Purebred dogs show higher levels of genomic damage compared to mixed breed dogs.

Inbreeding is a common phenomenon in small, fragmented or isolated populations, typical conditions of many threatened species. In the present paper, we used a new non-invasive approach based on the buccal micronucleus assay to evaluate the possible relationships between inbreeding and genomic damage using the dog as model species. In particular, we assessed the frequencies of micronuclei and other nuclear aberrations in a group of purebred dogs (n = 77), comparing the obtained data with those from a control group represented by mixed breed dogs (n = 75). We found a significant increase of micronuclei, nuclear buds and total nuclear aberrations frequencies in purebred dogs compared to mixed-bred dogs. The absence of significant differences in the frequency of micronuclei and other nuclear aberrations amongst different breeds reinforces the hypothesis that the observed increased genomic damage amongst purebred dogs may not be due to a different genomic instability typical of a particular breed, but to inbreeding itself. This hypothesis is further confirmed by the fact that other endogen confounding factors, such as sex, age and weight, do not contribute significantly to the increase of genomic damage observed amongst purebred dogs. In conclusion, results presented in this study showed that, in purebred dogs, inbreeding may increase the levels of genomic damage. Considering that genomic damage is associated with increased physiological problems affecting animal health, the results we obtained may represent a stimulus to discourage the use of intensive inbreeding practices in captive populations and to reduce the fragmentation of wild populations.

Adaptation to bioinvasions: When does it occur?

The presence of alien species represents a major cause of habitat degradation and biodiversity loss worldwide, constituting a critical environmental challenge of our time. Despite sometimes experiencing reduced propagule pressure, leading to a reduced genetic diversity and an increased chance of inbreeding depression, alien invaders are often able to thrive in the habitats of introduction, giving rise to the so-called "genetic paradox" of biological invasions. The adaptation of alien species to the new habitats is therefore a complex aspect of biological invasions, encompassing genetic, epigenetic, and ecological processes. Albeit numerous studies and reviews investigated the mechanistic foundation of the invaders' success, and aimed to solve the genetic paradox, still remains a crucial oversight regarding the temporal context in which adaptation takes place. Given the profound knowledge and management implications, this neglected aspect of invasion biology should receive more attention when examining invaders' ability to thrive in the habitats of introduction. Here, we discuss the adaptation mechanisms exhibited by alien species with the purpose of highlighting the timing of their occurrence during the invasion process. We analyze each stage of the invasion separately, providing evidence that adaptation mechanisms play a role in all of them. However, these mechanisms vary across the different stages of invasion, and are also influenced by other factors, such as the transport speed, the reproduction type of the invader, and the presence of human interventions. Finally, we provide insights into the implications for management, and identify knowledge gaps, suggesting avenues for future research that can shed light on species adaptability. This, in turn, will contribute to a more comprehensive understanding of biological invasions.

Genotoxic effects of particulate matter on larvae of a common and widespread butterfly along an urbanization gradient.

Biodiversity is currently declining worldwide. Several threats have been identified such as habitat loss and climate change. It is unknown if and how air pollution can work in addition or in synergy to these threats, contributing to the decline of current species and/or local extinction. Few studies have investigated the effects of particulate matter (PM), the main component of air pollution, on insects, and no studies have investigated its genotoxic effects through Micronucleus assay. Butterflies play an important role in the environment, as herbivores during larval stages, and as pollinators as adults. The aim of this study was to evaluate the genotoxic effects of PM10 from different sites along a gradient of population urbanization, on a common cabbage butterfly species (Pieris brassicae). PM10 was collected from April to September in an urban (Turin, Italy), a suburban (Druento, Italy) and a mountain site (Ceresole Reale, Italy) with different urbanization levels. P. brassicae larvae (n = 218) were reared in the laboratory under controlled conditions (26 °C, L:D 15:9) on cabbage plants (average 9.2 days), and they were exposed to PM10 organic extracts (20 and 40 m3/mL) or dimethyl sulfoxide (controls) through vaporization. After exposure, larvae were dissected and cells were used for the Micronucleus (MN) assay. Results showed that all PM extracts induced significant DNA damage in exposed larvae compared to controls, and that increasing the PM dose (from 20 to 40 m3/mL) increased genotoxic effects. However, we did not detect any significant differences between sites with different urbanization levels. In conclusion, PM at different concentrations induced genotoxic effects on larvae of a common butterfly species. More alarmingly, PM could work in addition to and/or in synergy with other compounds (e.g. pesticides) and, especially on species already threatened by other factors (e.g. fragmentation), thus affecting the vitality of populations, leading to local extinctions.

