72 results on '"Cannizzo FT"'
Search Results
2. Flow cytometry of non-hematopoietic cells in canine effusions.
- Author
-
Sini F, Melega M, Cannizzo FT, Miniscalco B, Valenti P, and Riondato F
- Abstract
The identification of non-hematopoietic cells in effusions is a diagnostic challenge in cytology. Biopsies from mesothelium or primary lesions are infrequently performed in clinical settings and immunochemistry on smears or immunohistochemistry on cell blocks are the most common ancillary test to refine the cytological diagnosis. Cavitary effusions are an ideal matrix for flow cytometry and the availability of a cytometric panel to describe non-hematopoietic cells would represent a useful tool. Here we present the results of the flow cytometric and immunohistochemical determination of cytokeratin (CK), vimentin (VIM) and desmin (DES) in 36 canine effusions. The concordance between the two methods was perfect for CK (100%), substantial for VIM (77.8%), and almost perfect for DES (97.2%). The panel was interpreted to define the epithelial (CK+VIM-DES-), mesothelial (CK+VIM+DES+), or mesenchymal (CK-VIM+DES-) origin of the cells. Unexpected profiles were considered doubtful and observed patterns were individually discussed. The concordance of the panel interpretation between two methods was 75%. The evaluation of discordant and doubtful cases suggests a lower sensitivity of flow cytometry in detecting VIM expression and revealed a high frequency of VIM+ epithelial cells, variable expression of VIM in mesothelial cells, and an important role of DES in excluding an epithelial origin when positive. Multicentric studies based on histopathological diagnoses are necessary to confirm these findings and evaluate the diagnostic utility of the panel to refine cytological diagnosis. Our results show that flow cytometry can be a timesaving alternative to IHC on cell blocks in clinical settings to detect CK, VIM and DES expression. The interpretation of the panel is similar in most cases; however, occasional discordant results, particularly for VIM, may occur., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Sini, Melega, Cannizzo, Miniscalco, Valenti and Riondato.)
- Published
- 2024
- Full Text
- View/download PDF
3. Extracellular vesicle miRNome during subclinical mastitis in dairy cows.
- Author
-
Cuccato M, Divari S, Giannuzzi D, Grange C, Moretti R, Rinaldi A, Leroux C, Sacchi P, and Cannizzo FT
- Subjects
- Animals, Cattle, Female, Biomarkers metabolism, Mastitis, Bovine diagnosis, Mastitis, Bovine microbiology, Mastitis, Bovine genetics, Extracellular Vesicles metabolism, MicroRNAs metabolism, MicroRNAs genetics, Milk
- Abstract
Bovine mastitis is one of the main inflammatory diseases that can affect the udder during lactation. Somatic cell counts and sometimes microbiological tests are routinely adopted during monitoring diagnostics in dairy herds. However, subclinical mastitis is challenging to identify, reducing the possibility of early treatments. The main aim of this study was to investigate the miRNome profile of extracellular vesicles isolated from milk as potential biomarkers of subclinical mastitis. Milk samples were collected from a total of 60 dairy cows during routine monitoring tests. Small RNA sequencing technology was applied to extracellular vesicles of milk samples collected from cows classified according to the somatic cell count to identify differences in the miRNome between mastitic and healthy cows. A total of 1997 miRNAs were differentially expressed between both groups. Among them, 68 miRNAs whose FDRs were < 0.05 were mostly downregulated, with only one upregulated miRNA (i.e., miR-361). Functional analysis revealed that miR-455-3p, miR-503-3p, miR-1301-3p and miR-361-5p are involved in the regulation of several biological processes related to mastitis, including immune system-related processes. This study suggests the involvement of extracellular vesicle-derived miRNAs in the regulation of mastitis. Moreover, these findings provide evidence that miRNAs from milk extracellular vesicles can be used to identify biomarkers of mastitis. However, further studies must be conducted to validate these miRNAs, especially for subclinical diagnosis., (© 2024. The Author(s).)
- Published
- 2024
- Full Text
- View/download PDF
4. Actinobacillus pleuropneumoniae Serotypes by Multiplex PCR Identification and Evaluation of Lung Lesions in Pigs from Piedmont (Italy) Farms.
- Author
-
Cuccato M, Divari S, Ciaramita S, Sereno A, Campelli D, Biolatti PG, Biolatti B, Meliota F, Bollo E, and Cannizzo FT
- Abstract
Porcine pleuropneumonia (PPP) is one of the main causes leading to massive losses in the pig industry, with high economic impacts. Among different etiological agents, Actinobacillus pleuropneumoniae (APP) is responsible for severe fibrinous-necrotizing pleuropneumonia. A total of 19 different APP serotypes are currently recognized. This study aimed to identify APP serotypes isolated from pneumonic lesions in naturally infected and dead pigs in the Piedmont Region and to describe lesions. A total of 107 dead pigs with a suspected PPP diagnosis were included in this study. Lungs were evaluated using gross-pathology scoring systems, histopathology, and APP isolation and serotypes identification by multiplex PCR were conducted. Gross lung lesions were mainly represented by fibrinous pneumonia and pleuropneumonia. APP was isolated in 20/107 (18.7%) samples. PCR indicated APP DNA presence in 53/107 (49.5%) of lung samples. The most observed serotypes were serotype 2 in 24/53 (45.3%) and serotype 6 in 13/53 (24.5%) samples. Moreover, multiplex PCR results suggested a coinfection of different serotypes in five samples. This study emphasizes the importance of an integrated approach, utilizing various techniques, such as gross- and histopathology, and bacteriological culture and PCR, to enhance the diagnosis of APP infections.
- Published
- 2024
- Full Text
- View/download PDF
5. Milk fat miRNome changes in response to LPS challenge in Holstein cows.
- Author
-
Leroux C, Cuccato M, Pawłowski K, Cannizzo FT, Sacchi P, Pires JAA, and Faulconnier Y
- Subjects
- Female, Cattle, Animals, Milk, Lipopolysaccharides pharmacology, Lactation, Diet veterinary, Escherichia coli genetics, Mastitis veterinary, MicroRNAs genetics, MicroRNAs metabolism, Cattle Diseases metabolism
- Abstract
Mastitis is an inflammatory disease in dairy cows, causing economic losses and reducing animal welfare. In order to contribute for the discovery of early and noninvasive indicators, our objective was to determine the effects of a lipopolysaccharide (LPS) challenge on the microRNA profile (miRNome) of milk fat, using microarray analyses in cows. Cows were fed a lactation diet at ad libitum intake (n = 6). At 27 ± 3 days in milk, cows were injected with 50 µg of LPS Escherichia coli in one healthy rear mammary quarter. Milk samples were collected just before LPS challenge (LPS-) and 6.5 h after LPS challenge (LPS +) from the same cows. Microarray analysis was performed using customized 8 × 60 K ruminant miRNA microarrays to compare LPS- to LPS + miRNome. In silico functional analyses were performed using OmicsNet and Mienturnet software. MiRNome comparison between LPS- and LPS + identified 37 differentially abundant miRNAs (q-value ≤ 0.05). The predicted target genes of the 37 differentially abundant miRNAs are mostly involved in cell life including apoptosis, cell cycle, proliferation and differentiation and in gene expression processes. MiRNome analyses suggest that miRNAs profile is related to the inflammation response of the mammary gland. In conclusion, we demonstrated that milk fat might be an easy and rapid source of miRNAs that are potential indicators of early mastitis in cows., (© 2023. The Author(s).)
- Published
- 2023
- Full Text
- View/download PDF
6. Evaluation of the biofilm-forming ability and molecular characterization of dairy Bacillus spp. isolates.
- Author
-
Catania AM, Di Ciccio P, Ferrocino I, Civera T, Cannizzo FT, and Dalmasso A
- Subjects
- Polystyrenes, Stainless Steel, Biofilms, Enterotoxins, Bacillus genetics
- Abstract
Food processing lines represents a suitable environment for bacterial biofilm formation. One of the most common biofilm-forming genera in dairy processing plants is Bacillus , which includes species that may have a negative impact on safety and/or quality of dairy products. In the current study, we evaluated the biofilm forming ability and molecular characteristics of dairy Bacillus spp. isolates ( B. cereus and B. subtilis ). Reference strains ( B. cereus ATCC 14579 and B. subtilis NCTC 3610) were also included in the experiment. All isolates were screened by micro-titer plate (96 wells) to assess their ability to form biofilm. Then, they were tested on two common food contact surfaces (polystyrene and stainless steel) by using 6-well plates and AISI 316 stainless steel coupons. Biofilm formation, expressed as biofilm production index (BPI), was higher on polystyrene than stainless steel (except for B. cereus ATCC 14579). These observations were further confirmed by scanning electron microscopy, which allowed the microscopy observation of biofilm structure. Moreover, a possible correlation among total viable cell counts (CFU) and BPI was examined, as well as a connection among biofilm formation and bacterial cell hydrophobicity. Finally, whole genome sequencing was performed highlighting a genetic similarity among the strains belonging to the same species. The presence of selected genes involved in biofilm formation was also examined showing that strains with a greater presence of these genes were able to produce more biofilm in the tested materials. Additionally, for B. cereus strains enterotoxin genes were detected., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Catania, Di Ciccio, Ferrocino, Civera, Cannizzo and Dalmasso.)
- Published
- 2023
- Full Text
- View/download PDF
7. MALDI-TOF mass spectrometry profiling of bovine skim milk for subclinical mastitis detection.
- Author
-
Cuccato M, Divari S, Sacchi P, Girolami F, and Cannizzo FT
- Abstract
Introduction: Mastitis is one of most impacting health issues in bovine dairy farming that reduces milk yield and quality, leading to important economic losses. Subclinical forms of the disease are routinely monitored through the measurement of somatic cell count (SCC) and microbiological tests. However, their identification can be tricky, reducing the possibilities of early treatments. In this study, a MALDI-TOF mass spectrometry approach was applied to milk samples collected from cows classified according to the SCC, to identify differences in polypeptide/protein profiles., Materials and Methods: Twenty-nine raw milk samples with SCC >200,000 cell/ml (group H) and 91 samples with SCC lower than 200,000 (group L) were randomly collected from 12 dairy farms. Spectral profiles from skim milk were acquired in the positive linear mode within the 4,000-20,000 m/z mass acquisition range., Results and Discussion: Based on signal intensity, a total of 24 peaks emerged as significant different between the two groups. The most discriminant signals (4,218.2 and 4,342.98 m/z) presented a ROC curve with AUC values higher than 0.8. Classification algorithms (i.e., quick classifier, genetic algorithm, and supervised neural network) were applied for generating models able to classify new spectra (i.e., samples) into the two classes. Our results support the MALDI-TOF mass spectrometry profiling as a tool to detect mastitic milk samples and to potentially discover biomarkers of the disease. Thanks to its rapidity and low-cost, such method could be associated with the SCC measurement for the early diagnosis of subclinical mastitis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Cuccato, Divari, Sacchi, Girolami and Cannizzo.)
- Published
- 2022
- Full Text
- View/download PDF
8. Amoxicillin and thiamphenicol treatments may influence the co-selection of resistance genes in the chicken gut microbiota.
