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Queensland fruit fly (Q-fly), Bactrocera tryoni (Froggatt), presents a major threat to Australian fruit production and trade. The sterile insect technique (SIT) is increasingly employed to manage Q-fly. Quality of sterile males released in SIT programs, and hence program efficacy, can be affected by pre- and post-production processes, such as mass rearing, packing, irradiation, transportation, and release. Given long distances from rear-out facilities to release sites, adult flies are usually chilled to reduce metabolism and stress during transportation. To guide SIT procedures, it is important to understand the impact of such practices on performance of sterile Q-fly. The present study assesses the effect of chilling temperature and exposure period on quality parameters of sterile Q-fly. We considered the effects of two temperature regimes (4 and 6°C) and six exposure periods (0, 1, 2, 4, 6, and 12 h) on chill-coma recovery time, flight ability, survival under nutritional stress, and longevity of both males and females. Flies chilled at 4°C took longer to recover than that those chilled at 6°C. Flight ability, survival under nutritional stress, and longevity all decreased as chilling period increased but did not differ between the two tested temperatures. We recommend that periods of chilling during transportation from rear-out facilities to release sites be minimized in order to retain quality of sterile Q-fly and that increased release rates be considered when longer chilling periods are required.

In preclinical populations, binge consumption of a high-fat diet (HFD) initiated during either adolescence or adulthood increases the intravenous self-administration (IVSA) of cocaine, whereas ad lib HFD consumption initiated during adulthood reduces or fails to influence cocaine intake. From this, it appears that binge exposure is a sufficient condition to increase cocaine IVSA and that such effects occur independent of the exposure period. It is not clear, however, if ad lib exposure would be sufficient to affect the IVSA of cocaine if initiated during adolescence, a developmental period associated with high-risk behavior. To investigate this question, the present experiment evaluated the effects of consumption of a HFD given throughout adolescence and adulthood on cocaine IVSA (0.75 mg/kg/infusion). Specifically, male Sprague-Dawley rats were maintained on either a HFD (n = 24) or chow diet (n = 15) beginning on postnatal day (PND) 21 and as adults underwent cocaine IVSA [Fixed Ratio (FR) 1, FR 5, FR 10, FR 20, Progressive Ratio (PR) and cue- and drug + cue-induced responding] from PNDs 77-126. Under all of these conditions, animals maintained on the HFD displayed higher rates of cocaine IVSA than those given access to chow. The present data demonstrate that under these specific conditions long-term exposure during the risk period of adolescence and extended throughout adulthood is capable of impacting the subsequent likelihood of cocaine self-administration and suggest that diet type and the duration of exposure may be important factors influencing the vulnerability to drug intake. (PsycINFO Database Record (c) 2020 APA, all rights reserved).

Background There is an increased need to mitigate the emergence of insecticide resistance and incorporate new formulations and modes of application to control the urban vector Aedes aegypti. Most research and development of insecticide formulations for the control of Ae. aegypti has focused on their peridomestic use as truck-mounted ULV-sprays or thermal fogs despite the widespread knowledge that most resting Ae. aegypti are found indoors. A recent modification of indoor residual spraying (IRS), termed targeted IRS (TIRS) works by restricting applications to 1.5 m down to the floor and on key Ae. aegypti resting sites (under furniture). TIRS also opens the possibility of evaluating novel residual insecticide formulations currently being developed for malaria IRS. Methods We evaluated the residual efficacy of chlorfenapyr, formulated as Sylando 240SC, for 12 months on free-flying field-derived pyrethroid-resistant Ae. aegypti using a novel experimental house design in Merida, Mexico. On a monthly basis, 600 female Ae. aegypti were released into the houses and left indoors with access to sugar solution for 24 hours. After the exposure period, dead and alive mosquitoes were counted in houses treated with chlorfenapyr as well as untreated control houses to calculate 24-h mortality. An evaluation for these exposed cohorts of surviving mosquitoes was extended up to seven days under laboratory conditions to quantify “delayed mortality”. Results Mean acute (24-h) mortality of pyrethroid-resistant Ae. aegypti ranged 80–97% over 5 months, dropping below 30% after 7 months post-TIRS. If delayed mortality was considered (quantifying mosquito mortality up to 7 days after exposure), residual efficacy was above 90% for up to 7 months post-TIRS application. Generalized Additive Mixed Models quantified a residual efficacy of chlorfenapyr of 225 days (ca. 7.5 months). Conclusions Chlorfenapyr represents a new option for TIRS control of Ae. aegypti in urban areas, providing a highly-effective time of protection against indoor Ae. aegypti females of up to 7 months., Author summary Vector control (VC) for managing Aedes aegypti and reducing transmission of Aedes-borne diseases is largely focused on peridomestic insecticide applications. However, the indoor resting behavior of Ae. aegypti and the acceleration of insecticide resistance owed to reduced modes of action have diminished the effectiveness of many VC tools. A targeted Indoor residual spraying (TIRS) modality in experimental housing units was employed to investigate the potential of chlorfenapyr, a pyrrole-class insecticide with known effectiveness to resistant mosquito species. This was the first investigation for chlorfenapyr use against locally resistant Ae. aegypti (Merida, Mexico) with this approach. Two treatment arms were investigated in the present study: TIRS and a control house where only water was sprayed. A comparison of entomological efficacy for TIRS applied to interior perimeter walls below 1.5 m with chlorfenapyr (formulated as Sylando 240SC) at 250 mg/m2 over 12 months was assessed. TIRS chlorfenapyr treatments were highly efficacious and led to acute mortalities (after 24 exposure) above 80% up to 5 months; delayed mortalities (to Ae. aegypti) were monitored over seven days post exposures vs untreated controls. When delayed mortality was considered, residual efficacy of chlorfenapyr extended to 7 months. These data provide evidence that TIRS chlorfenapyr is an effective Aedes management tool that surpassed efficacy profiles for other TIRS insecticides that have been previously reported with this method. Further, Chlorfenapyr emerges as a novel addition to Ae. aegypti VC, and future studies should focus on its effectiveness and residual power as part of Phase II-III TIRS trials.

The purpose of this work was to determine the safety of inhaled hydrogen gas in healthy animals. Female mice were exposed to medical air with or without hydrogen gas (concentration 2.4%) for 72 hours (n = 25 mice/group). Mice underwent a standardized and validated neurobehavioral examination, SmithKline Beecham, Harwell, Imperial College, Royal London Hospital, Phenotype Assessment (SHIRPA) protocol, prior to and following the exposure period. Blood was withdrawn for serologic evaluation and all major organ tissues were evaluated histologically. The average hydrogen concentration within the chamber was 2.27%. Following exposure, there was no significant change in body weight in either group. Similarly, there was no significant change in the total SHIRPA score, although hydrogen-treated mice exhibited significantly lower spontaneous locomotor activity (P < 0.0001) in a subset of the test; all other aspects of the mouse neurologic exam were normal in hydrogen-treated animals. Brain histopathology was also normal in all mice, as was the histology of all other major organs. There were no significant differences in complete blood count, serum chemistry, or arterial blood gases between control and hydrogen-treated mice (P > 0.05 for all). Hydrogen gas did not appear to cause significant adverse effects when administered to healthy mice for 72 hours, with the possible exception of decreased spontaneous locomotor activity. The study was approved by the Institutional Animal Care and Use Committee at Boston Children’s Hospital, USA (approved number 18-01-3536) on January 25, 2018.

