of the order Cyprinodontiformes, has a suite of unique phenotypes and behaviors not observed in other fishes. Many of these phenotypes are discrete and highly plastic-varying over time within an individual, and in some cases reversible. Kmar and its interfertile sister species, K. hermaphroditus, are the only known self-fertile vertebrates. This unusual sexual mode has the potential to provide unique insights into the regulation of vertebrate sexual development, and also lends itself to genetics. Kmar is easily adapted to the lab and requires little maintenance. However, its internal fertilization and small clutch size limits its experimental use. To support Kmar as a genetic model, we compared alternative husbandry techniques to maximize recovery of early cleavage-stage embryos. We find that frequent egg collection enhances yield, and that protease treatment promotes the greatest hatching success. We completed a forward mutagenesis screen and recovered several mutant lines that serve as important tools for genetics in this model. Several will serve as useful viable recessive markers for marking crosses. Importantly, the mutant kissylips lays embryos at twice the rate of wild-type. Combining frequent egg collection with the kissylips mutant background allows for a substantial enhancement of early embryo yield. These improvements were sufficient to allow experimental analysis of early development and the successful mono- and bi-allelic targeted knockout of an endogenous tyrosinase gene with CRISPR/Cas9 nucleases. Collectively, these tools will facilitate modern developmental genetics in this fascinating fish, leading to future insights into the regulation of plasticity.

The African turquoise killifish Nothobranchius furzeri (N. furzeri) is a useful model organism for studying aging, age-related diseases, and embryonic diapause. CRISPR/Cas9-mediated gene knockout and Tol2 transposon-mediated transgenesis in N. furzeri have been reported previously. However, these methods take time to generate knockout and transgenic fish. In addition, knock-in technology that inserts large DNA fragments as fluorescent reporter constructs into the target gene in N. furzeri has not yet been established. Here, we show that triple-target CRISPR-mediated single gene disruption efficiently produces whole-body biallelic knockout and enables the examination of gene function in the F0 generation. In addition, we developed a method for creating the knock-in reporter N. furzeri without crossing by optimizing the CRISPR/Cas9 system. These methods drastically reduce the duration of experiments, and we think that these advances will accelerate aging and developmental studies using N. furzeri.

Studies of sex chromosome systems at early stages of divergence are key to understanding the initial process and underlying causes of recombination suppression. However, identifying signatures of divergence in homomorphic sex chromosomes can be challenging due to high levels of sequence similarity between the X and the Y. Variations in methodological precision and underlying data can make all the difference between detecting subtle divergence patterns or missing them entirely. Recent efforts to test for X-Y sequence differentiation in the guppy have led to contradictory results. Here, we apply different analytical methodologies to the same data set to test for the accuracy of different approaches in identifying patterns of sex chromosome divergence in the guppy. Our comparative analysis reveals that the most substantial source of variation in the results of the different analyses lies in the reference genome used. Analyses using custom-made genome assemblies for the focal population or species successfully recover a signal of divergence across different methodological approaches. By contrast, using the distantly related Xiphophorus reference genome results in variable patterns, due to both sequence evolution and structural variations on the sex chromosomes between the guppy and Xiphophorus. Changes in mapping and filtering parameters can additionally introduce noise and obscure the signal. Our results illustrate how analytical differences can alter perceived results and we highlight best practices for the study of nascent sex chromosomes.

Parasitic castration is a phenomenon where the parasite blocks or reduces the host reproductive output. This study explored the impact of Tetracotyle wayanadensis infection on the reproductive performance of the freshwater fish Aplocheilus lineatus. Heavily infected A. lineatus showed slow and sluggish movements with reduced food intake and feeding activities. Histopathological observations of the ovary revealed severe necrosis and degeneration of previtellogenic and vitellogenic oocytes and follicular atresia. The oogonial nests were absent; chromatin nucleolar oocytes irregularly shaped and vacuolated. Perinucleolar oocytes have a vacuolated appearance with the hypertrophied highly columnar follicular cells invading them. Vitellogenic oocytes had a shrunken appearance with folded zona radiata; yolk globules broken down into smaller granules together with vacuolar degeneration of the ooplasm. The hypertrophied highly columnar follicular cells invaded the early vitellogenic oocytes to phagocytize the degenerating material. Zona radiata was found broken and liquefaction of yolk globules was evident with advancement in follicular atresia. Seasonal variation of parasitic infection has also been studied. Metacercaria infecting the vital organs showed seasonal variation with the highest values for prevalence and abundance during monsoon and mean intensity during winter. The lowest values for these descriptors were documented during the post-monsoon period.

