Giulia Andreani

Comparative toxicity of CuO nanoparticles and CuSO4 in rainbow trout.

This study compared the toxicity and accumulation of two different Cu compounds, CuO nanoparticles (NPs) and soluble CuSO4, in erythrocytes and different tissues in rainbow trout (Oncorhynchus mykiss). The crystal structure of CuO NP analysed by XRD indicates that the NP are Tenorite, a monoclinic CuO. The in vitro toxicity results indicate that both Cu compounds increase the haemolysis rate in a dose-dependent way, but the effect was reduced treating cells with CuO NP. Moreover, both Cu compounds induce DNA damage and the entity of the damage, similarly to haemolysis, was more marked in cells treated with CuSO4. In vivo results, obtained after intraperitoneal injection, showed that Cu concentrations were significantly higher in gills (p<0.0001), kidney (p=0.007) and liver (p<0.05) of exposed fish with a significant increase in plasma Cu concentration 15h after CuSO4 treatment. Cu concentrations were significantly higher in fish exposed to CuSO4 than CuO in kidney (p<0.05) and gills (p<0.0001). Significant DNA damage with respect to controls was detected only when Cu was injected as CuSO4. The present data could serve to evaluate environmental Cu toxicity in fish depending on Cu speciation.

Cadmium accumulation and antioxidant responses in Sparus aurata exposed to waterborne cadmium.

Cadmium (Cd), a nonessential trace element, is rapidly accumulated by most living organisms and subsequently exerts its toxicity at different molecular levels. This study exposed gilthead sea bream (Sparus aurata) to waterborne 0.1xa0mg/l Cd for 11xa0days and investigated the Cd accumulation pattern, lipid oxidation, and response of antioxidant defences. At the end of the experiment, mean Cd concentrations in gills and liver, the organs most prone to metal accumulation, were 209.4 and 371.7xa0ng/g ww, respectively. Muscle did not show any Cd retention during the 11xa0days of exposure. In liver, the cytosolic fraction of the metal was chelated into the nontoxic form by metallothionein (MT), a specific Cd-inducible protein. Zn and Cu concentrations were not influenced by Cd exposure. Glutathione (GSH) concentrations and the antioxidant enzyme activities of GSH reductase and GSH peroxidase showed an overall decreasing trend. In addition, lipid and aqueous hydroperoxide levels did not show any significant variation. Oxidative stress indirectly generated by Cd seems to be compensated for by the different biochemical systems tailored to decrease cellular damage. In particular, the negative effects of Cd accumulation in tissues were probably counteracted by the induction of MT.

Trace elements in unconventional animals: A 40-year experience

The role of trace elements in animal health has attracted increasing interest in recent years. The essentiality and toxicity of these elements have been extensively investigated in humans, laboratory animal models and partially in domestic animals, whereas little is known about trace element metabolism in most living organisms. Forty years ago our research started on Cd metabolism in molluscs, thereafter expanding to Zn, Cu, and Fe metabolism in many unconventional animal species of veterinary interest. This review summarizes the main results obtained over this long period of time: some of the findings are original and have not been published to date. They are discussed in more detail and compared with data obtained in conventional animals, including man.