The formation of SCEs as an effect of occupational exposure to formaldehyde.

Formaldehyde (FA) is a ubiquitous toxic chemical employed worldwide due to its disinfectant and preservative properties. Despite being classified as a human carcinogen, FA is still employed as formalin in pathology wards as standard fixative. We evaluated its relationship with the formation of sister-chromatid exchanges (SCEs) in cultured peripheral blood lymphocytes on 57 pathologists and 48 controls and the risk/protective role played by several genetic polymorphisms. All subjects were assessed for SCEs and genotyped for the most common cancer-associated gene polymorphisms: CYP1A1 exon 7 (A > G), CYP1A1*2A (T > C), CYP2C19*2 (G > A), GSTT1 (presence/absence), GSTM1 (presence/absence), GSTP1 (A > G), XRCC1 (G399A), XRCC1 (C194T), XRCC1 (A280G), XPC exon 15 (A939C), XPC exon 9 (C499T), TNFα - 308 G > A), IL10 - 1082 (G > A), and IL6 - 174 (G > C). Air-FA concentration was assessed through passive personal samplers. Pathologists, exposed to 55.2 µg/m3 of air-FA, showed a significantly higher SCEs frequency than controls, exposed, respectively, to 18.4 µg/m3. Air-FA was directly correlated with SCEs frequency and inversely with the replication index (RI). Regression models showed FA exposure as a significant predictor in developing SCEs, while did not highlight any role of the selected polymorphisms. Our study confirms the role of low air-FA levels as genotoxicity inductor, highlighting the importance to define exposure limits that could be safer for exposed workers.

Association of TGFß1 codon 10 (T>C) and IL-10 (G>C) cytokine gene polymorphisms with longevity in a cohort of Italian population.

Longevity is a complex process controlled by both environmental and genetic factors. We evaluated the association of four cytokine gene polymorphisms with longevity in an Italian cohort. A sample of 1019 subjects aged 10 to 100 and belonging to the North-Italian population was genotyped for IL-6 (G>C, rs1800796), IL-10-1082 (G>A, rs1800896), TNF-α-308 (G>A, rs1800629), and TGFß1 codon 10 (T>C, rs1800471) gene polymorphisms. The association between cytokine gene polymorphisms and longevity was evaluated by dividing the sample into four age groups: 10 to 24, 25 to 49, 50 to 85, and 86 to 100. We observed a significant decrease in the frequency of IL-10 A allele in the 25 to 49 (P = 1.1 × 10-3 ), 50 to 85 (P < 1 × 10-4 ), and 86 to 100 (P = 2 × 10-3 ) age groups compared to that in the youngest age group. Similarly, we found a significant decrease (P < 1 × 10-4 ) in the frequency of TGFß1 C allele in the 50 to 85 and 86 to 100 age groups compared to that in the 10 to 24 and 25 to 49 age groups. Previously, high levels of TGFß1 were detected in elderly subjects, suggesting that this cytokine could counterbalance the harmful effects of inflammation. Similarly, IL-10 has strong anti-inflammatory properties and can inhibit the production of proinflammatory cytokines. In the literature, the lowest levels of functional cytokines were found to be associated with TGFß1 (T>C) and IL-10 (G>A) gene polymorphisms, with consequent increase in the duration of inflammation and cancer risk. For these reasons, it is plausible to observe low rates of these mutations in elderly subjects, as found in our study.

GSTT1, GSTP1 and XPC genes are associated with longevity in an Italian cohort.

