Halogenated flame retardants in stranded sperm whales (Physeter macrocephalus) from the Mediterranean Sea.

In recent years, decline of marine mammals' populations and increased frequency of strandings have arised the interest on the role that pollution may have in these events. The present work aimed at quantifying levels of brominated flame retardants (BFRs) and dechloranes (DECs) in tissues of 3 adult females and one foetus of sperm whales stranded in the Southern Adriatic Sea coasts (Italy). Results proved the presence of different flame retardants (FRs) in tissues of sperm whales, including various polybrominated diphenyl ethers (PBDE) congeners (47, 99, 100, 154, entering the composition of PentaBDE mixture), hexabromocyclodecanes (HBCDs), Dec 602 and methoxylated polibrominated diphenyl ethers (MeO-BDEs). In blubber, a target tissue for contaminant accumulation, ΣPBDEs reached values of 160, 158 and 183 ng/g lw, α-HBCD of 5.75 ng/g lw, Dec 602 of 1632 ng/g lw and MeO-BDEs of 563 ng/g lw. The availability of foetal tissues allowed evaluating the potential maternal transfer on many of these compounds, and to discuss the potential adverse effects on foetal health. To the best of our knowledge, obtained data are the first reporting placental transfer of FRs in sperm whales. PBDE levels detected in foetus suggested a potentially long-term exposure to BFRs, which could cause severe damages to the developing organism, likely at the cerebral, endocrine and immunologic levels. Dec 602, which was detected at the highest concentrations among all FRs considered, could potentially cause dysfunctional effects on the immune system of adult females.

Role of sentinel node in differentiated thyroid cancer: a prospective study comparing patent blue injection technique, lymphoscintigraphy and the combined technique.

PURPOSE: The purpose of the present study was to evaluate the feasibility and reproducibility of the sentinel lymph node (SLNs) biopsy in differentiated thyroid cancer using patent blue injection, lymphoscintigraphy and the combined techniques. METHODS: Between January 2011 and January 2013, 82 consecutive patients were enrolled in our prospective multicentre study. Inclusion criteria were 18 years of age, preoperative diagnosis of differentiated thyroid carcinoma, no evidence of lymph node enlargement and multifocal neoplasm. To investigate the benefits of each procedure, all patients underwent total thyroidectomy plus central compartment lymphadenectomy, and in all cases, the SLN was identified via one of three techniques using the same protocol. RESULTS: Lymphoscintigraphy was used in five patients, patent blue injection was used in 40 patients, and a combined technique was used in 40 patients to identify sentinel lymph nodes (SLN). SLNs were identified in 61 cases. In the patent blue injection technique, the sensitivity, specificity and false negative rates were 88.9, 94.4 and 3.8%, respectively. In the lymphoscintigraphy technique, the percentages of sensitivity and specificity were 100%, and the percentage false negative was 0%. For the combined techniques, the corresponding values were, respectively, 69.2, 90, and 17.4%. Metastases were detected in nine cases of lateral-cervical nodes, ipsilateral tumour metastases were observed in eight cases, and contralateral tumour metastasis was observed in one case. CONCLUSION: Additional well-designed randomized studies are needed to validate and further optimize the SLN biopsy in patients with differentiated thyroid cancer.

Role of BRAF molecular analysis in the management of papillary thyroid carcinoma: analysis of cytological and histological samples.

BACKGROUND: Although fine needle aspiration (FNA) is the standard diagnostic test for the characterization of a suspicious thyroid nodule, in some cases cytological evaluation is inconclusive. The aim of this study was to determine the role of BRAF mutation in aiding diagnosis and to verify whether archival cytological samples could be suitable for molecular analysis. METHODS: Eighty-five patients with suspicious (Thy4) or follicular (Thy3) lesions on cytology were resubmitted to a second FNA for BRAF mutation analysis. Of these, 56 subsequently underwent surgery. The usefulness of archival samples for molecular analysis was also studied in a second cohort of 42 patients with a confirmed diagnosis of papillary thyroid carcinoma for whom both archived paraffin-embedded histological samples and cytological smears were available. A further 15 patients with paired fresh FNA and archived cytological and histological samples were recruited. RESULTS: BRAF mutation was found in the fresh FNA samples from 10 of 56 patients who had surgery with previous inconclusive cytology (4/45, 9%, Thy3 and 6/11, 55%, Thy4). The BRAF test showed a specificity and positive predictive value of 100% (26/26 and 10/10, respectively), sensitivity of 33% (10/30) and negative predictive value of 57% (26/46). There was absolute concordance between the BRAF results obtained with 42 histological and cytological archived samples. BRAF analysis on 15 archived cytological samples showed absolute concordance with histology, whereas there was one false negative on the matched fresh FNA. CONCLUSION: BRAF analysis is a highly specific test that can facilitate cytological diagnosis in some cases and can also be performed on archived cytological samples.

