Poisoning by Nerium oleander L. in Franconia Geese.

This study describes the acute poisoning of four 3-month-old Franconia geese (Anser anser) by oleander plants (Nerium oleander). After the accidental ingestion of oleander clippings, the geese exhibited a rapid onset of severe symptoms, leading to mortality within 15-90 min. Necropsy revealed cardiac and renal lesions. Specifically, interstitial edema, red blood cell infiltration, and myofibril loss were observed in the cardiac muscle, and tubular epithelial degeneration, interstitial edema, and hemorrhages were evident in the kidneys. Oleandrin, a glycoside with cardiac effects, was detected in the liver, kidneys, heart, brain, and muscles. The clinical implications underscore the urgency of veterinary intervention upon oleander ingestion, and the specific findings contribute valuable insights into the pathological effects of acute oleander poisoning in geese, aiding veterinarians in prompt diagnosis and treatment.

The Effect of By-Pass Linseed Oil Supplementation on the Maternal Antioxidant System during the Embryo-Maternal Recognition Period in Ewes.

This study analyzed the effects of dietary supplementation with by-pass linseed oil (LO; rich in α-linolenic acid) on maternal antioxidant systems at Days 14 and 16 of pregnancy in Sarda ewes. This trial used sixteen dry ewes. Eight ewes (CT group) were fed with a control diet without LO, and eight ewes (LO group) were fed with a diet supplemented with LO (10.8 g of α-linolenic acid/ewe/day). Both diets had similar crude protein and energy levels. The experiment included 10 days of an adaptation period and 31 days of a supplementation period. This supplementation period was divided into Period -2 (from Day -15 to -8), Period -1 (from Day -7 to -1; before synchronized mating period/Day 0), Period +1 (from Day +1 to + 7 after mating), and Period +2 (from Day +8 to +15 after mating). Estrous synchronization was induced in all the ewes using an intravaginal sponge (45 mg fluorgestone acetate) for 14 days and equine chorionic gonadotropin (350 UI/ewe) at the end of the treatment. On Days 14 (CT, N = 4; LO, N = 4) and 16 (CT, N = 4; LO, N = 4) after mating, the ewes were slaughtered. Samples of plasma, uterine, and luteal tissues were collected. Thiols, total antioxidant activity (TEAC), superoxide dismutase (SOD) activity, and malondialdehyde (MDA) content were measured. On Day 16, thiol and TEAC in luteal tissues were higher in the LO group when compared with the control one (p < 0.05). Moreover, TEAC was higher for the LO group in uterine tissues on Days 14 and 16 (p < 0.05). SOD activity was higher in the LO group in luteal and uterine tissues on Day 14 and Day 16, respectively (p < 0.001). On Day 16, uterine MDA content was lower for the LO group (p < 0.001). No differences were found between groups at the plasmatic level. However, the by-pass LO supplementation enhanced the analyzed antioxidant parameters in luteal and uterine tissues. In conclusion, these results demonstrate that by-pass LO supplementation exerted a positive effect on antioxidative defenses on maternal structures during the embryo-maternal recognition period in ewes. Thus, this could contribute to improving the maternal environment during the embryo-maternal recognition period in mammals.

An Overview on Assay Methods to Quantify ROS and Enzymatic Antioxidants in Erythrocytes and Spermatozoa of Small Domestic Ruminants.

The present review aims to provide an overview of the assay methods for the quantification of ROS and principal enzymatic antioxidants as biomarkers of oxidative stress in erythrocytes and spermatozoa of small domestic ruminants. A complete literature search was carried out in PubMed, Scopus and the World Wide Web using relevant keywords and focusing on the last five years (2018-2023). Among spectrophotometry, fluorometry and chemiluminescence, the most widely used method for ROS assay is fluorometry, probably because it allows to simultaneously assay several ROS, using different probes, with greater economic advantages. Regarding intracellular antioxidant enzymes, recent literature reports only spectrophotometric methods, many of which use commercial kits. The use of a less sensitive but cheapest method is suitable because both erythrocytes and spermatozoa samples are highly concentrated in domestic ruminant species. All methods considered in this review have been found to be appropriate; in general, the differences are related to their costs and sensitivity. Quantification of ROS and enzymatic antioxidant activity in erythrocytes and spermatozoa may find application in the study of the welfare and health status of small domestic ruminants for monitoring livestock production.

