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Simple Summary: This is the first case of an experimental study with a novel approach to analyze microsatellite instability (MSI) as a detection tool for genetic alterations in a wild population of birds, in this case for great tits (Parus major) being exposed to lead compounds. Although this preliminary study did not find any MSI in the analyzed microsatellite markers, it contributes to the examination of this molecular technique as a potentially applicable bioindication tool in ecotoxicological bird studies. We believe that certain issues should be considered in finding an explanation for our result. First, the exposure to lead was too low to induce genetic alterations. Second, the panel of microsatellite markers studied may have been unsusceptible to lead genotoxicity. Third, the time interval (5 days) between lead exposure and blood sampling for genetic analyses were too short to trigger genotoxicity. Further analyzes are needed to verify these findings and to evaluate the scope of application of the MSI analysis in wild bird population studies. Although many avian studies have investigated the toxic effects of lead on important biochemical and physiological processes, organ and system function, and behavior, studies evaluating the specific genotoxic effects of exposure to lead are scarce. Nowadays, rapid technological advances can provide new molecular techniques in this regard. In this study, as a novel approach in bird studies, we used a panel of ten microsatellite loci to investigate the microsatellite instability (MSI) in response to experimental lead intoxication in a common hole-nesting species, the great tit Parus major. For this purpose, an experiment based on an intentional single supplementation of a lead (II) acetate trihydrate compound was conducted, with the use of two different doses, applied to randomly chosen great tit nestlings from randomly selected broods, being at the stage of intensive erythropoiesis. Although this preliminary study did not find any MSI in the seven microsatellite markers retained for the final comparison, it contributes to the examination of this molecular technique in field conditions as being potentially applicable in ecotoxicological bird studies. We believe that certain issues should be considered in finding an explanation for our result. First, the single doses of lead used in this study may have been too weak to induce genetic instability. Second, the panel of microsatellite markers studied may have been unsusceptible to lead genotoxicity in general. Third, the relatively short time interval (5 days) between the experimental procedure (lead exposure) and the sampling of post-exposure material (blood) for genetic analyses could have limited the effect of lead genotoxicity. Further analyzes are needed to verify these findings and to evaluate the scope of application of the MSI analysis in wild bird population studies. [ABSTRACT FROM AUTHOR]

Dechlorane plus (DP) has been detected in a variety of environmental media and in human. Measurement of DPs in hair, urine, and house dust across different habitats allows for the assessment of short-term spatial changes in human exposure to DPs, as well as their excretion in urine. This offers a significant reference point for further research on the behavior of persistent pollutants within organisms. We measured and analyzed the concentrations of DP in the hair and urine of 32 students from a university in Beijing during school and home phases, and in indoor dust from dormitories and some home environments. The results indicated that the concentrations of DP in three types of samples were higher during the home phase compared to the school phase. We compared the f anti values and identified selective enrichment of syn-DP in hair, along with selective excretion of syn-DP in urine. Utilizing molecular docking technique, we simulated the binding effect between DP and the Megalin protein. The results demonstrated that the binding energy of anti-DP to Megalin was higher than that of syn-DP, suggesting that anti-DP has a greater propensity to bind to Megalin and be reabsorbed. This results in higher levels of syn-DP excretion in urine. Finally, we categorized students based on their participation in the organic exposure experiment and their BMI. The results indicated that the concentrations of DP in hair and urine were higher in the exposed group compared to the non-exposed group during the school year. After excluding the effect of exposure, habitat changes were more likely to affect the accumulation and excretion of DP in normal-weight students (BMI ≤24 kg/m2, n = 28), while overweight students (BMI >24 kg/m2, n = 4) were less affected by the effect of habitat because of their higher body fat percentage and their greater ability to accumulate DP. [Display omitted] • Dechlorane Plus (DP) concentration levels in dust, hair, and urine vary significantly between school and home environments. • Hair samples can effectively reflect changes in DP levels in the human body due to short-term variations in exposure environments. • Anti-DP is selectively reabsorbed by the body and more likely to bind to Megalin proteins, resulting in lower concentrations in urine compared to syn-DP. • Multiple exposure pathways primarily influence the overall DP levels in the body, with minimal impact on the selective enrichment and excretion capacities. [ABSTRACT FROM AUTHOR]

Anthropogenic sound has increasingly become part of the marine soundscape and may negatively affect animals across all taxa. Invertebrates, including bivalves, received limited attention even though they make up a significant part of the marine biomass and are very important for higher trophic levels. Behavioural studies are critical to evaluate individual and potentially population-level impacts of noise and can be used to compare the effects of different sounds. In the current study, we examined the effect of impulsive sounds with different pulse rates on the valve gape behaviour and phytoplankton clearance rate of blue mussels (Mytilus spp.). We monitored the mussels' valve gape using an electromagnetic valve gape monitor and their clearance rate using spectrophotometry of phytoplankton densities in the water. We found that the mussels' valve gape was positively correlated with their clearance rate, but the sound exposure did not significantly affect the clearance rate or reduce the valve gape of the mussels. They did close their valves upon the onset of a pulse train, but the majority of the individuals recovered to pre-exposure valve gape levels during the exposure. Individuals that were exposed to faster pulse trains returned to their baseline valve gape faster. Our results show that different sound exposures can affect animals differently, which should be taken into account for noise pollution impact assessments and mitigation measures. [ABSTRACT FROM AUTHOR]

Although many avian studies have investigated the toxic effects of lead on important biochemical and physiological processes, organ and system function, and behavior, studies evaluating the specific genotoxic effects of exposure to lead are scarce. Nowadays, rapid technological advances can provide new molecular techniques in this regard. In this study, as a novel approach in bird studies, we used a panel of ten microsatellite loci to investigate the microsatellite instability (MSI) in response to experimental lead intoxication in a common hole-nesting species, the great tit Parus major. For this purpose, an experiment based on an intentional single supplementation of a lead (II) acetate trihydrate compound was conducted, with the use of two different doses, applied to randomly chosen great tit nestlings from randomly selected broods, being at the stage of intensive erythropoiesis. Although this preliminary study did not find any MSI in the seven microsatellite markers retained for the final comparison, it contributes to the examination of this molecular technique in field conditions as being potentially applicable in ecotoxicological bird studies. We believe that certain issues should be considered in finding an explanation for our result. First, the single doses of lead used in this study may have been too weak to induce genetic instability. Second, the panel of microsatellite markers studied may have been unsusceptible to lead genotoxicity in general. Third, the relatively short time interval (5 days) between the experimental procedure (lead exposure) and the sampling of post-exposure material (blood) for genetic analyses could have limited the effect of lead genotoxicity. Further analyzes are needed to verify these findings and to evaluate the scope of application of the MSI analysis in wild bird population studies.