Effects of two water disinfectants (chloramine T and peracetic acid) on the epidermis and gills of Garra rufa used in human ichthyotherapy.

Doctor fish (Garra rufa) have recently been used for aesthetic purposes and as a medical treatment in patients with psoriasis (ichthyotherapy). For this particular kind of human therapy it is essential to guarantee adequate hygienic conditions for both people and fish. The aim of this study was to test two concentrations of water disinfectants, chloramine T and peracetic acid, on Garra rufa to ascertain possible exposure damage to the epidermis and gills. Fish were exposed to 2 mg/l and 10 mg/l of chloramine T and to 15 microl/l and 45 microl/l of peracetic acid in a 40-minute static bath up to six times a day for one week. The epidermis and gills were checked for histological changes and the number of epidermal mucous cells, club cells and taste buds were quantified; mucous cells were also characterized histochemically to detect alterations in mucin production. No mortality or severe histological changes were found in treated or control fish. Cell count showed a significant increase (p < 0.05) in mucous cells (mean 49.1 +/- 6.7 vs 37.0 +/- 13.1 of controls) in animals treated with peracetic acid independently of the dose. Club cell number showed a significant (p < 0.05) decrease in fish treated with 2 mg/l of chloramine T (mean 74.3 +/- 15.6) and with 45 microl/1 of peracetic acid (mean 78.17 +/- 10.5) compared to controls (mean 107.0 +/- 19.2). Histochemical evaluation of mucous cells did not reveal changes in mucin type in fish exposed to the two disinfectants. The results suggest a good tolerability of Garra rufa to the two disinfectants at the concentrations tested.

Potential adverse effects of inorganic pollutants on clinical parameters of loggerhead sea turtles (Caretta caretta): results from a nesting colony from Cape Verde, West Africa.

A large number of nesting loggerhead sea turtles (n = 201) were sampled to establish the blood levels of 11 elements (Cu, Mn, Pb, Zn, Cd, Ni, Cr, As, Al, Hg, and Se). Almost all of the samples showed detectable levels of these 11 elements, and Zn and Se exhibited the highest concentrations (median values as high as 6.05 and 2.28 µg/g, respectively). The median concentrations of the most toxic compounds, As, Cd, Pb, and Hg, were relatively low (0.38, 0.24, 0.06, and 0.03 µg/g, respectively). We also determined the haematological and biochemical parameters in a subsample of 50 turtles to evaluate the potential effects of these contaminants on clinical parameters and found several associations. Our study reinforces the usefulness of blood for the monitoring of the levels of contaminating elements and their adverse effects on blood parameters in sea turtles.

Cerebral toxoplasmosis in striped dolphins (Stenella coeruleoalba) stranded along the Ligurian Sea coast of Italy.

This article reports the results of necropsy, parasitologic, microbiologic, histopathologic, immunohistochemical, indirect immunofluorescence, biomolecular, and serologic investigations on 8 striped dolphins (Stenella coeruleoalba) found stranded from August to December 2007 on the Ligurian Sea coast of Italy. Severe, nonsuppurative meningoencephalitis was found in 4 animals, as characterized by prominent perivascular mononuclear cell cuffing and macrophage accumulations in neuropil. These lesions were associated with mild lymphocytic-plasmacytic infiltration of choroid plexuses in 1 dolphin. Toxoplasma gondii cysts and zoites, confirmed by immunohistochemical labeling, were scattered throughout the brain parenchyma of 2 of the 4 dolphins. No viral inclusions were seen in the brain of any animal. Other findings included severe bronchointerstitial pneumonia and pulmonary atelectasis, consolidation, and emphysema. Parasites were identified in a variety of organs, including lung (Halocerchus lagenorhynchi). Microbiologic and serologic examinations for Brucella spp were negative on all 8 dolphins. The 4 animals with meningoencephalitis had serum antibodies against T gondii (titers ranging from 1:80 to 1:320) but not against morbillivirus. In contrast, the other 4 dolphins were seropositive for morbillivirus (with titers ranging from 1:10 to 1:40) but seronegative for T gondii. No morbillivirus antigen or nucleic acid was detected in the tissues of any dolphin. It is concluded that the severe lung and brain lesions were the cause of death and that T gondii was the likely etiologic agent of the cerebral lesions. Morbillivirus infection was not considered to have contributed to death of these animals.