- Author
-
Laconi A, Tolosi R, Mughini-Gras L, Cuccato M, Cannizzo FT, and Piccirillo A
- Subjects
- Animals, Amoxicillin pharmacology, Chickens genetics, Drug Resistance, Microbial genetics, RNA, Ribosomal, 16S genetics, Anti-Bacterial Agents pharmacology, Anti-Bacterial Agents analysis, beta-Lactams, Thiamphenicol pharmacology, Gastrointestinal Microbiome genetics
- Abstract
The aim of this study was to assess the dynamics of microbial communities and antimicrobial resistance genes (ARGs) in the chicken gut following amoxicillin and thiamphenicol treatments and potential co-selection of ARGs. To this purpose, the microbial community composition, using 16S rRNA NGS, and the abundance of ARGs conferring resistance to β-lactams and phenicols, using qPCRs, were determined. Results revealed that the administered antimicrobials did not significantly reduce the gut microbiota diversity, but changed its composition, with taxa (e.g. Gallibacterium and Megamonas) being enriched after treatment and replacing other bacteria (e.g. Streptococcus and Bifidobacterium). Positive correlations were found between ARGs (e.g. cmlA, bla
CMY-2 , and blaSHV ) and the relative abundance of specific taxa (e.g. Lactobacillus and Subdoligranulum). The selective pressure exerted by both amoxicillin and thiamphenicol resulted in an increased abundance of ARGs conferring resistance to β-lactams (e.g. blaTEM-1 , blaSHV, and blaCTX-M1-like ) and phenicols (e.g. floR and cmlA). These findings, together with the co-occurrence of genes conferring resistance to the two antimicrobial classes (e.g. blaTEM-1 and cmlA), suggest a possible interaction among antimicrobials on resistance emergence, possibly due to the presence of mobile genetic elements (MGEs) carrying multiple resistance determinants., (© 2022. The Author(s).)- Published
- 2022
- Full Text
- View/download PDF
9. Removal of antibiotics from milk via ozonation in a vortex reactor.
- Author
-
Liu P, Wu Z, Cannizzo FT, Mantegna S, and Cravotto G
- Subjects
- Animals, Anti-Bacterial Agents, Fertilizers, Hydrogen Peroxide chemistry, Milk chemistry, Oxidation-Reduction, Sodium, Oxytetracycline, Ozone chemistry, Sulfamonomethoxine, Water Pollutants, Chemical chemistry, Water Purification
- Abstract
Antibiotics (ABX) residues occur frequently in milk, causing considerable wastage of medicated milk and serious economic losses, and making the issue a burden for the dairy industry. Improper disposal of medicated milk harms dairy production, animal welfare, and the environment. This work studies the use of ozonation in a vortex reactor for removing ceftiofur hydrochloride (CEF), sulfamonomethoxine sodium (SMM), marbofloxacin (MAR) and oxytetracycline (OTC) from milk. In terms of residual concentration, O
3 efficiency and the degradation kinetics of the various O3 -involving processes in the vortex reactor, ABX removal via ozonation is better using stronger vortexing, which induces hydrodynamic cavitation. CEF undergoes the fastest degradation, followed by SMM, MAR, and OTC. High ABX hydrophobicity favors ABX degradation via ozonation, O3 /H2 O2 , and O3 /Na2 S2 O8 . ABX oxidation by• OH at the O3 gas-bubble/milk interface is the principle degradation pathway, except for MAR. ABX degradation follows pseudo-first-order kinetics and is affected by initial ABX concentration, O3 concentration/flow rate, reaction temperature, and milk components to varying degrees. Under optimal ozonation conditions, ABX residues meet the maximum limits as set by the European Commission and no antimicrobial activity was observed. The decontaminated milk was therefore suggested to be reused as calf food, animal feed, organic fertilizer, etc., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)- Published
- 2022
- Full Text
- View/download PDF
10. A Practical Application of Genomic Predictions for Mastitis Resistance in Italian Holstein Heifers.
- Author
-
Moretti R, Chessa S, Sartore S, Soglia D, Giaccone D, Cannizzo FT, and Sacchi P
- Abstract
Heifers are a fundamental resource on farms, and their importance is reflected in both farm management and economy. Therefore, the selection of heifers to be reared on a farm should be carefully performed to select only the best animals. Genomic selection is available nowadays to evaluate animals in a fast and economic way. However, it is mainly used on the sire line and on performance traits. Ten farms were selected based on their 5-year records of average somatic cell count and evenly classified into high (>300,000 cells/mL) and low somatic cell count (<150,000 cells/mL). Genomic indexes (regarding both wellness and productive traits) were evaluated in 157 Italian Holstein heifers reared in the selected ten farms (90 from high-cells farms and 67 from low-cells ones). Linear mixed models were fitted to analyze the effects of the abovementioned genomic indexes on related phenotypes. Results have shown that farms classified into low somatic cell count had an overall better animal genomic pool compared to high somatic cell count ones. Additionally, the results shown in this study highlighted a difference in wellness genomic indexes in animals from farms with either a high or a low average somatic cell count. Applying genomic tools directly to heifer selection could improve economic aspects related to herd turnover.
- Published
- 2022
- Full Text
- View/download PDF
11. Development of a droplet digital PCR assay to detect illicit glucocorticoid administration in bovine.
- Author
-
Divari S, Cuccato M, Fanelli A, and Cannizzo FT
- Subjects
- Animals, Biological Assay, Cattle, Male, Polymerase Chain Reaction, Prednisolone pharmacology, Dexamethasone pharmacology, Glucocorticoids analysis
- Abstract
Glucocorticoids are often used illegally in food-producing animals for the growth promotion of livestock animals. In accordance to official chemical methods for glucocorticoid detection, an animal is declared as non-compliant when a residue is identified in the sample. Neverthless, growth promoting molecules can often escape identification due to their rapid elimination or due to the use of non-detectable new generation drugs. Therefore, an indirect screening method able to detect the biological effect of long-term administration of low doses of dexamethasone and prednisolone on livestock has been developed to support official methods. As already described, FKBP5 (FKBP prolyl isomerase 5) expression in bovine thymus is regulated by glucocorticoids, and this specific regulation can be exploited in an indirect screening assay. In the present study, male veal calves and young bulls were considered in three different trials in which estradiol, dexamethasone, and prednisolone were administered alone or in combination with Revalor-200 subcutaneous pellets. Thoracic thymus was sampled from all animals and molecular analysis was performed. A duplex droplet digital PCR assay with EvaGreen® was employed to detect the target gene expression using absolute quantification. The developed droplet digital PCR assay was precise, showing intra- and inter-assay mean coefficient of variation values of about 6.16% and 3.17%, respectively. It was also highly specific (100%) with Youden's index of 76.92% and 53.57% applied to veal calves and young bulls, respectively. The lowest detection limit in which the target gene expression level was kept constant, was 0.05 ng/μl of cDNA with 1 copies/μL and 0.5 copies/μL for target and reference gene, respectively. This study establishes the basis for using a digital PCR-based assay as an efficient test to identify animals illegally treated with glucocorticoids., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2022
- Full Text
- View/download PDF
12. Assessment of Antimicrobial Effects on Broiler Gut Barrier Through Histopathology and Immunohistochemistry of Tight-Junction Proteins.
- Author
-
Cuccato M, Scaglione FE, Centelleghe C, Divari S, Biolatti B, Pregel P, and Cannizzo FT
- Abstract
In recent years, antimicrobial (AM) use in poultry farming has been attracting attention worldwide mainly due to AM resistance spreading. The role of AM prophylaxis in the modulation of gut microbiota, as well as of gut health, is still not clearly understood. Therefore, this study aimed to investigate the role of different prophylaxis protocols in the modulation of the gut barrier in broilers by applying a histopathological approach. Intestinal tissue samples were collected from a total of 240 male broilers (Ross 306), reared and treated with different AM protocols. Haematoxylin and Eosin (HE) staining and a multiple scoring system were used to evaluate the presence of lesions in ileum, cecum and colon of treated broilers. Moreover, immunohistochemistry (IHC) was performed to assess the expression of claudin-3 and ZO-1 proteins in intestinal tissues. The application of a semi-quantitative scoring system was used in IHC stained samples. HE results revealed that intestinal tissues were mainly characterized by epithelial detachment and fusion of the intestinal villi, but also by the presence of lymphocytic infiltrate in the mucosa and submucosa of AM-treated broilers. However, the IHC approach for the evaluation of claudin-3 and ZO-1 proteins showed that their expression was not affected by the different AM treatments. Nevertheless, the presence of intestinal lesions highlighted by histopathology suggests that AM treatments could harm the gut health of broilers, inducing an inflammatory response and consequent epithelial lesions. In order to clarify the role of AM treatments in the modulation of gut barrier in broilers, further studies are needed., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Cuccato, Scaglione, Centelleghe, Divari, Biolatti, Pregel and Cannizzo.)
- Published
- 2022
- Full Text
- View/download PDF
13. Coping with Tissue Sampling in Suboptimal Conditions: Comparison of Different Tissue Preservation Methods for Histological and Molecular Analysis.
- Author
-
Nicoletti A, Pregel P, Starvaggi Cucuzza L, Cannizzo FT, Sereno A, and Scaglione FE
- Abstract
A high quality of samples is crucial for the success of the analysis and diagnostic purposes, and therefore the right method of conservation is vitally important for an optimal preservation of tissues. Indeed, the time to deliver the sample to the laboratory could be remarkably long, especially under suboptimal conditions, and the use of specific fixatives or cold storage may not be possible. Moreover, the portability and cost of storage equipment, their toxicity, and their ease of use play a central role when choosing the correct preservation method. The aim of this study was the identification of a reliable and economic method for tissue preservation, to be used in "in-field" sampling, suitable for both histological and molecular analysis. Punch biopsies were collected from six cattle livers. Comparisons among methods of preservation using RNAlater, silica beads, and under-vacuum was carried out. These methods were tested through considering different times and temperatures, assuming three days as a maximum time interval from sampling to laboratory and choosing 4 °C and 24 °C as references for refrigeration temperature and room temperature, respectively. Histologically, the integrity of nucleus, cytoplasm, preservation of liver structure, and easiness of recognition of inflammatory infiltrate were evaluated. The integrity of the extracted DNA and RNA was evaluated through PCR and by means of an automated electrophoresis station, respectively. RNAlater and silica beads poorly preserved the histological parameters evaluated, independently from the temperature. Conversely, the vacuum-sealed samples showed a good grade of preservation until 48 h. DNA quality was acceptable for each sample. RNA integrity showed promising results only for samples preserved with silica beads.
- Published
- 2021
- Full Text
- View/download PDF
14. 16S rRNA Sequencing Analysis of the Gut Microbiota in Broiler Chickens Prophylactically Administered with Antimicrobial Agents.
- Author
-
Cuccato M, Rubiola S, Giannuzzi D, Grego E, Pregel P, Divari S, and Cannizzo FT
- Abstract
In poultry production, gut microbiota (GM) plays a pivotal role and influences different host functions related to the efficiency of production performances. Antimicrobial (AM) use is one of the main factors affecting GM composition and functions. Although several studies have focused their attention on the role of AMs as growth promoters in the modulation of GM in broilers, the consequences of higher AM concentrations administered during prophylactic treatments need to be better elucidated. For this purpose, 16S rRNA gene sequencing was performed to evaluate the impact of different prophylactic AM protocols on the composition and diversity of the broiler GM. Diversity analysis has shown that AM treatment significantly affects alpha diversity in ileum and beta diversity in both ileum and caecum. In ileal samples, the Enterobacteriaceae family has been shown to be particularly affected by AM treatments. AMs have been demonstrated to affect GM composition in broiler. These findings indicate that withdrawal periods were not enough for the restoral of the original GM. Further studies are needed for a better elucidation of the negative effects caused by an altered GM in broilers.
- Published
- 2021
- Full Text
- View/download PDF
15. Application of RNA-sequencing to identify biomarkers in broiler chickens prophylactic administered with antimicrobial agents.