Analysis of 2,5-dimethoxy-amphetamines and 2,5-dimethoxy-phenethylamines aiming their determination in biological matrices: a review.

PURPOSE: The present review aims to provide an overview of methods for the quantification of 2,5-dimethoxy-amphetamines and -phenethylamines in different biological matrices, both traditional and alternative ones. METHODS: A complete literature search was carried out with PubMed, Scopus and the World Wide Web using relevant keywords, e.g., designer drugs, amphetamines, phenethylamines, and biological matrices. RESULTS: Synthetic phenethylamines represent one of the largest classes of "designer drugs", obtained through chemical structure modifications of psychoactive substances to increase their pharmacological activities. This practice is also favored by the fact that every new synthetic compound is not considered illegal by existing legislation. Generally, in a toxicological laboratory, the first monitoring of drugs of abuse is made by rapid screening tests that sometimes can occur in false positive or false negative results. To reduce evaluation errors, it is mandatory to submit the positive samples to confirmatory methods, such as gas chromatography or liquid chromatography combined to mass spectrometry, for a more specific qualitative and quantitative analysis. CONCLUSIONS: This review highlights the great need for updated comprehensive analytical methods, particularly when analyzing biological matrices, both traditional and alternative ones, for the search of newly emerging designer drugs.

Cross-reactivity of commercial immunoassays for screening of new amphetamine designer drugs. A review.

The chemical modification of the molecular structure of psychoactive substances is a very common practice in the illicit drugs market, to by-pass current regulations; this lead to the production of compounds, known as "designer drugs", with the same or greater pharmacological effects of the parent drug. The phenomenon is also favored by the fact that the new synthetic compounds are not considered illegal by existing legislation. Amphetamine derivatives represent one of the largest classes of designer drugs. Generally, in toxicological laboratories, rapid screening tests are used for a first monitoring of drugs abuse. However, the available immunoassays for this class of substances are designed for amphetamine, methamphetamine and methylenedioxymethamphetamine, and generally they are unable to detect various amphetamine analogues. This can constitute a disadvantage because it can generate a great number of false-negative results. The present review aims to provide an overview of the cross-reactivity studies carried out on commercially available immunoassays to identify the presence of amphetamine derivatives in biological samples. The knowledge of cross-reactivity data makes it easier to interpret analytical results by demonstrating that a negative result does not always indicate the non-consumption of an amphetamine derivative. This review highlights the great need for more comprehensive screening immunoassays to use when analyzing biological matrices for drugs of abuse search, specifically for the more recent designer drugs..

Non-invasive Assay for Measurement of Fecal Triiodothyronine (T3) Metabolite Levels in European Mouflon (Ovis aries musimon).

Thyroid hormones (THs) are important indicators of metabolism and animal health. Traditionally, they have been determined from blood or urine samples. However, as their collection may be stressful and requires ethical approval, alternative non-invasive matrices are preferred when dealing with wild animals. Triiodothyronine (T3) is the active form of THs in blood and the major metabolite excreted in feces. This creates the ideal conditions for its assay in fecal samples. Fecal sampling eliminates the stress of the animals and the need to physically capture them. However, in wild species it is rare to find species-specific kits for the hormone assay. So, the objective of this work was to validate a method for the quantification of T3 metabolite (FTM) levels in feces of European mouflon by using an economic and easily available ELISA kit designed to quantify T3 in human plasma. Analytical and biological validations were performed in feces collected from 10 mouflons (5 ewes and 5 rams). An efficient liquid-extraction method was optimized. Precision, dilution linearity, parallelism, recovery and stability of T3 in fecal samples were calculated. Obtained data were considered acceptable according to international guidelines. The reliability of the results was verified comparing human plasma and mouflon fecal samples fortified with the same T3 standard solutions. The biological validation showed higher FTM levels in March compared to June, and no differences between mouflon ewes and rams. The validation of the present method provides a non-invasive and affordable tool for the quantification of FTM in European mouflon.

