Two-Generation Toxicity Study of the Antioxidant Compound Propyl-Propane Thiosulfonate (PTSO).

Propyl-propane thiosulfonate (PTSO), an antioxidant organosulfur compound present in the genus Allium, has become a potential natural additive for food and feed, as well as a possible biopesticide for pest control in plants. A toxicological assessment is necessary to verify its safety for livestock, consumers, and the environment. As part of the risk assessment of PTSO, this study was designed to explore its potential reproductive toxicity in mice following the OECD 416 guideline. The investigation spans two generations to comprehensively evaluate potential reproductive, teratogenic, and hereditary effects. A total of 80 CD1 mice per sex and generation were subjected to PTSO exposure during three phases (premating, gestation, and lactation). This evaluation encompassed three dose levels: 14, 28, and 55 mg PTSO/kg b.w./day, administered through the feed. No clinical changes or mortality attributed to the administration of PTSO were observed in the study. Some changes in the body weight and food consumption were observed, but not related to sex or in a dose-dependent manner. The two parental generations (F0, F1) exhibited normal reproductive performance, and the offspring (F1 and F2) were born without any abnormalities. The serum sexual hormone levels (progesterone -P-, testosterone -T-, estradiol -E2-, follicular stimulating hormone -FSH-, and luteinizing hormone -LH-) were in a normal range. Although significant changes were observed in the sperm analysis in the case of F0 group, no variation was found for F1 group, and no alterations in fertility were recorded either. The absolute organ weights and relative organ weight/body weight and organ weight/brain weight ratios, and the complete histopathological study, showed no significant alterations in males and females for all the generations considered. Considering all the results obtained, PTSO is not considered a reproductive or developmental toxicant in mice under the assayed conditions. These results support the good safety profile of PTSO for its potential application in the agrifood sector.

Aliphatic hydrocarbons in fin spines of adult sturgeon (Acipenser stellatus) and their relationship with potentially toxic elements in the northern and southern regions of the Caspian Sea.

Currently, the pollution of the Caspian Sea by the oil industry is one of the highest problems in this area. Critically endangered species inhabit this sea, such as sturgeons, whose ecological value is incalculable. Thus, we aimed to evaluate the level of contamination of aliphatic hydrocarbons of petroleum and its relation with several toxic elements directly on sturgeons spines. A total of 40 adult starry sturgeons (Acipenser stellatus) were obtained within a repopulation programme in the northern and southern coastal waters of the Caspian Sea. The marginal pectoral fin was extracted from each fish to determine aliphatic hydrocarbons, arsenic, cadmium, mercury, nickel, lead, and vanadium. Subsequently, the sturgeons were released. Clearly, the presence of hydrocarbons was evidenced in all the sampled areas finding higher concentrations in the northern areas (N1 = 1.35 ± 0.4; N2 = 1.65 ± 0.46; N3 = 1.27 ± 0.40; S1 = 0.61 ± 0.22; S2 = 0.85 ± 0.43 mg/kg). Furthermore, to a greater or lesser extent, some toxic elements, mainly Hg and As, have been linked to aliphatic hydrocarbons.

An Overview of the Health Effects of Bisphenol A from a One Health Perspective.

Bisphenol A (BPA) is a chemical compound, considered as an "emerging pollutant", that appears ubiquitously, contaminating the environment and food. It is an endocrine disruptor, found in a multitude of consumer products, as it is a constituent of polycarbonate used in the manufacture of plastics and epoxy resins. Many studies have evaluated the effects of BPA, using a wide range of doses and animal models. In this work, we carried out a review of relevant research related to the effects of BPA on health, through studies performed at different doses, in different animal models, and in human monitoring studies. Numerous effects of BPA on health have been described; in different animal species, it has been reported that it interferes with fertility in both females and males and causes alterations in their offspring, as well as being associated with an increase in hormone-dependent pathologies. Similarly, exposure to BPA has been related to other diseases of great relevance in public health such as obesity, hypertension, diabetes, or neurodevelopmental disorders. Its ubiquity and nonmonotonic behavior, triggering effects at exposure levels considered "safe", make it especially relevant when both animal and human populations are constantly and inadvertently exposed to this compound. Its effects at low exposure levels make it essential to establish safe exposure levels, and research into the effects of BPA must continue and be focused from a "One Health" perspective to take into account all the factors that could intervene in the development of a disease in any exposed organism.