- Author
-
Giannuzzi D, Biolatti B, Longato E, Divari S, Starvaggi Cucuzza L, Pregel P, Scaglione FE, Rinaldi A, Chiesa LM, and Cannizzo FT
- Subjects
- Animals, Biomarkers, Meat analysis, RNA, Anti-Infective Agents, Chickens genetics
- Abstract
Antimicrobial (AM) resistance is largely acknowledged as one of the biggest global health and food safety challenges and the overuse of AMs is known to generate resistance in bacteria that may affect both animals and humans. Poultry meat is the second most-produced meat in the European Union and in recent years consumers are becoming more concerned about food safety, traceability, and animal welfare in poultry rearing system, increasingly requiring meats from broilers reared without AMs. In the present study, we performed RNA sequencing to analyze 64 liver and 54 muscle transcriptomic profiles in broilers reared without treatment or treated with different classes of AMs. Moreover, we validated the most differentially expressed genes among the treated groups to detect putative novel biomarkers able to discriminate meats of broilers reared without AMs. The PDK4, IGFBP1, and RHOB genes were identified as putative novel hepatic biomarkers, discriminating broilers treated with AMs compared to broilers reared without treatments. The whole transcriptome changes revealed the liver as a valuable target organ for AM administration screening. In addition, our results suggest a leading effect of the coccidiostat when associated with AMs, influencing several biological processes. Our study showed that RNA sequencing is a powerful and valuable method to detect aberrant regulated genes and to identify biomarker candidates for AM misuse detection in farm animals. Further validation on larger sample size and a wider spectrum of AMs are needed to confirm the viability of the aforementioned biomarkers in poultry population., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2021
- Full Text
- View/download PDF
16. FKBP5 gene expression in skeletal muscle as a potential biomarker for illegal glucocorticoid treatment in veal calves.
- Author
-
Starvaggi Cucuzza L, Pregel P, Biolatti B, and Cannizzo FT
- Subjects
- Animals, Biomarkers analysis, Cattle, Dexamethasone pharmacology, Estradiol pharmacology, Glucocorticoids pharmacology, Muscle, Skeletal chemistry, Prednisolone pharmacology, Receptors, Glucocorticoid genetics, Food Analysis methods, Gene Expression drug effects, Gene Expression Regulation drug effects, Muscle, Skeletal drug effects, Tacrolimus Binding Proteins genetics
- Abstract
For the current European legislation, the chemical analysis of drug residues is the exclusive accepted method to identify animals illicitly treated with growth promoters. Glucocorticoids and their metabolites are no detectable by LC/MS-MS methods in biological fluids when the growth promoter administration is discontinued several days prior to the slaughtering. The aim of this study was to elucidate the effect on the expression of genes belonging to the glucocorticoid pathway in three types of skeletal muscle of calves treated with prednisolone or dexamethasone in combination with estradiol. A gene expression change of glucocorticoid receptors (NR3C1 and NR3C2), their chaperones molecules (FKBP prolyl isomerase 4 and 5, FKBP4 and 5) and pre-receptor system (hydroxysteroid 11-beta dehydrogenases 1 and 2, HSD11B1 and 2) may indicate potential biomarkers of glucocorticoid treatment. In the biceps brachii muscle, the administration of dexamethasone with estradiol increased HSD11B2 (P < 0.01) and NR3C2 (P < 0.01) gene expression, whereas prednisolone administration increased HSD11B1 transcript levels (P < 0.05). In the longissimus lumborum muscle, NR3C2 gene expression decreased following prednisolone administration (P < 0.05). FKBP5 gene expression decreased in all considered muscles of calves administered with dexamethasone and estradiol (P < 0.01), whereas increased in the longissimus lumborum (P < 0.01) and vastus lateralis (P < 0.05) muscle of prednisolone-treated group (P < 0.05). The opposite effect of dexamethasone and prednisolone appears very promising to develop a low-cost screening test, because the expression analysis of a unique gene in a given tissue may distinguish the dispensed molecules., (Copyright © 2020 Elsevier Ltd. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
17. Characterization of the ablation zones produced by three commercially available systems from a single vendor for radiofrequency thermoablation in an ex vivo swine liver model.
- Author
-
Bullone M, Garberoglio R, Pregel P, Cannizzo FT, Gagliardo A, Martano M, Bollo E, and Scaglione FE
- Subjects
- Animals, Dogs, Models, Animal, Radiofrequency Ablation instrumentation, Reproducibility of Results, Electrodes statistics & numerical data, Liver surgery, Radiofrequency Ablation veterinary, Sus scrofa surgery
- Abstract
Background: Radiofrequency Ablation (RFA) is rarely performed in veterinary medicine. A rationale exists for its use in selected cases of canine liver tumours. RFA induces ablation zones of variable size and geometry depending on the technique used and on the impedance of the targeted organ., Objectives: (a) to describe the geometry and reproducibility of the ablation zones produced by three commercially available systems from a single company, using isolated swine liver parenchyma as a model for future veterinary applications in vivo; (b) to study the effects of local saline perfusion into the ablated parenchyma through the electrode tip and of single versus double passage of the electrode on size, geometry and reproducibility of the ablation zones produced., Methods: Size, and geometry of ablation zones reproduced in six livers with one cooled and perfused (saline) and two cooled and non-perfused systems, after single or double passage (n = 6/condition), were assessed macroscopically on digitalized images by a blinded operator. Longitudinal and transverse diameters, equivalent diameter, estimated volume and roundness index were measured. Reproducibility was assessed as coefficient of variation., Results and Conclusions: Ablation zone reproducibility was higher when expressed in terms of ablation zone diameters than estimated volume. Local saline perfusion of the parenchyma through the electrode tip during RFA increased the ablation zone longitudinal diameter. Ablation zone estimated volume increased with saline perfusion only when double passage was performed. These data may provide useful information for those clinicians who intend to include RFA as an additive tool in veterinary interventional radiology., (© 2020 The Authors. Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)
- Published
- 2020
- Full Text
- View/download PDF
18. LC-MS/MS analyses of bile and histological analyses of thymus as diagnostic tools to detect low dose dexamethasone illicit treatment in beef cattle at slaughterhouse.
- Author
-
Sebastianelli M, Forte C, Galarini R, Gobbi M, Pistidda E, Moncada C, Cannizzo FT, Pezzolato M, Bozzetta E, Cenci-Goga BT, and Manuali E
- Subjects
- Administration, Oral, Animals, Cattle, Chromatography, Liquid, Dexamethasone administration & dosage, Dose-Response Relationship, Drug, Male, Tandem Mass Spectrometry, Abattoirs, Bile chemistry, Dexamethasone analysis, Illicit Drugs analysis, Thymus Gland chemistry
- Abstract
Dexamethasone (DXM) is a synthetic adrenal corticosteroid with anti-inflammatory properties used for therapeutic purposes in a wide range of pathologies and of the most common corticosteroids used for anabolic purposes in beef cattle. It is proven that DXM induces histological changes, traceable as increasing fatty infiltration of the thymus associated with a concurrent decrease of the cortex-medulla ratio, so the histological examination of the thymus gland has been established as an indirect morphological biomarker. The aim of the present study is to compare thymus histology and DXM concentrations in biological fluids collected at slaughterhouse after 1 month of DXM treatment. Our findings demonstrate that a low dosage of DXM administered to 12 months-old-Chianina beef cattle induces severe thymic atrophy with concurrent reduction of the cortex/medulla ratio, demonstrable even when DXM residues are not found in serum and urine samples. It is worth to note that, at the slaughterhouse, DXM residues are detectable in bile samples, indicating the ability of this biological fluid to bio-concentrate the administered drug if compared to serum and urine. Therefore, bile could be candidates as new liquid matrix for the screening programs planned to contrast the illegal use of anabolic substances., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
19. Tissue and species identification in minced meat and meat products from Italian commercial markets by DNA microarray and histological approach.
- Author
-
Sohrabi H, Cannizzo FT, Pregel P, Scaglione FE, Beltramo C, Acutis PL, Dalmasso A, and Biolatti B
- Subjects
- Animals, Cattle, Italy, Meat Products standards, Food Contamination statistics & numerical data, Food Labeling standards, Meat standards, Oligonucleotide Array Sequence Analysis veterinary
- Abstract
Adequate testing and adulterant detection of food products are required to assure its safety and avoid fraudulent activities. Adulteration/substitution of costlier meat with a cheaper or inferior meat is one of the most common fraudulence in meat industry. Aim of this study was to check the correct labelling of meat and ready to cook bovine meat products, combining the DNA microarray approach to identify the animal species with the histological examination, to check the composition and safety of meat. One hundred and one samples of bovine minced meat (Group 1) and ready to cook meat products (Group 2) were collected from supermarkets in Turin, Italy. DNA microarray revealed that 25.7% of samples were positive for species not declared on the label, swine being the most common. Histology showed the presence of cartilage, bone and glandular tissue. A higher presence of bacteria and inflammatory cells was detected in Group 1. Bacterial cells associated to inflammatory cells were detected with a higher score in Group 2. Sarcocystis spp. were present in 83.3% samples of Group 1 and 49.1% of Group 2. This study confirmed that the mislabelling of meat products is not uncommon. The combination of DNA microarrays and histology can increase the monitoring capacity in bovine meat industry.
- Published
- 2020
- Full Text
- View/download PDF
20. Expression of corticosteroid hormone receptors, prereceptors, and molecular chaperones in hypothalamic-pituitary-adrenal axis and adipose tissue after the administration of growth promoters in veal calves.
- Author
-
Starvaggi Cucuzza L, Divari S, Biolatti B, and Cannizzo FT
- Subjects
- Adrenal Glands drug effects, Adrenal Glands metabolism, Animals, Dexamethasone administration & dosage, Dexamethasone pharmacology, Estradiol administration & dosage, Estradiol analogs & derivatives, Estradiol pharmacology, Gene Expression Regulation drug effects, Glucocorticoids pharmacology, Hypothalamus drug effects, Hypothalamus metabolism, Male, Pituitary Gland drug effects, Pituitary Gland metabolism, Adipose Tissue metabolism, Cattle physiology, Hypothalamo-Hypophyseal System metabolism, Molecular Chaperones metabolism, Pituitary-Adrenal System metabolism, Receptors, Steroid metabolism
- Abstract
The action of glucocorticoids on target tissues is regulated by the glucocorticoid and mineralocorticoid receptors (codified by the NR3C1 and NR3C2 gene, respectively). Moreover, the prereceptor system, represented by the hydroxysteroid 11-beta dehydrogenases (HSD11Bs), catalyzes the interconversion from active glucocorticoids into inactive compounds. This study aimed to determine whether the expression of the prereceptor system, the corticosteroid receptors, and the molecules regulating their intracellular trafficking (FKBP prolyl isomerase 4 and FKBP prolyl isomerase 5) could be regulated in the hypothalamic-pituitary-adrenal axis and in different type of adipose tissue of calves by the administration of dexamethasone in combination with estradiol or prednisolone. Research about the glucocorticoid effects on bovine target tissues may allow development of new diagnostic methods that use potential molecular biomarkers of glucocorticoid treatment. The administration of dexamethasone in combination with estradiol increased the gene expression of HSD11B1 (P < 0.01), HSD11B2 (P < 0.05), NR3C1 (P < 0.01), and NR3C2 (P < 0.01) in the adrenal glands; NR3C2 in the intramuscular adipose tissue (P < 0.01), and HSD11B1 in the subcutaneous adipose tissue (P < 0.01). Prednisolone administration increased the gene expression of HSD11B1 (P < 0.01), NR3C1 (P < 0.05), and NR3C2 (P < 0.05) in the adrenal glands and HSD11B1 (P < 0.01) in the subcutaneous adipose tissue. Interestingly, most of the examined tissues/organs showed a significant variation of FKBP5 gene expression after the administration of dexamethasone in combination with estradiol. So, these changes suggest that the FKBP5 gene expression could be a possible biomarker of the illegal dexamethasone administration in calves., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
21. Dexamethasone and prednisolone treatment in beef cattle: influence on glycogen deposition and gene expression in the liver.