Evaluation of curcuma and ginger mixture ability to prevent ROS production induced by bisphenol S: an in vitro study.

The use of bisphenol S (BPS) as a substitute of Bisphenol A is increasing in several products and it can be found in different environmental and biological matrices. Its toxicity has been studied at different levels and one of BPS toxic mechanisms at high concentrations seems to be the induction of oxidative stress through the generation of reactive oxygen species (ROS). This study evaluates the ability of a curcuma and ginger (CG) mixture to exert an antioxidant effect on rat hepatocytes treated with BPS. The effects of the mixture were compared to those of a well-known antioxidant (Trolox). Three different BPS concentrations were used in order to verify ROS production. 70 µg/mL and 150 µg/mL of BPS generated a significant ROS increase (p < 0.01) as compared to control, while CG mixture was able to decrease this ROS production in hepatic cells, as compared to cells treated with 70 µg/ml of BPS (p < 0.01) restoring control levels. BPS 70 µg/mL was tested for total antioxidant capacity (TEAC), superoxide dismutase (SOD) and total thiols. TEAC and SOD significant decreased (p < 0.05 and p < 0.01, respectively) as compared to controls and CG mixture was able to restore control values. Given the widespread BPS use, results obtained in this study can be of high impact for the community, demonstrating the ability of a mixture of natural products to prevent BPS-induced oxidative stress.

Measurement of progesterone in sheep using a commercial ELISA kit for human plasma.

Determination of serum or plasma progesterone (P4) concentrations is important to recognize pregnant and non-pregnant ewes, and also to predict the number of carried lambs. The 2 most common methodologies for the detection of plasma P4 are radioimmunoassay (RIA) and enzyme immunoassay (EIA). RIA is very expensive, and not all laboratories are equipped to perform this test; EIA is commercially available for human use, but only a few companies produce species-specific kits, which are expensive. We verified for ovine plasma a less expensive and easily available ELISA kit (DiaMetra) designed to quantify P4 in humans. Pools of ovine and human plasma were used to compare repeatability, accuracy, sensitivity, and stability of P4 measured by the DiaMetra kit. Repeatability data were within 15%, and accuracy values were ~90% for both plasma matrices. Stability data showed a loss of <20% for freeze-thaw and <30% for 30-d storage. All parameters were acceptable under international guidelines for method validation. The human ELISA kit was used successfully to quantify plasma P4 in 26 ewes during pregnancy until delivery. P4 concentrations were also correlated with the number of carried lambs.

An Overview of Antibiotics as Emerging Contaminants: Occurrence in Bivalves as Biomonitoring Organisms.

Antibiotics are used for therapeutic and prophylactic purposes in both human and veterinary medicine and as growth promoting agents in farms and aquaculture. They can accumulate in environmental matrices and in the food chain, causing adverse effects in humans and animals including the development of antibiotic resistance. This review aims to update and discuss the available data on antibiotic residues, using bivalves as biomonitoring organisms. The current research indicates that antibiotics' presence in bivalves has been investigated along European, American and Asian coasts, with the majority of studies reported for the last. Several classes of antibiotics have been detected, with a higher frequency of detection reported for macrolides, sulfonamides and quinolones. The highest concentration was instead reported for tetracyclines in bivalves collected in the North Adriatic Sea. Only oxytetracycline levels detected in this latter site exceeded the maximum residual limit established by the competent authorities. Moreover, the risk that can be derived from bivalve consumption, calculated considering the highest concentrations of antibiotics residues reported in the analyzed studies, is actually negligible. Nevertheless, further supervisions are needed in order to preserve the environment from antibiotic pollution, prevent the development of antimicrobial resistance and reduce the health risk derived from seafood consumption.