Longevity is a complex process controlled by environmental and genetic factors. We evaluated the association of seven drug metabolising and DNA-repair gene polymorphisms with longevity in an Italian cohort. A sample of 756 subjects aged 18-98 was genotyped for CYP1A1 (rs1048943, A>G), GSTM1 (rs 1183423000, presence/absence), GSTT1 (rs1601993659, presence/absence), GSTP1 (rs1695, A>G), XRCC1 (rs1799782, C>T), XRCC1 (rs25489, A>G) and XPC (rs2228001, A>C) gene polymorphisms. The association between the studied gene polymorphisms and longevity was evaluated by dividing the sample into three age groups: 18-50, 51-85, and 86-98. We observed a significant decrease in the frequency of the GSTT1 null, GSTP1 G and XPC C alleles in the oldest group with respect to the youngest one. We also obtained the same results when dividing the sample into 18-85 and 86-98 age groups. The general linear model analyses confirmed a significant decreasing trend with age of the above mentioned alleles. We hypothesised that these minor alleles, being important in the sensitivity against the development of different types of cancer, may reflect a reduced life-expectancy in carrier subjects and may explain their significantly lower frequency observed among subjects belonging to the oldest age group.

A micronucleus assay detects genotoxic effects of herbicide exposure in a protected butterfly species.

Lycaena dispar Hawort (Lepidoptera: Lycaenidae), a protected butterfly, is declining in Europe, but it thrives in rice fields in northern Italy. Here, agrochemical usage could threaten its long-term survival. We investigated, by micronucleus (MN) assay, the genotoxic effect of glyphosate, a common herbicide, on L. dispar larvae. Micronuclei (MNi) are DNA fragments separated from the main nucleus and represent the result of genomic damage that has been transmitted to daughter cells. In a control/treatment experiment, we extracted epithelial cells from last-instar larvae fed with Rumex spp. plants sprayed with a solution containing 3.6 g/L of glyphosate, and from larvae fed with unsprayed plants. MNi and other chromosomal aberrations-nuclear buds (NBUDs) and bi-nucleated cells-were then scored in 1000 cells/subject. Significant differences were found between glyphosate-exposed and control groups in terms of MNi and total genomic damage, but not in terms of NBUDs or bi-nucleated cells. We reported a possible genomic damage induced by glyphosate on larvae of L. dispar. For the first time, a MN assay was used in order to evaluate the genomic damage on a phytophagous invertebrate at the larval stage. Increased levels of MNi reflect a condition of genomic instability that can result in reduced vitality and in an increased risk of local extinction. Therefore, farmland management compatible with wildlife conservation is needed.

Micronuclei frequency in peripheral blood lymphocytes of healthy subjects living in turin (North-Italy): contribution of body mass index, age and sex.

Background: Increased micronuclei (MNi) frequencies in human lymphocytes are an indicator of chromosome instability and could be influenced by different exogenous and endogenous factors. The increased exposure to environmental pollutants has led to the awareness of the necessity for constant monitoring of urban human populations.Aim: We evaluated the MNi frequency in a sample belonging to the non-occupationally exposed population of Turin (North-Western Italy). A possible effect of body mass index, age and sex on the genomic damage levels was also investigated.Subjects and Methods: The study included 150 subjects. MNi, nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) were scored in 1,000 lymphocytes per subject.Results: The MNi, NPBs and NBUDs average frequencies (‰ ± S.D.) were 7.19 ± 2.51, 1.65 ± 1.54 and 2.07 ± 1.76, respectively. Turin shows one of the highest MNi frequencies with respect to other Italian cities and European regions. A significant correlation was found between MNi, NPBs, NBUDs frequencies, age and body mass index.Conclusion: Baseline MNi frequency was established in a sample of a city, like Turin, exposed to high levels of environmental pollutants. We hope that the results of this study can be used as a stimulus for future biomonitoring programmes in other Italian and globally distributed cities.

Induction of chromosomal aberrations and micronuclei by 2-hydroxy-4-methoxybenzophenone (oxybenzone) in human lymphocytes.