Risk assessment and environmental consequences of the use of the Allium-derived compound propyl-propane thiosulfonate (PTSO) in agrifood applications.

The organosulfur compound propyl-propane thiosulfonate (PTSO), mainly found in Allium cepa, has a promising use in the agrifood industry. To confirm its safety for livestock, consumers, and environment, toxicological assessment is needed. In this regard, endocrine-disrupting chemicals (EDCs) are in the spotlight of research. Therefore, as part of the risk assessment of PTSO, in the present work, an in vivo study was performed in mice exposed to PTSO to investigate its potential reproductive toxicity considering fertility, genetic and endocrine endpoints. Five-weeks-old CD1 mice (80 males, 80 females) were exposed for 11 or 16 weeks (males or females, respectively) to different doses of PTSO (0, 14, 28 and 55 mg PTSO/kg b.w./day; 20 animals per group and sex) through the food pellets. No clinical observations or mortality and no changes in absolute organ weights and relative organ weights/body weight or brain ratios occurred during the study. The estrous cycle did not undergo any significant toxicologically relevant change. Most of the sex hormones displayed normal values. Some alterations in the expression of some genes related to reproduction is only observed in females, but they do not appear to have consequences in the development of sex organs. Docking results showed the impossibility of stable binding to estrogen and androgen receptors. Considering all the results obtained, the safe profile of PTSO can be confirmed for different agrifood applications at the conditions assayed.

Multivariate analysis of trace elements in starry sturgeon (Acipenser stellatus) spine in different areas of the Caspian Sea.

Sturgeons are one of the most valuable species in the Caspian Sea. There, habitation of the seabed and feeding on benthic organisms makes this species a good indicator of trace element status. Thus, we aimed to determine the concentrations of 31 trace elements in the pectoral fin spine of starry sturgeons, and to evaluate the relationships between the different chemical elements. For this, a total of 40 starry sturgeons were obtained in a repopulation programme from the coastal waters north and south of the Caspian Sea. First, we used a multivariate analysis of variance to establish the differences between zones. Later, to assess relationships between trace elements, we used principal component analysis and cluster analysis. In general, the concentration of many trace elements did not vary between zones. However, some elements, including mercury or arsenic, were found in the north areas at higher concentrations.

Bioaccumulation of rare earth elements and trace elements in different tissues of the golden grey mullet (Chelon auratus) in the southern Caspian Sea.

Rare earth elements are essential for modern life, although they are also classified as emerging pollutants. Currently, fish studies on these elements are very limited in general, but, with regard to the Caspian Sea, there is no reference to them at all. For this reason, our objective was to determine the concentrations of these elements in the golden grey mullet (Chelon auratus) and to contrast its bioaccumulation patterns with those of arsenic, cadmium, mercury and lead. For that purpose, 20 fish were caught in the southern part of the Caspian Sea. Heavy rare earth element concentrations were higher than light ones and the terbium levels were very high, probably due to anthropogenic contamination. The intestine tissue gave the highest concentrations, which could be indicative of a very low gastrointestinal absorption. For both rare earth and trace elements, muscle was the tissue that accumulated the least, despite which, cadmium and lead levels in muscle were of concern.

Effects of chronic low-dose aflatoxin B1 exposure in lactating Florida dairy goats.