A controlled feeding experiment was conducted to investigate the toxicokinetic of polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/Fs) in laying hens. The laying hens were fed with fly ash contaminated diets to replicate the typical environment-feed-chicken transfer chain in China. Levels of PCDD/Fs rapidly increased in the pectoralis, adipose tissue, liver and blood of laying hens with daily ingestion of dioxin-associated diets during the 14-days exposure period, and then a gradual decrease was observed in the 28-days depletion period. The depletion rates (kd) of the toxic equivalent of PCDD/Fs (TEQPCDD/Fs) were 0.043, 0.031 and 0.030 day−1 for pectoralis, liver and adipose tissue in the high-exposure group, respectively. The kd of individual PCDD/Fs in liver increased with the numbers of chlorine and n-octanol/water partition coefficient (logKOW), indicating that lower chlorinated congeners had higher half-lives in liver. Decreasing ratios of liver to adipose tissue for PCDD/Fs (L/AT) throughout the experiment suggested a tendency of equilibrium partitioning between liver and adipose tissue. Congener-specific sequestration of PCDD/Fs in liver was revealed by the positive correlation between L/AT ratios and logKOW. Physiological bioconcentration factors of PCDD/Fs were estimated at the end of exposure, indicating the preferential accumulation of hexachlorinated congeners in most tissues. Furthermore, maternal transfer of PCDD/Fs was positively correlated with logKOW, implying that more lipophilic congeners were transferred to egg along with the lipid circulation.

The group of per- and polyfluoroalkyl substances (PFAS) comprises thousands of chemicals, which are used in various industrial applications and consumer products. In this study, a feeding experiment with laying hens and feed grown on a contamination site was conducted, and PFAS were analyzed in the feed and eggs to assess the transfer of PFAS into eggs. A targeted analysis of perfluoroalkyl acids (PFAAs) and different sulfonamides was performed. Additionally, the total oxidizable precursor (TOP) assay was modified by fully oxidizing small amounts of the samples instead of oxidizing their extracts in order to overcome potential losses during extraction. Targeted analysis showed the presence of known PFAAs and four sulfonamides in the feed and egg yolk samples. In the plant-based feed, short-chain PFAAs, methyl and ethyl perfluorooctane sulfonamidoacetic acid (Me- and EtFOSAA), and perfluorooctane sulfonic acid (PFOS) were the most abundant PFAS. In the eggs, PFOS, FOSAA, and its alkylated homologues showed the highest concentrations. The TOP assay revealed the presence of substantial amounts of precursors with different chain lengths from C4 to C8. The highest relative increase of PFOA after oxidation was observed in egg yolk from the end of the exposure period (828%). The results of this study demonstrate the transfer of PFAAs and their precursors into hens' eggs and emphasize the contribution of (known and unidentified) precursors to the overall PFAS burden in edible products. The modified TOP assay approach was shown to be a powerful tool to better assess the total burden of samples with PFAS.

Cannabis and alcohol co-use is prevalent in adolescence, but the long-term behavioural effects of this co-use remain largely unexplored. The aim of this study is to investigate the effects of adolescent alcohol and Δ9-tetrahydracannabinol (THC) vapour co-exposure on cognitive- and reward-related behaviours. Male Sprague-Dawley rats received vapourized THC (10 mg vapourized THC/four adolescent rats) or vehicle every other day (from post-natal day (PND) 28–42) and had continuous voluntary access to ethanol (10% volume/volume) in adolescence. Alcohol intake was measured during the exposure period to assess the acute effects of THC on alcohol consumption. In adulthood (PND 56+), rats underwent behavioural testing. Adolescent rats showed higher alcohol preference, assessed using the two-bottle choice test, on days on which they were not exposed to THC vapour. In adulthood, rats that drank alcohol as adolescents exhibited short-term memory deficits and showed decreased alcohol preference; on the other hand, rats exposed to THC vapour showed learning impairments in the delay-discounting task. Vapourized THC, alcohol or their combination had no effect on anxiety-like behaviours in adulthood. Our results show that although adolescent THC exposure acutely affects alcohol drinking, adolescent alcohol and cannabis co-use may not produce long-term additive effects.

The current understanding of nanoplastics (NPLs) toxicity to freshwater biota, especially the potential toxic effects of polymethylmethacrylate (PMMA), remains limited. Thus, the present work intended to add knowledge about the ecotoxicity of ∼40 nm PMMA-NPLs focusing on lethality, morphology, feeding and regeneration capacity of the freshwater cnidarian Hydra viridissima, after an exposure period of 96 h. Results showed that high concentrations of PMMA-NPLs can impair the survival of H. viridissima, with an estimated 96 h-LC50 of 84.0 mg PMMA-NPLs/L. Several morphological alterations were detected at concentrations below 40 PMMA-NPLs mg/L, namely partial or total loss of tentacles, which, however, did not induce significant alterations on the feeding rates. Morphological alterations not previously reported in the literature were also found after the 96 h exposure, such as double or elbow-like tentacles. Exposure to 40 mg PMMA-NPLs/L significantly impacted hydra regeneration, with organisms exposed to PMMA-NPLs presenting significant slower regeneration rates comparatively to controls, but with no impacts on the feeding rates. Overall, this work highlights the need to assess the effects of NPLs in freshwater biota. Hydra viridissima species was sensitive in a wide range of endpoints showing its value as a biological model to study the effects of small plastic particles. published

The primary objective of this experiment was to assess the effect of mouthpiece chamber vacuum on teat-end congestion. The secondary objective was to assess the interactive effects of mouthpiece chamber vacuum with teat-end vacuum and pulsation setting on teat-end congestion. The influence of system vacuum, pulsation settings, mouthpiece chamber vacuum, and teat-end vacuum on teat-end congestion were tested in a 2×2 factorial design. The low-risk conditions for teat-end congestion (TEL) were 40 kPa system vacuum (Vs) and 400-ms pulsation b-phase. The high-risk conditions for teat-end congestion (TEH) were 49 kPa Vs and 700-ms b-phase. The low-risk condition for teat-barrel congestion (TBL) was created by venting the liner mouthpiece chamber to atmosphere. In the high-risk condition for teat-barrel congestion (TBH) the mouthpiece chamber was connected to short milk tube vacuum. Eight cows (32 quarters) were used in the experiment conducted during 0400 h milkings. All cows received all treatments over the entire experimental period. Teatcups were removed after 150 s for all treatments to standardize the exposure period. Calculated teat canal cross-sectional area (CA) was used to assess congestion of teat tissue. The main effect of the teat-end treatment was a reduction in CA of 9.9% between TEL and TEH conditions, for both levels of teat-barrel congestion risk. The main effect of the teat-barrel treatment was remarkably similar, with a decrease of 9.7% in CA between TBL and TBH conditions for both levels of teat-end congestion risk. No interaction between treatments was detected, hence the main effects are additive. The most aggressive of the 4 treatment combinations (TEH plus TBH) had a CA estimate 20% smaller than for the most gentle treatment combination (TEL plus TBL). The conditions designed to impair circulation in the teat barrel also had a deleterious effect on circulation at the teat end. This experiment highlights the importance of elevated mouthpiece chamber vacuum on teat-end congestion and resultant decreases in CA.

Agricultural use of neonicotinoid insecticides is increasing worldwide, posing a risk to nontarget organisms. The present study investigated developmental toxicity of a widely used neonicotinoid, acetamiprid, to zebrafish embryos. Sublethal (malformations, hatchability, heart rate, body length, alteration of spontaneous movement and touch responses) and lethal effects were monitored during exposure period from 6 h post fertilization (hpf) to 120 hpf. Zebrafish embryos exhibited significant mortality and teratogenic effects at acetamiprid concentration greater than 263 mg/L, with bent spine being the main malformation. Toxicity spectra were constructed to rank the sensitivity of individual end points to acetamiprid exposure and impaired spontaneous movement was the most sensitive end point of those tested. The present study provides the basis for understanding developmental toxicity of acetamiprid exposure to zebrafish embryos. This information is critical for future studies evaluating aquatic risk from neonicotinoids as little is known regarding adverse effects of neonicotinoids to aquatic vertebrate species.

