Biological invasions and continued salinization of freshwater are two global issues with largely serious ecological consequences. Increasing salinity in freshwater systems, as an environmental stressor, may negatively affect normal life activities in fish. It has been documented that salinity limits the invasive success of alien species by mediating physiological and life-history performances, however, there are few studies on how salinity affects its invasive process via altered behaviors. Using wild-caught invasive western mosquitofish (Gambusia affinis) as animal model, in this study, we asked whether gradual increasing salinity affects behaviors (personality and mate choice decision here), life-history traits, as well as the correlation between them by exposing G. affinis to three levels salinity (freshwater, 10 and 20‰). Results showed that, with increased salinity, male tended to be shyer, less active, less sociable, and reduced desire to mate, and female tended to be shyer, less active and lost preferences for the larger male. Furthermore, across salinity treatments, male exhibited reduced body fat content and rising reproduction allocation, however, pregnant female revealed diametrically opposed trends. In addition, the correlation between life-history traits and behaviors was only identified in pregnant female. It seems that either salinity or life-history traits directly affects mosquitofish behaviors. In summary, our results partially emphasize the harmful consequences of salinity on both life-history traits and behavioral performances. These findings provide a novel perspective on how salinity potentially affect fish fitness via altering personalities, mate choice decisions, as well as body condition, and hence supports the idea that salinity could affect the spread of invasive mosquitofish.

The first investigation of microplastics in G. affinis from the Brantas River was carried out in this study. Microplastics were found at higher concentrations in gambusia fish captured downstream (209.18 ± 48.85 particles/gram) than upstream (24.44 ± 0.14 particles/gram). Microplastic particle concentrations in G. affinis have a positive linear relationship with fish length. The fiber was the most prominent shape at Sites 1 and 2, whereas the fragment was dominant at Sites 3 and 4. With a value of 45–48%, black dominates the entire site, followed by blue (29–38%), transparent colors (7–11%), red (2–4%), purple (1–3%), and other colors (5–7%). Microplastics measuring <0.1 mm are commonly found in fish bodies. Cyclohexylmethyl octyl ester (phthalic acid) is the most abundant component found in microplastics, accounting for 30.11% of the total. This study provides evidence that G. affinis can be used to monitor the presence of microplastic pollution in the Brantas River but further studies are needed regarding the effects of microplastics and their health hazards on fish.

Silver nanoparticles (AgNPs) inevitably discharge into aquatic environments due to their abundant use in antibacterial products. It was reported that in laboratory conditions, AgNPs display dose-dependent toxicity to aquatic organisms, such as bacteria, algae, macrophytes, snails and fishes. However, AgNPs could behave differently in natural complex environments. In the present study, a series of microcosms were established to investigate the distribution and toxicity of AgNPs at approximately 500 μg L−1 in aquatic systems. As a comparison, the distribution and toxicity of the same concentration of AgNO3 were also determined. The results showed that the surface layer of sediment was the main sink of Ag element for both AgNPs and AgNO3. Both aquatic plant (Hydrilla verticillata) and animals (Gambusia affinis and Radix spp) significantly accumulated Ag. With short-term treatment, phytoplankton biomass was affected by AgNO3 but not by AgNPs. Chlorophyll content of H. verticillata increased with both AgNPs and AgNO3 short-term exposure. However, the biomass of phytoplankton, aquatic plant and animals was not significantly different between control and samples treated with AgNPs or AgNO3 for 90 d. The communities, diversity and richness of microbes were not significantly affected by AgNPs and AgNO3; in contrast, the nitrification rate and its related microbe (Nitrospira) abundance significantly decreased. AgNPs and AgNO3 may affect the nitrogen cycle and affect the environment and, since they might be also transferred to food web, they represent a risk for health.

The essential oil (EO) of Cupressus sempervirens was obtained by hydrodistillation and analysed using gas chromatography–flame ionization detection (GC–FID) and gas chromatography–mass spectrometry (GC–MS). Two monoterpenes, α-pinene (49.1%) and δ-3-carene (21.4%), and one sesquiterpene hydrocarbon, α-cedrol (5.1%), were isolated as the EO major terpenes. An oil-in-water nanoemulsion (particle size 71.2 nm) was produced from the EO through a low-energy method. The EO, its nanoemulsion and its main constituents showed mosquitocidal and biochemical effects against Culex quinquefasciatus Say, the common vector of lymphatic filariasis parasites. All treatments showed dose-dependent bioactivity, and adults were more susceptible to the EO products than the larvae. The nanoemulsion showed superior activity, followed by the crude EO and α-cedrol. At 40 μg/ml, the nanoemulsion caused 100% larval mortality, while the EO and α-cedrol required twice this concentration to achieve the same larval mortality. The LC₅₀ values were 8.4, 16.1, 15.1, 30.7 and 53.4 μg/ml at 24 h after exposure for the nanoemulsion, crude oil, α-cedrol, δ-3-carene and α-pinene, respectively. For adults, 20.0 μl/l nanoemulsion caused 100% mortality, while twice this concentration of the EO was required to achieve the same effect. The LC₅₀’s against adults ranged between 6.2 and 40.4 μl/l. EO products prominently repelled mosquitoes at concentrations between 0.75 and 6.0 μl/cm². The EO products caused remarkable inhibition of Cx. quinquefasciatus acetylcholinesterase activity but were safer towards the non-target aquatic species Gambusia affinis. These results recommend the use of C. sempervirens EO, its nanoemulsion and main terpenes as natural tools to control Cx. quinquefasciatus.