Validation of an electrophoretic method to detect albuminuria in cats

Objectives The aims of this study were to validate a semi-automated high-resolution electrophoretic technique to quantify urinary albumin in healthy and diseased cats, and to evaluate its diagnostic performance in cases of proteinuria and renal diseases. Methods Urine samples were collected from 88 cats (healthy; chronic kidney disease [CKD]; lower urinary tract disease [LUTD]; non-urinary tract diseases [OTHER]). Urine samples were routinely analysed and high-resolution electrophoresis (HRE) was performed. Within-assay and between-assay variability, linearity, accuracy, recovery and the lowest detectable and quantifiable bands were calculated. Receiver operating curve (ROC) analysis was also performed. Results All coefficients of variation were <10%, percentage recovery was between 97% and 109% with a high linearity (r = 0.99). HRE allowed the visualisation of a faint band of albumin and a diffused band between alpha and beta zones in healthy cats, while profiles from diseased cats were variable. Albumin (mg/dl) and urine albumin:creatinine ratio (UAC) were significantly (P <0.05) different between healthy and diseased cats. After ROC analysis, UAC values of 0.035 and 0.074 had a high sensitivity and high specificity, respectively, to classify proteinuria and identify borderline proteinuric cats. Moreover, a UAC of 0.017 had a high sensitivity in distinguishing between healthy and diseased cats. However, UAC was not able to distinguish between renal (CKD) and non-renal diseases (LUTD/OTHER), probably owing to the pathophysiology of CKD in cats, which is characterised by low-grade proteinuria and less glomerular involvement than in dogs. Conclusions and relevance HRE is an accurate and precise method that could be used to measure albuminuria in cats. UAC was useful to correctly classify proteinuria and to discriminate between healthy and diseased cats. HRE might also provide additional information on urine proteins with a profile of all proteins (albumin and globulins) to aid clinicians in the diagnosis of diseases characterised by proteinuria.

Total plasma magnesium in healthy and critically ill foals

Abnormalities in total Mg (tMg) concentration in plasma and/or serum are common in critically ill humans, and the association with increased mortality has been documented in several clinical studies in adults and newborns with hypoxic-ischemic encephalopathy. Abnormalities in tMg were studied in hospitalized dogs, cats, and adult horses. Newborn foals were scarcely studied with regard to Mg concentration. The aims of the present study were: (1) to compare two analytical methods for the determination of tMg in plasma: the automated colorimetric method and the atomic absorption spectrometry; (2) to measure plasma tMg in healthy foals during the first 72xa0hours after birth and in sick foals during the first 72xa0hours of hospitalization; (3) to compare total plasma Mg concentration among healthy foals, foals affected by perinatal asphyxia syndrome (PAS), prematurity and/or dismaturity, and sepsis; (4) to evaluate tMg plasma concentration in surviving and non-surviving foals. One hundred seventeen foals were included in the study: 20 healthy and 97 sick foals. The automated method used in clinical practice probably overestimates plasma tMg. Due to its higher sensitivity and specificity, the atomic absorption spectrometry should be considered the method of choice from an analytical point of view, but requires an instrumentation not easily available in any laboratory and specific technical skills and competencies. Plasma tMg in healthy foals were included in the range 0.52 to 1.01xa0mmol/L and did not show any time-dependent change during the first 72xa0hours of life. In sick foals, tMg evaluated at T0 was statistically higher than tMg measured at subsequent times. Foals affected by PAS had a tMg at T0 significantly higher (Pxa0<xa00.01) than healthy, septic, and premature and/or dysmature foals. The t test found significantly higher (Pxa0<xa00.01) plasma tMg measured at T0 in non-surviving than in surviving foals. Plasma tMg could be a useful parameter for the diagnosis of PAS and the formulation of the prognosis in critically ill foals.

Biochemical responses to cadmium exposure in Oncorhynchus mykiss erythrocytes

Cd is known for its carcinogenic effects, however its mechanism of toxicity and in particular its ability to promote oxidative stress is debated. In fact, although it is considered a redox-inactive metal, at high concentration Cd was shown to promote indirectly oxidative stress. In this study we investigated metal accumulation in ex vivo exposed trout (Oncorhynchus mykiss) erythrocytes and Cd dose-dependent effect in terms of RBC viability, cytosolic and mitochondrial ROS levels as well as its effects on mitochondrial membrane depolarization, hemoglobin stability and precipitation. In the concentration range used, Cd did not affect cell viability. However, metal accumulation was associated with an increase in all oxidative indexes evaluated, except mitochondrial superoxide anion production that, on the contrary, was significantly decreased, probably due to a lowered respiration rate associated with interference of Cd with complex I, II and III, as suggested by the observed Cd-dependent mitochondrial membrane depolarization. On the other hand, hemoglobin destabilisation seems to be the major trigger of oxidative stress in this cell type.