- Author
-
Divari S, De Lucia F, Berio E, Sereno A, Biolatti B, and Cannizzo FT
- Subjects
- Animals, Cattle, Glucocorticoids pharmacology, Liver drug effects, Male, Tissue Distribution, Dexamethasone pharmacology, Gene Expression Regulation drug effects, Glycogen metabolism, Liver metabolism, Prednisolone pharmacology
- Abstract
The illegal administration of glucocorticoids in livestock is problematic and identification of pathways in which these hormones are involved is critically important, and new direct or indirect biomarkers should be identified. In this work, glucocorticoid transcriptional effects on some genes involved in the glucose metabolism were studied in the bovine liver. This study was conducted on adult Charolais male cattle treated with long-term low dose dexamethasone or prednisolone. Gene expression analysis was conducted in the liver by qPCR, and the geNorm algorithm was applied to select optimal reference genes. In line with the literature, a significant overexpression of genes involved in the gluconeogenic pathway and glycogen synthesis was detected in the liver of dexamethasone-treated animals, but histological and biochemical examination showed hepatocyte glycogen depletion particularly in dexamethasone-treated animals. It possible to hypothesize that glucocorticoids or adrenal insufficiency due to glucocorticoids withdrawal inhibit the enzymatic activity of glycogen synthase and/or induce glycogen autophagy in bovine liver. In fact, markers of glycophagy as starch-binding domain-containing protein 1 and γ-aminobutyric acid receptor-associated protein-like 1 mRNAs were upregulated in the liver by glucocorticoids treatment. Furthermore, glycogen synthase kinase-3 beta gene was significantly overexpressed in dexamethasone-treated animals, and this protein is also implicated in liver autophagy modulation and glycogen synthesis inhibition. These results showed that glucocorticoids likley have dual roles in hepatic glycogen metabolism of cattle, and investigation of these pathways could help find treatment biomarkers., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
22. Meat quality traits and canonical discriminant analysis to identify the use of illicit growth promoters in Charolais bulls.
- Author
-
Barbera S, Biolatti B, Divari S, and Cannizzo FT
- Subjects
- Animals, Cattle, Dexamethasone pharmacology, Discriminant Analysis, Electronic Nose, Estradiol pharmacology, Farms, Food Analysis instrumentation, Food Analysis statistics & numerical data, Food Quality, Male, Prednisolone pharmacology, Food Analysis methods, Growth Substances pharmacology, Meat analysis, Muscle, Skeletal drug effects, Muscle, Skeletal growth & development
- Abstract
The administration of anabolic agents in farm animals to improve meat production has been prohibited in EU, due to the potential risks to human health. Meat quality was investigated to detect the effects of illegal administration of dexamethasone or prednisolone or 17β-estradiol on Charolais bulls. Three groups of 6 bulls were treated and 12 bulls were the control. Meat quality parameters were measured on live animals, carcasses and on samples of Longissimus thoracis and multivariate statistical data analysis was applied. In Charolais bulls, these parameters were affected by growth promoter administration and the multivariate canonical discriminant analysis was able to distinguish between treated and untreated animals mainly due to three electronic nose's parameters, 24 h carcass temperature and drip loss. Therefore, meat quality control and the multivariate analysis could be useful as a first screening to address targeted controls on farms suspected of illicit use of growth promoters., (Copyright © 2019 Elsevier Ltd. All rights reserved.)
- Published
- 2019
- Full Text
- View/download PDF
23. Fatal Leucocytozoon Infection in a Captive Grey-headed Parrot ( Poicephalus robustus suahelicus ).
- Author
-
Galosi L, Scaglione FE, Magi GE, Cork SC, Peirce MA, Ferraro S, Cucuzza LS, Cannizzo FT, and Rossi G
- Subjects
- Animals, Bird Diseases pathology, Fatal Outcome, Female, Protozoan Infections, Animal pathology, Bird Diseases parasitology, Haemosporida, Parrots, Protozoan Infections, Animal parasitology
- Abstract
A necropsy was conducted on a female grey-headed parrot ( Poicephalus robustus suahelicus ) that died following signs of depression, ruffled feathers, and inappetence. Microscopic examination revealed the presence of hemoprotozoa in the liver. A nested polymerase chain reaction (PCR), targeting the mitochondrial cytochrome b gene of Haemoproteus species, Plasmodium species, and Leucocytozoon species, was performed on frozen tissue samples collected at necropsy. The hemoprotozoa were identified by PCR analysis as Leucocytozoon species. Hemoprotozoa are rarely reported in African parrots, and this is the first report of a Leucocytozooon species infection in a Poicephalus robustus suahelicus ., (© 2019 by the Association of Avian Veterinarians.)
- Published
- 2019
- Full Text
- View/download PDF
24. A survey on zoo mortality over a 12-year period in Italy.
- Author
-
Scaglione FE, Biolatti C, Pregel P, Berio E, Cannizzo FT, Biolatti B, and Bollo E
- Abstract
Background: The zoo is a unique environment in which to study animals. Zoos have a long history of research into aspects of animal biology, even if this was not the primary purpose for which they were established. The data collected from zoo animals can have a great biological relevance and it can tell us more about what these animals are like outside the captive environment. In order to ensure the health of all captive animals, it is important to perform a post-mortem examination on all the animals that die in captivity., Methods: The causes of mortality of two hundred and eighty two mammals which died between 2004 and 2015 in three different Italian zoos (a Biopark, a Safari Park and a private conservation center) have been investigated., Results: Post mortem findings have been evaluated reporting the cause of death, zoo type, year and animal category. The animals frequently died from infectious diseases, in particular the causes of death in ruminants were mostly related to gastro-intestinal pathologies. pulmonary diseases were also very common in each of the zoos in the study. Moreover, death was sometimes attributable to traumas, as a result of fighting between conspecifics or during mating. Cases of genetic diseases and malformations have also been registered., Discussion: This research was a confirmation of how conservation, histology and pathology are all connected through individual animals. These areas of expertise are extremely important to ensure the survival of rare and endangered species and to learn more about their morphological and physiological conditions. They are also useful to control pathologies, parasites and illnesses that can have a great impact on the species in captivity. Finally, this study underlines the importance of a close collaboration between veterinarians, zoo biologists and pathologists. Necropsy findings can help conservationists to determine how to support wild animal populations., Competing Interests: The authors declare there are no competing interests.
- Published
- 2019
- Full Text
- View/download PDF
25. Gene expression profile associated with thymus regeneration in dexamethasone-treated beef cattle.
- Author
-
Cannizzo FT, Cucuzza LS, Divari S, Berio E, Scaglione FE, and Biolatti B
- Subjects
- Animals, Basic Helix-Loop-Helix Transcription Factors genetics, Cattle, Dexamethasone administration & dosage, Gene Expression drug effects, Genes, myc genetics, Keratin-5 genetics, Prednisolone administration & dosage, RNA, Messenger analysis, Red Meat, Regeneration drug effects, Transcription Factors genetics, Tumor Suppressor Proteins, Glucocorticoids administration & dosage, Regeneration genetics, Thymus Gland physiology, Transcriptome drug effects
- Abstract
Glucocorticoids (GCs) are illegally used as growth promoters in cattle, and the analytical methods officially applied most likely underestimate the precise frequency of the abuse. As a side effect, the administration of GCs causes fat infiltration, apoptosis, and atrophy of the thymus. However, gross and histological observations carried out previously showed that the thymus preserves an intrinsic ability to regenerate. The aim of this work was to study the transcriptional effects of GCs on genes likely involved in regeneration of the epithelial cell network in the cervical and thoracic thymus of beef cattle treated with dexamethasone (DEX) or prednisolone (PRD) in comparison with a control group. Moreover, the ratio of bax/bcl2 genes was examined to verify a possible antiapoptotic activity occurring at the same time. In the cervical thymus, DEX administration increased the gene expression of c-myc (P < 0.01), tcf3 (P < 0.05), tp63 (P < 0.01), and keratin 5 (krt5; P < 0.01). In the thoracic thymus of DEX-treated cattle, the gene expression of tcf3 (P < 0.01), tp63 (P < 0.01), and krt5 (P < 0.05) was increased. These results suggested that thymic regeneration is underway in the DEX-treated animals. However, the bax/bcl2 ratio was decreased in both cervical and thoracic thymus of DEX-treated cattle (P < 0.01 and P < 0.05, respectively), showing an antiapoptotic effect through the mitochondrial pathway. Conversely, PRD administration caused no change in the expression of all considered genes. These results sustain the hypothesis that regeneration occurs in the thymus parenchyma 6 d after the DEX treatment was discontinued. This hypothesis is also supported by the absence of alterations in the thymus of PRD-treated beef cattle. Indeed, previous studies showed the inability of PRD to induce macroscopic and microscopic lesions in the thymus. Therefore, in this context, it is not surprising that PRD induced no alteration of genes involved in the regeneration pathway., (Copyright © 2018 Elsevier Inc. All rights reserved.)
- Published
- 2018
- Full Text
- View/download PDF
26. A two-step immunomagnetic separation of somatic cell subpopulations for a gene expression profile study in bovine milk.
- Author
-
Divari S, Starvaggi Cucuzza L, Riondato F, Pregel P, Sacchi P, Rasero R, Biolatti B, and Cannizzo FT
- Subjects
- Animals, Cattle, Female, Lymphocytes chemistry, Lymphocytes immunology, Mastitis, Bovine microbiology, Mastitis, Bovine pathology, Milk chemistry, Milk immunology, Monocytes chemistry, Monocytes immunology, Neutrophils chemistry, Neutrophils immunology, RNA, Messenger analysis, Staphylococcal Infections immunology, Staphylococcal Infections pathology, Staphylococcal Infections veterinary, Immunity, Innate genetics, Immunomagnetic Separation veterinary, Mastitis, Bovine immunology, Milk cytology, Transcriptome
- Abstract
The objective of this study was to demonstrate the usefulness of an immunomagnetic method to purify subpopulations of milk somatic cells. The experiment was conducted on milk samples collected from healthy cows (n = 17) and from cows with clinical mastitis (n = 24) due to a Staphylococcus aureus natural infection. A two-step immunomagnetic purification was applied to simultaneously separate three somatic cell subpopulations from the same milk sample. Total RNA was extracted and qPCR was performed to determinate mRNA levels of innate immunity target genes in purified somatic cell subpopulations. Good quality and quantity of RNA allowed the reference gene analysis in each cell subpopulation. An up-regulation of the main genes involved in innate immune defence was detected in separated polymorphonuclear neutrophilic leucocytes-monocytes and lymphocytes of mastitic milk. These results and flow cytometric analysis suggest that the immunomagnetic purification is an efficient method for the isolation of the three populations from milk, allowing the cells to be studied separately.
- Published
- 2018
- Full Text
- View/download PDF
27. Role of FKBP51 in the modulation of the expression of the corticosteroid receptors in bovine thymus following glucocorticoid administration.