Twelve healthy dogs were included in this laboratory efficacy study. Six dogs were randomly allocated based on body weight to an untreated control group and six to an afoxolaner (NexGard®) treated group. In the treatment group, afoxolaner was administered orally on Day 0 in accordance with label instructions. On Days 1, 14 and 28, each dog was exposed to 60 unfed female and 10 male Phlebotomus perniciosus sandflies for 1 h. At the end of each exposure period, sandflies were counted and assessed for viability and feeding status. There was no statistical difference in mortality (0.0–5.4%), nor in feeding proportion (61.6–78%) between the control and the treated groups at all 1-h post-exposure assessments. After collection, live fed and unfed sandflies were kept for viability assessments at 48 and 72 h post-exposure. In the untreated control group, the average percentages of live, fed, female sandflies after exposure, on Days 1, 14 and 28, ranged from 51% to 74% at 48 h and from 46% to 57% at 72 h, demonstrating model robustness over the 28 days of the study. Significantly fewer live fed sandflies were recorded for the afoxolaner treated group (p ®) to dogs significantly killed P. perniciosus sandflies 48 and 72 h after blood feeding for 1 month.

On August 3rd, 2017, a Q fever outbreak alert was issued at a courier company that in addition to urgent freight transport offered pet delivery services. The epidemiological investigation set the exposition period between June 1 and August 8. In this period, 180 workers from two operational platforms for parcel distribution located in two provinces of the Basque Country (Bizkaia and Araba) were exposed; 64 filled a questionnaire and provided blood samples for serological testing, resulting in 10 confirmed cases (15.6%) and six (9.4%) probable cases. Nine workers (8 confirmed and 1 probable) showed Q fever symptoms, including pneumonia (five cases), and required medical care services, including one hospital admission. The attack rate was 25% (16/64), being higher among workers that visited the Bizkaia platform. This suggested that the origin of the outbreak was in the Bizkaia platform, where animals in transit waited at a pet holding site until being moved to their destination. Environmental samples consisting on 19 surface dust and two aerosol samples were collected at the Bizkaia platform to investigate the presence of C. burnetti DNA. All dust samples were positive by real time PCR, the lowest Ct values being found in dust collected at the pet holding facilities, and therefore suggesting that contamination originated at the pet holding site. The genotype identified in dust was SNP1/MST13, one of the most commonly identified genotypes in goats and sheep in the Basque Country. During the exposure period, two deliveries of miniature goats were made, of which only one could be investigated and tested negative. Although the contamination source could not be unequivocally identified, transport of ruminants was banned at the company, and Q fever was included among the occupational-associated health risks. This work was funded by the Spanish National Institute for Agricultural and Food Research and Technology (INIA) (RTA2017-00055-C02-00), the European Regional Development Funds (ERDF) and the Basque Government. RAA is beneficiary of a PhD contract funded by INIA (FPI-2015-014). The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Sí

Objective: Complete systems for laboratory-based inhalation toxicology studies are typically not commercially available; therefore, inhalation toxicologists utilize custom-made exposure systems. Here we report on the design, construction, testing, operation and maintenance of a newly developed in vivo rodent ozone inhalation exposure system. Materials and methods: Key design requirements for the system included large-capacity exposure chambers to facilitate studies with large sample sizes, automatic and precise control of chamber ozone concentrations, as well as automated data collection on airflow and environmental conditions. The exposure system contains two Hazelton H-1000 stainless steel and glass exposure chambers, each providing capacity for up to 180 mice or 96 rats. We developed an empirically tuned proportional-integral-derivative control loop that provides stable ozone concentrations throughout the exposure period (typically 3h), after a short ramp time (∼8 min), and across a tested concentration range of 0.2–2 ppm. Specific details on the combination of analog and digital input/output system for environmental data acquisition, control and safety systems are provided, and we outline the steps involved in maintenance and calibration of the system. Results: We show that the exposure system produces consistent ozone exposures both within and across experiments, as evidenced by low coefficients of variation in chamber ozone concentration and consistent biological responses (airway inflammation) in mice, respectively. Conclusion: Thus, we have created a large and robust ozone exposure system, facilitating future studies on the health effects of ozone in rodents.

International audience; Juvenile crabs of Carcinus maenas thrive in coastal waters reputed to be the receptacle of continental pollution. Amongst the many pollutants encountered, antidepressants, such as fluoxetine (FLX) and venlafaxine (VEN), often detected at the ng•L(-1) range, are particularly worrying because of their action on the levels of monoamines, such as serotonin, noradrenaline and dopamine. In crustaceans, those monoamines are involved in colour change through their action on neuropeptide hormones. In addition, they are known to have a role in different behaviours, such as locomotion. Both colour change and locomotion are strategies used by juvenile crabs to hide and escape from predators. To investigate if the presence of antidepressants may alter behaviours of ecological importance, juvenile crabs were exposed to environmentally realistic concentrations of either 5 ng•L(-1) of FLX alone or in combination with VEN at 5 ng•L(-1). The ability to change colour depending on the environment and the locomotor activity of juvenile crabs were monitored weekly over 25 days. Animals exposed to antidepressants displayed a different pattern of colour change than the controls, especially those exposed to the combination of FLX and VEN at 5 ng•L(-1) each, and were less efficient to adapt to their environment, i.e., they were not as pale and not as dark as controls or crabs exposed to FLX at 5 ng•L(-1). Moreover, juvenile crabs exposed to the combination of antidepressants exhibited an enhanced locomotor activity throughout the exposure period with a higher velocity and distance moved as well as more time spend moving. The alteration of cryptic behaviours, such as colour change and locomotion by antidepressants persistently present in marine environment at low concentrations may have an impact on the survival of juvenile of C. maenas on the long term.

The rat tapeworm Hymenolepis diminuta can bioconcentrate several elements to conspicuously higher concentrations than tissues of their definitive host. The main aim of this study was to locate parts of the tapeworm into which lead, cadmium, zinc, copper, manganese and iron are accumulated. Male Wistar rats were experimentally infected with H. diminuta and worms were exposed to two different forms of lead for 6 weeks through the oral exposure of their rat hosts. After the exposure period, the element levels were determined in the posterior and anterior proglottids of the tapeworm. In all cases, lead concentrations were higher in the anterior parts than the posterior parts. Concentrations of cadmium, copper, iron, manganese and zinc were also significantly higher in the anterior parts. Zinc concentrations showed an opposite trend, with higher zinc levels detected in the posterior part of the strobila, in the control group. The present study demonstrates that risk and essential elements are accumulated mainly into the anterior part of H. diminuta.

Recent human behavioral studies have shown that the position of a visual target is instantly represented relative to the background (e.g., a large square) and used for evaluating the error in reaching the target. In the present study, we examined whether the same allocentric mechanism is shared by the monkey. We trained two monkeys to perform a fast and accurate reaching movement toward a visual target with a square in the background. Then, a visual shift (20mm or 4.1°) was introduced by wedge prisms to examine the process of decreasing the error during an exposure period (30 trials) and the size of the error upon removal of the prisms (aftereffect). The square was shifted during each movement, either in the direction of the visual displacement or in the opposite direction, by an amount equal to the size of the visual shift. The ipsilateral shift of the background increased the asymptote during the exposure period and decreased the aftereffect, i.e., prism adaptation was attenuated by the ipsilateral shift. By contrast, a contralateral shift enhanced adaptation. We further tested whether the shift of the square alone could cause an increase in the motor error. Although the target did not move, the shift of the square increased the motor error in the direction of the shift. These results were generally consistent with the results reported in human subjects, suggesting that the monkey and the human share the same neural mechanisms for representing a target relative to the background.

Bacteria play an important role in the degradation of bone material. However, much remains to be learnt about the structure of their communities in degrading bone, and how the depositional environment influences their diversity throughout the exposure period. We genetically profiled the bacterial community in an experimental series of pig bone fragments (femur and humeri) deposited at different well-defined environments in Denmark. The bacterial community in the bone fragments and surrounding depositional environment were studied over one year, and correlated with the bioerosion damage patterns observed microscopically in the bones. We observed that the bacterial communities within the bones were heavily influenced by the local microbial community, and that the general bone microbial diversity increases with time after exposure. We found the presence of several known collagenase producing bacterial groups, and also observed increases in the relative abundance of several of these in bones with tunneling. We anticipate that future analyses using shotgun metagenomics on this and similar datasets will be able to provide insights into mechanisms of microbiome driven bone degradation. © 2020 Eriksen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Bioaccumulation of 2,4,6-trinitrotoluene (TNT) and its main metabolites 2-amino-4,6-dinitrotoluene (2-ADNT) and 4-amino-2,6-dinitrotoluene (4-ADNT) leaking from corroded munitions at a munitions dumping site (Kolberger Heide, Germany) was evaluated in transplanted blue mussels (Mytilus edulis). Six moorings with mussel bags were placed east and west at varying positions near the mine mound. In order to monitor any differences resulting from changing seasons, three exposure times were chosen. First exposure period: April–July 2016 (106 days); second exposure period: July–December 2016 (146 days); third exposure period: December 2016–March 2017 (92 days). We found amounts of 4-ADNT in mussel tissue ranging from 2.40 ± 2.13 to 7.76 ± 1.97 ng/(g mussel wet weight). Neither TNT nor 2-ADNT could be detected. Considering seasonal differences, orientation and distances of the moorings to the mine mound no correlation between levels in mussel tissue was evident.