Oxybenzone or benzophenone-3 (2-hydroxy-4-methoxybenzophenone; BP-3) is a filter used in a variety of personal care products for protection of human skin and hair from damage by ultraviolet radiation. BP-3 is suspected to exhibit endocrine disruptive properties. Indeed, it was found to be able to interact with the endocrine system causing alteration of its homeostasis, with consequent adverse health effects. Moreover, it is ubiquitously present in the environment, mostly in aquatic ecosystems, with consequent risks to the health of aquatic organisms and humans. In the present study, we analyzed the cytogenetic effects of BP-3 on human lymphocytes using in vitro chromosomal aberrations and micronuclei assays. Blood samples were obtained from five healthy Italian subjects. Lymphocyte cultures were exposed to five concentrations of BP-3 (0.20, 0.10, 0.05, 0.025, and 0.0125 µg/mL) for 24 and 48 h (for chromosomal aberrations and micronuclei tests, respectively). The concentration of 0.10 µg/mL represents the acceptable/tolerable daily intake reference dose established by European Union, whereas 0.20, 0.05, 0.025, and 0.0125 µg/mL represent multiple and sub-multiple of this concentration value. Our results reported cytogenetic effects of BP-3 on cultured human lymphocytes in terms of increased micronuclei and chromosomal aberrations' frequencies at all tested concentrations, including concentrations lower than those established by European Union. Vice versa, after 48-h exposure, a significant reduction of the cytokinesis-block proliferation index value in cultures treated with BP-3 was not observed, indicating that BP-3 does not seem to produce effects on the proliferation/mitotic index when its concentration is equal to or less than 0.20 µg/mL.

Evaluation of the possible association of body mass index and four metabolic gene polymorphisms with longevity in an Italian cohort: a role for APOE, eNOS and FTO gene polymorphisms.

Background: Longevity is considered the result of interactions between environmental and genetic factors.Aim: To investigate the possible association of body mass index and the frequencies of APOE, ACE, eNOS and FTO gene polymorphisms with longevity.Subjects and methods: In total, 1100 healthy volunteers aged 10-100 were recruited. Subjects were genotyped for APOE, ACE, eNOS and FTO gene polymorphisms. Data about height and weight were also collected. The sample was split into four age groups: 1-24, 25-49, 50-85 and 86-100 years.Results: Significant differences were found in BMI values between age groups. A significant decrease of the APOE4, eNOS 393 and FTO A and allele frequencies was observed in the 86-100 age group compared to the younger groups. For ACE gene, no significant differences were found in the allele frequencies between groups. A similar trend was also observed when the sample was subdivided into two main age groups: 1-85 and 86-100 years.Conclusion: This study provides evidence for a role of APOE, eNOS and FTO gene polymorphisms in longevity. It has been estimated that the number of centenarians worldwide will double each decade until 2100, making population data about gene polymorphisms relevant for further studies about longevity.

Genomic damage induced by the widely used fungicide chlorothalonil in peripheral human lymphocytes.

Chlorothalonil is an important broad spectrum fungicide widely used in agriculture, silviculture, and urban settings. As a result of its massive use, chlorothalonil was found in all environmental matrices, with consequent risks to the health of terrestrial and aquatic organisms, as well as for humans. We analyzed the effects of chlorothalonil on human lymphocytes using in vitro chromosomal aberrations (CAs) and micronuclei (MNi) assays. Lymphocytes were exposed to five concentrations of chlorothalonil: 0.600 µg/mL, 0.060 µg/mL, 0.030 µg/mL, 0.020 µg/mL, and 0.015 µg/mL, where 0.020 and 0.600 µg/mL represent the ADI and the ARfD concentration values, respectively, established by FAO/WHO for this compound; 0.030 and 0.060 µg/mL represent intermediate values of these concentrations and 0.015 µg/mL represents the ADI value established by the Canadian health and welfare agency. We observed cytogenetic effects of chlorothalonil on cultured human lymphocytes in terms of increased CAs and MNi frequencies at all tested concentrations, including the FAO/WHO ADI and ARfD values of 0.020 and 0.600 µg/mL, respectively, but with exception of the Canadian ADI value of 0.015 µg/mL. Finally, no sexes differences were found in the levels of CAs and MNi induced by different chlorothalonil concentrations. Similarly, the mitotic index and the cytokinesis-block proliferation index did not show any significant effect on the proliferative capacity of the cells, although at the chlorothalonil concentration of 0.600 µg/mL the P-values of both indices were borderline.