Chronic traffic noise is increasingly recognised as a potential hazard to wildlife. Several songbird species have been shown to breed poorly in traffic noise exposed habitats. However, identifying whether noise is causal in this requires experimental approaches. We tested whether experimental exposure to chronic traffic noise affected parental behaviour and reproductive success in zebra finches (Taeniopygia guttata). In a counterbalanced repeated-measures design, breeding pairs were exposed to continuous playback of one of two types of highway noise previously shown to be either neutral (control) or aversive. Parental nest attendance positively correlated with feeding effort and was higher for the aversive than the control sound and this effect was more pronounced for parents attending larger broods. However, neither noise condition affected offspring number, growth or body mass. The absence of an effect held when we combined our data with data from two other comparable studies into a meta-analysis. We discuss whether the increased nest attendance could be a compensatory strategy that alleviated detrimental noise effects on the chicks, and whether it could be caused by impaired parent-offspring or within-pair communication. Future work should test these hypotheses and investigate potential long-term costs of increased parental engagement.

Ranaviruses have been associated with rising numbers of mass die-offs in amphibian populations around the globe. However, most studies on ranaviruses to date focused on larval amphibians. To assess the role of postmetamorphic amphibians in the epidemiology of ranaviruses and to determine the role of viral immune-suppression genes, we performed a bath-exposure study on post-metamorphic wood frogs (Rana sylvatica) using environmentally relevant concentrations of wild-type Frog virus 3 (WT FV3), and a gene-knockout mutant (KO FV3), deficient for the putative immune-suppression gene vIF-2α. We observed a 42% infection rate and 5% mortality across the virus challenges, with infection rates and viral loads following a dose-dependent pattern. Individuals exposed to the knockout variant exhibited significantly decreased growth and increased lethargy compared with wild-type treatments. Although 85% of exposed individuals exhibited common signs of ranavirosis throughout the experiment, most of these individuals did not exhibit signs of infection by 40 d post-exposure. Overall, we showed that even a single short time exposure to environmentally relevant concentrations of ranavirus may cause sublethal infections in postmetamorphic amphibians, highlighting the importance of this life stage in the epidemiology of ranaviruses. Our study also supports the importance of the vIF-2α gene in immune-suppression in infected individuals.

Ciguatera poisoning is a food intoxication associated with the consumption of fish or shellfish contaminated, through trophic transfer, with ciguatoxins (CTXs). In this study, we developed an experimental model to assess the trophic transfer of CTXs from herbivorous parrotfish, Chlorurus microrhinos, to carnivorous lionfish, Pterois volitans. During a 6-week period, juvenile lionfish were fed naturally contaminated parrotfish fillets at a daily dose of 0.11 or 0.035 ng CTX3C equiv. g−1, as measured by the radioligand-receptor binding assay (r-RBA) or neuroblastoma cell-based assay (CBA-N2a), respectively. During an additional 6-week depuration period, the remaining fish were fed a CTX-free diet. Using r-RBA, no CTXs were detectable in muscular tissues, whereas CTXs were measured in the livers of two out of nine fish sampled during exposure, and in four out of eight fish sampled during depuration. Timepoint pooled liver samples, as analyzed by CBA-N2a, confirmed the accumulation of CTXs in liver tissues, reaching 0.89 ng CTX3C equiv. g−1 after 41 days of exposure, followed by slow toxin elimination, with 0.37 ng CTX3C equiv. g−1 measured after the 6-week depuration. These preliminary results, which need to be pursued in adult lionfish, strengthen our knowledge on CTX transfer and kinetics along the food web.

• Novel methods explored for examining how particle motion and sound pressure affect acoustic responses in zebrafish. • The ratio between particle motion and sound pressure in a small aquarium varied considerably from open water conditions. • Sound pressure and particle motion were predicted at the location of the fish during each exposure. • Particle ellipticity was measured as an indication of the strength of directional information in the soundfield. Underwater sound fields can be complex, both in open water and small tank environments. Here we measured 1) spatial variation in artificially elevated sound levels in a small fish tank for both particle motion and sound pressure. We confirmed that the ratio of pressure and particle motion deviated considerably from what would be expected in theoretical far field environments. We also tested 2) whether the acoustic response tendency of adult zebrafish (Danio rerio) was correlated to the sound field conditions at their position at the moment of sound on-set. We found no correlation between the intensity, quality, or directionality of the behavioural response and the sound pressure or the directivity and ellipticity of particle motion. There was a negative correlation, however, between the tendency to freeze and the particle velocity level. The data and experimental setup provided here may serve a basis to further explore the acoustic world of fish in complex environments and may contribute to the study of potential welfare and conservation issues related to anthropogenic noise. [ABSTRACT FROM AUTHOR]

Marine plastic pollution is a major environmental issue. Given their ubiquitous nature and small dimensions, ingestion of microplastic (MP) and nanoplastic (NP) particles and their subsequent impact on marine life are a growing concern worldwide. Transfers along the trophic chain, including possible translocation, for which the hazards are less understood, are also a major preoccupation. Effects of MP ingestion have been studied on animals through laboratory exposure, showing impacts on feeding activity, reserve depletion and inflammatory responses, with consequences for fitness, notably reproduction. However, most experimental studies have used doses of manufactured virgin microspheres that may not be environmentally realistic. As for most ecotoxicological issues, the environmental relevance of laboratory exposure experiments has recently been debated. Here we review constraints and priorities for conducting experimental exposures of marine wildlife to microplastics based on the literature, feedback from peer reviewers and knowledge gained from our experience. Priorities are suggested taking into account the complexity of microplastics in terms of (i) aggregation status, surface properties and interactions with organic and inorganic materials, (ii) diversity of encountered particles types and concentrations, (iii) particle bioavailability and distribution in experimental tanks to achieve reproducibility and repeatability in estimating effects, and (iv) strict experimental procedures to verify the existence of genuine translocation. Relevant integrative approaches encompass a wide spectrum of methods from -omics to ecophysiological approaches, including modeling, are discussed to provide novel insights on the impacts of MP/NP on marine ecosystems from a long-term perspective. Knowledge obtained in this way would inform stakeholders in such a way as to help them mitigate impacts of the micro- and nano-plastic legacy.