The aversive flavor of ethanol limits intake by many consumers. We asked whether intermittent consumption of ethanol increases its oral acceptability, using rats as a model system. We focused on adolescent rats because they (like their human counterparts) have a higher risk for alcohol overconsumption than do adult rats following experience with the drug. We measured the impact of ethanol exposure on 1) the oral acceptability of ethanol and surrogates for its bitter (quinine) and sweet (sucrose) flavor components in brief-access lick tests and 2) responses of the glossopharyngeal (GL) taste nerve to oral stimulation with the same chemical stimuli. During the exposure period, the experimental rats had access to chow, water and 10% ethanol every other day for 16 days; the control rats had access to chow and water over the same time period. The experimental rats consumed 7-14 g/day of 10% ethanol across the exposure period. This ethanol consumption significantly increased the oral acceptability of 3%, 6% and 10% ethanol, but had no impact on the oral acceptability of quinine, sucrose or NaCl. The ethanol exposure also diminished responses of the GL nerve to oral stimulation with ethanol, but not quinine, sucrose or NaCl. Taken together, these findings indicate that ethanol consumption increases the oral acceptability of ethanol in adolescent rats and that this increased oral acceptability is mediated, at least in part, by an exposure-induced reduction in responsiveness of the peripheral taste system to ethanol per se, rather than its bitter and sweet flavor components.

Long-term genotoxic effects of two auxinic herbicide formulations, namely, the 58.4% 2,4-dichlorophenoxyacetic acid (2,4-D)-based DMA® and the 57.7% dicamba (DIC)-based Banvel® were evaluated on Cnesterodon decemmaculatus. Primary DNA lesions were analyzed by the single-cell gel electrophoresis methodology. Two sublethal concentrations were tested for each herbicide corresponding to 2.5% and 5% of the LC5096h values. Accordingly, fish were exposed to 25.2 and 50.4 mg/L or 41 and 82 mg/L for 2,4-D and DIC, respectively. Fish were continuously exposed for 28 days with replacement of test solutions every 3 days. Genotoxicity was evaluated in ten individuals from each experimental point at the beginning of the exposure period (0 day) and at 7, 14, 21 and 28 days thereafter. Results demonstrated for first time that 2,4-D-based formulation DMA® induced primary DNA strand breaks after 7–28 days exposure on C. decemmaculatus regardless its concentration. On the other hand, DIC-based formulation Banvel® exerted its genotoxic effect after exposure during 7–14 days and 7 days of 2.5 and 5% LC5096h, respectively. The present study represents the first evidence of primary DNA lesions induced by two widely employed auxinic herbicides on C. decemmaculatus, namely 2,4-D and DIC, following long-term exposure. 2,4-D and dicamba exert primary DNA lesions in blood cells of C. decemmaculatus after long-term exposure. Fil: Ruiz de Arcaute, Celeste. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Facultad de Ciencias Naturales y Museo, Universidad Nacional de la Plata; Argentina Fil: Larramendy, Marcelo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Facultad de Ciencias Naturales y Museo, Universidad Nacional de la Plata; Argentina Fil: Soloneski, Sonia Maria Elsa. Facultad de Ciencias Naturales y Museo, Universidad Nacional de la Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

We investigated the olfactory toxicity of copper (Cu) to rainbow trout in low-hardness (27 mg/L as CaCO3 ) water formulated in the laboratory over a 120-h period using a flow-through design. The fish's response to an alarm cue (e.g., reduction in activity) was recorded to determine the exposure concentrations and durations that inhibited olfactory detection of the cue after 3, 24, 48, and 96 h of Cu exposure and after 24 h of clean water recovery following the 96-h exposure period. Exposures were conducted with a range of Cu concentrations from 0.13 (control) to 7.14 μg Cu/L (dissolved Cu). We observed a dose-dependent response in olfactory inhibition with a 20% reduction in the probability of responding to the alarm cue, relative to controls, at 2.7 and 2.4 μg Cu/L after 24 or 96 h of exposure, respectively. Olfactory inhibition manifested between 3 and 24 h of exposure. Our 24- and 96-h 20% olfactory inhibition estimates fell between the criteria derived using the biotic ligand model (BLM; criterion maximum concentration [CMC] and criterion continuous concentration [CCC] values were 0.63 and 0.39 μg Cu/L, respectively) and water hardness-based criteria (CMC and CCC values were 3.9 and 2.9 μg Cu/L, respectively). Therefore, the hardness-based criteria do not appear to be protective and the BLM-derived criteria do appear to be protective against Cu-induced olfactory inhibition given our test water chemistry. Neither the hardness-based criteria nor the BLM-derived criteria appear to be protective against our estimated Cu behavioral avoidance response concentrations at 24- and 96-h exposures (0.54 and 0.50 μg Cu/L, respectively). Environ Toxicol Chem 2019;38:198-209. © 2018 SETAC.

Our aim was to evaluate whether exposure to Bisphenol A (BPA) modifies the male rat mammary gland (MG) development and to compare the effects on male and female MG. From gestational day 9, pregnant rats were exposed either subcutaneously to 0, 25 or 250µg BPA/kgbw/day until parturition or orally to 0 and 64µg BPA/kgbw/day until weaning. MG development was analyzed on postnatal days (PND) 5, 15 and 30. On PND30, steroid hormone receptor expression and mammary growth were also evaluated. On PND30, the exposure to 64BPA and 250BPA induced a delay in male MG development, evidenced by reduced ductal growth, decreased number of terminal structures and lower expression of androgen receptor (AR). In contrast, female mammary ductal growth was altered only by 250BPA. Regardless of the administration route and length of the exposure period, BPA induced a delay in MG development and modified AR expression in prepubertal male rats. Fil: Kass, Laura. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Fisiología. Laboratorio de Endocrinología y Tumores Hormonodependientes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina Fil: Durando, Milena de Lourdes. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Fisiología. Laboratorio de Endocrinología y Tumores Hormonodependientes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina Fil: Altamirano, Gabriela Anahí. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Fisiología. Laboratorio de Endocrinología y Tumores Hormonodependientes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina Fil: Manfroni Ghibaudo, Gisela E.. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Fisiología. Laboratorio de Endocrinología y Tumores Hormonodependientes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina Fil: Luque, Enrique Hugo. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Fisiología. Laboratorio de Endocrinología y Tumores Hormonodependientes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina Fil: Muñoz de Toro, Monica Milagros. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Fisiología. Laboratorio de Endocrinología y Tumores Hormonodependientes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina

In the search for new molluscicidal plants for controlling the snail vectors of schistosomiasis, laboratory evaluation was made to assess the molluscicidal activity of Agave angustifolia and Pittosporum tobira plants against Biomphalaria alexandrina snails. Results indicated that both plants have promising molluscicidal activity as the LC90 of the dry powder of both plants was 120 ppm. Both plants showed marked cercaricidal and miracidicidal potencies against S. mansoni larvae. The LC90 of both plants (120 ppm) killed most B. alexandrina eggs within 24 h of exposure. The sub-lethal concentrations of both plants markedly suppressed the survival rate of B. alexandrina snails and the mortality increased with increasing the concentrations and the exposure period up to 10 successive weeks. The accumulative toxic effect of these concentrations was continuous during the recovery period. Also, the reproductive rates of exposed snails were greatly affected even through the recovery period. This depression in reproductive ability of snails was accompanied by histological damage in the hermaphrodite glands of exposed snails. Meanwhile, the growth of snails was estimated weekly and it showed great inhibition in exposed snails comparing with the control ones.