Bisphenols' occurrence in bivalves as sentinel of environmental contamination.

Bisphenols are massively used in several manufacture processes such that bisphenol A (BPA) is ubiquitous in environment worldwide. After the implementation of regulations about BPA use, manufacturers have moved their production toward alternative substances structurally similar to it. Unfortunately, BPA analogues, given their structural similarity, exert also similar adverse effects. This review aims to investigate the occurrence of bisphenols (BPs) in bivalve molluscs. In this way, valuable information on the amount of BPs released into the environment in different areas are given. The current research indicates that BPA presence in bivalve molluscs has been investigated in Asia (Indian Ocean and Pacific Ocean), Europe (Mediterranean Sea, Baltic Sea and Atlantic Ocean) and America (Lake Mead, Nevada) with the highest amount of studies reported in bivalves harvested in Asian Coasts. BPA analogues are frequently detected in several matrices and their levels will continuously increase in the environment. Nevertheless, there is a current lack of studies analysing BPs other than BPA in bivalves. Further investigations should be conducted in this direction, in order to assess environmental distribution and the hazard for animals and human health given that seafood consumption could be an important pathway of bisphenols intake.

Determination of Praziquantel in Sparus aurata L. after Administration of Medicated Animal Feed.

Praziquantel (PZQ) is an anthelmintic drug used in humans and animals against Platyhelminthes and in aquaculture in the Far East. Medicated feed is one of the most convenient forms of oral administration of drugs in aquaculture because it allows to treat a large population of fish in an easy way. However, this treatment may lead to residues in fish intended for human consumption. In this study, a liquid chromatography with tandem mass spectrometry (LC-MS/MS) method was developed in order to verify the presence of PZQ in samples of Sparus aurata after oral administration of feed treated with PZQ. The method was validated according to international guidelines. It showed good recoveries, selectivity and sensitivity (LOD and LOQ were 3.0 and 9.3 ng/g, respectively), with precision and matrix effect values ≤ 15%. This method could also be applied to determine PZQ residue in other fish species and thus to evaluate the appropriate withdrawal time in treated fish intended for human consumption.

LC-MS/MS analysis of two new designer drugs (FLY serie) in rat plasma and its application to a pharmacokinetic study.

The widespread diffusion of new psychoactive substances, requires a continuous update and development of new methods able to identify and quantify these new molecules in biological matrices. In this study an analytical method for the determination of two new benzodifuranyl derivatives, 1-(2,3,6,7-tetrahydrofuro[2,3-f][1]benzofuran-4-yl)propan-2-amine and 2-(2,3,6,7-tetrahydrofuro[2,3-f][1]benzofuran-4-yl)ethanamine, in rat plasma was developed. A solid phase extraction using C18 cartridges was carried out obtaining good recoveries with low matrix effect. Quantification was performed by a liquid chromatography tandem mass spectrometry (LC-MS/MS) method. Separation was carried out on a C18 reverse phase column with water/methanol containing 0.1% of formic acid as mobile phase. These conditions allowed to achieve adequate separation, resolution and signal-to-noise ratio for analytes and internal standard. Calibration curves were linear over the concentration range from 10 to 400 ng/ml with correlation coefficients that exceeded 0.995. Obtained precision, accuracy and recovery showed good reproducibility and selectivity. Finally, the validation method was successfully applied to an in vivo study in order to evaluate the pharmacokinetic profile of these new amphetamines.

Commercial human kits' applicability for the determination of biochemical parameters in sheep plasma.

