Sublethal Exposure Research Articles

Environmental Toxicant Exposure Paralyzes Human Placental Macrophage Responses to Microbial Threat.

Exposure to environmental toxicants (such as dioxins) has been epidemiologically linked to adverse reproductive health outcomes, including placental inflammation and preterm birth. However, the molecular underpinnings that govern these outcomes in gravid reproductive tissues remain largely unclear. Placental macrophages (also known as Hofbauer cells) are crucial innate immune cells that defend the gravid reproductive tract and help promote maternal-fetal tolerance. We hypothesized that exposure to environmental toxicants such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) could alter placental macrophage responses to inflammatory insults such as infection. To test this, placental macrophages were cultured in the presence or absence of TCDD and then infected with the perinatal pathogen Group B Streptococcus (GBS). Our results indicate that TCDD is lethal to placental macrophages at and above a 5 nM concentration and that sublethal dioxin exposure inhibits phagocytosis and cytokine production. Taken together, these results indicate that TCDD paralyzes placental macrophage responses to bacterial infection.

Mosquito larvae exposed to a sublethal dose of photosensitive insecticides have altered juvenile development but unaffected adult life history traits

BackgroundLarvicides are critical for the control of mosquito-borne diseases. However, even sublethal exposure to a larvicide can alter development and life history traits, which can then affect population density and disease transmission dynamics. Photosensitive insecticides (PSIs) are a promising class of larvicide that are toxic when ingested and activated by light. We investigated whether the time of day when exposure occurs, or the process of pupation, affects larval susceptibility to PSI phototoxicity in the mosquito Anopheles gambiae, and whether sublethal exposure to PSIs alters life history traits.MethodsLarvae were treated with lethal concentrations of the PSIs methylene blue (MB) and rose bengal (RB), and larval survival was measured at various times of day. Additionally, larvae were exposed to two concentrations of each PSI that resulted in low and medium mortality, and the life history traits of the surviving larvae were measured.ResultsPupation, which predominantly occurs in the evening, protected larvae from PSI toxicity, but the toxicity of PSIs against larvae that had yet to pupate was unaffected by time of day. Larval exposure to a sublethal concentration of MB, but not RB, shortened the time to pupation. However, larval exposure to a sublethal concentration of RB, but not MB, increased pupal mortality. Neither PSI had a meaningful effect on the time to eclosion, adult longevity, or adult melanization potential.ConclusionsPSIs are lethal larvicides. Sublethal PSI exposure alters mosquito development, but does not affect adult life history traits.Graphical

Effects of transfluthrin-treated jute and cotton clothing against resistant and susceptible Aedes aegypti (Diptera: Culicidae) in a semifield system.

Volatile pyrethroids exert a range of both lethal and behavioral effects on mosquitoes through the passive release of insecticides into the atmosphere. We investigated the protective efficacy (PE) of transfluthrin-treated jute (TI-jute) and cotton (TI-cotton) fabrics, worn at the back of a protective black vest, against laboratory-reared pyrethroid susceptible and resistant strains of Aedes aegypti (L.) in a semifield system (SFS). Each fabric (1,029 cm2) was treated with 1.79 mg/cm2 of transfluthrin as the intervention. Human landing collections were conducted by 2 collectors seated in designated treatment and control compartments of the SFS. The trials were conducted for 41 days, with 16 days partitioned into morning and evening phases. Furthermore, we examined blood feeding behavior and fecundity of the surviving mosquitoes post-exposure. Results showed that in the morning, the PE of TI-jute (49.4%) was higher than that of TI-cotton (36.8%). TI-jute demonstrated a lower PE of 9.6% against the transfluthrin-resistant strain. Remarkably, a significantly higher number of eggs were laid by the transfluthrin-resistant mosquitoes that survived the intervention (36.5 eggs/female) compared to the control group (11.8 eggs/female). These findings suggest that TI-jute can help protect against bites and alter the life traits of Ae. aegypti. The study highlights that the timing of the intervention during the day affected the efficacy of TI-jute and TI-cotton, while sublethal exposure to transfluthrin stimulated egg production in the resistant strain. These are critical challenges that warrant attention in vector control strategies. Investigating this phenomenon in mosquito reproduction necessitates future research at a molecular level.