Fish are capable of sensing water-borne chemicals at sub-lethal concentrations. Inadequate behavioral responses to physiological and environmental stimuli owing to adverse effects of aquatic toxicants can have serious implications for survival. In this study we exposed juvenile rainbow trout ( Oncorhynchus mykiss ) during 5 days to a low-concentration mixture of three co-occurring herbicides: atrazine, linuron and metolachlor, at maximum concentrations of 4.5, 4.9 and 13.4 μg L −1 , respectively. Our hypothesis was that fish behavior – swimming activity and interactions between individuals – would be modified due to exposure to the mixture. We studied these behaviors by observing fish twice-daily throughout the exposure period at 30-s intervals for 5 min, registering the vertical distribution of fish in the water column and the number of agoniztic acts between all individuals. Fish exposed to the mixture of herbicides were hypoactive and spent more time in the lower parts of the aquaria in comparison to non-exposed controls, reflecting inhibited swimming activity. Average swimming height of exposed fish decreased significantly with the number of agoniztic acts, whilst in control groups there was no significant relationship between the two behaviors. Overall, behavior of fish exposed for a short time to the herbicide mixture was altered in comparison to control-fish behavior. The behavioral endpoints chosen here were easily observed, simple to quantify, and of ecological relevance.

Traditional toxicological studies usually depict the dose–effect relationship at the given exposure time while ignore how toxic effects vary during and after the exposure. In the present study, feeding depression of the rotifer Brachionus calyciflorus under the impact of dimethoate was evaluated not only in the exposure time but also in the post-exposure time. Both the filtration rates ( F ) and the ingestion rates ( I ) of the rotifers varied during the exposure period under any concentration of dimethoate. A three-stepwise stress was observed when the rotifers were exposed to 0.2 and 1.0 mg L −1 of the pesticide respectively, whereas 1.8 mg L −1 dimethoate caused only one-step stress on the rotifers during the whole period. In addition, although the feeding behaviors of the rotifers recovered after the exposure, two feeding rates both got even more depressed at 8 h in the post-exposure time. The latent inhibition after exposure was related to the exposure concentrations and the exposure time. Our works indicated that the impact of dimethoate on the feeding behavior of the rotifers was highly process-dependent that caused the stepwise stress during the exposure period and the latent stress after the exposure.

Flow cytometry was used to investigate the immune parameters of haemocytes in thick-shell mussel Mytilus coruscus exposed to different concentrations of ZnO nanoparticles (NPs) (0, 2.5, and 10 mg l− 1) at two pH levels (7.3 and 8.1) for 14 days following a recovery period of 7 days. ZnO NPs significantly affected all of the immune parameters throughout the experiment. At high ZnO NPs concentrations, total haemocyte counting, phagocytosis, esterase, and lysosomal content were significantly decreased whereas haemocyte mortality and reactive oxygen species (ROS) were increased. Although low pH also significantly influenced all of the immune parameters of the mussels, its effect was not as strong as that of ZnO NPs. Interactive effects were observed between pH and ZnO NPs in most haemocyte parameters during the exposure period. Although a slight recovery from the stress of ZnO NPs and pH was observed for all immune parameters, significant carry-over effects of low pH and ZnO NPs were still detected. This study revealed that high concentration of ZnO NPs and low pH exert negative and synergistic effects on mussels, and these effects remain even after the mussels are no longer exposed to such stressors.

Diamide insecticides have become the fourth most commonly used insecticide class in the world. Chlorantraniliprole (CAP) is a first-generation diamide insecticide with broad application potential. In this experiment, the eco-toxicity of CAP in soil at 0.1, 1.0, 5.0 and 10.0mg/kg on earthworms (Eisenia fetida) was evaluated during a 42 d exposure. More specifically, the environmental fate and transport of CAP between soil and earthworms was monitored during the exposure period. The present results indicated that the CAP contents of 0.1, 1.0, 5.0 and 10.0mg/kg treatments decreased to no more than 20% in the soil after 42 d of exposure. The accumulation of CAP in earthworms was 0.03, 0.58, 4.28 and 7.21mg/kg earthworm (FW) at 0.1, 1.0, 5.0 and 10.0mg/kg after 42 d of exposure. At 0.1mg/kg and 1.0mg/kg, CAP had no effect on earthworms during the exposure period. The weight of earthworms was significantly reduced at 5.0 and 10.0mg/kg at 28 and 42 days after CAP application. After the 14th day, CAP induced excess production of reactive oxygen species (ROS) at 5.0 and 10.0mg/kg, resulting in oxidative damage to biomacromolecules. We believe that CAP has a high risk potential for earthworms when used at 5.0 and 10.0mg/kg.

Rock bream iridovirus (RBIV) is a member of the Megalocytivirus genus that causes severe mortality to rock bream. Water temperature is known to affect the immune system and susceptibility of fish to RBIV infection. In this study, we evaluated the time dependent virus replication pattern and time required to completely eliminate virus from the rock bream body against RBIV infection at different water temperature conditions. The rock bream was exposed to the virus and held at 7 (group A1), 4 (group A2) and 2 days (group A3) at 23 °C before the water temperature was reduced to 17 °C. A total of 28% mortality was observed 24–35 days post infection (dpi) in only the 7 day exposure group at 23 °C. In all 23 °C exposure groups, virus replication peaked at 20 to 22 dpi (10 6 –10 7 /μl). In recovery stages (30–100 dpi), the virus copy number was gradually reduced, from 10 6 to 10 1 with faster decreases in the shorter exposure period group at 23 °C. When the water temperature was increased in surviving fish from 17 to 26 °C at 70 dpi, they did not show any mortality or signs of disease and had low virus copy numbers (below 10 2 /μl). Thus, fish need at least 50 days from peaked RBIV levels (approximately 20–25 dpi) to inhibit the virus. This indicates that maintaining the fish at low water temperature (17 °C) for 70 days is sufficient to eradicate RBIV from fish body. Thus, RBIV could be eliminated slowly from the fish body and the virus may be completely eliminated under the threshold of causing mortality.

Objective To observe the effects of Shisha smoke on submucosal glands of trachea of mice; and compare it with tracheal glands of mice exposed to cigarette smoke. Study design Randomised controlled trial. Place and duration of study Department of Anatomy, Islamic International Medical College, Rawalpindi, in collaboration with National Institute of Health (NIH), Islamabad from October 2013 till April 2014. Methodology Sample comprised of 40 adult male mice of strain BALB/c. They were randomly divided into three groups. Control group was labelled as Group 'C'. The mice in this group were kept in a whole body smoke exposure chamber and were exposed to fresh air. Shisha group was labelled as Group 'SS', and the mice in this group were exposed to Shisha smoke. Mice in the third group labelled as Group CS were exposed to cigarette smoke. All the mice were dissected after an exposure period of eight weeks. Tracheal tissue was stained and examined microscopically for submucosal gland hypertrophy and compared with the control group, using Reid's Index. An Index of more than 0.4 is termed as hypertrophy. Results There was significant submucosal gland hypertrophy in groups CS and SS as compared to group C. There was also significant difference in the frequency of mucosal hypertrophy between SS (93.7%) and CS groups (53.3%), which was found statistically significant (p Conclusion Shisha smoking was significantly associated with mucosal hypertrophy when compared with cigarette smoking and controls. Shishasmoke contains higher level of toxicants as compared to cigarette smoke, and it causes more oxidative damage of tissues.

