31 results on '"Marogna G"'
Search Results
2. Identification of coagulase-negative staphylococci isolated from ovine milk samples by PCR–RFLP of 16S rRNA and gap genes
- Author
-
Onni, T., Sanna, G., Cubeddu, G.P., Marogna, G., Lollai, S., Leori, G., and Tola, S.
- Published
- 2010
- Full Text
- View/download PDF
3. Outbreak of mastitis in sheep caused by multi-drug resistant Enterococcus faecalis in Sardinia, Italy
- Author
-
SANCIU, G., MAROGNA, G., PAGLIETTI, B., CAPPUCCINELLI, P., LEORI, G., and RAPPELLI, P.
- Published
- 2013
4. Evaluation of tet genes distribution in bacteria isolated from milk samples of ovine herds: P1779
- Author
-
Lollai, S., Ziccheddu, M., Manunta, D., Zara, P., Marogna, G., and Leori, G.
- Published
- 2005
5. Genetic data on alghero population (sardinia): contrast between biological and cultural evidence
- Author
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Moral, P., Marogna, G., Salis, M., Succa, V., and Vona, G.
- Subjects
Genetic polymorphisms -- Research ,Human population genetics -- Research ,Anthropology/archeology/folklore - Abstract
Data on 20 genetic polymorphisms (61 alleles) in the Algehero population on the northwestern coast of Sardinia are presented and discussed in relation to its linguistic peculiarity inside the island. Since the Aragonese (Spain) conquest of Sardinia in the 13th century, the Catalan language, the same as that spoken in Northeastern Spain and certain districts of Southern France, has been used in Alghero even until today. Analysis for heterogeneity of gene frequency distributions indicates that the genetic information obtained on Alghero is adequate to discriminate Sardinians from other neighbouring populations. Genetic variation between populations measured through genetic distances and principal-component analysis shows that the present-day population of Alghero is much closer genetically to Sardinians than to Catalonians. Our genetic results do not support any interpretation of the linguistic affinities between Alghero and Catalonia at present as indicative of biological kinship.
- Published
- 1994
6. Glucose-6-Phosphate Dehydrogenase Deficiency and Blood Groups in Northern Sardinia
- Author
-
Veronese, F. Martuzzi, Stangoni, A., Cuccuru, G.B., Mulas, G., Marogna, G., Cossu, G., and Pettener, D.
- Published
- 1985
7. Variazioni dei livelli ematici di leptina, GH e del peso corporeo in agnelli di razza Sarda in accrescimento
- Author
-
Carcangiu V., Vacca G. M., Marogna G., Rapposelli E., Bini P. P., PARMEGGIANI, ALBAMARIA, Carcangiu V., Vacca G.M., Parmeggiani A., Marogna G., Rapposelli E., and Bini P.P.
- Subjects
LEPTIN ,SARDA BREED LAMBS ,GROWTH HORMONE ,BODY WEIGHT ,PHYSIOLOGY - Abstract
The growing period is very important in the life of the animal, since it may affect its productive and reproductive carreer. In this phase, many hormones are involved in regulating metabolism, namely GH, but also leptin could play an important role, considering its tissue of origin. In order to investigate leptin plasmatic levels and its correlations with GH and body weight, a research was carried out on 20 Sarda breed lambs. Blood samples were withdrawn fortnightly, from day 40° to day 210°, and the body weight of each animal was registered. Animals grazed during the day and in the night they were kept in the sheep pen where they received 300 g/head of concentrated feed containing 16% protein and 12.5% ME/kg of dry matter. Hay and water were ad libitum. Leptin and GH were analysed by RIA. The data were submitted to correlation and variance analysis. Leptin plasmatic levels displayed an increasing trend until 11th blood withdrawal, in correspondence with the end of the greatest body weight increase. GH blood levels showed a decreasing trend (P
- Published
- 2004
8. Characterization of size and composition of milk fat globules from Sarda and Saanen dairy goats
- Author
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Pisanu, S., primary, Marogna, G., additional, Pagnozzi, D., additional, Piccinini, M., additional, Leo, G., additional, Tanca, A., additional, Roggio, A.M., additional, Roggio, T., additional, Uzzau, S., additional, and Addis, M.F., additional
- Published
- 2013
- Full Text
- View/download PDF
9. Outbreak of mastitis in sheep caused by multi-drug resistantEnterococcus faecalisin Sardinia, Italy
- Author
-
SANCIU, G., primary, MAROGNA, G., additional, PAGLIETTI, B., additional, CAPPUCCINELLI, P., additional, LEORI, G., additional, and RAPPELLI, P., additional
- Published
- 2012
- Full Text
- View/download PDF
10. Identification of coagulase-negative staphylococci isolated from caprine milk samples by PCR-RFLP of groEL gene
- Author
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Onni, T., primary, Vidili, A., additional, Bandino, E., additional, Marogna, G., additional, Schianchi, S., additional, and Tola, S., additional
- Published
- 2012
- Full Text
- View/download PDF
