ABSTRACT
Turkey seminal plasma contains three serine proteinase inhibitors. Two of them, with low molecular masses (6 kDa), were identified as single-domain Kazal-type inhibitors responsible for regulating acrosin activity. Our experimental objective was to isolate and characterize the inhibitor with the high molecular weight from turkey seminal plasma. The inhibitor was purified using hydrophobic interaction and affinity chromatography. Pure preparations of the inhibitor were used for identification by mass spectrometry, for determination of physicochemical properties (molecular weight, pI, and content and composition of the carbohydrate component), for kinetic studies, and for antibacterial tests. Gene expression and immunohistochemical detection of the inhibitor were analyzed in the testis, epididymis, and ductus deferens. The inhibitor with a high molecular weight from turkey seminal plasma was identified as an ovoinhibitor, which was found in avian semen for the first time. The turkey seminal plasma ovoinhibitor was a six-tandem homologous Kazal-type domain serine proteinase inhibitor that targeted multiple proteases, including subtilisin, trypsin, and elastase, but not acrosin. Our results suggested that hepatocyte growth factor activator was a potential target proteinase for the ovoinhibitor in turkey seminal plasma. The presence of the ovoinhibitor within the turkey reproductive tract suggested that its role was to maintain a microenvironment for sperm in the epididymis and ductus deferens. The turkey seminal plasma ovoinhibitor appeared to play a significant role in an antibacterial semen defense against Bacillus subtilis and Staphylococcus aureus.
Subject(s)
Egg Proteins, Dietary/isolation & purification , Seminal Plasma Proteins/isolation & purification , Serine Proteinase Inhibitors/isolation & purification , Turkeys , Amino Acid Sequence , Animals , Egg Proteins, Dietary/analysis , Egg Proteins, Dietary/chemistry , Electrophoresis, Gel, Two-Dimensional , Male , Mass Spectrometry , Molecular Sequence Data , Molecular Weight , Protein Structure, Tertiary , Semen/chemistry , Seminal Plasma Proteins/analysis , Seminal Plasma Proteins/chemistry , Serine Proteinase Inhibitors/analysis , Serine Proteinase Inhibitors/chemistry , Turkeys/metabolismABSTRACT
BACKGROUND: The aim of our current study was to investigate the possible occurence of pyrethroid (taufluvalinate) resistant Varroa mites infestations in 24 randomly chosen apiaries of Warmia-Mazury province of northeast Poland. METHODS: The methodology used for the analysis of resistant Varroa strains strictly followed the protocol described by Milani. RESULTS: We identified 3 apiaries that were infested with high risk pyrethroid resistance mites and a further 9 apiaries that were free from this resitance. The brood samples collected from the remaining apiaries did not contain sufficient numbers of parasites to enable us to properly perform the assay. CONCLUSIONS: Our finding that 25% of the tested brood samples showed a high risk of fully pyrethroid resistant Varroa mite contamination indicates that resistant Varroa may become wide spread in apiaries in Poland. Interestingly these high risk resistant mites were found in honeybee colonies with low levels of Varroa infestation, with an average rate of 2.16%. We also discuss the role of amitraz (amidine) in the phenomenon of Varroa resistance to pyrethroids.