Currently, several commercially available biochemical kits are validated for their use in human but not in animals. The purpose of this work is to demonstrate the applicability of human kits for alanine-aminotransferase, aspartato-aminotransferase, albumin, total protein, total cholesterol, and triglycerides in ovine plasma. Assays were validated according to international guidelines and stability was explored. Accuracy values were between 67 and 100%, and intra and interday precisions (%RSD) were <15% for all studied parameters. These results confirm the suitability of the studied human kits for their use in ovine plasma and they were used in plasma collected from pregnant ewes.

Investigation of the effects of Lycium barbarum polysaccharides against cadmium induced damage in testis.

This study describes the effects of Lycium barbarum polysaccharides (LBP) on testicular damage induced by cadmium (Cd). Adult male rats were i.p. injected with CdCl2 (4mg/Kg, once) with or without LBP pretreatment (300mg/Kg orally, once a day, for 30days). Testis weight, morphological/histological structure and oxidative stress parameters were evaluated. Several adverse effects were observed after CdCl2 injection, with a significant decrease in body/testis weight ratio (P<0.05), gross morphological changes with hyperemia of the parenchyma, increased volume and alteration in the structure of the seminiferous tubules. Furthermore, Cd intoxication caused a significant decrease of glutathione (GSH) and Trolox equivalent antioxidant capacity (TEAC) in testis (P<0.05) together with a significant increase (P<0.01) of 3-nitro-l-tyrosine (3NT) while malondialdehyde (MDA) did not change. LBP pretreatment caused slight signs of improvement in the morphology of the seminiferous tubules. Our results confirm that Cd induces testicular damage and suggest the oxidative stress involvement. LBP could ameliorate Cd testicular damage but further investigations are needed.

Possible antioxidant effect of Lycium barbarum polysaccharides on hepatic cadmium-induced oxidative stress in rats.

The aim of this study was to investigate the potential protective effect of Lycium barbarum polysaccharides (LBP) pretreatment against cadmium (Cd)-induced hepatotoxicity in rats. Wistar rats were divided into control group, LBP group (300 mg/kg orally, once a day, for 30 days), Cd group (CdCl2 4 mg/kg i.p. once), and LBP + Cd group (LBP 300 mg/kg orally, once a day, for 30 days + CdCl2 4 mg/kg i.p. 24 h after the last treatment). Cd liver injury was examined by morphological/histological changes, transaminases, total protein concentration, and oxidative stress evaluated by MDA, 3NT, GSH, SOD, and TEAC activities. Cd intoxication caused gross morphological changes with hyperemia of the parenchyma, increased volume, and disappearance of the anatomical limits of the lobes associated with an increase of ALT, GSH, and TEAC in plasma and a decrease of MDA, GSH, and TEAC in liver, SOD, and total proteins in plasma. LBP pretreatment caused a slight improvement in the histological architecture and in the 3NT amount together with a significant improvement of hematic parameters. On the basis of the obtained results, we can affirm that LBP pretreatment can ameliorate liver conditions, but further studies are needed to better evaluate the protective antioxidant effects of LBP against Cd-induced toxicity.

Microcystins Presence in Mussels (M. galloprovincialis) and Water of Two Productive Mediterranean's Lagoons (Sardinia, Italy).

Microcystins (MCs) are hepatotoxins harmful for animal and human health. The most toxic type between them is MC-LR whose presence has been investigated in different reservoirs all around the world. In this work microcystins were monitored in spring and summer in water and mussels (Mytilus galloprovincialis) of two Sardinia lagoons: Cabras and Calich lagoons. A Solid Phase Extraction method was developed to clean and concentrate samples before the Enzyme Linked Immunosorbent Assay (ELISA) and the following Mass Spectrometry detection. MCs presence was detected using the screening ELISA test in both lagoons. MCs peak was revealed in July for water and mussels belonging to Cabras lagoon (0.75 ± 0.07 ng/L in water and 0.12 ± 0.04 ng/g ww in mussels). In water of Calich lagoon there was a constant trend in the concentration of MCs during the considered months, while there was a MCs peak in July (0.6 ± 0.5 ng/g ww) in mussels. The following LC-MS/MS analysis did not reveal MC-LR presence in all analyzed samples. These results can be useful to enrich knowledge on public health and consumer's safeguard.