In the past few years there has been a growing trend in the prevalence of aflatoxins, attributable to climate change, in substances destined for animal feeding, together with an increase in dairy product consumption. These facts have triggered great concern in the scientific community over milk pollution by aflatoxin M1. Therefore, our study aimed to determine the transfer of aflatoxin B1 from the diet into milk as AFM1 in goats exposed to different concentrations of AFB1, and its possible effect on the production and serological parameters of this species. For this purpose, 18 goats in late lactation were divided into 3 groups (n = 6) and exposed to different daily doses of aflatoxin B1 (T1 = 120 µg; T2 = 60 µg, and control = 0 µg), during 31 d. Pure aflatoxin B1 was administered 6 h before each milking in an artificially contaminated pellet. The milk samples were taken individually in sequential samples. Milk yield and feed intake were recorded daily, and a blood sample was extracted on the last day of exposure. No aflatoxin M1 was detected, either in the samples taken before the first administration, or in the control group ones. The aflatoxin M1 concentration detected in the milk (T1 = 0.075 µg/kg; T2 = 0.035 µg/kg) increased significantly on a par with the amount of aflatoxin B1 ingested. The amount of aflatoxin B1 ingested did not have any influence on aflatoxin M1 carryover (T1 = 0.066% and T2 = 0.060%), these being considerably lower than those described in dairy goats. Thus, we concluded that the concentration of aflatoxin M1 in milk follows a linear relationship with respect to the aflatoxin B1 ingested, and that the aflatoxin M1 carryover was not affected by the administration of different aflatoxin B1 doses. Similarly, no significant changes in the production parameters after chronic exposure to aflatoxin B1 were observed, revealing a certain resistance of the goat to the possible effects of that aflatoxin.

Comparative study of tricaine methanesulfonate (MS-222) and eugenol as euthanasia agents in zebrafish (Danio rerio) as an experimental model.

In experimental procedures inevitably leading to the sacrifice of animals, suitable measures should be taken to minimise their pain and suffering as much as possible, as well as to prevent any modification or masking in the experimental results obtained. An overdose of anaesthetic is the method of euthanasia most employed in fish, since it is effective and easy to apply. Our objectives were to compare the efficacy of eugenol and of tricaine methanesulfonate (MS-222) as euthanasia agents in zebrafish, and to make a histological evaluation of the possible effects derived from their application. The concentrations established for eugenol were 0.25 and 0.35 mg/mL, and those for MS-222 were 0.25 and 0.50 mg/mL, for both the buffered solution and the non-buffered one. Eugenol turned out to be a stronger euthanasia agent than MS-222 in zebrafish, presenting with significantly shorter euthanasia times. However, the exposure of the fish to euthanasia doses of eugenol triggered branchial alterations, in addition to serious lesions and changes in their nerve tissue. The results obtained with MS-222 also revealed a marked branchial alteration derived from its use. In this respect, the addition of a buffer to the MS-222 solution enhanced the effectiveness of the drug, with significantly shorter euthanasia times being achieved than with the non-buffered solution, and diminished the severity of the lesions described. We therefore determined that the buffered MS-222 solution is the most effective, reliable and safest method of euthanasia for use in research on zebrafish.

Health risk assessment of heavy metal concentration in muscle of Chelon auratus and Chelon saliens from the southern Caspian Sea.

Heavy metals are one of the most serious pollutants in aquatic ecosystems, and their accumulation in fish products causes harmful effects on human health. In this context, we set out to determine the concentrations of heavy metals in the muscle of two fish species of commercial interest, Chelon auratus and Chelon saliens on the south coast of the Caspian Sea. We aimed to assess the degree of environment contamination in this area and to estimate the potential risk to human health derived from the consumption of fish. The mean concentrations of the different metals analysed were very varied in both species. In fact, some concentrations exceeded the permissible limits for the protection of human health for Cd and Pb, and some values of As were above those referenced by other authors in same species in the Caspian Sea. None of the estimated daily intake values exceeded the tolerable intake based on the consumption under consideration. Nonetheless, the accumulative hazard values evidenced a potential risk to human health, Pb and Hg being those giving a higher target hazard quotient. The cancer risk from exposure to As from fish consumption in children was above the "acceptable" risk to life. Thus, in view of the accumulative nature of heavy metals, a moderate and non-abusive fish consumption in this area, particularly in children, would be recommendable.