Long-term sublethal exposure to polyethylene and tire wear particles: Effects on risk-taking behaviour in invasive and native fish

Anthropogenic polymeric particles pollute even the most remote ecosystems and may compromise organisms' behaviour and movement skills. It is expected that invasive species cope better with pollutants than native species (i.e., pollution resistance hypothesis). In this study, invasive gibel carp (Carassius gibelio) and native crucian carp (Carassius carassius) were used as model organisms. Specimens were fed daily with food pellets (1 % body weight) added with 0.1 % polyethylene (PE), tire wear particles (TWPs) and control. Their behavioural parameters were compared before and after 14 and 60 days of exposure. Additionally, we evaluated burst swimming capacity after 60 days of exposure to the treatments. The fishes exposed to the PE and TWPs treatments showed significant trends toward increased boldness scores and, in the PE treatment, higher utilization of the open field, and both behavioural changes are associated with higher risk-taking. Invasive gibel carp had substantially better swimming performance than crucian carp, but the expected trend in relation to the treatments was not found. Fish exposed to sublethal doses of PE and TWPs showed signs of behavioural changes after two months of exposure that may affect risk-taking behaviour, which might impact species interactions with predators.

Transgenerational sublethal pyrethroid exposure gives rise to insecticide resistance in a pest insect

The evolution of insecticide resistance has been attributed to strong directional selection by lethal concentrations of insecticides, but there is growing evidence that sublethal doses may also modify resistance through the hormetic effects. Hormesis is a beneficial effect caused by exposure to low doses. However, the role of parental (transgenerational) effects on hormesis, and through that on insecticide resistance, is still unclear. We investigated the effects of several sublethal pyrethroid insecticide (Decis) doses on survival, body mass, and reproduction within four generations (F0, F1, F2, and F3) of the Colorado potato beetle (Leptinotarsa decemlineata). We found that insecticide exposure had mostly linear adverse within-generation effects: decreased larva-to-adult survival, adult body mass, and egg hatching. However, transgenerational exposure led to hormetic effects: increased larva-to-adult survival and pre-diapause adult body mass. Moreover, transgenerational effects were even more positive for offspring exposed to insecticides, leading to decreased larva-to-adult survival, increased body mass, and egg hatching. Our results show that despite mostly negative within-generation effects, transgenerational sublethal exposure to insecticide can cause unwanted positive hormetic effects in their offspring, making them to resist or tolerate the insecticides better, even though the underlying mechanisms are still unclear.

Biosensor-guided detection of outer membrane-specific antimicrobial activity against Pseudomonas aeruginosa from fungal cultures and medicinal plant extracts.

Pseudomonas aeruginosa responds to sub-lethal antimicrobial exposure by inducing the expression of lipopolysaccharide (LPS) surface modifications that mask antibiotic binding sites and contribute to repair and resistance of the outer membrane (OM). We exploit these membrane damage-responsive operons in a biosensor approach used to discover new antimicrobials that specifically target the OM. Chromosomal transcriptional luxCDABE reporters from the pmr (polymyxin resistance; aminoarabinose LPS modification) and speD2E2 (spermidine synthesis) operons are induced by validated outer membrane-acting agents including cationic antimicrobial peptides, cation chelators, ascorbic acid, detergents, and cell wall synthesis inhibitors cycloserine and bacitracin. To identify novel sources of OM-disrupting antimicrobials, we used these OM damage-responsive biosensors to screen a panel of fungal culture supernatants for novel antimicrobial and biosensor activity. Biosensor activity was used to determine the optimal time point of antimicrobial production from fungal supernatants and to guide the purification of active fractions after size-exclusion chromatography. Water and ethanol extracts of Chinese medicinal plants also proved to be a source of biosensor activity. The pathogen box is a 400-member drug library of potential antimicrobials, but none of these compounds induced our OM damage biosensors. This novel, sensitive, cell-based screening assay has potential for future discovery of lead compounds that specifically target the outer membrane, which is a significant barrier to antibiotic entry into Gram-negative bacteria.IMPORTANCENew approaches are needed to discover novel antimicrobials, particularly antibiotics that target the Gram-negative outer membrane. By exploiting bacterial sensing and responses to outer membrane (OM) damage, we used a biosensor approach consisting of polymyxin resistance gene transcriptional reporters to screen natural products and a small drug library for biosensor activity that indicates damage to the OM. The diverse antimicrobial compounds that cause induction of the polymyxin resistance genes, which correlates with outer membrane damage, suggest that these LPS and surface modifications also function in short-term repair to sublethal exposure and are required against broad membrane stress conditions.