- Author
-
Starvaggi Cucuzza L, Biolatti B, Scaglione FE, and Cannizzo FT
- Subjects
- Animals, Cattle, Dexamethasone administration & dosage, Gene Expression Regulation drug effects, Glucocorticoids administration & dosage, Glucocorticoids pharmacology, Male, Prednisolone administration & dosage, RNA genetics, RNA metabolism, Tacrolimus Binding Proteins genetics, Thymus Gland metabolism, Dexamethasone pharmacology, Prednisolone pharmacology, Tacrolimus Binding Proteins metabolism, Thymus Gland drug effects
- Abstract
The aim of this work was to study the transcriptional effects of glucocorticoids on corticosteroid hormone receptors, prereceptors (11β-hydroxysteroid dehydrogenase 1 and 2, 11β-HSD1 and 2), and chaperones molecules regulating intracellular trafficking of the receptors (FKBP51 and FKBP52) in thymus of veal calves. Moreover, the expression of FKBP51 and FKBP52 gene were investigated in beef cattle thymus. In the cervical thymus of veal calves, dexamethasone administration in combination with estradiol decreased FKBP51 expression (P < 0.01). The same treatment increased mineralocorticoid receptor (MR) (P < 0.01) and 11β-HSD1 expression (P < 0.05) compared to control group in the cervical thymus of veal calves. The thoracic thymus of veal calves treated with dexamethasone and estradiol showed a decrease of FKBP51 (P < 0.05), FKBP52 (P < 0.05), glucocorticoid receptor (P < 0.05), and MR expression (P < 0.05) compared to control group in the thoracic thymus of veal calves. The gene expression of FKBP51 decreased both in cervical (P < 0.01) and thoracic thymus (P < 0.01) of beef cattle treated with dexamethasone and estradiol. In addition, also prednisolone administration reduced FKBP51 expression in the cervical thymus (P < 0.01) and in the thoracic thymus of beef cattle (P < 0.01). The gene expression of FKBP52 increased only in the cervical thymus following dexamethasone administration (P < 0.01). The decrease of FKBP51 gene expression in thymus could be a possible biomarker of illicit dexamethasone administration in bovine husbandry. Moreover, so far, an effective biomarker of prednisolone administration is not identified. In this context, the decrease of FKBP51 gene expression in thymus of beef cattle following prednisolone administration could play an important role in the indirect identification of animals illegally treated with prednisolone., (Copyright © 2017 Elsevier Inc. All rights reserved.)
- Published
- 2018
- Full Text
- View/download PDF
28. Reference Gene Selection and Prednisolone Target Gene Expression in Adipose Tissues of Friesian Cattle.
- Author
-
Divari S, Berio E, Biolatti B, and Cannizzo FT
- Subjects
- Adipose Tissue drug effects, Animals, Cattle metabolism, Male, Proteins genetics, Proteins metabolism, Adipose Tissue metabolism, Cattle genetics, Gene Expression drug effects, Prednisolone pharmacology
- Abstract
Corticosteroids are frequently used in livestock production, and their use is permitted by the European Union for therapeutic purposes only. However, small doses of corticosteroids are often administered in meat-producing animals to improve zootechnical performance. Prednisolone is one of the most commonly used corticosteroids with a growth-promoting purpose in animal husbandry. This study proposes to identify a gene whose expression is significantly regulated by prednisolone in visceral and subcutaneous adipose tissues. The analysis was conducted on Friesian cattle treated with prednisolone (30 mg day
-1 ). The reference gene expression stability and optimal number for gene expression normalization were calculated. Family with sequence similarity 107 member A (FAM107A) and pyruvate dehydrogenase kinase 4 are the prednisolone target genes identified in adipose tissue. FAM107A was downregulated by ∼2.9-fold by prednisolone in subcutaneous adipose tissue. This result suggests that FAM107A could be a possible indirect biomarker of prednisolone treatment in cattle and encourages a deeper investigation in this direction.- Published
- 2017
- Full Text
- View/download PDF
29. Development and Application of a Screening Method of Absolute Quantitative PCR To Detect the Abuse of Sex Steroid Hormone Administration in Male Bovines.
- Author
-
Starvaggi Cucuzza L, Biolatti B, Divari S, Pregel P, Scaglione FE, Sereno A, and Cannizzo FT
- Subjects
- Animals, Cattle growth & development, Consumer Product Safety, Gene Expression, Humans, Male, Testis drug effects, Testis growth & development, Cattle genetics, Gonadal Steroid Hormones administration & dosage, Intracellular Signaling Peptides and Proteins genetics, Real-Time Polymerase Chain Reaction methods, Veterinary Drugs administration & dosage
- Abstract
A methodology for the absolute quantification of regucalcin gene through quantitative PCR was set up to confirm that the decrease of regucalcin gene expression in the testis is an effective biomarker for tracing sex steroid hormone treatment in bovine husbandry. On the basis of TaqMan technology, an external standard curve was generated. Using in vivo experiments, a ROC curve was developed to calculate the criterion value, specificity, and sensitivity for this potential biomarker. Then, regucalcin gene expression was assessed in veal calves and beef intended for human consumption. In 11 of 54 calves and in 5 of 70 beef cattle the regucalcin gene was expressed under their respective cutoff. Additionally, a mild decrease of regucalcin protein expression was revealed by immunohistochemistry in subjects tested positive via qPCR. These preliminary results suggest that this transcriptomics test may be employed as a novel diagnostic screening tool, improving significantly the overall efficacy of food control.
- Published
- 2017
- Full Text
- View/download PDF
30. Effects and detection of Nandrosol and ractopamine administration in veal calves.
- Author
-
Divari S, Berio E, Pregel P, Sereno A, Chiesa L, Pavlovic R, Panseri S, Bovee TFH, Biolatti B, and Cannizzo FT
- Subjects
- Animals, Cattle, Gene Expression, Male, Nandrolone chemistry, Chromatography, Liquid methods, Mass Spectrometry methods, Nandrolone analogs & derivatives, Phenethylamines chemistry
- Abstract
The present study describes different effects of the selective androgen receptor modulator (SARM) nandrolone phenylpropionate (Nandrosol) and the β-agonist ractopamine administration in veal calves, and it investigates different strategies applied to trace these molecules. Morphological changes of gonads and accessory glands attributed to androgen effects, such as testicular atrophy, seminiferous tubule diameter reduction and hyperplasia of prostate epithelium, were detected, although SARMs are not described to cause these lesions. The gene expression analysis showed an anabolic activity of Nandrosol in Longissimus dorsi muscle, where myosin heavy chain (MYH) was significantly up-regulated. An IGF1 increase was weakly significant only in Vastus lateralis muscle. In conclusion, the anatomo-histopathological observations and the MYH mRNA up-regulation in Longissimus dorsi muscle confirm the androgenic treatment in experimental animals. The biosensor assay was not enough sensitive to detect residues in urines and only the direct chemical analysis of urine samples confirmed both β-agonist and SARM treatment., (Copyright © 2016 Elsevier Ltd. All rights reserved.)
- Published
- 2017
- Full Text
- View/download PDF
31. Evaluation of nandrolone and ractopamine in the urine of veal calves: liquid chromatography-tandem mass spectrometry approach.
- Author
-
Chiesa L, Panseri S, Cannizzo FT, Biolatti B, Divari S, Benevelli R, Arioli F, and Pavlovic R
- Subjects
- Animals, Chromatography, High Pressure Liquid methods, Food Safety, Limit of Detection, Male, Substance Abuse Detection methods, Anabolic Agents urine, Cattle urine, Growth Substances urine, Nandrolone urine, Phenethylamines urine, Tandem Mass Spectrometry methods
- Abstract
Under European legislation, the use of growth promoters is forbidden in food-producing livestock. The application of unofficial protocols with diverse combinations of veterinary drugs, administered in very low concentrations, hinders reliable detection and subsequent operative prevention. It was observed that nandrolone (anabolic steroid) and ractopamine (β-adrenergic agonist) are occasionally administered to animals, but little is known about their synergic action when they are administered together. Two specific analytical methods based on liquid chromatography-tandem mass spectrometry have been developed, both of which include hydrolysis of the corresponding conjugates. For the nandrolone method, solid-phase extraction was necessary for the complete elimination of the interferences, while employment of the Quantitation Enhanced Data-Dependent scan mode during MS acquisition of ractopamine enabled the utilization of simple liquid-liquid extraction. The nandrolone method was linear in the range of 0.5-25 ng/mL, while the ractopamine calibration curve was constructed from 0.5 to 1000 ng/mL. The corresponding coefficients of correlations were >0.9907. The lower limit of quantification for both methods was 0.5 ng/mL, followed by overall recoveries >81%. Precisions expressed as relative standard deviations were <17%, while matrix effects were minimal. Urine samples taken at the slaughterhouse from veal calves enrolled in an experimental treatment consisting of intramuscular administration of β-nandrolone-phenylpropionate accompanied with a ractopamine-enriched diet were analysed. Those methods might be useful for studying the elimination patterns of the administered compounds along with characterization of the main metabolic pathways. Copyright © 2016 John Wiley & Sons, Ltd., (Copyright © 2016 John Wiley & Sons, Ltd.)
- Published
- 2017
- Full Text
- View/download PDF
32. 17 β -estradiol upregulates oxytocin and the oxytocin receptor in C2C12 myotubes.
- Author
-
Berio E, Divari S, Starvaggi Cucuzza L, Biolatti B, and Cannizzo FT
- Abstract
Background: The endocrinology of skeletal muscle is highly complex and many issues about hormone action in skeletal muscle are still unresolved. Aim of the work is to improve our knowledge on the relationship between skeletal muscle and 17 β -estradiol., Methods: The skeletal muscle cell line C2C12 was treated with 17 β -estradiol, the oxytocin peptide and a combination of the two hormones. The mRNA levels of myogenic regulatory factors, myosin heavy chain, oxytocin, oxytocin receptor and adipogenic factors were analysed in C2C12 myotubes., Results: It was demonstrated that C2C12 myoblasts and myotubes express oxytocin and its receptor, in particular the receptor levels physiologically increase in differentiated myotubes. Myotubes treated with 17 β -estradiol overexpressed oxytocin and oxytocin receptor genes by approximately 3- and 29-fold, respectively. A decrease in the expression of fatty acid binding protein 4 (0.62-fold), a fat metabolism-associated gene, was observed in oxytocin-treated myotubes. On the contrary, fatty acid binding protein 4 was upregulated (2.66-fold) after the administration of the combination of 17 β -estradiol and oxytocin. 17 β -estradiol regulates oxytocin and its receptor in skeletal muscle cells and they act in a synergic way on fatty acid metabolism., Discussion: Oxytocin and its receptor are physiologically regulated along differentiation. 17 β -estradiol regulates oxytocin and its receptor in skeletal muscle cells. 17 β -estradiol and oxytocin act in a synergic way on fatty acid metabolism. A better understanding of the regulation of skeletal muscle homeostasis by estrogens and oxytocin peptide could contribute to increase our knowledge of muscle and its metabolism., Competing Interests: The authors declare there are no competing interests.
- Published
- 2017
- Full Text
- View/download PDF
33. Scrotal granulomatous aspergillosis in a dromedary camel (Camelus dromedarius).
- Author
-
Scaglione FE, Peano A, Piga S, Meda S, Bollo E, Cannizzo FT, Pasquetti M, and Jensen HE
- Subjects
- Animals, Animals, Zoo, Aspergillosis diagnosis, Aspergillosis microbiology, Granuloma diagnosis, Granuloma microbiology, Male, Scrotum microbiology, Subcutaneous Tissue pathology, Aspergillosis veterinary, Camelus, Granuloma veterinary, Scrotum pathology
- Abstract
Background: This report describes a case of primary subcutaneous aspergillosis in a 7-year-old neutered male dromedary camel (Camelus dromedarius)., Case Presentation: The animal developed a large nodular lesion in the right scrotum two years after surgical intervention for neutering. The mass had a firm consistency and was painful at palpation. Histopathology revealed dermal granulomatous inflammation with a necrotic centre, surrounded by plasma cells, macrophages, neutrophils, and sparse fungal hyphae characterised by parallel cell walls, distinct septa, and dichotomous branching. Fungal culture was not performed, but a panel of mono- and polyclonal antibodies specific for different fungal genera identified the hyphae as Aspergillus sp., Conclusions: The occurrence of subcutaneous lesions is a rare manifestation of aspergillosis in animals, and this appears to be the first case reported in the dromedary camel.