ELISA Detection of 30 New Amphetamine Designer Drugs in Whole Blood, Urine and Oral Fluid using Neogen® "Amphetamine" and "Methamphetamine/MDMA" Kits.

Amphetamine designer drugs are central nervous system stimulants that are widely disseminated in the illegal market. Generally, in forensic laboratories, immunoassay methods are the first line of screening for these types of drugs in a biological specimen (typically blood, urine or oral fluid). In this article, we describe the cross-reactivity profiles of 30 new amphetamine designer drugs, using the Neogen(®) [Amphetamine Specific and Methamphetamine/3,4-Methylenedioxymethamphetamine (MDMA) assays] drug tests. To assess the potential matrix influence on the response, each assay was tested on whole blood, urine and oral fluid. Concentrations of 10,000 ng/mL were not sufficient to produce a positive response for the majority of the analyzed amphetamines. This clearly demonstrates that, although these kits are extremely effective for the target drugs for which they are intended (amphetamine, methamphetamine and MDMA), they cannot be used to reliably identify the tested designer drugs in real cases, as these concentrations greatly exceed those expected to be found in forensic samples.

Determination of microcystin-LR in clams (Tapes decussatus) of two Sardinian coastal ponds (Italy).

The presence of microcystin-LR (MC-LR) was monitored in Tapes decussatus harvested in two Sardinian ponds (Cabras and Tortolì, Italy) in spring and summer. After solid phase extraction, samples were analyzed using a screening enzyme-linked immunosorbent assay (ELISA) followed by a liquid chromatographic coupled to tandem mass spectrometer (LC-MS/MS) analysis. Results obtained through the ELISA test showed the presence of microcystins with a maximum concentration in August for Cabras pond (0.55ng/g) and in September for Tortolì pond (0.85ng/g). The LC-MS/MS analysis did not confirm the presence of MC-LR suggesting that results obtained with the ELISA technique could be due to the presence of other microcystins. According to the tolerable daily intake suggested by the World Health Organization, these results hint that clams harvested in these ponds are safe for human health. These data can contribute to enrich the knowledge about the healthiness of Sardinian ponds and of their products.

Simultaneous Determination of 11 Illicit Phenethylamines in Hair by LC-MS-MS: In Vivo Application.

Existing phenethylamines are a class of synthetic compounds that differ from each other only in small changes to a largely conserved chemical structure. The recreational and illicit use of phenethylamines is a widespread problem. A simple procedure for the simultaneous quantitative determination in hair of 11 phenethylamines that are officially recognized as illicit by Italian legislation (p-methoxyamphetamine; p-methoxymethamphetamine; 3,4,5-trimethoxyamphetamine; 2,5-dimethoxyamphetamine; 2,5-dimethoxy-4-methylamphetamine; 2,5-dimethoxy-4-ethylamphetamine; 2,5-dimethoxy-4-bromoamphetamine; 2,5-dimethoxy-4-bromophenethylamine; 2,5-dimethoxy-4-iodophenethylamine; 2,5-dimethoxy-4-ethylthiophenethylamine and 2,5-dimethoxy-4-n-propylthiophenethylamine) has been developed and validated. Extraction from the matrix was performed after incubation in methanolic HCl and filtered reconstituted extracts were injected into a liquid chromatography/tandem mass spectrometry system (LC-MS-MS) without any further purification steps. This validated LC-MS-MS method has been used to determine the in vivo accumulation/retention of the above target analytes in hair after repeat oral administration to rats. This experiment further permitted investigation of the effect of pigmentation on the uptake of these phenethylamines by hair and the effect of hair pigmentation. The developed method could potentially be used for forensic and toxicological purposes, in the detection and quantitation of these illicit substances in human hair in workplace drug testing; drug-facilitated crime investigation; driver re-licensing; determining drug abuse history and postmortem toxicology.