International audience; A previous study claimed a differential behavioural resilience between spring or summer honey bees (Apis mellifera) and bumble bees (Bombus terrestris) after exposure to syrup contaminated with 125 μg L−1 imidacloprid for 8 days. The authors of that study based their assertion on the lack of body residues and toxic effects in honey bees, whereas bumble bees showed body residues of imidacloprid and impaired locomotion during the exposure. We have reproduced their experiment using winter honey bees subject to the same protocol. After exposure to syrup contaminated with 125 μg L−1 imidacloprid, honey bees experienced high mortality rates (up to 45%), had body residues of imidacloprid in the range 2.7–5.7 ng g−1 and exhibited abnormal behaviours (restless, apathetic, trembling and falling over) that were significantly different from the controls. There was incomplete clearance of the insecticide during the 10-day exposure period. Our results contrast with the findings reported in the previous study for spring or summer honey bees, but are consistent with the results reported for the other bee species

Bumblebees are facultative endotherms, having the ability to elevate thorax temperature above ambient temperature by elevating metabolism. Here, we investigated the influence of hypoxia on metabolic demands and thermoregulatory capabilities of the bumblebee Bombus terrestris. We measured thorax temperature, rates of oxygen consumption and carbon dioxide production, and abdominal pumping rates of bees randomly exposed to oxygen levels of 20, 15, 10 and 5 kPa at 26°C. Under normoxia, bumblebees maintained an elevated mean thorax temperature of 35.5°C. There was no significant change in thorax temperature at 15 kPa O2 (33.4°C). Mean thorax temperature decreased significantly at 10 kPa O2 (31.6°C) and 5 kPa O2 (27.3°C). Bees were able to maintain an elevated metabolic rate at 15 and 10 kPa O2. In normoxia, endothermic bees exhibited periods of rapid abdominal pumping (327 min−1) interspaced by periods of no abdominal pumping. At 10 kPa O2, abdominal pumping rate decreased (255 min−1) but became more continuous. Upon exposure to 5 kPa, metabolic rate and abdominal pumping rate (152 min−1) decreased, although the animals continued abdominal pumping at the reduced rate throughout the exposure period. Bumblebees are able to meet the energetic demands of endothermy at 15 kPa O2, but become compromised at levels of 10 kPa O2 and below.

Six, nonpregnant, Barbari goats aged 4–5 years were selected for the study. For the first 6 days, the animals were kept in psychrometric chamber at thermoneutral temperature for 6 h each day to make them acclimated to climatic chamber. On the 7th day, the animals were exposed to 41 °C temperature for 3 h and then to 45 °C for the next 3 h. Cardinal physiological responses were measured, and blood samples (3 ml) were collected at 1-h interval during the heat exposure period and then once after 6 h of the heat exposure. The rectal temperature (RT) and respiratory rate (RR) increased significantly (P

The insecticidal properties of Hoslundia opposita Vahl (Lamiaceae) leaves' essential oil (EO) against Tribolium castaneum were investigated using contact toxicity and repellency bioassays. Mortality progressed with exposure period and ranged from 61.13% observed at 24 h after treatment (HAT) to 88.86% at 168 HAT. The LT50 (lethal time for 50% of treated adults) of H. opposita EO against T. castaneum was 10.42 h. Application of EO at 20-30 μL/30 cm(2) caused significantly (P0.05) higher percentage repellency than what was observed in control, 10 and 15 μL/30 cm(2) at 1 and 3 HAT, with significant repellency at 24 HAT regardless of dosage. Repellency class increased with EO dosage, with class V observed at 30 μL/30 cm(2), regardless of exposure duration. The RD50 (repellency dose for 50% of treated adults) of 15.88 and 13.37 μL/30 cm(2) for 1 and 2 HAT, respectively, was significantly higher than 0.09 μL/30 cm(2) at 24 HAT.

This study was performed in water ditches under simulated natural conditions in Egypt to elucidate the effect of various environmental factors on Schistosoma mansoni cercarial host location and infection of the definitive host (using albino mice). Evaluation of these factors was dependent on both infection rate of exposed mice as well as the schistosome worm load under the same experimental conditions. The seasonal water temperature proved to be a very important factor and this was proven by the infection rate of mice and the worm load recovered were lower in January and April (16 degrees C and 22 degrees C midday water temperature) and much higher in July and October (29 degrees C and 25 degrees C). The daytime factor is similarly important as temperature illustrated by the schistosome infection of mice groups exposed at 8-10 am was much higher than in groups exposed between 1 pm and 3pm (p < 0.001). The greatest infection rate of mice and worm load were obtained when the shedding snails were close to the exposed group of mice. Both criteria increased with the increase of cercarial density in the water. The length of exposure period is also an extremely important factor for schistosome infection, being highest 87.5% (p < 0.001) in3 hours exposure period. Infection rate was found to be 88.2% and 55.6% of shedding snails were located at water surface and midway to the bottom, respectively, and no infection occurred when located at the bottom. The schistosome infection of mice decreased in presence of increasing density of the floating plant Eichhornia crassipes in the ditch water, but low condensation of the submersed plant Ceratophyllum demersum appeared to have stimulating effect.

A 120-day toxicity experiment was conducted to investigate the effect of dietary cyanobacteria on the growth and liver histopathology of yellow catfish, and subsequent recovery when the fish were free of cyanobacteria. Three experimental diets were formulated: the control (cyanobacteria-free diet), low-cyanobacteria diet (LCD, 32.3 mu g microsystins/g) and high-cyanobacteria diet (HCD, 71.96 mu g microsystins/g). Each diet was fed to fish for 60 days and then all fish were free of cyanobacteria for a further 60 days. The results showed that a significant decrease in the specific growth rate (SGR) was observed in both fish fed with the LCD and HCD after a 1st 30-day exposure period, however, no significant difference in the SGR between the LCD and control groups was observed after a 2nd 30-day exposure period. At the end of the 60 days exposure, all examined liver tissues in both doses exhibited what appeared as dose-dependent histopathological modifications. After a 60-day recovery, there were no significant differences in the SGR among groups, while no obvious histopathological alteration was observed in livers of fish previously fed with the LCD. The results indicate that the LCD-treated fish have a full recovery after a 60-day recovery, but the HCD-treated fish did not. (C) 2012 Elsevier Ltd. All rights reserved.

Carbamazepine (CBZ) is an antiepileptic drug commonly detected in aquatic systems, with toxic effects to inhabiting organisms. Limited information is known on stress response biomarkers associated to bioconcentration and depuration of CBZ in aquatic organisms. Moreover, few studies addressed if the response and recovery of organisms to a contaminant can change when they are collected in a contaminated site. This study intended to understand the bioconcentration and depuration of CBZ combined with its toxicological impact in Scrobicularia plana clams collected from two contrasting areas (MIRA, Mira channel, non-contaminated and LAR, Laranjo bay, anthropogenically impacted) from the Ria de Aveiro (Portugal). The clams were exposed for 14 days to environmentally relevant CBZ concentrations (0.0, 4.0 and 8.0 μg/L), followed by a 14 day depuration period. CBZ concentrations in S. plana tissues were rapidly bioconcentrated during the exposure period. In the depuration period CBZ was eliminated, in some extent. The main toxic effects occurred at the highest concentration (8.0 μg/L) after 14 days of exposure in which the clams from LAR accumulated a higher CBZ concentration (LAR: ~ 10 ng/g FW) than clams from MIRA (MIRA: ~ 7 ng/g FW). LAR clams exhibited higher oxidative damage at this concentration, demonstrated by higher LPO levels over time (increase of ~ 1.4% relative to control) and, in comparison with MIRA clams (LAR: 17.7 nmol/g FW; MIRA: 11.4 nmol/g FW). After the depuration period, LAR clams recovered from the stress induced by CBZ. A decrease in LPO for LAR (decrease of ~ 40% in relation to the end of the exposure period) was accompanied by a decrease in CBZ tissue concentrations (decrease of ~ 61% relative to the end of the exposure period). MIRA clams were not oxidatively injured (low LPO levels remained unchanged after the depuration and CBZ decreased ~ 80% relative to the end of the exposure period).