- Published
- 2017
- Full Text
- View/download PDF
34. Bovine teeth as a novel matrix for the control of the food chain: liquid chromatography-tandem mass spectrometry detection of treatments with prednisolone, dexamethasone, estradiol, nandrolone and seven β 2 -agonists.
- Author
-
Chiesa LM, Nobile M, Panseri S, Biolatti B, Cannizzo FT, Pavlovic R, and Arioli F
- Subjects
- Animals, Cattle, Adrenergic beta-2 Receptor Agonists blood, Chromatography, Liquid methods, Dexamethasone analysis, Estradiol analysis, Food Chain, Nandrolone analysis, Prednisolone analysis, Tandem Mass Spectrometry methods
- Abstract
Veterinary drugs usually have rapid clearance rates in the liver and kidney, hampering their detection in conventional matrices such as the liver or urine. Pharmacological principles such as esterification may be applied to facilitate the administration of veterinary drugs and increase drug half-life. Prednisolone, whose therapeutic administration is regulated for food producing animals in the EU, is available in its acetate form as well as nandrolone, a banned anabolic steroid, which may be obtained as nandrolone phenylpropionate and estradiol as a benzoyl ester. While the distribution and accumulation of lipophilic and hydrophilic substances in human teeth have been well documented, studies on residues in bovine teeth are lacking. We hypothesised that analysis of bovine teeth could be used to detect both regulated and banned veterinary drugs. Steroids may be illegally used as growth promoters in food producing animals, alone or combined with β2-agonists; therefore, we developed, and validated, in accordance with the Commission Decision 2002/657/EC, two analytical confirmatory LC-MS/MS methods to detect these classes of compounds following a unique liquid extraction procedure. Finally, we analysed teeth from three male Friesian veal calves treated with intramuscular estradiol benzoate, oral prednisolone acetate or intramuscular nandrolone phenylpropionate in combination with oral ractopamine, respectively, and from seven bovines from the food chain. Teeth from treated animals were positive for their respective drugs, with the exception of nandrolone phenylpropionate. One sample from a food chain bovine was positive for isoxsuprine, one of the seven β2-agonists studied. Non-esterified forms of the steroids were not found. These results demonstrate that bovine teeth are a suitable matrix for the determination of pseudoendogenous substances or illicit administration of veterinary drugs.
- Published
- 2017
- Full Text
- View/download PDF
35. Morphological Examination and Transcriptomic Profiling To Identify Prednisolone Treatment in Beef Cattle.
- Author
-
Cannizzo FT, Pegolo S, Pregel P, Manuali E, Salamida S, Divari S, Scaglione FE, Bollo E, Biolatti B, and Bargelloni L
- Subjects
- Animals, Apoptosis drug effects, Body Weight drug effects, Cattle, Cell Proliferation drug effects, Epididymis physiology, Epididymis ultrastructure, Leukocytes, Mononuclear drug effects, Male, Muscle, Skeletal physiology, Protein Serine-Threonine Kinases genetics, Red Meat, Testis physiology, Testis ultrastructure, Epididymis drug effects, Muscle, Skeletal drug effects, Prednisolone pharmacology, Testis drug effects, Transcriptome drug effects
- Abstract
In livestock production corticosteroids are licensed only for therapy; nevertheless, they are often illegally used as growth promoters. The aim of this study was to identify morphological or biomolecular alterations induced by prednisolone (PDN) in experimentally treated beef cattle, because PDN and its metabolites are no longer detectable by LC-MS/MS methods in biological fluids. Moreover, PDN does not induce any histological alterations in the thymus, different from dexamethasone treatments. Therefore, a marker of illicit treatment for this growth promoter could be useful. Eight male Italian Friesian beef cattle were administered prednisolone acetate 30 mg day
-1 per os for 35 days, and seven beef cattle represented the control group. Six days after drug withdrawal, the animals were slaughtered. Morphological and morphometric modifications were evaluated in the epididymis and testis, whereas transcriptomic changes induced by PDN administration were investigated in peripheral blood mononuclear cells (PBMCs) at different sampling times and in skeletal muscle and testis sampled at slaughtering. In the epididymis, spermatozoa number decreased in PDN-treated animals, and in some cases they were totally absent. Correspondingly, in the testis of treated animals, down-regulation for serine/threonine kinase 11 (STK11) gene expression was detected (p < 0.01). DNA microarray analysis revealed a total of 133 differentially expressed genes in skeletal muscle and testis, and 907 and 1416 in PBMCs after 33 days of treatment and at slaughtering, respectively. Histological investigations on epididymal content could represent a promising marker for PDN treatment in beef cattle and could be used as a screening method to identify animals worthy of further investigation with official methods. Moreover, the clear transcriptomic signature of PDN treatment evidenced in PBMCs supported the possibility of using this matrix to monitor the illicit treatment in vivo during ranching.- Published
- 2016
- Full Text
- View/download PDF
36. Blood parasites in hooded crows (Corvus corone cornix) in Northwest Italy.
- Author
-
Scaglione FE, Cannizzo FT, Pregel P, Perez Rodriguez AD, and Bollo E
- Subjects
- Animals, Female, Italy, Male, Apicomplexa isolation & purification, Crows blood, Crows parasitology, Plasmodium isolation & purification
- Abstract
Haemoparasites and their effects on hooded crows (Corvus corone cornix) are poorly studied. The aims are to evaluate the prevalence of Haemoproteus spp./Plasmodium spp. or Leucocytozoon spp., to correlate this with gross and histopathological findings, and to investigate the association among infection and geographical origin, age, gender, parasite distribution and prevalence among organs. Hooded crows (n = 47) were collected within a regional culling programme from 3 districts in the province of Turin (Italy) and subjected to necropsy. Histological and molecular analyses were carried out on some tissues. Leucocytozoon spp. was detected in 46 crows (97.9%) by polymerase chain reaction (PCR), whereas 28 birds (59.6%) were found to be positive for Haemoproteus spp./Plasmodium spp. The distribution of parasites in several organs varied significantly, showing that Leucocytozoon spp. is ubiquitous in organs in contrast with Haemoproteus spp./Plasmodium spp., which have a specific predilection for spleen and lungs. The prevalence of Haemoproteus spp./Plasmodium spp. also differed significantly among the crows captured in the areas of the study. The high prevalence of haemoparasites emphasizes the success of ornithophilic vectors and the susceptibility of this species to infection. Differences in prevalence among the sites are probably due to orographic features of the areas, variations in vector species and density, or to crow population size or structure. In spite of the high infection rate, no gross and histological lesions were found. This finding further suggests an evolutionary adaptation between crows and avian blood parasites.
- Published
- 2016
- Full Text
- View/download PDF
37. HPLC-ESI-MS/MS assessment of the tetrahydro-metabolites of cortisol and cortisone in bovine urine: promising markers of dexamethasone and prednisolone treatment.
- Author
-
Chiesa L, Panseri S, Pavlovic R, Cannizzo FT, Biolatti B, Divari S, Villa R, and Arioli F
- Subjects
- 11-beta-Hydroxysteroid Dehydrogenases antagonists & inhibitors, 11-beta-Hydroxysteroid Dehydrogenases urine, Animals, Biomarkers urine, Biotransformation, Cattle, Chromatography, High Pressure Liquid, Dexamethasone pharmacology, Male, Oxidoreductases Acting on CH-CH Group Donors antagonists & inhibitors, Oxidoreductases Acting on CH-CH Group Donors urine, Prednisolone pharmacology, Spectrometry, Mass, Electrospray Ionization, Stereoisomerism, Tandem Mass Spectrometry, Tetrahydrocortisol urine, Cortisone urine, Dexamethasone urine, Hydrocortisone urine, Prednisolone urine, Tetrahydrocortisol analogs & derivatives, Tetrahydrocortisone urine
- Abstract
The effects of long-term administration of low doses of dexamethasone (DX) and prednisolone (PL) on the metabolism of endogenous corticosteroids were investigated in veal calves. In addition to cortisol (F) and cortisone (E), whose interconversion is regulated by 11β-hydroxysteroid dehydrogenases (11βHSDs), special attention was paid to tetrahydrocortisol (THF), allo-tetrahydrocortisol (aTHF), tetrahydrocortisone (THE) and allo-tetrahydrocortisone (aTHE), which are produced from F and E by catalytic activity of 5α and 5β-reductases. A specifically developed HPLC-ESI-MS/MS method achieved the complete chromatographic separation of two pairs of diastereoisomers (THF/aTHF and THE/aTHE), which, with appropriate mass fragmentation patterns, provided an unambiguous conformation. The method was linear (r(2) > 0.9905; 0.5-25 ng ml(-1)), with LOQQ of 0.5 ng ml(-1). Recoveries were in range 75-114%, while matrix effects were minimal. The experimental study was carried out on three groups of male Friesian veal calves: group PL (n = 6, PL acetate 15 mg day(-1) p.o. for 31 days); group DX (n = 5, 5 mg of estradiol (E2) i.m., weekly, and 0.4 mg day(-1) of DX p.o. for 31 days) and a control group (n = 8). Urine was collected before, during (twice) and at the end of treatment. During PL administration, the tetrahydro-metabolite levels decreased gradually and remained low after the suspension of treatment. DX reduced urinary THF that persisted after the treatment, while THE levels decreased during the experiment, but rebounded substantially after the DX was withdrawn. Both DX and PL significantly interfered with the production of F and E, leading to their complete depletion. Taken together, the results demonstrate the influence of DX and PL administration on 11βHSD activity and their impact on dysfunction of the 5-reductase pathway. In conclusion, profiling tetrahydro-metabolites of F and E might serve as an alternative, indirect but reliable, non-invasive procedure for assessing the impact of synthetic glucocorticosteroids administration.
- Published
- 2016
- Full Text
- View/download PDF
38. A Liquid Chromatography-Tandem Mass Spectrometry Method for the Detection of Antimicrobial Agents from Seven Classes in Calf Milk Replacers: Validation and Application.
- Author
-
Chiesa LM, Nobile M, Panseri S, Biolatti B, Cannizzo FT, Pavlovic R, and Arioli F
- Subjects
- Amoxicillin analysis, Amoxicillin chemistry, Animals, Anti-Infective Agents chemistry, Cattle, Cephalosporins analysis, Cephalosporins chemistry, Chromatography, High Pressure Liquid, Fluoroquinolones analysis, Fluoroquinolones chemistry, Lincosamides analysis, Lincosamides chemistry, Molecular Structure, Penicillins analysis, Penicillins chemistry, Sulfonamides analysis, Sulfonamides chemistry, Tetracyclines analysis, Tetracyclines chemistry, Anti-Infective Agents analysis, Chromatography, Liquid methods, Food, Formulated microbiology, Milk microbiology, Tandem Mass Spectrometry methods
- Abstract
Calf milk replacers are low-cost feeds that contain available, digestible protein. During their reconstitution, however, the addition of drugs, such as antibiotics, could make them a very simple route for illicit treatment for therapeutic, preventive, or growth-promoting purposes. We developed an HPLC-MS/MS method, preceded by a unique extraction step, able to identify 17 antibiotics from seven classes (penicillins, tetracyclines, fluoroquinolones, sulfonamides, cephalosporins, amphenicols, and lincosamides) in this matrix. Prior to solid phase extraction (SPE), the sample underwent deproteinization and defatting. The method was fully validated according to Commission Decision 2002/657/EC. Decision limits (CCα) and detection capability (CCβ) were in the ranges of 0.13-1.26 and 0.15-1.47 ng/mL, respectively. Thirty-eight samples were finally analyzed, showing the occasional presence of marbofloxacin (six samples) and amoxicillin (one sample).