11. Comparison of clinical findings, microbiological results, and farming parameters in goat herds affected by recurrent infectious mastitis
- Author
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Marogna, G., primary, Pilo, C., additional, Vidili, A., additional, Tola, S., additional, Schianchi, G., additional, and Leori, S.G., additional
- Published
- 2012
- Full Text
- View/download PDF
12. Antibiotics mastitis therapy: drug residue in lambs
- Author
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Testa, C., primary, Marogna, G., additional, Secchi, L., additional, Rubattu, N., additional, Leori, G., additional, and Calaresu, G., additional
- Published
- 2010
- Full Text
- View/download PDF
13. Ovine embryos viability after serum replacement with polyvinyl alcohol in vitrification solution
- Author
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Naitana, S., primary, Ledda, S., additional, Bogliolo, L., additional, Marogna, G., additional, Leoni, G., additional, Cappai, P., additional, and Loi, L., additional
- Published
- 1997
- Full Text
- View/download PDF
14. Heat shock protein 70 in vitrified ovine embryo
- Author
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Leoni, G., primary, Ledda, S., additional, Bogliolo, L., additional, Marogna, G., additional, and Naitana, S., additional
- Published
- 1997
- Full Text
- View/download PDF
15. Glucose-6-Phosphate Dehydrogenase Deficiency and Blood Groups in Northern Sardinia.
- Author
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Martuzzi Veronesi, F., Stangoni, A., Cuccuru, G.B., Mulas, G., Marogna, G., Cossu, G., and Pettener, D.
- Published
- 1985
- Full Text
- View/download PDF
16. Paesaggi urbani dell'Ottocento
- Author
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Riccini, Raimonda and Marogna, G.
- Subjects
grandi magazzini ,città-capitale ,esposizioni ,merce - Published
- 1981
17. Glucose-6-Phosphate Dehydrogenase Deficiency and Blood Groups in Northern Sardinia
- Author
-
Veronesi, Martuzzi, primary, Stangoni, A., additional, Cuccuru, G.B., additional, Mulas, G., additional, Marogna, G., additional, Cossu, G., additional, and Pettener, D., additional
- Published
- 1985
- Full Text
- View/download PDF
18. Antibiotics mastitis therapy: drug residue in lambs
- Author
-
Testa, C., Marogna, G., Secchi, L., Rubattu, N., Leori, G., and Calaresu, G.
- Abstract
AbstractMeat coming from suckling lambs (max 12 Kg BW) is a typical Sardinian taste dish, normally consumed during the religious linked feasts. The aim of this work is to evaluate drug residues in suckling lambs meat as consequence of antibiotics mastitis therapy in their mothers during lactation.The study was performed on twelve Sardinian ewes, that had lambed within two days of one another, with suckling lambs from a single flok of 150 animals. Milk bacteriological screening showed that 10 ewes out of 12 were positive and Coagulase-Negative Staphylococci were identified. The ten positive sheep were divided into two groups A and B, and each of them were given two successive intramuscularconsecutive intramuscolar injection of 6 ml of oxytetracycline OTC (Terralon 20% LA, Virbac) within 72 hours; the two negative sheep were used as control (C group). With the two ewes Group two trials were conducted: to the A treatment started 17 days after delivery, while group of 6 ewes, drug administration was given when lambs were 17 days old, to the B has been treated 28 days after delivery. During the experimental period milk was sampled twice a week; 5 milk samples/ewe for group A and 2 samples/ewe for group B were collected. Lambslambs were regularly slaughtered at about 35 days old and muscle tissue has been collected.sampled. All samples were immediately frozen until analysis. Oxytetracycline milk residues were measured by High Performance Liquid Chromatography with diode array detector while, for OTC tissue levels, LC/MS-MS technique was used. OTC concentration in milk, as observed in our own previous study, decreasedranged from 3,500 to 0,050 µ g/ml over three weeks. OTC residues were detected in both groups of lambs at levels below Maximum Residue Limit (MRL 0.100 µg/g). In order to avoid any drug residue in food chain, and an increase of drug resistance, national legislation should pay attention to avoid use of antibiotics in ewes feeding lambs that will be slaughtered.
- Published
- 2007
- Full Text
- View/download PDF
19. Production and Release of Antimicrobial and Immune Defense Proteins by Mammary Epithelial Cells following Streptococcus uberis Infection of Sheep
- Author
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Carla Cacciotto, Maria Filippa Addis, Salvatore Pisanu, Daniela Pagnozzi, Stefano Rocca, Sergio Uzzau, Franca Campesi, Gavino Marogna, Tiziana Cubeddu, Giuseppe Martino Schianchi, Addis M.F., Pisanu S., Marogna G., Cubeddu T., Pagnozzi D., Cacciotto C., Campesi F., Schianchi G., Rocca S., and Uzzau S.