Although animal cloning is becoming increasingly practicable, cloned embryos possess many abnormalities and so there has been a low success rate for producing live animals. This is most likely due to incomplete reprogramming of somatic cell nuclei before they start to develop as the donor nuclei are usually only exposed to the oocyte cytoplasm for 1-2 hours before reconstructed oocytes are activated to avoid oocyte aging. Therefore, in this study, we attempted to extend the exposure period of somatic cell nuclei to the oocyte cytoplasm to determine whether this could enhance reprogramming of donor nuclei. Donor nuclei were transferred into oocytes, following which pseudo-MII spindles (pMIIs) derived from these were extracted and injected into newly collected enucleated oocytes 24 hours after the first nuclear transfer (NT). These serial NT oocytes were then activated and their developmental potential was examined to full term. There was no obvious difference in the pMIIs of reconstructed oocytes at 6 and 24 hours after donor nucleus injection; however, in both of these, the chromosomes were more widely spread inside the spindle than in fresh intact oocytes. Furthermore, a few chromosomes remained in 25% and 47% of these enucleated oocytes at 6 and 24 hours after donor nucleus injection, respectively. When these pMIIs were injected into fresh enucleated oocytes, the developmental rate to blastocyst was significantly lower, but we could still obtain several healthy cloned offspring. Thus, serial NT at intervals of 24 hours using fresh oocytes is possible, but the success rate could not be improved due to loss of chromosomes during the second NT.

We recently reported that increased glucocorticoid (GC) levels in immobilized mice were suppressed by exposure to a 50-Hz electric field (EF) in kV/m-dependent and exposure duration-dependent manners. Here, we characterized time-dependent changes in the effect of EF exposure in immobilized mice. Using control, EF-alone, immobilization-alone, and co-treated groups, plasma GC levels, and blood properties were first measured (0-60 min) to observe changes induced by each treatment and measured again (60-120 min) to assess recovery from each treatment. The 50-Hz, 10-kV/m EF was formed in a parallel plate electrode. Co-treated mice were exposed to the EF for 60 min for the first measurement and were immobilized for the second half (30-60 min) of the EF exposure period. Plasma GC levels did not change significantly over time in the control and EF-alone groups. GC levels in the immobilization-alone and co-treated groups increased after immobilization, peaking 30 min after the start of immobilization and then decreasing gradually; however, the GC peak was lower in the co-treated group than in the immobilization-alone group (P < 0.05 at 50 and P < 0.001 at 60 min). Red blood cell counts, hemoglobin levels, and hematocrit values increased after immobilization but were not affected by the EF. Our findings indicate that the EF did not shift the peak of the time-dependent increase in plasma GC levels in immobilized mice but simply reduced it. Bioelectromagnetics. 38:272-279, 2017. © 2017 Wiley Periodicals, Inc.

The present study was designed to determine the effect of N'-[6,7-dichloro-4-(4-methoxy-phenyl)-3-oxo-3,4-dihydroquinoxalin-2-yl] hydrazide (SJA 7019), a selective nonpeptide inhibitor of calpain, and sodium orthovanadate, a selective inhibitor of tyrosine phosphatase, on the development of physiological dependence, as assessed by precipitated morphine withdrawal behavior in mice. Subchronic morphine administration (5 mg/kg, intraperitoneally, twice daily for 5 days), followed by a single injection of naloxone was used to precipitate the opioid withdrawal syndrome in mice. Behavioral observations were made immediately after naloxone treatment. Withdrawal syndrome was assessed quantitatively in terms of the withdrawal severity score and the frequency of jumping, rearing, forepaw licking, and circling. Daily single administration of SJA 7019 (1.5, 3, and 6 mg/kg, intraperitoneally) or sodium orthovanadate (5, 10, and 20 mg/kg, intraperitoneally) was continued during the morphine treatment procedure. Administration of SJA 7019 as well as the sodium orthovanadate dose-dependently attenuated the naloxone-induced morphine withdrawal syndrome. Neither SJA 7019 nor sodium orthovanadate significantly affected locomotor activity or morphine-induced antinociception. Therefore, it may be concluded that treatment with SJA 7019 or sodium orthovanadate during the morphine exposure period attenuated the development of physiological dependence on morphine, possibly through mechanisms linked to activation of tyrosine phosphatase and calpain.

The study compared the ability of various chemical methods (XAD, β-hydroxypropylcyclodextrin - HPCD) and solid phase micro-extraction (SPME)) to mimic earthworm uptake from two similar soils containing either spiked or aged p,p´-DDE, thus representing two extreme scenarios with regard to the length of pollutant-soil contact time and the way of contamination. The extent of bioaccumulation was assessed at fixed exposure periods (10 and 21 days) and at equilibrium derived from uptake curves by multiple-point comparison or kinetic modeling. The decision on the best chemical predictor of biological uptake differed. The degree of bioaccumulation at equilibrium was best predicted by XAD while HPCD rather reflected the extent of accumulation derived after 21 days when, however, steady-state was not reached for spiked p,p´-DDE. SPME seemed to underestimate the uptake of aged p,p´-DDE, probably of the fraction taken up via soil particles. Thus, the degree of predictability seems to be associated with the capability of the chemical method to mimic the complex earthworm uptake via skin and intestinal tract as well as with the quality of biological data where the insufficient length of exposure period appears to be the major concern.

The harmful alga Chattonella marina has caused massive fish kills and economic losses worldwide. Suffocation is generally believed to be the major cause of fish death by C. marina. However, the specific mechanisms leading to respiratory disorder in fish and subsequent fish kills by C. marina remain unknown. The goldlined seabream, highly susceptible to C. marina, was employed to investigate temporal changes of physiological, histopathological and biochemical parameters related to respiratory function at different stages of exposure to C. marina. Hemoglobin oxidation and blood lyses were not found in goldlined seabream exposed to C. marina, which could not be the key reasons accounting for pO(2) drop in the stressed fish. Gill histopathology such as irregular organization of lamellae, mucous with algal cells trapped in interfilamental spaces, were typical in C. marina exposed fish. A surge of plasma lactate occurred in goldlined seabream shortly after exposure to C. marina (0.5h) and sustained throughout the exposure period, indicating rapid onset of and persistent anaerobic respiration in C. marina exposed fish. Depletion of plasma glucose was clearly evident in goldlined seabream showing stress symptoms and near death. Yet, fish alive in the C. marina bloom did not exhibit plasma glucose depletion. The results suggest that availability of fermentable fuel as indicated by glucose level is critical to determine fish survival in C. marina exposure. Overall, our findings have rebuked the involvement of hemolysins and/or nitric oxide as the culprits for C. marina toxicity to fish. This study is the first to demonstrate the pathway of respiratory toxicity induced by the harmful alga C. marina in fish.

The coefficients of partitioning (Kpew) between low-density polyethylene (LDPE) film (50-µm thickness) and water for 23 polybrominated diphenyl ether (PBDE) congeners were determined based on a regression analysis of sorption kinetics over an extended exposure period (up to 365 d). A curvilinear relationship between log Kpew and log KOW (octanol–water partition coefficient) was obtained for the target BDE congeners with the turning point at log KOW approximately 8. Previously obtained dietary uptake efficiencies of BDE congeners in common carp (Cyprinus carpio) were also found to relate curvilinearly to log KOW. In addition, field-measured relative abundances of BDE-209 compiled from previous investigations conducted in the Pearl River Delta of South China were significantly (p

The objective of this study was to develop an estimate of the percent dermal absorption of permethrin in humans to provide more accurate estimates of potential systemically absorbed dose associated with dermal exposure scenarios. Piperonyl butoxide (PBO) was used as a reference compound. The human percutaneous absorption estimate was based on the assumption that the ratio of in vivo dermal absorption (expressed as a percentage during a given time period) of permethrin through rat skin to in vitro dermal absorption through rat skin was the same as the ratio of in vivo dermal absorption in humans to in vitro dermal absorption with human skin, known as the parallelogram method. The ratio of dermal absorption by in vitro rat skin to absorption by in vitro human skin ranged from 6.7 to 15.4 (for a 24-h exposure period) with an average of 11. Data suggest in vivo human dermal absorption values for permethrin ranging from 1.4 to 3.3% when estimated based on 24-h in vivo rat values, and 2.5 to 5.7% based on 5-d in vivo rat values. The parallelogram method used to estimate dermal absorption of permethrin and PBO is supported by results from several other compounds for which in vivo and in vitro rat and human dermal absorption data exist. Collectively, these data indicate that estimating human dermal absorption from in vitro human and rat plus in vivo rat data are typically accurate within ±3-fold of the values measured in human subjects.