The present study supports the usefulness of ancillary techniques, such as immunohistochemistry, as a valid diagnostic tool in the field of fish oncology. The immunohistochemical patterns observed in four neoplasms on four individual teleosts belonging to different species are described. Cytokeratin, vimentin, actin, S100, calretinin, and Melan-A antibodies were used. Diagnoses of papilloma in a Bream Abramis brama, fibroma in a Sand Steenbras Lithognathus mormyrus, schwannoma in a Crucian Carp Carassius carassius, and melanoma in a spontaneously inbred Xiphophorus hybrid were made. Diagnosis of tumors in fish is not always easy to carry out, and the tool provided by antibodies used on mammalian tissue is essential for obtaining definitive, unambiguous, and inexpensive identification.

The natural history of model organisms is often overlooked despite its importance to correctly interpret the outcome of laboratory studies. Ageing is particularly understudied in natural populations. To address this gap, we present lifetime demographic data from wild populations of an annual species, the turquoise killifish, Nothobranchius furzeri, a model species in ageing research, and two other species of coexisting annual killifishes. Annual killifish hatch synchronously, have non-overlapping generations, and reproduce daily after reaching sexual maturity. Data from 13 isolated savanna pools in southern Mozambique demonstrate that the pools supporting killifish populations desiccated 1-4 months after their filling, though some pools persisted longer. Declines in population size over the season were stronger than predicted, because they exceeded the effect of steady habitat shrinking on population density that, contrary to the prediction, decreased. Populations of N. furzeri also became more female-biased with progressing season suggesting that males had lower survival. Nothobranchius community composition did not significantly vary across the season. Our data clearly demonstrate that natural populations of N. furzeri and its congeners suffer strong mortality throughout their lives, with apparent selective disappearance (condition-dependent mortality) at the individual level. This represents selective force that can shape the evolution of lifespan, and its variation across populations, beyond the effects of the gradient in habitat persistence.

Electrocatalytic treatment of real textile wastewater was investigated in continuous electrochemical reactor using dimensionally stable Ti/RuO2 anode. Effects of various parameters such as: elapsed time, current, pH, retention time on the COD removal, color removal and specific energy consumed were evaluated. Central Composite Design under RSM was used for experimental design, data analysis, optimization, interaction analysis between the various electrochemical parameters and steady state time analysis. GC-MS and UV spectrophotometric analysis of the untreated and treated wastewater were conducted to identify the oxidized and transformed/degraded compounds during the oxidation process, and a suitable degradation mechanism was proposed. Treated wastewater may contain toxic chlorinated compounds due to mediated oxidation by various hydrolyzed chlorine species. Therefore, disposability of treated wastewater was assessed by conducting toxicity bioassay test. The optimal set of operating parameters were found to be elapsed time = 124 min, current = 1.37 A, pH = 5.54 and retention time = 157.6 min to simultaneously achieve COD removal, color removal and specific energy consumed as 86.22%, 94.74% and 0.012 kW h, respectively. GC-MS analysis showed presence of chlorinated compounds in the treated wastewater. The toxicity bioassay test resulted acute toxicity with 100% mortality rate within one minute and one hour exposure with untreated and treated textile wastewater, respectively.

The mosquitofish (Gambusia affinis) naturally inhabits freshwater (FW; 1-3‰) and seawater (SW; 28-33‰) ponds in constructed wetland. To explore the physiological status and molecular mechanisms for salinity adaptation of the mosquitofish, cytoprotective responses and osmoregulation were examined. In the field study, activation of protein quality control (PQC) mechanism through upregulation of the abundance of heat shock protein (HSP) 90 and 70 and ubiquitin-conjugated proteins was found in the mosquitofish gills from SW pond compared to the individuals of FW pond. The levels of aggregated proteins in mosquitofish gills had no significant difference between FW and SW ponds. Furthermore, the osmoregulatory responses revealed that the body fluid osmolality and muscle water contents of the mosquitofish from two ponds were maintained within a physiological range while branchial Na+/K+-ATPase (NKA) expression was higher in the individuals from SW than FW ponds. Subsequently, to further clarify whether the cellular stress responses and osmoregulation were mainly induced by hypertonicity, a laboratory salinity acclimation experiment was conducted. The results from the laboratory experiment were similar to the field study. Branchial PQC as well as NKA responses were induced by SW acclimation compared to FW-acclimated individuals. Taken together, induction of gill PQC and NKA responses implied that SW represents an osmotic stress for mosquitofish. Activation of PQC was suggested to provide an osmoprotection to prevent the accumulation of aggregated proteins. Moreover, an increase in branchial NKA responses for osmoregulatory adjustment was required for the physiological homeostasis of body fluid osmolality and muscle water content.