- Subjects
Proteomics ,medicine.medical_treatment ,Immunology ,Mammary gland ,Inflammation ,Microbiology ,Cathelicidin ,Mammary Glands, Animal ,Immune system ,Anti-Infective Agents ,Cathelicidins ,Streptococcal Infections ,medicine ,Animals ,Lactation ,Mastitis, antimicrobial peptides, proteomics ,Udder ,Glycoproteins ,Streptococcus uberis ,Host Response and Inflammation ,Sheep ,Innate immune system ,biology ,Streptococcus ,Epithelial Cells ,Lipid Droplets ,biology.organism_classification ,Milk ,Infectious Diseases ,medicine.anatomical_structure ,Parasitology ,Glycolipids ,Calprotectin ,medicine.symptom ,Leukocyte L1 Antigen Complex ,Antimicrobial Cationic Peptides - Abstract
Investigating the innate immune response mediators released in milk has manifold implications, spanning from elucidation of the role played by mammary epithelial cells (MECs) in fighting microbial infections to the discovery of novel diagnostic markers for monitoring udder health in dairy animals. Here, we investigated the mammary gland response following a two-step experimental infection of lactating sheep with the mastitis-associated bacterium Streptococcus uberis . The establishment of infection was confirmed both clinically and by molecular methods, including PCR and fluorescent in situ hybridization of mammary tissues. Proteomic investigation of the milk fat globule (MFG), a complex vesicle released by lactating MECs, enabled detection of enrichment of several proteins involved in inflammation, chemotaxis of immune cells, and antimicrobial defense, including cathelicidins and calprotectin (S100A8/S100A9), in infected animals, suggesting the consistent involvement of MECs in the innate immune response to pathogens. The ability of MECs to produce and release antimicrobial and immune defense proteins was then demonstrated by immunohistochemistry and confocal immunomicroscopy of cathelicidin and the calprotectin subunit S100A9 on mammary tissues. The time course of their release in milk was also assessed by Western immunoblotting along the course of the experimental infection, revealing the rapid increase of these proteins in the MFG fraction in response to the presence of bacteria. Our results support an active role of MECs in the innate immune response of the mammary gland and provide new potential for the development of novel and more sensitive tools for monitoring mastitis in dairy animals.
- Published
- 2013
20. Neutrophil extracellular traps in sheep mastitis
- Author
-
Carla Cacciotto, Stefano Rocca, Sergio Uzzau, Daniela Pagnozzi, Tiziana Cubeddu, Alberto Alberti, Maria Filippa Addis, Gavino Marogna, Salvatore Pisanu, Pisanu S., Cubeddu T., Pagnozzi D., Rocca S., Cacciotto C., Alberti A., Marogna G., Uzzau S., and Addis M.F.
- Subjects
Proteases ,Extracellular Traps ,Neutrophils ,[SDV]Life Sciences [q-bio] ,Sheep Diseases ,Mastitis ,Biology ,Microbiology ,03 medical and health sciences ,chemistry.chemical_compound ,Mammary Glands, Animal ,Streptococcal Infections ,Extracellular ,Animals ,Humans ,Mastitis, neutrophils, amps, immunity ,030304 developmental biology ,0303 health sciences ,Sheep ,General Veterinary ,030302 biochemistry & molecular biology ,Citrullination ,Streptococcus ,Neutrophil extracellular traps ,veterinary(all) ,Histone ,Milk ,Biochemistry ,chemistry ,Nucleic acid ,biology.protein ,Female ,DNA ,Research Article - Abstract
Neutrophil extracellular traps (NETs) are structures composed of DNA, histones, and antimicrobial proteins that are released extracellularly by neutrophils and other immune cells as a means for trapping and killing invading pathogens. Here, we describe NET formation in milk and in mammary alveoli of mastitic sheep, and provide a dataset of proteins found in association to these structures. Nucleic acid staining, immunomicroscopy and fluorescent in-situ hybridization of mastitic mammary tissue from sheep infected with Streptococcus uberis demonstrated the presence of extranuclear DNA colocalizing with antimicrobial proteins, histones, and bacteria. Then, proteomic analysis by LTQ-Orbitrap Velos mass spectrometry provided detailed information on protein abundance changes occurring in milk upon infection. As a result, 1095 unique proteins were identified, of which 287 being significantly more abundant in mastitic milk. Upon protein ontology classification, the most represented localization classes for upregulated proteins were the cytoplasmic granule, the nucleus, and the mitochondrion, while function classes were mostly related to immune defence and inflammation pathways. All known NET markers were massively increased, including histones, granule proteases, and antimicrobial proteins. Of note was the detection of protein arginine deiminases (PAD3 and PAD4). These enzymes are responsible for citrullination, the post-translational modification that is known to trigger NET formation by inducing chromatin decondensation and extracellular release of NETs. As a further observation, citrullinated residues were detected by tandem mass spectrometry in histones of samples from mastitic animals. In conclusion, this work provides novel microscopic and proteomic information on NETs formed in vivo in the mammary gland, and reports the most complete database of proteins increased in milk upon bacterial mastitis. Electronic supplementary material The online version of this article (doi:10.1186/s13567-015-0196-x) contains supplementary material, which is available to authorized users.