Association of GSTT1 null, XPD 751 CC and XPC 939 CC genotypes with increased levels of genomic damage among hospital pathologists.

CONTEXT: Hospital workers are at risk for genotoxic damage following occupationally exposure to xenobiotics. Pathologists are exposed to chemicals during their use in health care environments, particularly throughout inhalation of airborne agents, absorption through skin or contact with the patient's body fluids. OBJECTIVE: We evaluated the level of genomic damage in a sample of 61 hospital pathologists (occupationally exposed to antineoplastic drugs and sterilizing agents) and 60 control subjects. MATERIALS AND METHODS: Lymphocytes were analyzed by SCEs and CAs assays and genotyped for GSTT1, GSTM1, CYP1A1 Ile/Val, XPD (A751C) and XPC (A939C) gene polymorphisms. RESULTS: Pathologists showed significantly higher frequencies of SCEs and CAs with respect to control subjects. GSTT1 null genotype was found to be associated with higher SCEs and CAs frequencies, whereas XPD 751 CC and XPC 939 CC genotypes only with a higher level of SCEs. DISCUSSION AND CONCLUSIONS: The SCEs and CAs results are consistent with other published data, placing hospital workers as a category at risk for genotoxic damage caused by chronic exposure to xenobiotics. The higher levels of cytogenetic damage observed among GSTT1 null, XPD 751 and XPC 939 CC homozygote subjects confirm the importance of the genetic polymorphisms analysis associated to genotoxicological studies.

Evaluation of baseline frequency of sister chromatid exchanges in an Italian population according to age, sex, smoking habits, and gene polymorphisms.

OBJECTIVES: Increased SCEs frequencies in human lymphocytes are an indicator of spontaneous chromosome instability and could be influenced by different exogenous and endogenous factors. In this study, we evaluated the influence of age, sex, smoking habits, and genetic polymorphisms on the background levels of SCEs in peripheral blood lymphocytes. METHODS: Two hundred-thirty healthy Italian subjects were recruited. Data about age, sex and smoking habits were recorded. Subjects were also genotyped for GSTT1, GSTM1, GSTP1 A/G, CYP1A1 Ile/Val, CYP2C19 G/A, ERCC2/XPD Lys751Gln, XRCC1 Arg194ATrp, XRCC1 Arg399Gln, and XRCC1Arg208His gene polymorphisms. RESULTS: The frequency of SCEs/cell was 5.15 ± 1.87, with females showing a significantly higher SCEs value with respect to males (5.36 ± 2.10 and 4.82 ± 1.39, respectively). Smokers showed significantly increased levels of SCEs with respect to nonsmokers (5.93 ± 1.75 and 4.70 ± 1.79, respectively) whereas no differences were observed between heavy and light smokers. Age correlated with the RI value (P = .01) but not with the SCEs frequency (P = 07), although the 31-40 age group showed a significantly lower SCEs frequency with respect to the other age groups. A significant association was also found between GSTP2C19-AA, GSTT1-null, GSTM1-null, ERCC2/XPD Gln751Gln, and XRCC1 His208His genotypes, and higher frequencies of SCEs. CONCLUSION: We describe the association between some phase I, phase II, and DNA-repair gene polymorphisms with increased SCEs frequencies, reinforcing the importance of genetic analysis in biomonitoring studies. Sex and age were found to be important endogenous factors that affect the level of genomic damage and the replicative capacity of cells, respectively.

Baseline frequency of chromosomal aberrations and sister chromatid exchanges in peripheral blood lymphocytes of healthy individuals living in Turin (North-Western Italy): assessment of the effects of age, sex and GSTs gene polymorphisms on the levels of genomic damage.