Failure of Cronobacter sakazakii to acquire direct and cross-protection against lethal stresses after exposure to sublethal ethanol

Abstract Cronobacter sakazakii may encounter subinhibitory concentrations of ethanol stress over its lifecycle. Bacterial tolerance to homologous or heterologous stress agents may be altered as a result of ethanol adaptive responses. Therefore, the tolerance of ethanol-exposed and control cells to subsequent lethal stresses was evaluated in the current work. It was discovered that sublethal ethanol exposure increased the susceptibility to lethal ethanol stress in C. sakazakii as determined by the Weibull model. Furthermore, sublethal ethanol concentration exposure in C. sakazakii did not lead to any cross-tolerance against other stressors such as benzalkonium chloride (120 mg/L), heat (55 °C), cold (4 °C), simulated gastric fluid (pH 3.0), osmotic stress (sorbitol, 0.75 g/mL), and desiccation stress. Analysis of zeta potential, scanning electron microscope, and surface-enhanced Raman spectroscopy spectra revealed that cellular injury and changes in cellular chemical composition may contribute to the reduced resistance of C. sakazakii after ethanol exposure. Furthermore, sublethal ethanol exposure resulted in an elevated proportion of unsaturated fatty acids (USFA), while reducing the proportion of saturated fatty acids (SFA) and the ratio of SFA to USFA. The developed inactivation models can serve as a valuable source of data to support quantitative microbial risk assessment. Moreover, a better understanding of the response of C. sakazakii to sublethal ethanol exposure may provide valuable insights into the prevention and control of C. sakazakii.

Phenotypic and Transcriptional Responses of Pseudomonas aeruginosa Biofilms to UV-C Irradiation via Side-Emitting Optical Fibers: Implications for Biofouling Control.

Biofilms give rise to a range of issues, spanning from harboring pathogens to accelerating microbial-induced corrosion in pressurized water systems. Introducing germicidal UV-C (200-280 nm) irradiation from light-emitting diodes (LEDs) into flexible side-emitting optical fibers (SEOFs) presents a novel light delivery method to inhibit the accumulation of biofilms on surfaces found in small-diameter tubing or other intricate geometries. This work used surfaces fully submerged in flowing water that contained Pseudomonas aeruginosa, an opportunistic pathogen commonly found in water system biofilms. A SEOF delivered a UV-C gradient to the surface for biofilm inhibition. Biofilm growth over time was monitored in situ using optical conference tomography. Biofilm formation was effectively inhibited when the 275 nm UV-C irradiance was ≥8 μW/cm2. Biofilm samples were collected from several regions on the surface, representing low and high UV-C irradiance. RNA sequencing of these samples revealed that high UV-C irradiance inhibited the expression of functional genes related to energy metabolism, DNA repair, quorum sensing, polysaccharide production, and mobility. However, insufficient sublethal UV-C exposure led to upregulation genes for SOS response and quorum sensing as survival strategies against the UV-C stress. These results underscore the need to maintain minimum UV-C exposure on surfaces to effectively inhibit biofilm formation in water systems.

Sub-lethal exposure to chlorfenapyr reduces the probability of developing Plasmodium falciparum parasites in surviving Anopheles mosquitoes