- Published
- 2015
21. Proteomics and pathway analyses of the milk fat globule in sheep naturally infected by Mycoplasma agalactiae provide indications of the in vivo response of the mammary epithelium to bacterial infection
- Author
-
Stefania Ghisaura, Alessandro Tanca, Sergio Uzzau, Grazia Biosa, Carla Cacciotto, Daniela Pagnozzi, Salvatore Pisanu, Marco Pittau, Tonina Roggio, Maria Filippa Addis, Gavino Marogna, Alberto Alberti, Addis M.F., Pisanu S., Ghisaura S., Pagnozzi D., Marogna G., Tanca A., Biosa G., Cacciotto C., Alberti A., Pittau M., Roggio T., and Uzzau S.
- Subjects
Proteomics ,Virulence Factors ,Difference gel electrophoresis ,Mycoplasma agalactiae ,Immunology ,ved/biology.organism_classification_rank.species ,Blotting, Western ,Sheep Diseases ,Mastitis ,Biology ,Microbiology ,Epithelium ,Tandem Mass Spectrometry ,Heat shock protein ,Animals ,Lactation ,Secretion ,Mycoplasma Infections ,Cytoskeleton ,Heat-Shock Proteins ,Glycoproteins ,Milk fat globules mastitis biomarker Mycoplasma agalactiae CA ,chemistry.chemical_classification ,Host Response and Inflammation ,Sheep ,ved/biology ,Endoplasmic reticulum ,Lipid Droplets ,Milk Proteins ,Molecular biology ,Oxidative Stress ,Infectious Diseases ,Milk ,chemistry ,Proteome ,Parasitology ,Electrophoresis, Polyacrylamide Gel ,Female ,Glycolipids ,Glycoprotein ,Chromatography, Liquid - Abstract
Milk fat globules (MFGs) are vesicles released in milk as fat droplets surrounded by the endoplasmic reticulum and apical cell membranes. During formation and apocrine secretion by lactocytes, various amounts of cytoplasmic crescents remain trapped within the released vesicle, making MFGs a natural sampling mechanism of the lactating cell contents. With the aim of investigating the events occurring in the mammary epithelium during bacterial infection, the MFG proteome was characterized by two-dimensional difference gel electrophoresis (2-D DIGE), SDS-PAGE followed by shotgun liquid chromatography-tandem mass spectrometry (GeLC-MS/MS), label-free quantification by the normalized spectral abundance factor (NSAF) approach, Western blotting, and pathway analysis, using sheep naturally infected by Mycoplasma agalactiae . A number of protein classes were found to increase in MFGs upon infection, including proteins involved in inflammation and host defense, cortical cytoskeleton proteins, heat shock proteins, and proteins related to oxidative stress. Conversely, a strikingly lower abundance was observed for proteins devoted to MFG metabolism and secretion. To our knowledge, this is the first report describing proteomic changes occurring in MFGs during sheep infectious mastitis. The results presented here offer new insights into the in vivo response of mammary epithelial cells to bacterial infection and open the way to the discovery of protein biomarkers for monitoring clinical and subclinical mastitis.