We assessed the suitability of two nonnative poeciliid fishes—western mosquitofish (Gambusia affinis) and sailfin mollies (Poecilia latipinna)—for monitoring selenium exposure in desert pupfish (Cyprinodon macularius). Our investigation was prompted by a need to avoid lethal take of an endangered species (pupfish) when sampling fish for chemical analysis. Total selenium (SeTot) concentrations in both poeciliids were highly correlated with SeTot concentrations in pupfish. However, mean SeTot concentrations varied among fish species, with higher concentrations measured in mosquitofish than in mollies and pupfish from one of three sampled agricultural drains. Moreover, regression equations describing the relationship of selenomethionine to SeTot differed between mosquitofish and pupfish, but not between mollies and pupfish. Because selenium accumulates in animals primarily through dietary exposure, we examined fish trophic relationships by measuring stable isotopes (δ 13C and δ 15N) and gut contents. According to δ 13C measurements, the trophic pathway leading to mosquitofish was more carbon-depleted than trophic pathways leading to mollies and pupfish, suggesting that energy flow to mosquitofish originated from allochthonous sources (terrestrial vegetation, emergent macrophytes, or both), whereas energy flow to mollies and pupfish originated from autochthonous sources (filamentous algae, submerged macrophytes, or both). The δ 15N measurements indicated that mosquitofish and mollies occupied similar trophic levels, whereas pupfish occupied a slightly higher trophic level. Analysis of gut contents showed that mosquitofish consumed mostly winged insects (an indication of terrestrial taxa), whereas mollies and pupfish consumed mostly organic detritus. Judging from our results, only mollies (not mosquitofish) are suitable for monitoring selenium exposure in pupfish.

The mosquito vectors of the genera Aedes and Anopheles present resistance to several commercial insecticides, which are also toxic to non-predator targets. On the other hand, essential oils are a promising source of insecticides. Thus, in this work, the essential oil from the leaves of Piper purusanum was characterized by gas chromatography–based approaches and evaluated as biodefensive against malaria and dengue vectors. The main compounds of P. purusanum essential oil were β-caryophyllene (57.05%), α-humulene (14.50%), and germacrene D (8.20%). The essential oil inhibited egg hatching (7.6 ± 1.5 to 95.6 ± 4.5%), caused larval death (LC₅₀ from 49.84 to 51.60 ppm), and inhibited the action of acetylcholinesterase (IC₅₀ of 2.29 µg/mL), which can be related to the mechanisms of action. On the other hand, the biological activities of β-caryophyllene, α-humulene, and germacrene D were higher than that of essential oil. In addition, these sesquiterpenes and essential oil did not show a lethal effect on Toxorhynchites splendens, Anisops bouvieri, Gambusia affinis, and Diplonychus indicus (LC₅₀ from 2098.80 to 7707.13 ppm), although D. indicus is more sensitive (SI/PSF from 48.56 to 252.02 ppm) to essential oil, representing a natural alternative against these relevant vectors.

Effluent discharge from wastewater treatment plants can be a substantial source of microplastics in receiving water bodies including rivers. Despite growing concern about microplastic pollution in freshwater habitats, the literature has not yet addressed effluent-dependent rivers, which derive 100% of their baseflow from effluent. The objective of this study was to document and explore trends in microplastic pollution within the effluent-dependent lower Santa Cruz River near Tucson, Arizona (USA). We examined microplastic concentrations in the water column and benthic sediment and microplastic consumption by mosquitofish (Gambusia affinis) at 10 sites along a ~40 km stretch of the lower Santa Cruz River across two time periods: baseflow (effluent only) and post-flood (effluent immediately following urban runoff). In total, across both sampling periods, we detected microplastics in 95% of water column samples, 99% of sediment samples, and 6% of mosquitofish stomachs. Flow status (baseflow vs post-flood) was the only significant predictor of microplastic presence and concentrations in our models. Microplastic fragment concentrations in the water column were higher post-flood, microplastic fiber concentrations in benthic sediment were lower post-flood, and mosquitofish were more likely to have consumed microplastics post-flood than during baseflow. The additional microplastics detected after flooding was likely due to a combination of allochthonous material entering the channel via runoff and bed scour that exhumed microplastics previously buried in the riverbed. Effluent-dependent urban streams are becoming increasingly common; more work is needed to identify microplastic pollution baselines and trends in effluent rivers worldwide.