Anti-Inflammatory Activity of Boswellia serrata Extracts: An In Vitro Study on Porcine Aortic Endothelial Cells

This study is aimed at investigating the cytotoxicity, anti-inflammatory, and angiogenic activities of two Boswellia serrata extracts on primary culture of porcine aortic endothelial cells (pAECs). Chemical characterization of a dry extract (extract A) and a hydroenzymatic extract (extract G) of B. serrata was performed by HPLC using pure boswellic acids (BAs) as standard. In cultured pAECs, extract G improved cell viability, following LPS challenge, in a dose-dependent manner and did not show any toxic effect. On the other hand, extract A was toxic at higher doses and restored pAEC viability after LPS challenge only at lower doses. Pure BAs, used at the same concentrations as those determined in the phytoextracts, did not contrast LPS-induced cytotoxicity. Extract A showed proangiogenic properties at the lowest dose, and the same result was observed using pure AKBA at the corresponding concentration, whereas extract G did not show any effect on the migration capacity of endothelial cells. In conclusion, an anti-inflammatory activity of B. serrata extracts on endothelial cells was reported, though cytotoxicity or proliferative stimulation can occur instead of a protective effect, depending on the dose and the formulation.

EFFECT OF Boswellia serrata SUPPLEMENTATION IN ADDITION TO INSULIN ON GLYCEMIC CONTROL IN A DIABETIC DOG

Diabetes mellitus (DM) is a common disorder in middle-aged to older dogs. Treatment options, similar to those for humans, include insulin injection, dietary changes and exercise. Since some diabetic dogs may develop humoral immune responses to exogenous insulin resulting in treatment failure, the use of alternative medicine could represent an interesting new therapeutic strategy for DM in addition to the traditional insulin therapy. The main objective of this report was to evaluate a new therapeutic strategy for DM, based on the association of insulin injections with an orally administered extract of Boswellia serrata to improve glycemic control in a diabetic dog. A nine year old female neutered mixed breed dog diagnosed with DM was treated with increasing doses of a porcine insulin zinc suspension starting from 0.2 U/kg up to 0.6 U/kg q 12 hours. Twenty weeks after the start of insulin therapy the duration of effect and glucose nadir were ideal and clinical symptoms had disappeared but hyperglycemia, although less severe, persisted. Supplementation with a dry extract from the gum- resin of Boswellia serrata was initiated, at a dosage of 15 mg/kg q 12 hours, to improve insulin sensitivity or possible remaining endogenous insulin secretion. B. serrata supplementation led to good glycemic control. No side-effect or adverse reaction were observed during the study. The present case report provides the first evidence in veterinary medicine of a positive effect of dietary supplementation with boswellic acids associated with traditional insulin therapy on glycemic control in a diabetic dog. A major pitfall of the study is the lack of a control. More extensive clinical trials are required to provide definitive evidence of B. serrata efficacy. Key words: diabetes mellitus; dog; boswellic acids; glycemic control VPLIV DODAJANJA BOSVELIJE ( Boswellia serrata ) OB ZDRAVLJENJU Z INZULINOM NA UREJANJE RAVNI GLUKOZE V KRVI PRI PSU S SLADKORNO BOLEZNIJO Sladkorna bolezen oz. diabetes mellitus (DM) je pogosta motnja pri psih v srednjih letih in starejsih psih. Možnosti zdravljenja, ki so podobne kot pri ljudeh, vkljucujejo dodajanje inzulina, spremembe v prehrani in telesno aktivnost. Ker lahko nekateri psi s sladkorno boleznijo razvijejo imunski odziv na dodajanje inzulina, kar lahko vodi do neuspesnega zdravljenja, lahko uporaba alternativne medicine predstavlja zanimivo novo metodo zdravljenja DM poleg tradicionalnega zdravljenja z inzulinom. Glavni cilj raziskave je bil ovrednotiti novo metodo zdravljenja DM, ki temelji na hkratnem dodajanju inzulina in peroralnem dodajanju izvlecka bosvelije ( Boswellia serrata ) za izboljsanje urejanja ravni glukoze v krvi pri psu s sladkorno boleznijo. Devet let stara sterilizirana samica mesane pasme, ki je bila diagnosticirana s sladkorno boleznijo, je bila zdravljena z narascajocimi odmerki suspenzije prasicjega inzulina in cinka (od 0,2 E/kg do 0,6 U/kg), ki jih je dobivala vsakih 12 ur. Dvajset tednov po zacetku zdravljenja z inzulinom so bili ucinki in najnižja vrednost izmerjene glukoze v normalnih mejah vrednosti, klinicni simptomi so izginili, le hiperglikemija je bila se vedno prisotna, vendar pa je bila manj resna. Dodatek suhega izvlecka iz gumijeve smole bosvelije Boswellia serrata v odmerku 15 mg/kg vsakih 12 ur je povecal obcutljivost na inzulin oz. morebitne preostanke endogenega izlocanja inzulina. Dodatek B. serrata je pozitivno vplival na nadzorovanje sladkorne bolezni. Med zdravljenjem niso bili opaženi stranski ali nezaželeni ucinki. Porocilo o opisanem primeru vsebuje prve dokaze o pozitivnem ucinku prehranskega dopolnila z bosvelicnimi kislinami v veterinarski medicini, povezanimi z obicajnim zdravljenjem z inzulinom, pri nadzoru glukoze v krvi pri psu s sladkorno boleznijo. Najbolj neugoden del te studije je odsotnost kontrole. Za zagotovitev dokoncnega dokaza o ucinkovitosti B. serrata so potrebni obsežnejsi klinicni poskusi. Kljucne besede: diabetes mellitus ; pes; bosvelicna kislina; nadzor sladkorne bolezn