- Published
- 2011
22. Extrusive and Lateral Luxation Injuries in permanent teeth: Literature Review and Treatment Complications Update.
- Author
-
Spinas E, Zangani A, Mallus T, Marogna G, and Carboni L
- Subjects
- Humans, Dentition, Permanent, Orthodontic Extrusion, Tooth Avulsion therapy
- Published
- 2024
- Full Text
- View/download PDF
23. Streptococcus ruminantium-associated sheep mastitis outbreak detected in Italy is distinct from bovine isolates.
- Author
-
Rosa MN, Vezina B, Marogna G, Canu A, Molotzu MR, and Tola S
- Subjects
- Sheep, Animals, Cattle, Female, Lactation, Streptococcus genetics, Anti-Bacterial Agents, Milk, Disease Outbreaks veterinary, Mastitis, Bovine, Cattle Diseases, Sheep Diseases epidemiology
- Abstract
Streptococcus ruminantium is the causative agent of several bovine and ovine diseases, however reports are uncommon and application of whole genome sequencing to identify is rare. We report for the first time, a severe ovine mastitis outbreak caused by S. ruminantium in Italy, 2022. S. ruminantium was isolated from 12 adult lactating ewes with diffuse nodules in the mammary parenchyma and predominantly serous and clotted milk. All outbreak isolates, along with five additional historical Italian isolates (between 2011 and 2017), were genomically characterised and then analysed in the context of all publicly available S. ruminantium genomes. Antimicrobial susceptibility testing was performed to determine the MICs of 16 antibiotics. The results showed that all isolates were susceptible to all antimicrobials tested except kanamycin. Single Nucleotide Variant analysis confirmed this as a clonal outbreak across 10 sheep (≤ 15 SNVs), while the two others were colonised by more distantly related clones (≤ 53 pairwise SNVs), indicating the presence of multiple infecting lineages. The five historical S. ruminantium isolates were comprised of genetically-distant singletons (between 1259 and 5430 pairwise SNVs to 2022 outbreak isolates). Ovine isolates were found to be genetically distinct to bovine isolates, forming monophyletic groups. Bovine isolates were similarly made up of singleton clones in all but two isolates. Taken together, our genomic analysis using all globally available genomes is consistent with general opportunistic pathogenesis of S. ruminantium. We encourage future genomic surveillance efforts to facilitate outbreak detection, as well as improve our understanding of this poorly-understood, multi-host, zoonotic pathogen., (© 2023. The Author(s).)
- Published
- 2023
- Full Text
- View/download PDF
24. Preliminary Study on the Host Response to Bivalent and Monovalent Autogenous Vaccines against Mycoplasma agalactiae in Dairy Sheep.
- Author
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Hussein HA, Tolone M, Condorelli L, Galluzzo P, Puleio R, Vazzana I, Scatassa ML, Marogna G, Barreca S, Loria GR, Galuppo L, and Migliore S
- Abstract
In Italy, dairy sheep farming represents a vital agro-industry sector, but it is still challenged by contagious agalactia (CA), which is endemic there, and vaccination is the most economical and sustainable tool for control. This study aimed to evaluate the combined Mycoplasma agalactiae (Ma)-Staphylococcus aureus ( Sa ) vaccine (Ma-Sa ) against the Ma monovalent vaccine in ewes. Twelve primiparous Ma -free ewes were randomly grouped into three equal groups: first, the control group injected with placebo, second, the group vaccinated with the Ma monovalent vaccine, and third, the group vaccinated with Ma-Sa combined vaccine, with two S/C doses at 45-day intervals. The animals were examined for serological, hematological, and somatic cell count (SCC) changes for 17 successive weeks. A significant increase in anti- Ma antibody mean titers, leukocytes, and platelets was observed in the vaccinated animals, with the highest values in those who received the combined vaccine. Neutrophils were high only in the animals who received the combined vaccine. SCC was lower in the vaccinated animals during the first six weeks. This study concludes that the combined Ma-Sa vaccines enhance immune response and potentiate its efficacy against Ma . This improvement might be attributed to the sensitization/activation effect of S. aureus on platelets, which are recoded to act as a key regulator for the coordination of all components of the innate immune system. Even though this study included a small number of animals, its findings about the potentialities of this inactivated vaccine in the control of CA are strongly encouraging. Further confirmation might be needed through additional replicates and a challenge study is needed before proceeding with widespread use.
- Published
- 2022
- Full Text
- View/download PDF
25. Identification of secreted and cellular antigens of Staphylococcus aureus causing dairy sheep mastitis and their potential for vaccine development.