Widely distributed western mosquitofish (Gambusia affinis) has been used as a new model species for hazard assessment of environmental stressors such as polycyclic aromatic hydrocarbons (PAHs). However, most of the PAH studies using G. affinis rely on targeted biomarker-based analysis, and thus may not adequately address the complexity of the toxic mechanisms of the stressors. In the present study, the whole transcriptional sequencing of G. affinis liver after exposure to a PAH model, benzo[a]pyrene (BaP) (100 μg/L), for 20 days was performed by using the HiSeq XTen sequencers. In total, 58,156,233 and 51,825,467 clean nucleotide reads were obtained in the control and BaP-exposed libraries, respectively, with average N50 lengths of 1419 bp. In addition, after G. affinis was exposed for 20 days, 169 genes were upregulated, and 176 genes were downregulated in liver. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were applied to all the genes to determine the genes’ biological functions and processes. The results clearly showed that the differentially expressed genes were mainly related to immune pathways and metabolic correlation pathways. Interestingly, almost all the pathways related with the immunity were upregulated, while the metabolism pathways were downregulated. Lastly, quantitative real-time PCR (qRT-PCR) was performed to measure expressional levels of twelve genes confirmed through the DGE analysis. These results demonstrate that BaP damages immunity and enhances the consumption of all available energy storage to activate mechanisms of the detoxification in G. affinis. Up until now, the present study is the first time that a whole transcriptome sequencing analysis in the liver of G. affinis exposed to BaP has been reported.

Recently, the biofabrication of metal nanoparticles has gained wide interest owing to its inherent features such as swift, simplicity, eco-friendliness, and cheaper costs. Different green-reducing agents led to the production of nanoparticles with varying toxicity on insects. In the current study, silver nanoparticles (AgNPs) were successfully synthesized using Habenaria plantaginea leaf extract. Ag nanoparticles were studied by UV–Vis spectroscopy (UV-Vis), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), atomic force microscopy (AFM), scanning electron microscopy (SEM) coupled with energy-dispersive spectroscopy (EDS), and transmission electron microscopy (TEM). H. plantaginea extract and AgNPs were tested for mosquito larvicidal activity on Anopheles stephensi, Aedes aegypti, Culex quinquefasciatus, An. subpictus, Ae. albopictus, and Cx. tritaeniorhynchus. LC₅₀ values were 102.51, 111.99, 123.47, 123.96, 136.56, 149.42 μg/ml and 12.23, 13.38, 14.78, 14.37, 15.39, 16.89 μg/ml, respectively. Moreover, H. plantaginea aqueous extract and AgNPs were tested against the non-target species Anisops bouvieri, Diplonychus indicus, Poecilia reticulata, and Gambusia affinis obtaining LC₅₀ values ranging from 831.82 to 36,212.67 μg/ml. Overall, this study showed the effectiveness of H. plantaginea-fabricated nanoparticles on a wide range of important mosquito vectors, highlighting their scarce toxicity on four natural enemies predating mosquito larvae and pupae.

The recent outbreaks of dengue, chikungunya, and Zika virus highlighted the pivotal importance of mosquito vector control in tropical and subtropical areas worldwide. However, mosquito control is facing hot challenges, mainly due to the rapid development of pesticide resistance in Culicidae and the limited success of biocontrol programs on Aedes mosquitoes. In this framework, screening botanicals for their mosquitocidal potential may offer effective and eco-friendly tools in the fight against mosquitoes. In the present study, the essential oil (EO) obtained from the medicinal plant Origanum scabrum was analyzed by GC-MS and evaluated for its mosquitocidal and repellent activities towards Anopheles stephensi, Aedes aegypti, Culex quinquefasciatus, and Culex tritaeniorhynchus. GC-MS analysis showed a total of 28 compounds, representing 97.1 % of the EO. The major constituents were carvacrol (48.2 %) and thymol (16.6 %). The EO was toxic effect to the A. stephensi, A. aegypti, C. quinquefasciatus, and C. tritaeniorhynchus larvae, with LC₅₀ of 61.65, 67.13, 72.45, and 78.87 μg/ml, respectively. Complete ovicidal activity was observed at 160, 200, 240, and 280 μg/ml, respectively. Against adult mosquitoes, LD₅₀ were 122.38, 134.39, 144.53, and 158.87 μg/ml, respectively. In repellency assays, the EOs tested at 1.0, 2.5, and 5.0 mg/cm² concentration of O. scabrum gave 100 % protection from mosquito bites up to 210, 180, 150, and 120 min, respectively. From an eco-toxicological point of view, the EO was tested on three non-target mosquito predators, Gambusia affinis, Diplonychus indicus, and Anisops bouvieri, with LC₅₀ ranging from 4162 to 12,425 μg/ml. Overall, the EO from O. scabrum may be considered as a low-cost and eco-friendly source of phytochemicals to develop novel repellents against Culicidae.