Analysis of Blood Biochemistry and Pituitary-Gonadal Histology after Chronic Exposure to Bisphenol-A of Mice.

Bisphenol-A is an emerging pollutant that is widespread in the environment, and to which live beings are continuously and inadvertently exposed. It is a substance with an endocrine-disrupting capacity, causing alterations in the reproductive, immunological, and neurological systems, among others, as well as metabolic alterations. Our study aimed to assess its clinical signs, and effects on the most relevant blood biochemical parameters, and to evaluate pituitary and gonadal histology after a chronic exposure of adult mice to different BPA doses (0.5, 2, 4, 50 and 100 µg/kg BW/day) through their drinking water. The biochemical results showed that a marked significant reduction (p < 0.05) was produced in the levels of serum glucose, hypoproteinaemia and hypoalbuminemia in the groups exposed to the highest doses, whereas in the group exposed to 50 µg/kg BW/day the glucose and total protein levels dropped, and the animals exposed to 100 µg/kg BW/day experienced a diminution in albumin levels. In the case of the group exposed to 50 µg/kg BW/day, however, hypertriglyceridemia and hypercholesterolemia were determined, and the blood parameters indicating kidney alterations such as urea and creatinine experienced a significant increase (p < 0.05) with respect to the controls. Regarding the pituitary and gonads, none of the animals exposed presented histological alterations at the doses tested, giving similar images to those of the control group. These results suggest that continuous exposure to low BPA doses could trigger an inhibition of hepatic gluconeogenesis, which would result in a hypoglycaemic state, together with an induction of the enzymes responsible for lipidic synthesis, a mechanism by which the increase in the lipid and serum cholesterol levels could be explained. Likewise, the decline in the protein and albumin levels would be indicative of a possible hepatic alteration, and the increase in urea and creatinine would point to a possible renal perturbation, derived from continuous exposure to this xenobiotic. Based on our results, it could be said that chronic exposure to low BPA doses would not produce any clinical signs or histological pituitary-gonadal effects, but it could cause modifications in some blood biochemical parameters, that could initially indicate a possible hepatic and renal effect.

Determination of the Heavy Metal Bioaccumulation Patterns in Muscles of Two Species of Mullets from the Southern Caspian Sea.

Although fish is a food that supplies nutrients of a high biological value, they can also be a source of some harmful substances, such as heavy metals. In the same context, some human activities in the Caspian Sea have contaminated this ecosystem during the past few years. For those reasons, our objective consisted of determining the concentrations of heavy metals and evaluating their bioaccumulation patterns in the different types of musculature in two species of mullets of commercial interest, Chelon auratus and Chelon saliens, from the southern coast of this sea. For this purpose, 20 C. auratus and 29 C. saliens were caught off this coastline and the metal concentrations in 3 different muscle locations were analyzed: the ventral, dorsal and caudal muscles of each fish. The caudal muscle had higher concentrations of Cd, Cu, Pb, and Zn, whereas As, Hg and Ni accumulation seemed to be independent of the musculature type. Overall, the Cd, Hg, and Pb concentrations exceeded the maximum levels permitted in fish by the European Union. In addition, the relationships between pairs of metals were positive and elevated in all the cases, which could be a sign of heavy metal pollution in the region sampled. Therefore, it will be necessary to continue monitoring and evaluating the degree of pollution in the Caspian Sea.

Analysis of Indirect Biomarkers of Effect after Exposure to Low Doses of Bisphenol A in a Study of Successive Generations of Mice.