- Author
-
Longheu CM, Azara E, Marogna G, Addis MF, and Tola S
- Subjects
- Animals, Antigens, Bacterial administration & dosage, Bacterial Toxins immunology, Cattle, Cattle Diseases immunology, Cattle Diseases microbiology, Cattle Diseases prevention & control, Exotoxins immunology, Female, Immunity, Humoral, Leukocidins immunology, Mastitis, Bovine prevention & control, Proteomics methods, Sheep, Sheep Diseases immunology, Sheep Diseases prevention & control, Staphylococcal Infections immunology, Staphylococcal Vaccines, Staphylococcus aureus genetics, Staphylococcus aureus pathogenicity, Tandem Mass Spectrometry, Antigens, Bacterial blood, Antigens, Bacterial immunology, Mastitis, Bovine microbiology, Sheep Diseases microbiology, Staphylococcal Infections veterinary, Staphylococcus aureus chemistry, Staphylococcus aureus immunology
- Abstract
Staphylococcus aureus is the leading cause of clinical mastitis and is associated with persistent subclinical infections in ewes, significantly compromising the quality and quantity of milk productions. To date, vaccines intended for use in sheep have been mainly focused on biofilm production traits, but many S. aureus pathogenic isolates do not produce biofilm, including those circulating in Sardinia, one of the leading sheep milk producers in Europe. The aim of this work was to identify suitable immunodominant, alternative candidates to biofilm components for vaccine and diagnostic development. An immunoproteomics study was carried out by testing sera from naturally infected sheep with a prevalent S. aureus lineage against cellular and secreted antigens, followed by tandem mass spectrometry identification of the most prominent immunogens. Four cellular and three secreted S. aureus antigens elicited a strong humoral host immune response. The four cellular antigens were the housekeeping proteins pyruvate kinase, elongation Factor Tu, dihydrolipoyl dehydrogenase, and alpha-keto acid dehydrogenase. The three secreted antigens were the bifunctional autolysin (Atl) and the two components of the Panton-Valentine leukocidin, lukF-PV/lukM, demonstrating the carriage of prophage phiPV83 in a sheep isolate and the strong response of the sheep host against them. In consideration of the key role played by these secreted proteins in S. aureus replication and immune evasion, these antigens may represent suitable candidates for developing vaccines eliciting a more successful immunological protection in areas where non-biofilm forming Staphylococcus spp. are the most widespread intramammary pathogens., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
26. Neutrophil extracellular traps in sheep mastitis.
- Author
-
Pisanu S, Cubeddu T, Pagnozzi D, Rocca S, Cacciotto C, Alberti A, Marogna G, Uzzau S, and Addis MF
- Subjects
- Animals, Extracellular Traps microbiology, Female, Humans, Mammary Glands, Animal microbiology, Mammary Glands, Animal pathology, Mastitis immunology, Mastitis microbiology, Milk cytology, Milk microbiology, Neutrophils microbiology, Sheep, Sheep Diseases microbiology, Streptococcal Infections immunology, Streptococcal Infections microbiology, Extracellular Traps metabolism, Mastitis veterinary, Neutrophils metabolism, Sheep Diseases immunology, Streptococcal Infections veterinary, Streptococcus physiology
- Abstract
Neutrophil extracellular traps (NETs) are structures composed of DNA, histones, and antimicrobial proteins that are released extracellularly by neutrophils and other immune cells as a means for trapping and killing invading pathogens. Here, we describe NET formation in milk and in mammary alveoli of mastitic sheep, and provide a dataset of proteins found in association to these structures. Nucleic acid staining, immunomicroscopy and fluorescent in-situ hybridization of mastitic mammary tissue from sheep infected with Streptococcus uberis demonstrated the presence of extranuclear DNA colocalizing with antimicrobial proteins, histones, and bacteria. Then, proteomic analysis by LTQ-Orbitrap Velos mass spectrometry provided detailed information on protein abundance changes occurring in milk upon infection. As a result, 1095 unique proteins were identified, of which 287 being significantly more abundant in mastitic milk. Upon protein ontology classification, the most represented localization classes for upregulated proteins were the cytoplasmic granule, the nucleus, and the mitochondrion, while function classes were mostly related to immune defence and inflammation pathways. All known NET markers were massively increased, including histones, granule proteases, and antimicrobial proteins. Of note was the detection of protein arginine deiminases (PAD3 and PAD4). These enzymes are responsible for citrullination, the post-translational modification that is known to trigger NET formation by inducing chromatin decondensation and extracellular release of NETs. As a further observation, citrullinated residues were detected by tandem mass spectrometry in histones of samples from mastitic animals. In conclusion, this work provides novel microscopic and proteomic information on NETs formed in vivo in the mammary gland, and reports the most complete database of proteins increased in milk upon bacterial mastitis.
- Published
- 2015
- Full Text
- View/download PDF
27. Production and release of antimicrobial and immune defense proteins by mammary epithelial cells following Streptococcus uberis infection of sheep.