Highlights • Canyon towhees time to avoid conspecific song but do not avoid anthropogenic noise. • Four species with variable singing style do not adjust spectrum or timing to noise. • Singing style in general does not appear to correlate with noise-dependent flexibility. • Song consistency may have signal value and therefore persist under noisy conditions. Abstract Many bird species adjust their songs to noisy urban conditions by which they reduce masking and counteract the detrimental impact on signal efficiency. Different species vary in their response to level fluctuations of ambient noise, but it remains unclear why they vary. Here, we investigated whether noise-dependent flexibility may relate to singing style and signal function of the flexible acoustic trait. Species with highly variable songs may generally be more flexible and strongly repetitive singers may be more limited to stray from their stringent patterns. We exposed males of four passerine species with contrasting singing styles (repertoire size, immediate or eventual variety singing and syllable diversity) to three experimental sound conditions: 1) continuous urban noise; 2) intermittent white noise and 3) conspecific song playback. We found no spectral or temporal changes in response to experimental noise exposure in any of the four species, but significant temporal adjustment to conspecific playback in one of them. We argue that the consistency in song frequency and timing may have signal value, independent of singing style, and therefore be an explanation for the general lack of noise-dependent flexibility in the four species of the current study. [ABSTRACT FROM AUTHOR]

Abstract A lack of well-established parameters and assessment values currently impairs biomonitoring of n-heptane exposure. Using controlled inhalation experiments, we collected information on urinary n-heptane metabolite concentrations and the time course of metabolite excretion. Relationships between external and internal exposure were analysed to investigate the suitability of selected metabolites to reflect n-heptane uptake. Twenty healthy, non-smoking males (aged 19–38 years, median 25.5) were exposed for 3 h to 167, 333 and 500 ppm n-heptane, each. Spot urine samples of the volunteers, collected before exposure and during the following 24 h, were analysed for heptane-2-one, 3-one, 4-one, 2,5-dione, 1-ol, 2-ol, 3-ol, and 4-ol using headspace solid phase dynamic extraction gas chromatography/mass spectrometry (HS-SPDE-GC/MS). Starting from median pre-exposure concentrations between <0.5 (3-one) and 82.9 μg/L (4-one), exposure increased the concentrations for all parameters except for 4-one. Median post-exposure concentrations ranged up to 840.4 μg/L (2-ol) and decreased with half-lifes <3 h after exposure. Non-parametric correlation analyses (n = 47, p < 0.05) revealed weak to moderate associations of volume related metabolite excretion with external exposure for 2-one, 3-one and 2,5-dione (R = 0.332–0.753). Heptanol excretion was moderately associated with exposure (R ≥ 0.509) only after creatinine adjustment. Lacking association with external exposure impedes the use of 4-one as heptane biomarker, whereas 2-ol and 3-ol turned out to be sensitive indicators of exposure if creatinine correction is applied. By providing fundamental data on a panel of eight potential heptane metabolites, our study can help to promote biological monitoring of n-heptane exposure. [ABSTRACT FROM AUTHOR]

The invasive nematode Anguillicoloides crassus parasitizes the swim bladder of eels, including that of the American eel, Anguilla rostrata. Despite its potential devastating effects on eel populations, however, its intermediate host(s) is/are unknown in North America. The goal of this study was to determine whether An. crassus could infect copepods collected in the wild from the Goose Creek Reservoir in South Carolina, U.S.A., where An. crassus is known to occur. Cyclopoid copepods. identified morphologically as belonging to the Acanthocyclops robustus-americanus-vemalis species complex, were successfully experimentally infected by second-stage larvae (L2S) of An. crassus from eels captured in South Carolina, U.S.A. One infected copepod was identified as Ac. americanus via sequencing of a 251-base pair (bp) portion of mitochondrial cytochrome oxidase I DNA. For each infection experiment, 30 to 60 copepods were exposed for 24 hr to L2s at either L2S or 26°C and maintained at their respective exposure temperatures to monitor larval development until larvae molted to the L3 stage. More than 85% of the exposed copepods showed L-s in their intestine or hemocoel at 24 hr postexposure. Larvae molted into the L3 stage in 7 d at 26°C and 12 d at 21C. Based on these results, species within the Acanthocyclops vernalis-robustus-americanus complex are implied to be natural intermediate hosts of An. crassus in North America. [ABSTRACT FROM AUTHOR]

Many animals live in or near urban areas that have become increasingly widespread and noisy over the last century. Especially those species that rely heavily on acoustics for communication may be affected by these elevated anthropogenic noise levels. Many bird species that sing to defend their territories and to attract mates may have to exploit specific noise coping strategies to persist in such acoustically challenging conditions. Eurasian wrens ( Troglodytes troglodytes ), like several other bird species, have been shown in a previous experiment to time their vocalizations such that they avoid overlap with other singing birds. Here, we tested whether Eurasian wrens also time their songs to avoid overlap with fluctuating anthropogenic noise. However, we did not find any evidence in favor of this potential phenomenon. Territorial wrens persisted in singing without temporal adjustments in noisy territories with ‘natural’ fluctuations of traffic noise levels as well as during experimental exposure to intermittent white noise. [ABSTRACT FROM AUTHOR]

Studies have linked air pollution exposure to cardiovascular health effects, but it is not clear which components drive these effects. We examined the associations between air pollution exposure and circulating white blood cell (WBC) counts in humans. To investigate independent contributions of particulate matter (PM) characteristics, we exposed 31 healthy volunteers at five locations with high contrast and reduced correlations amongst pollutant components: two traffic sites, an underground train station, a farm and an urban background site. Each volunteer visited at least three sites and was exposed for 5 h with intermittent exercise. Exposure measurements on-site included PM mass and number concentration, oxidative potential (OP), elemental- and organic carbon, metals, O3 and NO2. Total and differential WBC counts were performed on blood collected before and 2 and 18 h post-exposure (PE). Changes in total WBC counts (2 and 18 h PE), number of neutrophils (2 h PE) and monocytes (18 h PE) were positively associated with PM characteristics that were high at the underground site. These time-dependent changes reflect an inflammatory response, but the characteristic driving this effect could not be isolated. Negative associations were observed for NO2 with lymphocytes and eosinophils. These associations were robust and did not change after adjustment for a large suite of PM characteristics, suggesting an independent effect of NO2. We conclude that short-term air pollution exposure at real-world locations can induce changes in WBC counts in healthy subjects. Future studies should indicate if air pollution exposure-induced changes in blood cell counts results in adverse cardiovascular effects in susceptible individuals. [ABSTRACT FROM AUTHOR]