BackgroundPyrethroid resistance in the key malaria vectors threatens the success of pyrethroid-treated nets. To overcome pyrethroid resistance, Interceptor® G2 (IG2), a ‘first-in-class’ dual insecticidal net that combines alpha-cypermethrin with chlorfenapyr, was developed. Chlorfenapyr is a pro-insecticide, requiring bio-activation by oxidative metabolism within the insect’s mitochondria, constituting a mode of action preventing cross-resistance to pyrethroids. Recent epidemiological trials conducted in Benin and Tanzania confirm IG2’s public health value in areas with pyrethroid-resistant Anopheles mosquitoes. As chlorfenapyr might also interfere with the metabolic mechanism of the Plasmodium parasite, we hypothesised that chlorfenapyr may provide additional transmission-reducing effects even if a mosquito survives a sub-lethal dose.MethodsWe tested the effect of chlorfenapyr netting to reduce Plasmodium falciparum transmission using a modified WHO tunnel test with a dose yielding sub-lethal effects. Pyrethroid-resistant Anopheles gambiae s.s. with L1014F and L1014S knockdown resistance alleles and expression levels of pyrethroid metabolisers CYP6P3, CYP6M2, CYP4G16 and CYP6P1 confirmed by quantitative reverse transcription polymerase chain reaction (RT-qPCR) prior to conducting experiments were exposed to untreated netting and netting treated with 200 mg/m3 chlorfenapyr for 8 h overnight and then fed on gametocytemic blood meals from naturally infected individuals. Prevalence and intensity of oocysts and sporozoites were determined on day 8 and day 16 after feeding.ResultsBoth prevalence and intensity of P. falciparum infection in the surviving mosquitoes were substantially reduced in the chlorfenapyr-exposed mosquitoes compared to untreated nets. The odds ratios in the prevalence of oocysts and sporozoites were 0.33 (95% confidence interval; 95% CI 0.23–0.46) and 0.43 (95% CI 0.25–0.73), respectively, while only the incidence rate ratio for oocysts was 0.30 (95% CI 0.22–0.41).ConclusionWe demonstrated that sub-lethal exposure of pyrethroid-resistant mosquitoes to chlorfenapyr substantially reduces the proportion of infected mosquitoes and the intensity of the P. falciparum infection. This will likely also contribute to the reduction of malaria in communities beyond the direct killing of mosquitoes.Graphical

Does prior exposure to larvicides influence dengue virus susceptibility in Aedes aegypti (Diptera: Culicidae)?

Control of mosquito vector populations is primarily intended to reduce the transmission of pathogens they transmit. Use of chemical controls, such as larvicides, can have unforeseen consequences on adult traits if not applied properly. The consequences of under application of larvicides are little studied, specifically the impacts on pathogen infection and transmission by the vectors that survive exposure to larvicides. We compared vector susceptibility of Aedes aegypti (L.) for dengue virus, serotype 1 (DENV-1) previously exposed as larvae to an LC50 of different classes of insecticides as formulated larvicides. Larval exposure to insect growth regulators (methoprene and pyriproxyfen) significantly increased susceptibility to infection of DENV-1 in Ae. aegypti adults but did not alter disseminated infection or transmission. Larval exposure to temephos, spinosad, and Bti did not increase infection, disseminated infection, or transmission of DENV-1. Our findings describe a previously under observed phenomenon, the latent effects of select larvicides on mosquito vector susceptibility for arboviruses. These data suggest that there are unintended consequences of sublethal exposure to select larvicides that can influence susceptibility of Ae. aegypti to DENV infection, and indicates the need for further investigation of sublethal effects of insecticides on other aspects of mosquito biology, especially those parameters relevant to a mosquitoes ability to transmit arboviruses (life span, biting behavior, extrinsic incubation period).

Effect of Sublethal Concentrations of Nuvan on Enzymatic Profile of <I>Labeo rohita </I>(ham.)

Toxic effects of sublethal concentrations of nuvan on acid phosphatase (ACP), alkaline phosphatase (ALP), adenosine triphosphatase (ATPase), succinic dehydrogenase (SDH), lactic dehydrogenase (LDH) and acetyl cholinestrase (AChE) activities were studied in fingerlings of Labeo rohita. Two sublethal concentrations i.e. 10.3 mg/I and 2.06 mg/l were selected basing on the LCso value of nuvan in rohu for this experiment continuing for a period of 45 days. Sublethal exposures at two concentrations reduced the brain AChE, and brain, liver and kidney LDH and ATPase (P < 0.05) over a period of 45 days. The reduction of SDH was detected in all the organs examined except brain on 30 day at 2.06 mg/I. While ALP and AChE activities of brain were decreased, the ACP activity was increased.