- Author
-
Addis MF, Pisanu S, Marogna G, Cubeddu T, Pagnozzi D, Cacciotto C, Campesi F, Schianchi G, Rocca S, and Uzzau S
- Subjects
- Animals, Anti-Infective Agents metabolism, Antimicrobial Cationic Peptides immunology, Antimicrobial Cationic Peptides metabolism, Epithelial Cells microbiology, Glycolipids immunology, Glycolipids metabolism, Glycoproteins immunology, Glycoproteins metabolism, Lactation immunology, Lactation metabolism, Leukocyte L1 Antigen Complex immunology, Leukocyte L1 Antigen Complex metabolism, Lipid Droplets, Mammary Glands, Animal metabolism, Mammary Glands, Animal microbiology, Milk immunology, Milk metabolism, Milk microbiology, Proteomics methods, Sheep metabolism, Sheep microbiology, Streptococcal Infections prevention & control, Streptococcal Infections veterinary, Cathelicidins, Anti-Infective Agents immunology, Epithelial Cells immunology, Mammary Glands, Animal immunology, Sheep immunology, Streptococcal Infections immunology, Streptococcus immunology
- Abstract
Investigating the innate immune response mediators released in milk has manifold implications, spanning from elucidation of the role played by mammary epithelial cells (MECs) in fighting microbial infections to the discovery of novel diagnostic markers for monitoring udder health in dairy animals. Here, we investigated the mammary gland response following a two-step experimental infection of lactating sheep with the mastitis-associated bacterium Streptococcus uberis. The establishment of infection was confirmed both clinically and by molecular methods, including PCR and fluorescent in situ hybridization of mammary tissues. Proteomic investigation of the milk fat globule (MFG), a complex vesicle released by lactating MECs, enabled detection of enrichment of several proteins involved in inflammation, chemotaxis of immune cells, and antimicrobial defense, including cathelicidins and calprotectin (S100A8/S100A9), in infected animals, suggesting the consistent involvement of MECs in the innate immune response to pathogens. The ability of MECs to produce and release antimicrobial and immune defense proteins was then demonstrated by immunohistochemistry and confocal immunomicroscopy of cathelicidin and the calprotectin subunit S100A9 on mammary tissues. The time course of their release in milk was also assessed by Western immunoblotting along the course of the experimental infection, revealing the rapid increase of these proteins in the MFG fraction in response to the presence of bacteria. Our results support an active role of MECs in the innate immune response of the mammary gland and provide new potential for the development of novel and more sensitive tools for monitoring mastitis in dairy animals.
- Published
- 2013
- Full Text
- View/download PDF
28. Proteomics and pathway analyses of the milk fat globule in sheep naturally infected by Mycoplasma agalactiae provide indications of the in vivo response of the mammary epithelium to bacterial infection.
- Author
-
Addis MF, Pisanu S, Ghisaura S, Pagnozzi D, Marogna G, Tanca A, Biosa G, Cacciotto C, Alberti A, Pittau M, Roggio T, and Uzzau S
- Subjects
- Animals, Blotting, Western, Chromatography, Liquid, Cytoskeleton, Electrophoresis, Polyacrylamide Gel, Epithelium microbiology, Epithelium pathology, Female, Heat-Shock Proteins biosynthesis, Lactation, Lipid Droplets, Mastitis immunology, Mastitis metabolism, Mastitis microbiology, Milk chemistry, Milk Proteins analysis, Mycoplasma Infections immunology, Mycoplasma Infections metabolism, Mycoplasma Infections microbiology, Mycoplasma agalactiae genetics, Oxidative Stress, Proteomics, Sheep, Sheep Diseases metabolism, Sheep Diseases microbiology, Tandem Mass Spectrometry, Virulence Factors metabolism, Glycolipids chemistry, Glycolipids metabolism, Glycoproteins chemistry, Glycoproteins metabolism, Mastitis veterinary, Mycoplasma Infections veterinary, Mycoplasma agalactiae metabolism, Sheep Diseases immunology
- Abstract
Milk fat globules (MFGs) are vesicles released in milk as fat droplets surrounded by the endoplasmic reticulum and apical cell membranes. During formation and apocrine secretion by lactocytes, various amounts of cytoplasmic crescents remain trapped within the released vesicle, making MFGs a natural sampling mechanism of the lactating cell contents. With the aim of investigating the events occurring in the mammary epithelium during bacterial infection, the MFG proteome was characterized by two-dimensional difference gel electrophoresis (2-D DIGE), SDS-PAGE followed by shotgun liquid chromatography-tandem mass spectrometry (GeLC-MS/MS), label-free quantification by the normalized spectral abundance factor (NSAF) approach, Western blotting, and pathway analysis, using sheep naturally infected by Mycoplasma agalactiae. A number of protein classes were found to increase in MFGs upon infection, including proteins involved in inflammation and host defense, cortical cytoskeleton proteins, heat shock proteins, and proteins related to oxidative stress. Conversely, a strikingly lower abundance was observed for proteins devoted to MFG metabolism and secretion. To our knowledge, this is the first report describing proteomic changes occurring in MFGs during sheep infectious mastitis. The results presented here offer new insights into the in vivo response of mammary epithelial cells to bacterial infection and open the way to the discovery of protein biomarkers for monitoring clinical and subclinical mastitis.
- Published
- 2011
- Full Text
- View/download PDF
29. Virulence attenuation and live vaccine potential of aroA, crp cdt cya, and plasmid-cured mutants of Salmonella enterica serovar Abortusovis in mice and sheep.