Background: Specific characteristics of particulate matter (PM) responsible for associations with respiratory health observed in epidemiological studies are not well established. High correlations among, and differential measurement errors of, individual components contribute to this uncertainty. Objectives: We investigated which characteristics of PM have the most consistent associations with acute changes in respiratory function in healthy volunteers. Methods: We used a semiexperimental design to accurately assess exposure. We increased exposure contrast and reduced correlations among PM characteristics by exposing volunteers at five different locations: an underground train station, two traffic sites, a farm, and an urban background site. Each of the 31 participants was exposed for 5 hr while exercising intermittently, three to seven times at different locations during March-October 2009. We measured PM10, PM2.5, particle number concentrations (PNC), absorbance, elemental/organic carbon, trace metals, secondary inorganic components, endotoxin content, gaseous pollutants, and PM oxidative potential. Lung function [FEV1 (forced expiratory volume in 1 sec), FVC (forced vital capacity), FEF25-75 (forced expiratory flow at 25-75% of vital capacity), and PEF (peak expiratory flow)] and fractional exhaled nitric oxide (FENO) were measured before and at three time points after exposure. Data were analyzed with mixed linear regression. Results: An interquartile increase in PNC (33,000 particles/cm3) was associated with an 11% [95% confidence interval (CI): 5, 17%] and 12% (95% CI: 6, 17%) FENO increase over baseline immediately and at 2 hr postexposure, respectively. A 7% (95% CI: 0.5, 14%) increase persisted until the following morning. These associations were robust and insensitive to adjustment for other pollutants. Similarly consistent associations were seen between FVC and FEV1 with PNC, NO2 (nitrogen dioxide), and NOx (nitrogen oxides). Conclusions: Changes in PNC, NO2, and NOx were associated with evidence of acute airway inflammation (i.e., FENO) and impaired lung function. PM mass concentration and PM10 oxidative potential were not predictive of the observed acute responses. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was to determine the toxicokinetics of inhaled 1,1-difluoroethane (HFC-152a) in humans. Healthy volunteers were exposed to 0, 200 or 1000ppm 1,1-difluoroethane for 2h at light exercise in an exposure chamber. Capillary blood, urine and exhaled air were sampled up to 22h post-exposure and analyzed for 1,1-difluoroethane. Fluoride and other potential metabolites were analyzed in urine. Symptoms of irritation and central nervous system effects were rated and inflammatory markers were analyzed in blood. Within a few minutes of exposure to 200 and 1000ppm, 1,1-difluoroethane increased rapidly in blood and reached average levels of 7.4 and 34.3μM, respectively. The post-exposure decreases in blood were fast and parallel to those in exhaled air. The observed time courses in blood and breath agreed well with those obtained with the PBPK model. The PBPK simulations indicate a net uptake during exposure to 1000ppm of 6.6mmol (6.7%) which corresponds to the amount exhaled post-exposure. About 20μmol excess fluoride (0.013% of inhaled 1,1-difluoroethane on a molar basis) was excreted in urine after exposure to 1000ppm, compared to control. No fluorine-containing metabolites were detected in urine. Symptom ratings and changes in inflammatory markers revealed no exposure-related effects. [Copyright &y& Elsevier]

Sound plays an important role in the life of many animals, including many bird species. Typically, male birds sing to defend a territory and to attract mates. Ambient noise may negatively affect the signal efficiency of their songs, which may be critical to reproductive success. Consequently, anthropogenic noise may be detrimental to individual birds and to populations in cities and along highways. Several bird species that are still common in urban areas have been shown to sing at higher frequency at locations where there is more low-frequency traffic noise. Here we show that chiffchaffs along noisy highways also sing with a higher minimum frequency than chiffchaffs nearby at a quiet riverside. Furthermore, through experimental exposure to highway noise we show that these birds are capable of making such adjustments over a very short time scale. The first 10 songs sung during the noise exposure revealed an immediate shift to higher frequencies, with a return to pre-exposure levels in recordings without noise the following day. In a transmission re-recording experiment we tested the impact of a potential measurement artifact by recording playback of the same songs repeatedly under different controlled noise conditions. We found an upward shift in the minimum frequency measurement associated with more noisy recordings of the same song, but this artifact was not of a scale that it could explain the noise-dependent spectral shifts in chitfchatfs. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the toxicokinetics of inhaled 1,1,1,3,3-pentafluoropropane (HFC-245fa) in humans. Five healthy volunteers of each sex were exposed in random order to 0, 100, or 300 ppm HFC-245fa for 2 h at light exercise (50 W) in an exposure chamber. Capillary blood, urine, and exhaled air were sampled up to 22 h postexposure and analyzed for HFC-245fa. In addition, the metabolites fluoride, 3,3,3-trifluoropropionic acid (TFPA), and trifluoroacetic acid (TFAA) were analyzed in urine. Symptoms of irritation and central nervous system effects were rated in visual analogue scales. Various biochemical (aspartate-amino transferase, alanine-amino transferase, alkaline phosphate, glutamyl transferase, urate, creatine kinase [CK], and CK muscle brain) and inflammatory markers (serum amyloid A protein, fibrinogen, D-dimer, C-reactive protein, and interleukin-6) in plasma were analyzed. The initial increase in blood was fast and an apparent steady state was reached within a few minutes at both exposure levels. The postexposure decrease in blood was equally fast and parallel to that in exhaled air. Only minor amounts of unchanged HFC-245fa were excreted in breath (0.7% of inhaled) and urine (0.001%). The observed time courses in blood and breath agreed reasonably well those obtained by physiologically based pharmacokinetic (PBPK) modeling. The PBPK simulations indicate a relative uptake during exposure of 2.1%. TFPA was not detected in urine, and no increase in TFAA or fluoride above background was seen, suggesting little or no metabolism, the calculated minimum detectable metabolism being 0.001% of the inhaled amount. The symptom ratings revealed no HFC-245fa–related effects. None of the biochemical markers was affected. The changes in inflammatory markers, some of which are statistically significant, were not consistent with an inflammatory response. [ABSTRACT FROM PUBLISHER]

