Heavy Metal Assessment in Feathers of Eurasian Magpies (Pica pica): A Possible Strategy for Monitoring Environmental Contamination?

In the present study, the Eurasian magpie (Pica pica), was evaluated as a possible bioindicator of environmental pollution by heavy metals (HMs). Levels of Ni, Pb, Cd, and Hg in feathers of 64 magpies (31 males and 33 females) were measured by ICP-MS technique. Plasmatic biomarkers of oxidative stress (OS) were also assessed. The birds were captured in the province of Parma (Italy), in different capture sites within 1 km from urban area (UZ), and farther than 5 km from urban area (RZ). Median HM levels were 0.68 mg/kg (0.18-2.27), 2.80 mg/kg (0.41-17.7),

Lead Levels in Wild Boar Meat Sauce (Ragù) Sold on the Italian Market.

Game meat is endowed with excellent nutritional value, but it may also be a possible source of harmful substances, such as mycotoxins and heavy metals. In particular, several studies showed that lead fragments from hunting ammunition are able to represent a residual contaminant in the meat of wild boars or deer, representing a possible source of lead absorption. Even though wild boar meat consumption in Italy is rather limited, this meat could also be present in very popular Italian recipes, such as the typical meat sauce called ragù. We evaluated the lead levels in 48 samples (three different batches for each of the 16 brands) of ready-to-eat wild boar meat ragù sold on the Italian market in food stores and online distribution with the inductively coupled plasma-mass spectrometry (ICP-MS) technique. A high variability was found in the lead levels detected in the samples, with a median lead level of 0.10 mg/kg (0.01-18.3 mg/kg) and some of the samples showing very high lead concentrations. Since no intake level of lead is considered completely safe, and maximum levels for game meat have so far not been established, a greater attention on the risks to consumers' health related to the presence of this heavy metal in game meat is recommended.

Evaluation of Oxidative Stress in Blood of Domestic Chickens and Eurasian Magpies (Pica pica).

A physiological equilibrium exists between pro- and antioxidant factors. When the oxidant factors exceed the capacity of their removal or inactivation, oxidative stress (OS) occurs. The OS levels were assayed in plasma obtained from 2 bird species. Blood samples were collected from 20 healthy domestic chicken hens, 10 living in an intensive farming environment and 10 free-range, and from 18 healthy Eurasian magpies (Pica pica; 7 females and 11 males, with an estimated age of >1 year of age). For OS biomarker assessment, the determinable reactive oxygen metabolites (d-ROMs) were measured, and the plasmatic antioxidant test (PAT) was performed; the OS index (OSI) was then calculated (d-ROMs/PAT × 1000) as a parameter of overall oxidative stress. Moreover, lipid peroxidation was assessed by measuring plasmatic malondialdehyde (MDA) levels. A hematological evaluation was also performed on each bird with a hemocytometer, on which a blood sample was placed to obtain both a total and differential white blood cell (WBC) count. In hens, OSI and MDA levels were significantly higher (P = .04, and P = .004) in subjects from intensive farming (14.7 ± 7.1 and 27.2 ± 10.4 nmol/mL) than in those bred in rural conditions (5.6 ± 10.3 and 8.2 ± 13.3 nmol/mL). In magpies, a positive correlation between the total WBC count and OS was found, and both d-ROMs and OSI were significantly higher (P = .03) in subjects with a total WBC count greater than the median value (20.4 × 103 cells/µL) with respect to those with a total WBC count less than the median value. The results generated from this study indicate that higher OS levels occurred in hens bred in an intensive indoor farm environment compared with outdoor free-range conditions. Possibly the higher OS levels could be related to the higher stocking density and dust levels found in the indoor facility. Additionally, the correlation between OS biomarker levels in magpies and total WBC count suggests that OS level is influenced by immune response, in agreement with previous studies. Collectively, present data seem to be promising for the application of OS measurement in avian medicine for health and animal welfare monitoring.

Differential ABC transporter gene expression in adult Dirofilaria immitis males and females following in vitro treatment with ivermectin, doxycycline or a combination of both.

BACKGROUND: Combination doxycycline/macrocyclic lactone (ML) protocols have been shown to provide a more rapid adulticidal and microfilaricidal effect than either MLs or doxycycline alone, although female worms were reported to have a higher tolerance to treatments compared to male worms. The present study aimed to evaluate how ABC transporters may be involved in the synergic effect of the combination treatment. Adult worms of D. immitis were treated in vitro for 24 hours with doxycycline (DOXY), ivermectin (IVM) and a combination of both, and changes in the modulation of ABC transporter genes were measured. Levels of doxycycline inside different treatment media, post-treatment, were determined through HPLC analysis. RESULTS: Quantitative RT-PCR analysis showed the presence of changes in the modulation of ABC transporter genes evaluated in this study. In particular, in female worms, the combination treatment induced a substantial increase in gene expressions, especially of Dim-pgp-10 and Dim-haf-4; whereas in male worms, the greatest increase in gene expression was observed for Dim-pgp-10 and Dim-pgp-11 when treated with DMSO + IVM and DMSO + DOXY/IVM. HPLC analysis of the DOXY concentrations in the media after in vitro treatments of male worms showed a slight difference between the DMSO + DOXY samples and the combination (DMSO + DOXY + IVM), while no difference was observed among females. CONCLUSIONS: Further studies are required to explain whether the modulation of cellular efflux plays a role, even partially, in the adulticide effect of doxycycline/macrocyclic lactone combinations in heartworm-infected dogs. To the authors' knowledge, this is the first study to evaluate P-gp expression in adult D. immitis.

Role of muscarinic receptors in the contraction of jejunal smooth muscle in the horse: An in vitro study.

Nonselective antimuscarinic drugs are clinically useful in several pathologic conditions of horses, but, blocking all muscarinic receptor (MR) subtypes, may cause several side effects. The availability of selective antimuscarinic drugs could improve therapeutic efficacy and safety. We aimed to enlighten the role of different MR subtypes by evaluating the effects of nonselective, and selective M1, M2 and M3 MR antagonists on the contractions of horse jejunum. Segments of circular muscle of equine jejunum, were put into organ baths, connected to isotonic transducers, and the effects on ACh concentration-response curves, and on electrical field stimulation (EFS)-evoked contractions of intestinal preparations, induced by nonselective or selective MR antagonists, compared to pre-drug level, were studied. Atropine (nonselective MR antagonist), pirenzepine (selective M1 antagonist), and p-FHHSiD (selective M3 antagonist) competitively antagonized ACh (pA2=9.78±0.21; 7.14±0.25 and 7.56±0.17, respectively). Methoctramine (selective M2 antagonist) antagonized ACh in a concentration-unrelated fashion; however, it competitively antagonized carbachol, a nonselective muscarinic agonist (pA2=6.42±0.23). Atropine dose-dependently reduced EFS-evoked contractions, reaching a maximal effect of -45.64±6.54%; the simultaneous block of neurokinin receptors, almost completely abolished the atropine-insensitive contractions. p-FHHSiD dose-dependently reduced EFS-induced contractions, while pirenzepine caused a minor decrease. Methoctramine, ineffective up to 10-7M, enhanced the contractions at 10-6M; the block of neurokinin receptors abolished the increase of contraction. Cholinergic contractions of horse jejunum are mainly mediated by M3 receptors; M2 selective antagonists seem to scarcely affect cholinergic, and to enhance neurokininergic contractions of equine jejunum, thus their use entails a lower risk of causing intestinal hypomotility, compared to nonselective drugs.