Influence of cheese making process on STEC bacteriophage release.

Shiga toxin-producing Escherichia coli (STEC) are foodborne pathogens implicated in diseases including hemolytic uremic syndrome (HUS) and hemorrhagic colitis (HC). The main virulence factor are Shiga toxins; their production and secretion are by-products of the expression of late genes of prophages upon sub-lethal environmental stimuli exposure. Hence, the lysogenic prophage after a stress switch to lytic cycle spreading the Stx phages. In the present study, 35 STEC were screened for the presence and the ability to release Shiga toxin-encoding bacteriophages. Three bacterial strains showed signals of prophage presence both in plate and in PCR. Subsequently, these bacterial strains were subjected to stressors that simulate cheese manufacturing conditions: NaCl (1, 1.5 and 2% w/v), lactic acid (0.5, 1.5 and 3% v/v), anaerobic growth, pasteurization (72°C for 15 s), UV irradiation. The ability to release prophage was evaluated by Real Time qPCR. Induction of the prophages showed that the addition of NaCl at 1.5 and 2% significantly increased viral release compared to control. Conversely, the addition of lactic acid had a significant repressive effect. The other applied stressors had no significant effect in phage release according to the experimental conditions adopted.

Insights into sublethal effects of Metarhizium anisopliae on the biotic potentials of Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) on maize

BackgroundEntomopathogenic fungi are an important biological agent in integrated pest management, playing a critical role in controlling insect populations. In the present study, the sublethal effects of the entomopathogenic fungus, Metarhizium anisopliae (Ascomycota: Hypocreales), were investigated on the biotic potential of Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), which is a major pest of economically important crops.ResultsObtained results revealed that exposure to LC50 concentration of M. anisopliae resulted in a significantly longer larval developmental time (3.25–6.45 days) than the control group (2.42–5.49 days). Similarly, pupal duration was longer in the LC50 treatment (8.24 days) than in the control (6.91 days). Adult longevity was also significantly reduced in the LC50 (9.64 days) and LC30 (10.49 days) treatments compared to the control group (11.7 days). The number of eggs laid by female S. frugiperda exposed to LC50 value of M. anisopliae during the immature stages was significantly lower (464.79 eggs) than that in the control groups (696.93 eggs). Furthermore, all population and age-stage-specific parameters were significantly affected by the sublethal exposure to M. anisopliae at LC30 and LC50 values.ConclusionThese results suggest that sublethal exposure to M. anisopliae negatively impacts the life table parameters of S. frugiperda. However, the use of M. anisopliae at sublethal levels may have potential benefits for integrated pest management strategies seeking to reduce the use of chemicals.

Swimming under Pressure: The Sub-Lethal Effects of a Pesticide on the Behaviour of Native and Non-Native Cypriniformes Fish

River ecosystems are exposed to a multitude of stressors, including increasing pesticide run-off driven by precipitation and irrigation. Pyrethroids are the fourth major group of insecticides in use worldwide and have extremely negative effects on aquatic fauna. In this study, we aimed to assess the effects of an acute 2 h sub-lethal exposure to different levels of the pyrethroid esfenvalerate on the swimming behaviour of two Cypriniformes species: the native Iberian barbel (Luciobarbus bocagei) and the non-native invasive bleak (Alburnus alburnus). The experimental set-up consisted of previous exposure to three esfenvalerate concentrations (control, 1.2 (low), and 2.0 (high) μg/L) before being stocked in a three-artificial-flume-channel mesocosm for behavioural trials through direct observation. Monitored behaviours included (i) routine activity, (ii) shoal cohesion, and iii) boldness. Significant differences in fish behaviour were detected for the native species (barbel), as individuals spent significantly more time holding position (i.e., resting) in the control (44.9%) than in the high esfenvalerate concentration (25.2%). Concordantly, control barbels were also found to perform more directional changes than the ones exposed to high esfenvalerate concentrations. Behavioural changes were also found for boldness, measured by the proportion of fish attempts to negotiate the upstream ramp, which were significantly higher in the control (37.4%) and in the high concentration (41.5%) compared to the low one (21.1%). Finally, regarding shoal cohesion of the barbel, it was tighter in the control (81.3%) than in the low- (70.5%) and high- (71.1%) esfenvalerate treatments. For the invasive bleak, there were no significant differences in any of the behavioural traits upon previous exposure to an increasing esfenvalerate concentration. This experimental study demonstrated that even short-term exposure to the pyrethroid esfenvalerate was sufficient to alter the behaviour of a native Cypriniformes fish species while not affecting the non-native species. This may confer greater competitive advantages to non-native fish species in the context of global changes.