Bisphenol A (BPA) is considered as being an emerging pollutant, to which both animal and human populations are continuously and inadvertently exposed. The identification of indirect biomarkers of effect could be a key factor in determining early adverse outcomes from exposure to low doses of BPA. Thus, this study on mice aims to evaluate and identify indirect biomarkers of effect through the analysis of their blood biochemistry, and of certain reproduction parameters after exposure to different BPA concentrations (0.5, 2, 4, 50, and 100 µg/kg BW/day) in drinking water over generations. Our results showed that there were no modifications in the reproductive parameters evaluated, like estrous cycle duration, litter size, or the percentage of the young alive at reaching the weaning stage, at the exposure levels evaluated. However, there were modifications in the biochemical parameters, e.g., alterations in the glucose levels, that increased significantly (p < 0.05) in the breeders at the higher exposure doses (50 and 100 µg/kg BW/day in F1; 50 µg/kg BW/day in F2 and 100 µg/kg BW/day in F3), that would suggest that the BPA could induce hyperglycemia and its complications in adult animals, probably due to some damage in the pancreas cells; albumin, that increased in the breeders exposed to the highest dose in F1 and F3, inferring possible hepatic alterations. Further, total proteins showed a diminution in their values in F1 and F2, except the group exposed to 100 µg/kg BW/day, whereas in F3 the values of this parameter increased with respect to the control group, this aspect likely being related to a possible hepatic and renal alteration. Based on these results, glucose, albumin, and total proteins could initially be considered as early indicators of indirect effect after prolonged exposure to low BPA doses over generations.

Evaluation of the Toxicity of Bisphenol A in Reproduction and Its Effect on Fertility and Embryonic Development in the Zebrafish (Danio rerio).

Bisphenol A (BPA) is a chemical substance commonly used in the manufacture of plastic products. Its inhalation or ingestion from particles in suspension, water, and/or polluted foods can trigger toxic effects related to endocrine disruption, resulting in hormonal, reproduction, and immunological alterations in humans and animals. The zebrafish (Danio rerio) is an ideal experimental model frequently used in toxicity studies. In order to assess the toxic effects of BPA on reproduction and embryonic development in one generation after parental exposure to it, a total of 80 zebrafish, males and females, divided into four groups in duplicate (n = 20) were exposed to BPA concentrations of 500, 50, and 5 µg L-1, along with a control group. The fish were kept in reproduction aquariums for 21 days. The embryos obtained in the crosses were incubated in a BPA-free medium and observed for signs of embryotoxicity. A histopathological study (under optical and electron microscopes) was performed of adult fish gonads. The embryos of reproducers exposed to BPA were those most frequently presenting signs of embryotoxicity, such as mortality and cardiac and musculoskeletal malformations. In the histopathological studies of adult individuals, alterations were found in ovocyte maturation and in spermatazoid formation in the groups exposed to the chemical. Those alterations were directly related to BPA action, affecting fertility in both sexes, as well as the viability of their offspring, proportionally to the BPA levels to which they were exposed, so that our results provide more information by associating toxic effects on the offspring and on the next generation.

Tremorgenic mycotoxicosis in cattle, caused by Claviceps paspali.

Claviceps paspali is a fungus that mainly parasitises Paspalum dilatatum, generating a structure denominated sclerotium, in which indole-diterpenoid alkaloids are isolated. Its action mechanism is related to the inhibition of the gamma-aminobutyric acid receptor. It basically affects bovines, triggering the tremorgenic syndrome, the prevalence of this intoxication being relatively low in Europe. This work describes a clinical case on a cattle farm in Seville (southern Spain), composed of 91 bovines, 60% of which were principally affected with nervous clinical signs. The diagnosis was based on a clinical inspection of the animals, as well as of the presence of paspalum seed heads containing the sclerotia of Claviceps paspali in the plants present in the pastures. The causal agent was identified as being Claviceps paspali, which had parasitised numerous examples of Paspalum paspaloides. The indole-diterpenoid alkaloids produced by Claviceps paspali were identified using ultraviolet-visible spectrophotometry and mass spectrometry. At present, no effective aetiological treatment has been described for poisoning caused by this mycotoxin, so a supportive treatment was administered, and different handling methods were applied, resulting in the complete recovery of the animals. Finally, it was concluded that unusually high humidity and temperature levels for the region triggered the development of the sclerotium generated by this fungus.