Abstract: The chemosensory effects of propionic acid (PA) in humans have not been conclusively studied and there is no established occupational exposure limit (OEL) in Germany. In addition to sensory irritation caused by PA, it was hypothesized that the annoying odor of PA might interfere with the performance results. There were 23 consenting healthy participants (12 female, 11 male) in the group studied. They were exposed for 4h to PA in concentrations of 0.3, 5 and 10ppm in a cross-over design. During these exposures, performance was recorded with four cognitive tests measuring response-inhibition, working memory, set-shifting, and divided attention. Odor annoyance, other chemosensory sensations, and acute symptoms were assessed before, during, and after exposure with standardized rating scales. Moderate odor annoyance and weak sensory irritation were reported during 5 and 10ppm exposure conditions. The different levels of exposure to PA had no impact upon the results of three out of the four behavioral tests. The difficulties of the task were reflected in the results. However in the fourth, which was the response-inhibition task, there was significant increase in the error rates which corresponded to the exposure levels. Results from previous experiments suggested high odor annoyance at the investigated concentrations. Our findings showed that odor annoyance and reported sensory irritations were low. In conclusion, the hypothesis of a distractive effect due to the malodor of PA could not be confirmed. Only in concentrations as high as 10ppm acute PA exposure affected the response accuracy of one of the four neurobehavioral task. For other more annoying substances, a neurobehavioral effect influenced by an indirect mechanism of resources competition is still conceivable. [Copyright &y& Elsevier]

Thirty-six volunteers (in three susceptibility groups: 11 subjects were non-allergic with nasal histamine hypersensitivity, 13 were non-allergic with normal sensitivity, and 12 were pollen allergic with or without nasal hypersensitivity) were exposed for three and a half hours in a climate chamber. Each subject was exposed to clean air (dust 45 ± 38 μg/m3 total suspended particle, TSP), house dust at 357 ± 180 μg/m3 TSP, house dust 382 ± 175 μg/m3 TSP with added glucan (50 ng/m3) and house dust 394 ± 168 μg/m3 TSP with added aldehydes corresponding to a gaseous phase of 300 μg/m3 in the air. The study was explorative by nature. No significant effects of exposures as such were seen on break-up time, conjunctival epithelial damage score and Trolox Equivalent Antioxidant Capacity (TEAC) in tear film and subjective ratings. However, in TEAC a significant different time course was seen during exposures to aldehyde-containing dust indicating a subacute and late response to the exposures. Perceived eye irritation increased significantly during exposures to normal dust. The perception ratings were highly correlated, whereas no correlation was found between the subjective responses and the objective measurements. Practical Implications The findings indicate that measurement effects on the eyes are rather insensitive measures of short time effects of office dust exposures. [ABSTRACT FROM AUTHOR]

Abstract: In mussels impacted by the V- and Ni-rich oil dispersed in the environment after the « Erika » wreck, a significant and positive correlation was observed between metallothionein (MT) and V concentrations whereas no correlation was observed between MT and Ni concentrations. Mussels were exposed separately to V and Ni in the laboratory. The highest concentrations tested (≥2000 ng V per individual; ≥50 ng Ni per individual) were responsible for V and Ni bioaccumulation. In the case of V, the metal was preferentially stored in the insoluble fraction of the soft tissues but with increasing body burden, the part of soluble V increased from 19 to 41%. Metallothionein was induced in mussels exposed to V but not in those exposed to Ni. In addition, in mussels originating from a shellfish farming area which showed abnormally high levels of V, depuration showed concomitant decreases of V and MT. It is the first report in the literature of a functional link between MT and V. When interpreting surveys of metal pollution based upon the use of MT, one must be aware that V may be responsible for MT synthesis, explaining likely high levels of this biomarker in the absence of metals known as MT inducers. [Copyright &y& Elsevier]

A human experimental study was carried out with 16 volunteers to examine the elimination of N-methyl-2-pyrrolidone (NMP) after exposure to the solvent under simulated workplace conditions. The NMP concentrations were 10, 40 and 80 mg/m3 for 2 × 4 h with an exposure-free interval of 30 min. Additionally, a peak exposure scenario (25 mg/m3 baseline, 160 mg/m3 peaks for 4 × 15 min, time-weighted average: 72 mg/m3) was tested. The influence of physical activity on the uptake and elimination of NMP was studied under otherwise identical exposure conditions but involving moderate workload on a bicycle ergometer (75 W for 6 × 10 min). The peak times and biological half-lives of urinary NMP and its main metabolites 5-hydroxy- N-methyl-2-pyrrolidone (5-HNMP) and 2-hydroxy- N-methylsuccinimide (2-HMSI) in urine were analysed as well as the interrelationships between exposure and biomarkers. All analytes showed a close correlation between their post-shift peak concentrations and airborne NMP. An exposure to the current German workplace limit value of 80 mg/m3 under resting conditions resulted in urinary peak concentrations of 2,400 μg/L NMP, 117 mg/g creatinine 5-HNMP and 32 mg/g creatinine 2-HMSI (workload conditions: 3,400 μg/L NMP, 150 mg/g creatinine 5-HNMP, 44 mg/g creatinine 2-HMSI). Moderate workload enhanced the total uptake of NMP by approximately one third. Differences between the estimated and the observed total amount of urinary metabolites point to a significant contribution of dermal absorption on the uptake of NMP. This aspect, together with the influence of physical workload, should be considered for the evaluation of a biological limit value for NMP. [ABSTRACT FROM AUTHOR]

This study investigated the hypotheses that humans are affected by air pollution caused by ozone and house dust, that the effect of simultaneous exposure to ozone and dust in the air is larger than the effect of these two pollutants individually, and that the effects can be measured as release of cytokines and changes of the respiratory function. Experimental exposures of eight atopic but otherwise healthy subjects were performed in a climate chamber under controlled conditions. The three controlled exposures were about 75 μg/m3 total suspended particulate matter, 0.3 p.p.m. ozone, and the combination of these. The exposure duration was 3 h. The outcome measures were interleukins and cells in nasal lavages (NAL), respiratory function, bronchial metacholine responsiveness, rhinometry symptoms and general well-being in a questionnaire and time course of general irritation on a visual analogue scale. Indications of interactions between exposure types were demonstrated for peak expiratory flow (PEF) ( P < 0.05) and for discomfort symptoms ( P < 0.03). Non-significant interactions were found for the concentration of interleukin-8 in NAL. The combined exposure was found to cause significantly more effects than either ozone exposures or dust exposures. This is interpreted as indications of a potentiation caused by the combined exposures to dust and ozone. The findings in this study are based on a limited number of subjects and thus should not be over-interpreted. However, they support the hypothesis that ozone at relatively high concentrations interacts with dust exposures to cause decrements in PEF and increase in discomfort measures. Practical Implications If confirmed at lower Ozone and dust concentrations this finding could help to explain many problems with indoor air quality reported in offices throughout the world. [ABSTRACT FROM AUTHOR]