Alteration in Metabolic Enzyme Activity of Muscle Tissue in the Fish <i>Cyprinus carpio</i> (l.) on Chronic Exposure to Some Pesticides

Studies were conducted to assess the effects of sub-lethal exposure of pesticides endosulfan, chlorpyrifos and malathion for 40 days on freshwater teleost, Cyprinus carpio. Jerky and whirling movements, surfacing, exodus trials, engulfing air, excess mucus secretion and poor response to feed were the commonly observed behavioral changes in the treated fish. A dose dependent reduction in the activity levels of selected dehydrogenase enzymes (glutamate dehyrogenase, succinate dehyrogenase, malate dehyrogenase and pyruvate dehyrogenase) was evident in the muscle tissues of the test animal. A reduction of 30.2 and 70.2 percent in activity levels of succinate dehydrogensae (SDH) was evident in 0.0001 and 0.001 ppm endosulfan treatments, respectively. The combirmtion of pesticides appeared consequently additive in nature. The toxic effects of endosulfan in combination with chlorpyrifos and malathion inhibited the SDH activity to a further low of 71.6 percent. Lactate dehydrogenase activity in the muscle tissue was higher in pesticide-exposed fish in comparison to the control fish.

Acute and sub-acute toxic effects of cadmium to freshwater tropical oligochaete Tubifex tubifex with special reference to oxidative stress and behavioural biomarkers

ABSTRACT Current environmental issues include heavy metal contamination. Cadmium pollution in aquatic environments harms aquatic creatures and can pass to people through food chains. Cadmium poisoning damages bones, kidneys and causes cancer. Tubifex tubifex is a well-known water pollution indicator because of its good adaptation power in environmental pollution. Tubifex sp. is chosen as the test animal in this study since it is an indicator species and also a model non-target organism in ecotoxicology. The aim of this present work is to assess the toxicity of cadmium nitrate on Tubifex tubifex as a biomarker. Acute toxicity of cadmium nitrate was analysed by measuring the 96 h LC50 value. Physical observations revealed that cadmium induced autotomy of the caudal region of the worm and induced more mucus secretion. Behavioural alterations like changes in mucous secretion, clumping tendency and wrinkling effect were observed in cadmium-treated worms. Antioxidant enzymes level (MDA, CAT and SOD) increased significantly on cadmium nitrate exposure. In general, biomarker data show that cadmium exposure has stress-related consequences at the biochemical and physiological levels, reducing the overall health and survival of such animals. Highlights The toxicity of the heavy metal cadmium in acute and sub-acute levels was investigated in Tubifex tubifex. The goal of this study was to look at the effects of cadmium on oxidative stress and behavioural biomarkers in T. tubifex Sublethal cadmium exposure can impact the physiological functioning of annelids in the wild.

Effect of Sub-Lethal, Carbaryl and Cartap Exposures on Lipid and Cholesterol Contents in Liver, Kidney and Gills of Channa Punctatus (BLOCH)

Channa punctatus were exposed for 24, 48, 72 and 96 h to sub-lethal concentration (30% of LC50 for 96h) of carbaryl and cartap (carbamate pesticides) in order to record their effect on total lipid and cholesterol contents in liver, kidney and gills. The normal lipid content in liver, kidney and gills ranged from 17.95±0.41 to18.21±0.38 mg/g, 15.55i-0.51 to16.21±0.35 mg/g and 8.53±0.39 to 9.58±0.33 mg/g, whereas cholesterol contents varied between 3.60±0.06 to 3.81±0.04 mg/g, 1.61±0.05 to 1.72±0.02 mg/g and 0.71±0.02 to 0.77±0.10 mg/g, respectively. The treatment elicited significant decline in lipid and cholesterol contents throughout the exposure period suggesting response of the fish to the pesticide intoxication.