- Author
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Uzzau S, Marogna G, Leori GS, Curtiss R 3rd, Schianchi G, Stocker BA, and Rubino S
- Subjects
- Abortion, Veterinary prevention & control, Animals, Female, Mice, Mice, Inbred BALB C, Mutation, Plasmids, Pregnancy, Sheep, Typhoid-Paratyphoid Vaccines immunology, Vaccines, Attenuated immunology, Virulence, Salmonella Infections, Animal prevention & control, Salmonella Vaccines immunology, Salmonella enterica immunology
- Abstract
Three live vaccine candidates of Salmonella enterica subspecies I serotype Abortusovis (aroA, cya crp cdt, and plasmid-cured strains) have been developed, and their efficacies in inducing humoral antibodies and protecting against abortion after challenge with wild-type strain SS44 were evaluated in sheep. Following estrus synchronization, animals were immunized 3 weeks after fertilization and boosted once 3 weeks later. Following challenge with wild-type SS44, pregnancy failure of vaccinated ewes was reduced compared to that of nonimmunized controls. Attenuation of each vaccine was also assessed in challenge experiments with nonimmunized pregnant ewes and in BALB/c mice. All three vaccine candidates appear to be safe for use in sheep and provide a model for the development of live vaccine candidates against naturally occurring ovine salmonellosis.
- Published
- 2005
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30. Transposition of the heat-stable toxin astA gene into a gifsy-2-related prophage of Salmonella enterica serovar Abortusovis.
- Author
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Bacciu D, Falchi G, Spazziani A, Bossi L, Marogna G, Leori GS, Rubino S, and Uzzau S
- Subjects
- Animals, Bacterial Proteins genetics, Bacterial Proteins metabolism, Colony Count, Microbial, DNA, Bacterial chemistry, DNA, Bacterial isolation & purification, Escherichia coli genetics, Gene Deletion, Gene Dosage, Gene Transfer, Horizontal, Genes, Bacterial, Genes, Viral, Intestines microbiology, Liver microbiology, Lymph Nodes microbiology, Molecular Sequence Data, Prophages isolation & purification, Salmonella Infections, Animal microbiology, Salmonella enterica isolation & purification, Salmonella enterica pathogenicity, Sheep Diseases microbiology, Spleen microbiology, Viral Tail Proteins genetics, Bacterial Toxins genetics, DNA Transposable Elements, Prophages genetics, Salmonella enterica genetics, Salmonella enterica virology, Virulence Factors genetics
- Abstract
The horizontal transfer and acquisition of virulence genes via mobile genetic elements have been a major driving force in the evolution of Salmonella pathogenicity. Serovars of Salmonella enterica carry variable assortments of phage-encoded virulence genes, suggesting that temperate phages play a pivotal role in this process. Epidemic isolates of S. enterica serovar Typhimurium are consistently lysogenic for two lambdoid phages, Gifsy-1 and Gifsy-2, carrying known virulence genes. Other serovars of S. enterica, including serovars Dublin, Gallinarum, Enteritidis, and Hadar, carry distinct prophages with similarity to the Gifsy phages. In this study, we analyzed Gifsy-related loci from S. enterica serovar Abortusovis, a pathogen associated exclusively with ovine infection. A cryptic prophage, closely related to serovar Typhimurium phage Gifsy-2, was identified. This element, named Gifsy-2AO, was shown to contribute to serovar Abortusovis systemic infection in lambs. Sequence analysis of the prophage b region showed a large deletion which covers genes encoding phage tail fiber proteins and putative virulence factors, including type III secreted effector protein SseI (GtgB, SrfH). This deletion was identified in most of the serovar Abortusovis isolates tested and might be dependent on the replicative transposition of an adjacent insertion sequence, IS1414, previously identified in pathogenic Escherichia coli strains. IS1414 encodes heat-stable toxin EAST1 (astA) and showed multiple genomic copies in isolates of serovar Abortusovis. To our knowledge, this is the first evidence of intergeneric transfer of virulence genes via insertion sequence elements in Salmonella. The acquisition of IS1414 (EAST1) and its frequent transposition within the chromosome might improve the fitness of serovar Abortusovis within its narrow ecological niche., (Copyright 2004 American Society for Microbiology)
- Published
- 2004
- Full Text
- View/download PDF
31. [Incidence of healthy carriers of Australia antigen in various Italian regions].
- Author
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Sinicco A, Scalise G, Marogna G, Guarini L, and Bencini M
- Subjects
- Blood Donors, Complement Fixation Tests, Hepatitis B Antigens, Humans, Immunodiffusion, Italy, Carrier State epidemiology, Hepatitis B virus immunology
- Published
- 1972
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