BACKGROUND: The increased exposure to environmental pollutants has led to the awareness of the necessity for constant monitoring of human populations, especially those living in urban areas. AIM: This study evaluated the background levels of genomic damage in a sample of healthy subjects living in the urban area of Turin (Italy). The association between DNA damage with age, sex and GSTs polymorphisms was assessed. SUBJECTS AND METHODS: One hundred and one individuals were randomly sampled. Sister Chromatid Exchanges (SCEs) and Chromosomal Aberrations (CAs) assays, as well as genotyping of GSTT1 and GSTM1 genes, were performed. RESULTS: Mean values of SCEs and CAs were 5.137 ± 0.166 and 0.018 ± 0.002, respectively. Results showed age and gender associated with higher frequencies of these two cytogenetic markers. The eldest subjects (51-65 years) showed significantly higher levels of genomic damage than younger individuals. GSTs polymorphisms did not appear to significantly influence the frequencies of either markers. CONCLUSION: The CAs background frequency observed in this study is one of the highest reported among European populations. Turin is one of the most polluted cities in Europe in terms of air fine PM10 and ozone and the clastogenic potential of these pollutants may explain the high frequencies of chromosomal rearrangements reported here.

Evidence of genotoxicity in lymphocytes of non-smoking alcoholics.

Alcohol abuse is a significant public health issue. Epidemiological studies conducted on different populations consistently showed that consumption of alcoholic beverages is associated with cytogenetic damages and higher risk for several types of cancer. However, the interpretation of many cytogenetic studies resulted complicated because some confounding factors, such as smoking habit, are not always taken into account. In the present study, the frequency of sister chromatid exchanges (SCEs), chromosome aberrations (CAs) and micronuclei (MNs) in cultured human lymphocytes was assessed on 15 alcoholic and 15 non-alcoholic control male subjects. Moreover, considering the implication of the Glutathione S-transferases gene polymorphisms in the genetic susceptibility to alcoholic liver diseases, we considered an important issue to evaluate the relationship between these gene polymorphisms and the cytogenetic damage. In our sample we exclusively considered individuals that did not smoke nor consume drugs for a period of at least 2 years prior to the analysis. Statistically significant differences were found between alcoholics and controls in the frequency of SCEs/cell (P = 0.001), RI value (P = 0.001), CAs (P = 0.002) and CAB (P = 0.002). Vice versa, no significant differences were found between alcoholics and controls in terms of MNs frequency and CBPI value. In both samples, no statistically significant association was found between the analysed GSTs gene polymorphisms and the frequencies of MNs, SCEs and CAs. Finally, among alcoholics we found a positive correlation between SCEs and CAs frequencies and the duration of alcohol abuse.

Evaluation of Genomic Damage in Peripheral Lymphocytes from Occupationally Exposed Anesthetists: Assessment of the Effects of Age, Sex, and GSTT1 Gene Polymorphism.

Occupational exposure to anaesthetic gases is one of the major hazards to healthcare personnel. We evaluated the cytogenetic effects of chronic exposure to low concentrations of anaesthetic gases in operating theatres. The study included 21 anesthetists and 21 control subjects who matched in age and gender. Chromosome aberrations (CAs) and sister chromatid exchanges (SCEs) assays were performed. All subjects were also genotyped for glutathione S-transferase T1 (GSTT1) gene polymorphisms. Significant differences were found between exposed and controls in terms of SCEs frequency (P = 0.001) and replication index value (P = 0.005), but not in terms of CAs (P = 0.201) and aberrant cells (P = 0.227) frequencies. Regression analyses indicated that age and the years of employment did not influence the level of chromosomal damage in both groups. Finally, among anesthetists, GSTT1 null individuals showed a significant higher frequency of SCE with respect to GSTT1-positive subjects.

Cellular and Genomic Instability Induced by the Herbicide Glufosinate-Ammonium: An In Vitro and In Vivo Approach.

Glufosinate-ammonium (GLA), an organophosphate herbicide, is released at high concentrations in the environment, leading to concerns over its potential genotoxic effects. However, few articles are available in the literature reporting the possible cellular and nuclear effects of this compound. We assessed, by in vitro and in vivo micronucleus assays, the genotoxicity of GLA on cultured human lymphocytes and Lymnaea stagnalis hemocytes at six concentrations: 0.010 (the established acceptable daily intake value), 0.020, 0.050, 0.100, 0.200, and 0.500 µg/mL. In human lymphocytes, our results reveal a significant and concentration-dependent increase in micronuclei frequency at concentrations from 0.100 to 0.500 µg/mL, while in L. stagnalis hemocytes, significant differences were found at 0.200 and 0.500 µg/mL. A significant reduction in the proliferation index was observed at all tested concentrations, with the only exception of 0.010 µg/mL, indicating that the exposure to GLA could lead to increased cytotoxic effects. In L. stagnalis, a significant reduction in laid eggs and body growth was also observed at all concentrations. In conclusion, we provided evidence of the genomic and cellular damage induced by GLA on both cultured human lymphocytes and a model organism's hemocytes; in addition, we also demonstrated its effects on cell proliferation and reproductive health in L. stagnalis.