Thyroid hormones as a potential early biomarker of exposure to 4-nonylphenol in adult male shubunkins (Carassius auratus).

Nonylphenol, an estrogenic-like compound, can induce vitellogenin synthesis in males and immature Teleostean species, but little is known about its effects on thyroid hormones balance. The present study evaluated the potential effects of a single acute exposure to nonylphenol (i.p. injected) on the thyroid and reproductive axis of 250 shubunkins (Carassius auratus). Plasma levels of thyroid hormones were quantified immunoenzymatically by ELISA assay. Nonylphenol induced a significant decrease of thyroxin levels, whereas no effect on triiodothyronine concentrations was detected. No histopathological changes were detected in thyroid or testes. The toxicological data confirmed that nonylphenol exerts an estrogenic effect on male fish. In addition, nonylphenol was suspected to inhibit the thyroid hormones balance, suggesting the thyroid should be included among the other endocrine glands susceptible to endocrine disruption.

Persistent organochlorine pollutants in liver of birds of different trophic levels from coastal areas of Campania, Italy.

Liver samples of 12 species of birds of different trophic levels, collected during the period 1998-2000 from coastal areas of the Campania region, Southern Italy, were analyzed for organochlorine pesticides (OCs), such as dichlorodiphenyltrichloroethane (DDT) and its metabolites, hexachlorobenzene (HCB), gamma-hexachlorocycloexane (gamma-HCH), aldrin, dieldrin, endrin, and the seven PCB "target" congeners, IUPAC Nos. 28, 52, 101, 118, 138, 153 and 180. p,p'-DDE was present in all the samples analyzed, at concentrations ranging from 4 to 4504 ng/g wet wt, which were much higher than those found for HCB, dieldrin, and p,p'-DDD. The concentrations of the others OCs were below the detection limit in all the samples. PCBs were found in all the bird species at levels ranging between 6 and 8431 ng/g wet wt. The hepta-, hexa-, and penta-chlorinated congeners 180, 153, 138, and 118 were predominant since, in almost all the species, they contributed to more than 98% of the total seven determined PCBs. No significant differences in mean concentrations of organochlorine pesticides are detected between single species or between species grouped according to their feeding habits (p > 0.05). However, p,p'-DDE levels were higher in carnivorous species than in omnivorous and insectivorous ones (carnivorous > omnivorous > insectivorous). Concentrations of total PCBs were significantly higher in omnivorous birds than in carnivorous (p < 0.01) and insectivorous ones (p < 0.001), whereas carnivorous birds exhibited significantly higher total PCB levels than insectivorous ones (p < 0.01). Marked differences in total PCB concentrations were found also between single species (from p < 0.05 to p < 0.001). Levels of OCs and PCBs were generally below the thresholds known to affect reproduction, however, mean hepatic concentrations of total PCBs in the yellow-legged herring gull (Larus cachinnans), black-headed gull (Larus ridibundus), and kestrel (Falcus tinnunculus) were far higher than those estimated to elicit immunosuppressive effects and possibly increase susceptibility to parasitoses.

Cadmium, chromium and lead contamination of Athene noctua, the little owl, of Bologna and Parma, Italy.

A study was conducted to determine cadmium, chromium and lead concentrations in liver and brain of 52 little owls (Athene noctua) from two provinces of Emilia Romagna region, with the aim of furnishing indirect information concerning contamination of their habitat, also considering possible environmental dispersion of the metals. Metal analysis was performed by atomic absorption spectrophotometry with graphite furnace. Variance analysis with sampling area, gender and age shows that no statistical difference was found for gender, while a significant difference (P<0.05) was found for cadmium and lead, but not for chromium, when sampling areas and age were of concern. For all metals highest mean concentrations were found in liver (170 ppb for cadmium, 297 ppb for chromium and 312 ppb for lead). These levels can be considered as indicative of chronic exposure to low and "background" amounts of pollutants and they are of no toxicological concern, as they are always well below the toxic thresholds defined for each metal. The present study can be considered as a starting point for further analyses, aimed to the definition of any possible subtle effect (e.g. effects on enzymes activity) and of any possible correlation between levels of pollutants and appearance of possible adverse effects. It also furnished useful data for diagnostic cases and potentially for monitoring local contamination.