Two experimental studies were conducted with the intention to simulate exposure characteristics of work places with styrene exposure and to investigate the risk for neurobehavioral impairments. In experiment I 16 volunteers (8 in the morning, 8 in the afternoon) were exposed to 0.5 and 20 ppm styrene on a constant level for 3 h. In experiment II 24 volunteers (12 in the morning, 12 in the afternoon) were exposed for 4 h to 0.5 and 20 ppm styrene on a constant level as well as to a changing exposure between 0.5 and 40 ppm with a TWA of 14 ppm. Simple reaction, choice reaction, attention, acute symptoms, and ratings for well-being were measured.Exposure related performance effects could not be detected. However, 6 h time change resulted in delayed choice reactions in the morning hours. Analysing acute symptoms and the state of well-being the impact of styrene did not reach adverse extents of impaired well-being. [Copyright &y& Elsevier]

Objectives: For risk assessments of solvents the knowledge on chemosensory irritation effects is important, but the methodological base for that is incomplete. The psychological approach measuring chemosensory irritations leans on perceived symptoms and self-reported changes of well being. Characteristics assessing the validity of such psychological approaches are presented. Methods: The article is based on 14 experimental inhalation studies with (mostly) 4-h exposures to acetone, 2-butanone, ethanol, ethyl acetate, ethyl benzene, iso-propanol, 1-octanol, and styrene. The profiles of exposure include constant and changing concentrations using the range of the German maximum concentrations at the workplace (MAK) list. Irritations (eyes and nose), olfactory symptoms (odour), and annoyance are the dependent variables measured by ratings. Young and healthy subjects (n=160), – partially, subjects with self-reported odour sensitivity (measured by items from the questionnaire on chemical and general environmental sensitivity) – were investigated. Results: The reliability of ratings is sufficient. Dose–response relationships for perceived odour and annoyance are stronger than those for irritations. A ranked order of the size of effect (related to the values before exposure) for the substances investigated shows correspondence between odour and annoyance; that for irritation differs. Within the limits of the MAK list, perceived irritations are not correlated to annoyance, whereas perceived bad smell correlates significantly to annoyance. Reversibility of the self-reported effects to approximately the pre-exposure level can be shown 1 h after cessation of the experimental exposure for the "normal" subjects. Influences of trait anxiety and chemical sensitivity on reports of annoyance, bad odour or irritation are only weak. Conclusion: The psychological approach of repeated measurements for self-reported irritation includes distinctive advantages compared with other methods, the simple and repeated availability during exposure, the sufficient reliability and dose–response relationship, and the comparability between substances by means of effect size. The extension of the concept of "chemosensory irritations" on reports for annoyance and bad smell can be recommended. [ABSTRACT FROM AUTHOR]

Objective: N-Methyl-2-pyrrolidone (NMP) is a selective and powerful organic solvent. The aim of this study was to investigate whether the NMP metabolite N-methylsuccinimide (MSI) in plasma and urine can be used as a biomarker of exposure to NMP. Methods: Six healthy subjects were exposed to 10, 25, and 50 mg NMP/m3 in an exposure chamber for 8 h. The air levels were monitored by XAD-7 solid sorbent sampling, and analysed by gas chromatography (GC). Plasma and urine were sampled for two days following the exposure, and the levels of MSI were analysed by GC with mass spectrometric detection. Results: The concentration of MSI in plasma and urine rose during the exposure, and reached a peak at about 4 h after the end of the exposure. The concentration then decayed according to a one-compartment model with a half-time of approximately 8 h. About 1% of the inhaled NMP was excreted in urine as MSI. There were very close correlations between the NMP air levels and, on the one hand, the MSI concentrations in plasma collected at the end of exposure (r=0.98), or the urinary MSI concentration collected during the last 2 h of exposure (r=0.96), on the other. Conclusions: MSI in plasma or urine is applicable as a biomarker of exposure to NMP. The concentration in plasma and urine mainly reflects the exposure over one day. [ABSTRACT FROM AUTHOR]

Objectives: The aim of the study was to investigate the acute effects of experimental methylformate exposure on the nervous system. Methods: In an exposure chamber, 20 subjects were exposed to methylformate at 100 ppm [Swiss maximum allowable concentration (MAC)] for 8 h. The same number of subjects with the same ages (between 20 and 30 years), gender and education level (university) were examined by the same procedure as a control group. The subjects did not know if they were exposed or not. Three times (morning, noon, evening) during these 8 h, mood [Profile of Mood States (POMS)], neurobehavioral performance (reaction, Stroop, nonverbal learning, determination, tracking; Wiener Test System), vision (visual acuity, contrast sensitivity, color sensitivity) and postural sway were tested. During an undemanding test (POMS) and a demanding performance task (determination test), pulse, electromyography (EMG) of the forehead and of the neck were recorded. In the morning and evening spirometry [forced vital capacity (FVC), forced one-second expiration volume (FEV), medium expiration flow (MEF) and peak expiration flow (PEF)] and the odor perception threshold were measured. Results: In the evening, in the exposed group, fatigue was significantly increased and the EMG of the forehead during a demanding task showed a different development during exposure. The other tests showed no significant solvent effect, but 16 of 43 test parameters showed a significant effect of time. Conclusions: The results of this study indicate a possible effect of methylformate exposure on the subjective feeling of fatigue after 8 h exposure at 100 ppm in young and healthy subjects, without measurable impairment of neurobehavioral performance. We assume that a similar effect in normal work, combined with a heavy workload and shift work, can lead to an impairment of productivity, and increase the risk of accidents. [ABSTRACT FROM AUTHOR]

Objectives: The study investigated the acute effects of isopropanol exposure at the maximum allowable concentration (MAC) level on the performance of neurobehavioural functions. Methods: In an exposure chamber 20 healthy subjects aged between 21 and 30 years were exposed to isopropanol at a concentration of 400 ppm (Swiss MAC value) for 8 h. A control group of another 20 subjects of the same age range, gender and education as used in a previous study with methylformate were subjected to the same test procedures without exposure. Mood was measured with the profile of mood states (POMS) in the morning, at noon and in the evening. Neurobehavioural performance tests were administered using the Wiener Test System (Wiener reaction test, Stroop, nonverbal learning, Wiener determination test, “Konturtracking” test), and the digit span test of the Neurobehavioural evaluation system (NES2). A test for postural sway was also conducted. During the POMS and the Wiener determination test the electromyography of the forehead and the left neck muscle as well as the pulse were registered. In the morning and evening spirometry (FVC, FEV, MEF and PEF) and odour threshold were measured. Results: Only postural sway in bipedal standing at noon showed stronger deterioration in the exposed than in the control group when compared with the morning values. The evening values of monopedal standing were also more impaired in the exposed than the control group. In the other tests, no solvent effect could be established. Conclusions: It can be assumed, that, similar to other alcohols, isopropanol affects postural balance. Our results point to such a disturbance, but because it is the only study so far using the MAC value, any conclusions about safety risks would be premature. [ABSTRACT FROM AUTHOR]