Over the last three decades there has been a significant decline in the number of marine pollution monitoring-related studies in South Africa. Thus, the current study was conducted to assess the current state of metal contamination within the South African marine environment through the validation of the artificial mussel (AM). Indigenous reference mussels (Perna perna) were deployed alongside the passive device within the South African marine environment for a 6 week exposure period. Analysis of metal uptake (Cd, Cr, Cu, Pb and Zn) was determined by filtration and elution of the AM chelex resins, microwave digestion of the transplanted mussels, and determination of their metal concentrations by ICP-MS and ICP-OES analysis. Uptake patterns between the AM and transplanted mussels showed significant temporal and spatial correlation for the majority of the elements analysed. While the AM provided relevant and complementary information on the dissolved metal concentrations, limitations were also observed.

To assess the developmental toxicity of trichloroacetate (TCA), zebrafish embryos were exposed to 8 to 48 mM of TCA and evaluated for developmental milestones from 8- to 144-hour postfertilization (hpf). All developmental toxicities are reported in this paper. Embryos were found to have developed edema in response to 16 to 48 mM of TCA exposure at 32- to 80-hpf, experienced delay in hatching success in response to 24 to 48 mM at 80-hpf. Lordosis was observed in developing embryos exposed to 40 to 48 mM at 55- to 144-hpf. The observed toxic effects of TCA exposure were found to be concentration and exposure period independent. Effects were found to be associated with increases in superoxide anion production, but these increases were also found to be concentration and time independent. TCA resulted in concentration-dependent increases in embryonic lethality at 144-hpf, with an LC50 determined to be 29.7 mM.

Factorial design plan was experimented in Perna perna in order to find out the contribution of day exposure, Cd and Cu concentrations in water, and their interactions on metallothioneins (MT), Cd, and Cu (the dependent measured variables) in the gills. The picture obtained is more adequate than by studying the factor effect separately. Compared with the control group, the MT concentration after 22 days exposure period in the mixture of 100 μg/mL Cu and Cd is increased almost two times, showing that P. perna might be used as a biomonitor. Cd showed stronger effect than Cu on MT inducing.

The objectives were to determine if exposure of 12 mo old peri-puberal beef heifers to androgenized steers for 35 d hastened puberty, and if the response was related to the physical proximity between males and females and to heifer's initial body weight. Hereford x Aberdeen Angus heifers (n = 131), 12 mo old, were assigned to two treatments: 1) Exposed group, exposed to androgenized steers from Day 0 to Day 35 (E, n = 66); or 2) Control group, isolated from the steers and other males (C, n = 65). Cyclic activity was determined through estrous behavior detection (twice daily) and weekly ultrasound imaging to detect a CL. For each Exposed heifer, an association index (with males) was determined thrice weekly, based on the distance to androgenized steers every 10 min for 4 h (until cyclic activity began). Data were analyzed according to heifer's initial body weight, which was categorized into three ranges (low, medium, and high, designated LW, MW, and HW, respectively). The cumulative proportion of cyclic heifers was greater for Exposed than Control heifers as of Day 21. By the end of the exposure period, more Exposed than Control heifers had attained puberty (16/66 vs. 2/65; P0.001). Within the HW classification, more Exposed than Control heifers became puberal (11/20 vs. 2/21; P = 0.002). However, there were no differences between MW and LW in the proportions of heifers that reached puberty. Association index in Exposed heifers was greater in HW than in MW and LW (0.10 +/- 0.09 vs. 0.06 +/- 0.03 and 0.06 +/- 0.04; P0.05), and in heifers that began cyclic activity compared to those that did not in the MW heifers (0.09 +/- 0.05 vs. 0.05 +/- 0.02; P = 0.01) and tended to be different in the HW treatment (0.12 +/- 0.10 vs. 0.06 +/- 0.02; P = 0.09). In conclusion, exposure of peri-puberal beef heifers to androgenized steers for 35 d advanced puberty in heavier heifers; an earlier response occurred in heifers with greater proximity to androgenized steers.

In order to evaluate the chronic effect of polymerized toner particles on the lung, inflammation- and fibrosis-related genes were analyzed and 8-hydroxydeoxyguanosine (8-OHdG) was examined by using the lung tissue of rats subjected to 24 months of toner inhalation exposure. Wistar female rats were divided into four groups (5 weeks old, 30 rats in each): the high concentration exposure group (16.3 +/- 0.6 mg/m(3)), the medium concentration exposure group (4.4 +/- 0.3 mg/m(3)), the low concentration exposure group (1.6 +/- 0.2 mg/m(3)), and the control group (clean air). The material used was black toner, and its aerodynamic diameter in the exposure chamber was 3.0 microm. The rats were exposed to the material for 24 months (6 hours/day, 5 days/week) and dissected after the exposure period. RNA was extracted from one lung and the gene expression related to inflammation and fibrosis. Matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-2 (TIMP-2), and type I collagen were analyzed according to the ratio of each gene/beta-actin. Also, 8-OHdG level in the lung tissue was measured by HPLC with an electrochemical detector. Small fibrotic foci were found in the toner exposed groups; however, progressive or irreversible fibrosis was not found. The incidence of small fibrotic foci and cell aggregation increased in a dose-dependent manner. There were no significant differences of expression of MMP-2, TIMP-2, and type I collagen between the control group and each exposed group. Lung tumors did not develop in each group. A significant production of 8-OHdG was not observed in the toner exposed groups. In conclusion, toner produced by polymerization was not associated with evidence of carcinogenesis in this experiment.

In order to obtain basic risk assessment data on human health exposure to volatile organic compound (VOC) vapor by inhalation, a whole-body inhalation exposure system which allows blood collection during the exposure period was designed. The system was tested using chloroform as a model VOC. Chloroform vapor, sampled from the supply-header, animal-chambers and exhaust-header, remained constant in this system with variations in its concentration being less than 2%; flow rate of the vapor through the system was also constant. Rats were exposed to chloroform vapor and blood collected from the tail during exposure to the chloroform vapor. The chloroform concentration in the blood increased during the initial 60 min of exposure, and afterwards its concentration remained at about 2 microg/ml from 60 to 360 min. In conclusion, our design allows blood to be collected from individual rats during exposure by inhalation to test VOCs and changes in the blood concentration of the VOC during exposure to be assessed.

The effect of acrolein vapors against carefully aged eggs of Indian meal moth at 27 +/- 1 and 17 +/- 1 degrees C at different dosage levels of acrolein over various exposure times was determined. Considerable variation in the susceptibility of different age groups of eggs was apparent in the fiducial limits of the LD50 values. At both temperatures and 24 h exposure period, eggs aged 1-2 day-old were more tolerant to acrolein than other age groups. In all bioassays, eggs exposed to higher dosages of acrolein developed at smaller rate. This was significant for the eggs, which were exposed to the highest dosage for 24 h. Increasing the temperature from 17 +/- 1 to 27 +/- 1 degrees C greatly increased the efficacy of acrolein. Overall, at 27 +/- 1 degrees C eggs of P. interpunctella were killed by less than one-fourth of the dosage required for control at 17 +/- 1 degrees C. Acrolein achieved 50% mortality with a dosage of 3.80 mg L(-1) in 1-2 day-old eggs at 27 +/- 1 degrees C. At this temperature hatching was retarded and greatly reduced when eggs aged 1-2 day-old were exposed to 32 mg L(-1) of acrolein for the 24 h exposure period. There was no evidence of a hatch delay longer than the time spent under vapors for eggs exposed at 17 +/- 1 or 27 +/- 1 degrees C, indicating that some development must have occurred under fumigation.