In vitro genomic damage caused by glyphosate and its metabolite AMPA.

Glyphosate is the most widely used systemic herbicide. There is ample scientific literature on the effects of this compound and its metabolite aminomethylphosphonic acid (AMPA), whereas their possible combined genotoxic action has not yet been studied. With the present study, we aimed to determine the level of genomic damage caused by glyphosate and AMPA in cultured human lymphocytes and to investigate the possible genotoxic action when both compounds were present at the same concentrations in the cultures. We used a micronuclei assay to test the genotoxicity of glyphosate and AMPA at six concentrations (0.0125, 0.025, 0.050, 0.100, 0.250, 0.500 µg/mL), which are more realistic than the highest concentrations used in previous published studies. Our data showed an increase in micronuclei frequency after treatment with both glyphosate and AMPA starting from 0.050 µg/mL up to 0.500 µg/mL. Similarly, a genomic damage was observed also in the cultures treated with the same concentrations of both compounds, except for exposure to 0.0065 and 0.0125 µg/mL. No synergistic action was observed. Finally, a significant increase in apoptotic cells was observed in cultures treated with the highest concentration of tested xenobiotics, while a significant increase in necrotic cells was observed also at the concentration of 0.250 µg/mL of both glyphosate and AMPA alone and in combination (0.125 + 0.125 µg/mL). Results of our study indicate that both glyphosate and its metabolite AMPA are able to cause genomic damage in human lymphocyte cultures, both alone and when present in equal concentrations.

Genotoxicological and physiological effects of glyphosate and its metabolite, aminomethylphosphonic acid, on the freshwater invertebrate Lymnaea stagnalis.

Aminomethylphosphonic acid (AMPA) is the main metabolite in the degradation of glyphosate, a broad-spectrum herbicide, and it is more toxic and persistent in the environment than the glyphosate itself. Owing to their extensive use, both chemicals pose a serious risk to aquatic ecosystems. Here, we explored the genotoxicological and physiological effects of glyphosate, AMPA, and the mixed solution in the proportion 1:1 in Lymnaea stagnalis, a freshwater gastropod snail. To do this, adult individuals were exposed to increasing nominal concentrations (0.0125, 0.025, 0.050, 0.100, 0.250, 0.500 µg/mL) in all three treatments once a week for four weeks. The genotoxicological effects were estimated as genomic damage, as defined by the number of micronuclei and nuclear buds observed in hemocytes, while the physiological effects were estimated as the effects on somatic growth and egg production. Exposure to glyphosate, AMPA, and the mixed solution caused genomic damage, as measured in increased frequency of micronuclei and nuclear buds and in adverse effects on somatic growth and egg production. Our findings suggest the need for more research into the harmful and synergistic effects of glyphosate and AMPA and of pesticides and their metabolites in general.

Influence of Nutritional Stress on Female Allocation and Somatic Growth in the Simultaneously Hermaphroditic Polychaete Worm Ophryotrocha diadema.

Hermaphrodites are characterized by plastic sex allocation, by which they adjust their allocation of reproductive resources according to mating opportunities. However, since the plasticity of sex allocation is influenced by environmental conditions, it may also be affected by species-specific life-history traits. In this study, we explored the trade-off between nutritional stress due to food deficiency and the investment of resources in female allocation and somatic growth in the simultaneously hermaphroditic polychaete worm, Ophryotrocha diadema. To achieve this, we exposed adult individuals to three food supply levels: (1) ad libitum-100% food supply, (2) intense food deficiency-25% food resources, and (3) extreme food deficiency-0% food resources. Our findings show a progressive decrease in female allocation in the numbers of cocoons and eggs and in body growth rate of O. diadema individuals as the level of nutritional stress increased.