- Published
- 2016
- Full Text
- View/download PDF
39. Plasmodium spp. In a captive raptor collection of a Safaripark in northwest Italy.
- Author
-
Scaglione FE, Cannizzo FT, Chiappino L, Sereno A, Ripepi M, Salamida S, Manuali E, and Bollo E
- Subjects
- Animals, Antimalarials therapeutic use, Chloroquine therapeutic use, Italy epidemiology, Malaria, Avian drug therapy, Malaria, Avian parasitology, Animals, Zoo, Falconiformes, Malaria, Avian mortality, Plasmodium isolation & purification, Strigiformes
- Abstract
Blood parasites infect all vertebrates (Clayton and Moore 1997). Avian malaria parasites (Plasmodium spp., Plasmodiidae) are cosmopolitan in their distribution and are responsible for severe diseases in domestic and wild birds.In September 2009, nine raptorial birds that either arrived recently or were maintained as permanent residents at the Safaripark Pombia (northwest Italy) showed loss of stamina, developing listlessness, anorexia and regurgitation. Within one month three animals died and were necropsied.Following the diagnosis of Plasmodium infection all other raptorial birds were treated: clinical improvement was observed in all birds, and blood smears made after one month resulted negative for parasites.
- Published
- 2016
- Full Text
- View/download PDF
40. Regucalcin Expression as a Diagnostic Tool for the Illicit Use of Steroids in Veal Calves.
- Author
-
Starvaggi Cucuzza L, Biolatti B, Sereno A, and Cannizzo FT
- Subjects
- Androgens administration & dosage, Androgens metabolism, Animals, Bulbourethral Glands drug effects, Bulbourethral Glands metabolism, Calcium-Binding Proteins metabolism, Cattle, Gonadal Steroid Hormones administration & dosage, Male, Prostate drug effects, Prostate metabolism, Testis drug effects, Testis metabolism, Calcium-Binding Proteins genetics, Gonadal Steroid Hormones metabolism, Substance Abuse Detection veterinary
- Abstract
It has been previously demonstrated that sex steroid hormone treatment down-regulates regucalcin gene expression in the accessory sex glands and testis of prepubertal and adult male bovines. The aim of this study was to investigate whether low doses of sex steroid hormones combined with other drugs significantly affect regucalcin gene expression in the accessory sex glands and testis of veal calves. The regucalcin expression was down-regulated in the bulbo-urethral glands of estrogen-treated calves, whereas it was up-regulated in the prostate of estrogen-treated calves. Only the testis of androgen-treated calves showed a down-regulation of the regucalcin expression. Thus, the administration of sex steroid hormones, even in low doses and combined with other molecules, could affect regucalcin expression in target organs. Particularly, the specific response in the testis suggests regucalcin expression in this organ as a first molecular biomarker of illicit androgen administration in bovine husbandry.
- Published
- 2015
- Full Text
- View/download PDF
41. Proteomic identification of plasma proteins as markers of growth promoter abuse in cattle.
- Author
-
Kinkead RA, Elliott CT, Cannizzo FT, Biolatti B, and Mooney MH
- Subjects
- Anabolic Agents administration & dosage, Animals, Biomarkers analysis, Contraceptive Agents administration & dosage, Dexamethasone administration & dosage, Dexamethasone analogs & derivatives, Electrophoresis, Gel, Two-Dimensional methods, Estradiol administration & dosage, Estradiol analogs & derivatives, Glucocorticoids administration & dosage, Male, Prednisolone administration & dosage, Prednisolone analogs & derivatives, Biomarkers blood, Blood Proteins analysis, Cattle blood, Proteomics methods, Substance Abuse Detection methods, Tandem Mass Spectrometry methods
- Abstract
Growth-promoting agents are continually misused for increasing animal growth and fraudulent gain in the meat industry, yet detection rates from conventional targeted testing for drug residues do not reflect this. This is because testing currently relies on direct detection of drugs or related metabolites and administrators of such compounds can take adaptive measures to avoid detection through the use of endogenous or unknown drugs, and low dose or combined mixtures. New detection methods are needed which focus on the screening of biological responses of an animal to such growth-promoting agents as it has been demonstrated that genomic, proteomic and metabolomics profiles are altered by xenobiotic intake. Therefore, an untargeted proteomics approach using comparative two-dimensional gel electrophoresis (2DE) was carried out to identify putative proteins altered in plasma after treatment with oestradiol, dexamethasone or prednisolone. Twenty-four male cattle were randomly assigned to four groups (n = 6) for experimental treatment over 40 days, namely a control group of non-treated cattle, and three groups administered 17β-oestradiol-3-benzoate (0.01 mg/kg, intramuscular), dexamethasone sodium phosphate (0.7 mg/day, per os) or prednisolone acetate (15 mg/day, per os), respectively. Plasma collected from each animal at day 25 post study initiation was subjected to proteomic analysis by 2DE for comparison of protein expression between treated and untreated animals. Analysis of acquired gel images revealed 22 plasma proteins which differed in expression by more than 50% (p < 0.05) in treated animals compared to untreated animals. Proteins of interest underwent identification by LC-MS/MS analysis and were found to have associated roles in transport, blood coagulation, immune response and metabolism pathways. In this way, seven proteins are highlighted as novel biomarker candidates including transthyretin which is shown to be significantly increased in all treatment groups compared to control animals and potentially may find use as global markers of suspect anabolic practice.
- Published
- 2015
- Full Text
- View/download PDF
42. Prevalence of new and known species of haemoparasites in feral pigeons in northwest Italy.
- Author
-
Scaglione FE, Pregel P, Cannizzo FT, Pérez-Rodríguez AD, Ferroglio E, and Bollo E
- Subjects
- Age Factors, Animals, Coinfection, Female, Haemosporida classification, Italy epidemiology, Malaria, Avian epidemiology, Malaria, Avian parasitology, Male, Plasmodium classification, Plasmodium isolation & purification, Polymerase Chain Reaction veterinary, Prevalence, Sex Factors, Spleen parasitology, Bird Diseases epidemiology, Bird Diseases parasitology, Columbidae, Haemosporida isolation & purification, Protozoan Infections epidemiology, Protozoan Infections parasitology
- Abstract
Background: Haemoparasites in feral pigeons have been studied in several countries but no data are available from Italy. The aim of this work was to evaluate the prevalence and diversity of Haemoproteus spp./Plasmodium spp. and Leucocytozoon spp. in feral pigeons from northwest Italy, as well as the association between infection and host age or sex., Methods: Feral pigeons were collected during a regional culling programme from the Piedmont region (northwest Italy) and subjected to necropsy. Infections were detected from DNA extracted from the spleen following a nested PCR protocol. The association between sex or age and infection status was evaluated using the chi-squared test for independence or Fisher's exact test., Results: Out of 51 animals, 15 were positive for Haemoproteus/Plasmodium spp. and eight for Leucocytozoon spp., with a significant difference between haemoparasites prevalence. There was no significant association between age or sex and infection status. The coinfection with different haemoparasites was very significant (p < 0.01), showing a greater relative risk to be infected by a second haemoparasite in birds already infected, in particular in male and in adult pigeons. DNA sequencing of Leucocytozoon spp. showed six different lineages in pigeons, and one of Haemoproteus and Plasmodium, respectively., Conclusions: Blood parasites are continuously circulating around the world, and the results presented in the paper suggest that cross infection of feral pigeons with haemoparasites typical of other migratory or nonmigratory bird species is possible. Moreover, the geographical location of Italy along the main migratory routes is a crucial factor to be considered for migratory birds, because they can be affected by blood parasites detected in feral pigeons, and vice versa.
- Published
- 2015
- Full Text
- View/download PDF
43. Toxicogenomic markers for corticosteroid treatment in beef cattle: integrated analysis of transcriptomic data.
- Author
-
Pegolo S, Di Camillo B, Montesissa C, Cannizzo FT, Biolatti B, and Bargelloni L
- Subjects
- Administration, Oral, Animals, Breeding, Cattle, Male, Models, Molecular, Muscle, Skeletal drug effects, Oligonucleotide Array Sequence Analysis, Support Vector Machine, Transcriptome, Adrenal Cortex Hormones pharmacology, Gene Expression Profiling methods, Genetic Markers, Muscle, Skeletal metabolism, Toxicogenetics methods
- Abstract
In the present work, an integrated analysis was performed on DNA-microarray data of bovine muscle samples belonging to controls, animals treated with various growth promoters (GPs) and unknown commercial samples. The aim was identify a robust gene expression signature of corticosteroid treatment for the classification of commercial samples, despite the effects of biological variation and other confounding factors. DNA-Microarray data from 5 different batches of bovine skeletal muscle samples were analyzed (146 samples). After preprocessing, expression data from animals treated with corticosteroids and controls from the different batches (89 samples) were used to train a Support Vector Machines (SVMs) classifier. The optimal number of gene probes chosen by our classification framework was 73. The SVMs with linear kernel built on these 73 biomarker genes was predicted to perform on novel samples with a high classification accuracy (Matthew's correlation coefficient equal to 0.77) and an average percentage of false positive and false negative equal to 5% and 6%, respectively. Concluding, a relatively small set of genes was able to discriminate between controls and corticosteroid-treated animals, despite different breeds, animal ages, and combination of GPs. The results are extremely promising, suggesting that integrated analysis provides robust transcriptomic signatures for GP abuse., (Copyright © 2014 Elsevier Ltd. All rights reserved.)
- Published
- 2015
- Full Text
- View/download PDF
44. Pseudoendogenous presence of β-boldenone sulphate and glucuronide in untreated young bulls from the food chain.
- Author
-
Chiesa L, Pasquale E, Panseri S, Cannizzo FT, Biolatti B, Pavlovic R, and Arioli F
- Subjects
- Abattoirs, Animals, Biomarkers, Cattle, Chromatography, High Pressure Liquid, Food Contamination analysis, Male, Tandem Mass Spectrometry, Testosterone urine, Anabolic Agents urine, Drug Residues analysis, Glucuronides urine, Testosterone analogs & derivatives, Veterinary Drugs analysis
- Abstract
The administration of boldenone (bold) to bovines, either for growth promotion or therapeutic purposes, has been banned in the EU since 1981. It is, however, a pseudoendogenous hormone, thus its detection in bovine urine, in the form of α-boldenone conjugates, is considered fully compliant up to 2 ng ml(-1). Greater attention has been placed on β-boldenone, the anabolic active epimer, whose conjugated form must be absent in urine. Recently, the identification of a biomarker representing unquestionable evidence of illicit treatment with bold or its precursor androstadienedione has been a major topic in the literature regarding the detection of residues in bovine urine, and β-boldenone sulphate is a candidate molecule. In this study, we used a method previously validated according to the European Commission Decision 2002/657/EC for the determination of sulphate and glucuronide conjugates of β-boldenone. We assessed the occurrence of these molecules in young bull urine, with the aim of understanding whether they could be of endogenous origin, and to check for a possible relationship with particular environmental and stress conditions. Urine samples from 56 young bulls were collected after transport stress, under non-stressful conditions and after transport and slaughter stress. Histopathological investigation of the hormone target organs, i.e. the bulbourethral and prostate glands, was also performed. The results indicate an inverse relationship between the presence and concentration of β-boldenone sulpho- and gluco-conjugates in urine, and stress conditions, expressed by the absence of detection at the slaughterhouse. No significant macroscopic and histologic lesions were detected. Our study indicates that β-boldenone sulphate could be a biomarker of treatment only at the slaughterhouse, while at the farm, in untreated animals (i.e. after a five-month period under the control of Official Veterinarians), sulphate and glucuronide metabolites were found with a frequency of 78% and 46%, respectively, showing the endogenous origin of boldenone.
