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Giovanna Gentilomi,
Rossella Lelli,
Mirella D'Angelo, Vincenzo Langella,
Federica Monaco,
Ottavio Portanti,
Mirella Luciani,
Mara Mirasoli,
Aldo Roda,
Marialuisa Zerbini,
Monica Musiani
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ABSTRACT: The precise role of bovine interferon-gamma (BoIFN-gamma) in disease and therapy is still poorly defined. Clearly it is involved in defence against parasites, bacteria, viruses and possibly tumor cells. This paper reports the expression of BoIFN-gamma in a baculovirus system to generate a fully functional recombinant protein. Bovine interferon-gamma cDNA was cloned from mitogen stimulated peripheral blood mononuclear cell (PBMC) RNA utilizing the reverse transcription-polymerase chain reaction (RT-PCR). The cDNA open reading frame (ORF) encoding for a putative 166 amino acid protein (22KDa) was cloned and expressed into baculovirus transfer vector pBlueBac 4.5/V5 His. This vector was co-transfected with Autografa californica multiple nuclear polyhedrosis virus (AcMNPV) DNA into Spodoptera frugiperda cells (Sf9) and the recombinant virus, named AcBoIFN-gamma, was then recovered. Recombinant BoIFN-gamma (rBoIFN-gamma His) was accumulated in the serum-free medium of AcBoIFN-gamma-infected cells. The nickel affinity spin column purified rBoIFN-gamma His was shown to be a glycosylated 20-22 KDa protein as confirmed by SDS-PAGE glycan determination and showed antiviral activity in vitro against the bovine viral diarrhoea-mucosal disease virus (BVD/MD). The production of this bioactive rBoIFN-gamma His will allow us to explore this cytokine as a potential vaccine adjuvant or therapeutic agent for bovine diseases.
The new microbiologica: official journal of the Italian Society for Medical, Odontoiatric, and Clinical Microbiology (SIMMOC) 02/2006; 29(1):19-24. · 1.00 Impact Factor
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ABSTRACT: American foulbrood, caused by Paenibacillus larvae subsp. larvae (White 1906) is one of the most serious diseases of honey bees, causing beekeepers and health workers to make difficult, complex decisions and leading to the development of 'organic' treatments, such as grapefruit seed extract, with minor residue problems in the end product. This study evaluates the chemical composition of grapefruit seed extracts using gas chromatography/mass spectrometry for the detection of benzethonium chloride, cetrimonium bromide and decyltrimethylammonium chloride. The results obtained suggest a close correlation between the microbial effect and the presence of chemical additives in the samples analysed.
Veterinaria italiana 43(1):109-13. · 0.67 Impact Factor
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ABSTRACT: Macrophages are pivotal cells of the immune system and play a key role in the host defence mechanism against pathogens. To date, the importance of macrophages and the role of humoral response in eliciting macrophage activity against Mycoplasma mycoides subsp. mycoides small colony (Mmm-SC), the causative agent of contagious bovine pleuropneumonia (CBPP), have only been marginally elucidated or are almost unknown. The present study was undertaken to investigate the changes in surface morphology of macrophages after in vitro infection with Mmm-SC in the presence of bovine immune serum. Morphological analysis was performed on macrophage cultures at 6 h post infection using the three-dimensional vision of scanning electron microscopy. Non-infected macrophages in the presence of negative or immune serum and macro phages infected with Mmm-SC in the absence of serum showed only minor cell surface changes. In contrast, clear surface modifications, broad veils, fine philopodia highlighting cell activation and small aggregates of mycoplasma closely attached to the macrophage membrane, were observed in infected macrophage cultures in the presence of immune serum. Our results suggest that specific humoral response to Mmm-SC may contribute and support phagocytic activity of macrophages.
Veterinaria italiana 46(4):389-404. · 0.67 Impact Factor
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ABSTRACT: Riassunto La peste americana, sostenuta da Paenibacillus larvae subsp. larvae (White 1906) è una grave malattia delle api, che pone l'apicoltore e l'operatore sanitario di fronte a scelte difficili e complesse che portano a sviluppare presidi terapeutici "di tipo biologico", come gli estratti di semi di pompelmo, con scarsi problemi di residui nel prodotto finale. Il presente studio si propone di valutare la composizione chimica di estratti di semi di pompelmo utilizzando la gas cromatografia/spettrometria di massa per la ricerca di benzetonio cloruro, cetrimonio bromuro e trimetil-decanamonio cloruro. I risultati ottenuti ipotizzano una stretta correlazione tra l'effetto antimicrobico e la presenza di additivi chimici nei campioni analizzati. Parole chiave Additivi, Capacità di inibizione, Estratti di semi di pompelmo, Paenibacillus larvae, Peste americana. Introduzione La peste americana, sostenuta dal Paenibacillus larvae subsp. larvae (White 1906) è una grave malattia delle api, che pone l'apicoltore e l'operatore sanitario di fronte a scelte complesse quali la necessità di fronteggiare la malattia con appropriati presidi terapeutici e l'esigenza di ridurre al minimo i residui nel miele al fine di tutelare il consumatore finale. L'utilizzo, a volte indiscriminato, di antibiotici per combattere la forma vegetativa del batterio può portare alla commercializzazione di prodotti dell'alveare contenenti residui di sostanze inibenti (tetraciclina, ossitetraciclina e sulfatiazolo). I dati pubblicati recentemente da ricercatori del settore, mostrano la presenza di antibiotici in campioni di miele e pappa reale (2).
Veterinaria italiana 43:103-107. · 0.67 Impact Factor