Retrospective evaluation of circulating thyroid hormones in critically ill dogs with systemic inflammatory response syndrome.

Critical illness can be associated with transient alterations in circulating thyroid hormone concentrations, indicating the presence of non-thyroidal illness (NTI). NTI is well described in humans, but there are few reports on its occurrence and prognostic significance in dogs. This retrospective study assessed the occurrence of NTI in a population of dogs with systemic inflammatory response syndrome (SIRS) and investigated its association with disease severity (APPLEfast scores). A total of 41 SIRS dogs were included and were divided by SIRS origin (non-septic SIRS, n = 10; septic SIRS, n = 41) and final outcome (survivors, n = 37; non-survivors, n = 4). Healthy, age-matched dogs (n = 15) were included as controls. Serum thyroid hormone levels including total T3, free T3, total T4, and reverse T3 were measured upon admission. Compared to controls, there were significant changes in serum thyroid hormone concentrations in SIRS dogs, suggesting the presence of NTI. Septic SIRS dogs had higher APPLEfast scores and lower serum thyroid hormones concentrations than those in non-septic SIRS and control dogs. In conclusion, NTI was frequent in dogs with SIRS and may be associated with the presence of sepsis or high illness severity.

Antioxidant enzymes in canine mammary tumors

Abstract Spontaneous mammary tumors are very common in bitches. The involvement of oxidative stress and the function of antioxidant enzymes in cancerogenesis have been studied in depth in human medicine, while data in veterinary medicine are still fragmentary. The main aim of this study was to evaluate the activity and the expression of superoxide dismutases (Cu-ZnSOD and MnSOD) and the activity of catalase (CAT) in canine mammary tumors in comparison with the adjacent healthy tissue. Six female dogs (mean age 10.4 years) were included in this study. After surgery, fresh tumor and healthy tissue samples were immediately frozen in dry ice and stored at −80°C for biochemical analyses, while the remaining parts were used for histopathological analysis. Enzyme activity was measured by spectrophotometric assays and protein expression by western blotting. In canine mammary tumors, Cu-ZnSOD activity and expression increased significantly compared with healthy control tissues (p=0.03). MnSOD showed a significantly lower activity in tumoral tissues at stage 2 (p<0.05), while a significant increase of expression was measured in tumors. CAT activity was significantly higher in healthy tissues respect to tumors (p=0.015). These variations of antioxidant enzymes activities and expression could be related to an increase of oxidative stress in breast cancer tissues and could be considered as biomarker candidates for neoplastic transformation.