Chronic traffic noise is increasingly recognised as a potential hazard to wildlife. Several songbird species have been shown to breed poorly in traffic noise exposed habitats. However, identifying whether noise is causal in this requires experimental approaches. We tested whether experimental exposure to chronic traffic noise affected parental behaviour and reproductive success in zebra finches (Taeniopygia guttata). In a counterbalanced repeated-measures design, breeding pairs were exposed to continuous playback of one of two types of highway noise previously shown to be either neutral (control) or aversive. Parental nest attendance positively correlated with feeding effort and was higher for the aversive than the control sound and this effect was more pronounced for parents attending larger broods. However, neither noise condition affected offspring number, growth or body mass. The absence of an effect held when we combined our data with data from two other comparable studies into a meta-analysis. We discuss whether the increased nest attendance could be a compensatory strategy that alleviated detrimental noise effects on the chicks, and whether it could be caused by impaired parent-offspring or within-pair communication. Future work should test these hypotheses and investigate potential long-term costs of increased parental engagement., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)

• The effects of microplastic (MP) ingestion were examined in Sparus aurata. • Fish were fed for 35 days on natural prey pre-contaminated with 10–20 µm PE MP. • MP ingestion affected fish survival and brain primary metabolite profiles. • MP ingestion affected fish hepatocyte vacuolation and Zn assimilation. • MP ingestion did not affect fish growth nor C and N isotopic profiles in muscles. Plastic pollution has become a major environmental and societal concern in the last decade. From larger debris to microplastics (MP), this pollution is ubiquitous and particularly affects aquatic ecosystems. MP can be directly or inadvertently ingested by organisms, transferred along the trophic chain, and sometimes translocated into tissues. However, the impacts of such MP exposure on organisms' biological functions are yet to be fully understood. Here, we used a multi-diagnostic approach at multiple levels of biological organization (from atoms to organisms) to determine how MP affect the biology of a marine fish, the gilthead seabream, Sparus aurata. We exposed juvenile seabreams for 35 days to spherical 10–20 µm polyethylene primary MP through food (Artemia salina pre-exposed to MP) at a concentration of 5 ± 1 µg of MP per gram of fish per day. MP-exposed fish experienced higher mortality, increased abundance of several brain and liver primary metabolites, hepatic and intestinal histological defects, higher assimilation of an essential element (Zn), and lower assimilation of a non-essential element (Ag). In contrast, growth and muscle C/N isotopic profiles were similar between control and MP-exposed fish, while variable patterns were observed for the intestinal microbiome. This comprehensive analysis of biological responses to MP exposure reveals how MP ingestion can cause negligible to profound effects in a fish species and contributes towards a better understanding of the causal mechanisms of its toxicity. [ABSTRACT FROM AUTHOR]

Ciguatera poisoning is a food intoxication associated with the consumption of fish or shellfish contaminated, through trophic transfer, with ciguatoxins (CTXs). In this study, we developed an experimental model to assess the trophic transfer of CTXs from herbivorous parrotfish, Chlorurus microrhinos, to carnivorous lionfish, Pterois volitans. During a 6-week period, juvenile lionfish were fed naturally contaminated parrotfish fillets at a daily dose of 0.11 or 0.035 ng CTX3C equiv. g−1, as measured by the radioligand-receptor binding assay (r-RBA) or neuroblastoma cell-based assay (CBA-N2a), respectively. During an additional 6-week depuration period, the remaining fish were fed a CTX-free diet. Using r-RBA, no CTXs were detectable in muscular tissues, whereas CTXs were measured in the livers of two out of nine fish sampled during exposure, and in four out of eight fish sampled during depuration. Timepoint pooled liver samples, as analyzed by CBA-N2a, confirmed the accumulation of CTXs in liver tissues, reaching 0.89 ng CTX3C equiv. g−1 after 41 days of exposure, followed by slow toxin elimination, with 0.37 ng CTX3C equiv. g−1 measured after the 6-week depuration. These preliminary results, which need to be pursued in adult lionfish, strengthen our knowledge on CTX transfer and kinetics along the food web. [ABSTRACT FROM AUTHOR]

Six human volunteers were exposed to 10 and 20 ppm carbon disulfide at rest and to 3 and 10 ppm carbon disulfide under a 50 W level of physical exercise during four consecutive periods of 50 min. Every 5 min a sample was taken from the mixed exhaled air in which the concentration of carbon disulfide was determined. It was established that only an apparent steady state was reached during this exposure period. The retention values were established as 0.374 (SD = 0.106; n = 239) for exposure to 10 ppm carbon disulfide at rest and as 0.410 (SD = 0.103; n = 239) for exposure to 20 ppm carbon disulfide at rest. During exposure to 10 ppm and 3 ppm carbon disulfide, combined with a 50 W level of physical exercise, the retention values decreased to 0.286 (SD = 0.083; n = 239) and 0.277 (SD = 0.049; n = 239) respectively. Thereby, the measured individual retention values of carbon disulfide show considerable interindividual differences. The respiratory uptake of carbon disulfide (mg CS) proved significantly influenced by the amount of body fat estimated from skinfold thickness measurements. The respiratory elimination of carbon disulfide in the exhaled air can be described by means of a two-exponential decay. [ABSTRACT FROM AUTHOR]

In a climatic exposure chamber four healthy volunteers were exposed to 100ppm toluene, 100ppm toluene + ethanol, 100ppm toluene + cimetidine, and 100ppm toluene + propranolol for 7h each at random over four consecutive days. A control experiment and 3.5 h of exposure to 200 ppm toluene were also performed. Ethanol inhibited toluene metabolism by 0.5 as expressed by the urinary excretion of two of the metabolites of toluene, namely o-cresol and hippuric acid. In agreement with this, the mean alveolar concentration of toluene was greater by 1.7 during ethanol exposure; 45 min after discontinuation of exposure the increase was by 3.3. Neither cimetidine nor propranolol changed toluene metabolism significantly. The results indicate that ethanol may prolong the time interval in which toluene is retained in the human body in persons simultaneously exposed to ethanol and toluene. When using o-cresol or hippuric acid in biological monitoring of persons occupationally exposed to toluene, the consumption of ethanol should be considered. [ABSTRACT FROM AUTHOR]