- Published
- 2015
- Full Text
- View/download PDF
45. Regucalcin expression in bovine tissues and its regulation by sex steroid hormones in accessory sex glands.
- Author
-
Starvaggi Cucuzza L, Divari S, Mulasso C, Biolatti B, and Cannizzo FT
- Subjects
- Animals, Cattle, Male, Organ Specificity drug effects, Rats, Androgens pharmacology, Calcium-Binding Proteins biosynthesis, Estrogens pharmacology, Gene Expression Regulation drug effects, Prostate metabolism, Testis metabolism
- Abstract
Regucalcin (RGN) is a mammalian Ca2+-binding protein that plays an important role in intracellular Ca2+ homeostasis. Recently, RGN has been identified as a target gene for sex steroid hormones in the prostate glands and testis of rats and humans, but no studies have focused on RGN expression in bovine tissues. Thus, in the present study, we examined RGN mRNA and protein expression in the different tissues and organs of veal calves and beef cattle. Moreover, we investigated whether RGN expression is controlled through sex steroid hormones in bovine target tissues, namely the bulbo-urethral and prostate glands and the testis. Sex steroid hormones are still illegally used in bovine husbandry to increase muscle mass. The screening of the regulation and function of anabolic sex steroids via modified gene expression levels in various tissues represents a new approach for the detection of illicit drug treatments. Herein, we used quantitative PCR, western blot and immunohistochemistry analyses to demonstrate RGN mRNA and protein expression in bovine tissues. In addition, estrogen administration down-regulated RGN gene expression in the accessory sex glands of veal calves and beef cattle, while androgen treatment reduced RGN gene expression only in the testis. The confirmation of the regulation of RGN gene expression through sex steroid hormones might facilitate the potential detection of hormone abuse in bovine husbandry. Particularly, the specific response in the testis suggests that this tissue is ideal for the detection of illicit androgen administration in veal calves and beef cattle.
- Published
- 2014
- Full Text
- View/download PDF
46. Haematological and physiological responses of Piemontese beef cattle to different housing conditions.
- Author
-
Starvaggi Cucuzza L, Riondato F, Macchi E, Bellino C, Franco G, Biolatti B, and Cannizzo FT
- Subjects
- Animals, Biomarkers blood, Corticosterone blood, Disease Susceptibility blood, Disease Susceptibility physiopathology, Disease Susceptibility veterinary, Hydrocortisone blood, Male, Muramidase blood, Receptors, Glucocorticoid blood, Animal Welfare, Cattle blood, Cattle physiology, Housing, Animal, Stress, Physiological physiology
- Abstract
Public concern for animal welfare has progressively grown over the recent years. In this context, stress has a great economical impact on growth of animals and quality of animal products. The development and validation of methods to assess animal stress, particularly at the farm level, are desirable to evaluate animal production systems. Piemontese breed is traditionally tie-stall housed in the fattening period. Hence, the objective of this study was to characterise a profile of physiological and haematological changes of Piemontese beef cattle under different management conditions (tie-stall and loose housing). Our results suggest that the housing system is an important factor in animal welfare. Indeed, the values of the total protein, lysozyme, cortisol, serum and faecal corticosterone concentration and GR-α gene expression indicate that the tie-stall housing is more stressful than the loose system. All the alterations highlighted in this study considered together may be effective biomarkers of stress and disease susceptibility., (Copyright © 2014 Elsevier Ltd. All rights reserved.)
- Published
- 2014
- Full Text
- View/download PDF
47. Transcriptomic profiling as a screening tool to detect trenbolone treatment in beef cattle.
- Author
-
Pegolo S, Cannizzo FT, Biolatti B, Castagnaro M, and Bargelloni L
- Subjects
- Anabolic Agents administration & dosage, Animals, Body Weight drug effects, Male, Oligonucleotide Array Sequence Analysis, Polymerase Chain Reaction veterinary, RNA, Messenger chemistry, RNA, Messenger genetics, Random Allocation, Transcriptome genetics, Trenbolone Acetate administration & dosage, Anabolic Agents pharmacology, Cattle metabolism, Muscle, Skeletal metabolism, Trenbolone Acetate pharmacology
- Abstract
The effects of steroid hormone implants containing trenbolone alone (Finaplix-H), combined with 17β-oestradiol (17β-E; Revalor-H), or with 17β-E and dexamethasone (Revalor-H plus dexamethasone per os) on the bovine muscle transcriptome were examined by DNA-microarray. Overall, large sets of genes were shown to be modulated by the different growth promoters (GPs) and the regulated pathways and biological processes were mostly shared among the treatment groups. Using the Prediction Analysis of Microarray program, GP-treated animals were accurately identified by a small number of predictive genes. A meta-analysis approach was also carried out for the Revalor group to potentially increase the robustness of class prediction analysis. After data pre-processing, a high level of accuracy (90%) was obtained in the classification of samples, using 105 predictive gene markers. Transcriptomics could thus help in the identification of indirect biomarkers for anabolic treatment in beef cattle to be applied for the screening of muscle samples collected after slaughtering., (Copyright © 2014 Elsevier Ltd. All rights reserved.)
- Published
- 2014
- Full Text
- View/download PDF
48. Oxytocin precursor gene expression in bovine skeletal muscle is regulated by 17β-oestradiol and dexamethasone.
- Author
-
Divari S, Pregel P, Cannizzo FT, Starvaggi Cucuzza L, Brina N, and Biolatti B
- Subjects
- Animals, Cattle blood, Cattle metabolism, Dexamethasone metabolism, NIMA-Interacting Peptidylprolyl Isomerase, Oxytocin blood, Peptidylprolyl Isomerase metabolism, Cattle genetics, Estradiol metabolism, Gene Expression, Meat analysis, Muscle, Skeletal metabolism, Oxytocin biosynthesis, Peptidylprolyl Isomerase genetics, Up-Regulation
- Abstract
Growth promoter administration, in livestock, potentially poses a major threat to public health, due to the potential endocrine and carcinogenic activity of residues, accumulating in edible tissues, such as skeletal muscle. Therefore, development of new screening tests and methods for the detection of illicit treatments of food animals would be useful. In this study the serum concentrations of oxytocin peptide were measured in beef cattle receiving 17β oestradiol, dexamethasone or placebo over a period of 40 days. Changes in gene expression of oxytocin precursor in skeletal muscle were also examined in these animals. Serum analysis using an oxytocin EIA kit indicated a significant up-regulation of the biosynthesis of this nonapeptide only in cattle after 17β oestradiol, but not after dexamethasone or placebo treatment. Quantitative PCR (qPCR) analysis showed a significant overexpression of the oxytocin precursor gene by 33.5 and 13.3-fold in cattle treated with 17β oestradiol and dexamethasone, respectively, in comparison to placebo treated animals. Regulation of gene expression by some myogenic regulatory factors in skeletal muscle was also evaluated in these animal groups, confirming the activity of both growth promoters on this gene. To investigate the use of the oxytocin precursor gene as biomarker for 17β oestradiol and dexamethasone treatment in beef cattle, an absolute quantification of this gene by qPCR was developed. A standard curve was generated and developed with TaqMan® technology and optimal criterion value, sensitivity and specificity of this screening method were established through ROC analysis. This analysis suggested that the up-regulation of oxytocin precursor gene expression in skeletal muscle tissue is a valid marker for detection of illicit 17β oestradiol and/or dexamethasone use in beef cattle. This method may serve as a novel diagnostic tool in the screening phase, and, if introduced in routine testing, may significantly improve overall efficacy and success of the food screening process ordered by state authorities., (Copyright © 2013 Elsevier Ltd. All rights reserved.)
- Published
- 2013
- Full Text
- View/download PDF
49. Gene expression profiling of thymus in beef cattle treated with prednisolone.
- Author
-
Cannizzo FT, Pegolo S, Starvaggi Cucuzza L, Bargelloni L, Divari S, Franch R, Castagnaro M, and Biolatti B
- Subjects
- Animals, Gene Expression Regulation drug effects, Male, Cattle metabolism, Prednisolone pharmacology, Thymus Gland drug effects, Thymus Gland metabolism, Transcriptome
- Abstract
Glucocorticoids (GCs) are extensively used in livestock production, not only for their anti-inflammatory properties but also to improve the quality and quantity of meat in veal and beef production. In Italy, an increase in GC-positive cases has been observed in cattle since 2008, particularly prednisolone (PDN). Recent studies clearly demonstrate that both histopathological analysis and high-performance liquid chromatography tandem mass spectrometry (HPLC/MS-MS) were unable to detect PDN treatments. The aim of this study was to identify transcriptomic signatures of PDN administration in the thymus of experimentally treated animals by comparison with untreated controls, in order to identify gene expression changes or pathways alteration induced by the corticosteroid treatment. Microarray data analysis showed substantial modifications in thymus gene expression profiles after PDN treatment. Several of the 388 differentially expressed genes encoded pro-inflammatory and anti-inflammatory mediators or immune regulators which showed that PDN might have a role in the regulation of immunologic homeostasis, act on both innate and acquired components of the immunity and mainly induce the activation of immune tolerance and anti-inflammatory pathways. Thus, this study allowed to deepen the effects of PDN on the immune system and showed the potentiality of gene expression profiling by DNA-microarray as a powerful tool to complement the existing methods against the illegal use of growth promoting hormones, especially when working on samples collected after slaughtering., (Copyright © 2013 Elsevier Ltd. All rights reserved.)
- Published
- 2013
- Full Text
- View/download PDF
50. Potential of treatment-specific protein biomarker profiles for detection of hormone abuse in cattle.
- Author
-
Ludwig SK, Smits NG, Cannizzo FT, and Nielen MW
- Subjects
- Animals, Antibodies blood, Cattle, Dexamethasone blood, Estradiol blood, Feasibility Studies, Flow Cytometry, Immunoassay, Insulin-Like Growth Factor Binding Protein 2 blood, Insulin-Like Growth Factor I metabolism, Male, Osteocalcin blood, Prednisolone blood, Reproducibility of Results, Biomarkers blood, Growth Hormone blood
- Abstract
Targeted protein biomarker profiling is suggested as a fast screening approach for detection of illegal hormone treatment in meat production. The advantage of using biomarkers is that they mark the biological response and, thus, are responsive to a panel of substances with similar effects. In a preliminary feasibility study, a 4-plex protein biomarker flow cytometric immunoassay (FCIA) previously developed for the detection of recombinant bovine somatotropin (rbST) was applied to cattle treated with steroids, such as estradiol, dexamethasone, and prednisolone. Each treatment resulted in a specific plasma biomarker profile for insulin-like growth factor-1 (IGF-1), IGF binding protein 2, osteocalcin, and anti-rbST antibodies, which could be distinguished from the profile of untreated animals. In summary, the 4-plex biomarker FCIA is, apart from rbST, also capable of detecting treatment with other growth-promoting agents and therefore clearly shows the potential of biomarker profiling as a screening method in veterinary control. It is proposed to perform additional validation studies covering high numbers of treated and untreated animals to support inclusion or adaptation of protein biomarker approaches in future monitoring regulations.
- Published
- 2013
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.