Sublethal Exposure Enhances Suspectibility to Phoshine and Effects Certain Fitness Traits in Red Flour Beetle <i>Tribolium castaneum</i> (Herbst)

The insecticidal gaseous phosphine would fail to meet the required concentration owing to improper sealing in a majority of bulk storage units. The resulting sublethal concentration has toxicological and biological implications for the target pests. The red flour beetle Tribolium castaneum has developed considerable resistance to phosphine. In this study, the effect of sublethal phosphine (LC25 and LC40) dose exposure is seen in the first filial generation of T. castaneum, where it was found to increase the susceptibility of the F1 generation. The lethal concentration (LC50) of parent stock (0.076 mg/ l) was reduced to 0.059 mg/ l and 0.052 mg/ l in the F1 generation, respectively in the sample prior exposed to sublethal doses of LC25 and LC40. Surviving females of sublethal exposures (LC40 and LC25) produced fewer offspring with a cumulative fecundity reduction of 21 and 84% respectively, compared to parent stock. The variables tested in PCA highlight the negative effect of sublethal dose on the fecundity and fitness of female adults.

Bisphenol A effects on the host Biomphalaria alexandrina and its parasite Schistosoma mansoni.

Bisphenol A (BPA) is one of the most potent endocrine-disrupting chemicals (EDCs) that adversely affect aquatic organisms. The present investigation explored the effects of exposure to BPA at 0.1 and 1 mgL-1 concentrations on the fecundity of Biomphalaria alexandrina, snail's infection with Schistosoma mansoni, and histology of the ovotestis and topographical structure of S. mansoni cercariae emerged from exposed snails. The 24 h LC50 and LC90 values of BPA against B. alexandrina were 8.31 and 10.88 mgL-1 BPA, respectively. The exposure of snails to 0.1 or 1 mgL-1 BPA did not affect the snail's survival. However, these concentrations caused an increase in the reproductive rate (Ro) of infected snails. A slight decrease in egg production was observed in snails exposed to 0.1 mgL-1 BPA after being infected (infected then exposed). However, a significant increase in egg production was noted in snails exposed to 1 mgL-1 BPA after infection with S. mansoni. Histopathological investigations indicated a clear alteration in the ovotestis tissue structure of exposed and infected-exposed groups compared to the control snails. Chronic exposure to BPA caused pathological alterations in the gametogenic cells. SEM preparations of S. mansoni cercariae emerged from infected-exposed snails showed obvious body malformations. From a public health perspective, BPA pollution may negatively impact schistosomiasis transmission, as indicated by the disturbance in cercarial production and morphology. However, it has adverse effects on the reproduction and architecture of reproductive organs of exposed snails, indicating that B. alexandrina snails are sensitive to sublethal BPA exposure.

Environmental RNA as a Noninvasive Tool for Assessing Toxic Effects in Fish: A Proof-of-concept Study Using Japanese Medaka Exposed to Pyrene.

Although environmental RNA (eRNA) is emerging as a noninvasive tool to assess the health status of aquatic macroorganisms, the potential of eRNA in assessing chemical hazards remain largely untested. In this study, we investigated the ability of eRNA to detect changes in gene expression in Japanese medaka fish (Oryzias latipes) in response to sublethal pyrene exposure, as a model toxic chemical. We performed standardized acute toxicity tests and collected eRNA from tank water and RNA from fish tissue after 96 h of exposure. Our results showed that over 1000 genes were detected in eRNA and the sequenced read counts of these genes correlated with those in fish tissue (r = 0.50). Moreover, eRNA detected 86 differentially expressed genes in response to pyrene, some of which were shared by fish RNA, including the suppression of collagen fiber genes. These results suggest that eRNA has the potential to detect changes in gene expression in fish in response to environmental stressors without the need for sacrificing or causing pain to fish. However, we also found that the majority of sequenced reads of eRNA (>99%) were not mapped to the reference medaka genome and they originated from bacteria and fungi, resulting in low sequencing depth. In addition, eRNA, in particular nuclear genes, was highly degraded with a median transcript integrity number (TIN) of <20. These limitations highlight the need for future studies to improve the analytical methods of eRNA application.