The respiratory uptake rate of n-hexane showed considerable differences in six healthy male persons, exposed at rest to 360 mg/m and 720 mg/m of n-hexane in inspired air and to 360 mg/m under different levels of physical exercise. These differences could partly be explained by a positive correlation with the amount of body fat. At rest also a strong influence of the respiratory minute volume and respiratory frequency on the uptake rate could be proven. The average uptake rate remained virtually constant over a range of 20 to 60 W of continuous external physical load, indicating that under these circumstances the inspired n-hexane concentration alone predominantly determines the uptake rate. The respiratory elimination during the first hours after an exposure was also subject to important inter- and intraindividual fluctuations. The pulmonary ventilation rate at the moment of breath sampling had a pronounced influence on the measured exhaled concentration. On the other hand, there was no apparent effect of the amount of body fat. Generally, the correlation between the amount of n-hexane taken up and breath concentrations at different time intervals was rather poor. n-Hexane concentrations in peripheral venous blood reacted rapidly to changes in exposure conditions, but not in the same proportion as the uptake rate. The blood concentration proved more closely related to respiratory n-hexane retention than to the uptake rate, reflecting the state of saturation of different body tissues. At rest this parameter was clearly influenced by the amount of body fat. A decrease in relative blood perfusion of fatty tissue could explain why such relation was not found during exposure combined with physical effort. [ABSTRACT FROM AUTHOR]

The temporary threshold shift of vibratory sensation (TTSv) at 125 Hz after clasping a vibrating handle was investigated in relationship to four influencing factors: discrete frequency and acceleration of the vibration exposed to, the grip in clasping a handle and the time after exposure. Clasping the vibrating handle resulted in significantly larger TTSv than shown in clasping a non-vibrating handle. The TTSv after exposure to the 250 Hz vibration was significantly the largest in all frequencies at equal acceleration. The TTSv recovered exponentially as time elapsed after exposure. The TTSv also increased proportionally to the power of acceleration at each frequency and grip force. The larger grip force resulted in a significantly larger TTSv. A general form of regression equation of TTSv involving the four factors was established and its applicability was ascertained by a multiple regression analysis. The prediction equations obtained confirmed that a frequency around 200 Hz induced the largest TTSv among vibrations with equal acceleration and around 240 Hz among vibrations with equal velocity. [ABSTRACT FROM AUTHOR]

The relationship between the individual toluene uptake and the urinary hippuric acid excretion was studied under experimental conditions. Six healthy male subjects were exposed to various concentrations in inspired air (50, 100, 125, 150, and 200 ppm) at rest or under different levels of physical effort. The hippuric acid excretion near the end of the exposure appeared under all circumstances directly proportional to the time-weighted uptake rate of toluene. The correlation between respiratory uptake rate and the rate of metabolite excretion near the end of the exposure period proved not to be systematically influenced by personal factors such as body weight, amount of body fat, urine flow rate and urinary pH. The relatively pronounced differences in background excretion of hippuric acid and, perhaps, distribution phenomena of toluene between different tissues under heavy workload conditions, can partly explain the greater variability in metabolite excretions as compared to the individual uptake rates. The correlation between the individual uptake rate of toluene and the hippuric acid excretion proved substantially better when using the end exposure excretion rate as exposure parameter as compared with the end exposure hippuric acid concentration, even after correcting the latter for urine density. Reasonable biological limit values complying to an acceptable time-weighted toluene dose were found to be 3000-3500 mg/l and 2.0-2.5 mg/min, resp. for average hippuric acid concentrations and excretion rates in spot samples during the second half of a complete work shift. [ABSTRACT FROM AUTHOR]

The application as biologic exposure parameter of the toluene concentration in venous blood during and after an exposure to this solvent was experimentally studied. Under carefully controlled conditions 6 healthy male subjects were exposed to various concentrations of toluene in inspired air (50, 100, 125, 150 and 200 ppm) at rest or under different levels of physical effort. Peripheral venous concentrations (C) were followed in relation to the individual toluene uptake. A relatively constant relation was found between uptake rate of toluene and C under steady state conditions. Empirically, for lung clearances at rest and for differing inspired concentrations (C), this relation was given by: C (mg/l) = 0.303 CI (mg/1) x Lung clearance (1/min). Under constant C (50 ppm) and lung clearances varying from rest values to values under a continuous exercise of 50 W, the regression equation became: C (mg/l) = 0.328 C (mg/l) x Lung clearance (1/min). Under non-steady state conditions no simple relation existed between uptake rate and C, indicating that equally no simple connection could be made between C and the calculated mixed venous concentration, or, extrapolating, the expected toxic load of most inner organs. In relation to the individual dose, C always presented a much greater variability than the toluene uptake by itself. The observed differences, both intra-individually as between subjects, were mostly statistically significant. Apparently C was influenced by some host factors in another manner than the uptake rate. In this respect evidence was obtained that the local blood perfusion and the amount body fat were involved to a certain extent. For this reason C proved a somewhat less satisfactory individual dose-parameter. [ABSTRACT FROM AUTHOR]

Lead (Pb) is recognized as one of the most toxic trace elements that can induce a wide range of negative health effects in wildlife. Because the investigation of basic environment-sensitive physiological indicators is easy to perform in wild birds, they have been considered as valuable bioindicators of lead contamination. The harmfulness of lead poisoning can depend on the type of exposure, and in most studies of birds, an effect of chronic lead exposition has been considered. In this study, we investigated whether a single exposure to specific doses of lead affected such physiological indices as haematocrit and the heterophil-to-lymphocyte ratio (H/L ratio). For this purpose, we conducted an experiment based on intentional lead supplementation, with the use of two different doses, applied to randomly chosen great tit ( Parus major ) nestlings from randomly selected broods. A few days after the exposure (when the nestlings were 15 days old), we determined haematocrit and the H/L ratio as potentially sensitive physiological indicators of lead intoxication. We found that the treatment with lead resulted in a significant decline in haematocrit level. In addition, we found that the age when lead exposure occurred can be considered as an important factor influencing haematocrit variation. A decrease in haematocrit was observed on consecutive days of nestling life. In contrast, the H/L ratio did not show any significant response to lead exposure. These results clearly show that the assessment of haematocrit level for nestling great tits can provide a simple and rapid method of indirect evaluation of physiological effects of lead intoxication caused by a single exposure., (© The Author(s) 2019. Published by Oxford University Press and the Society for Experimental Biology.)