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Aims: To test the suitability of the Agrobacterium tumefaciens-mediated transformation (AMT) method with Paecilomyces fumosoroseus, a fungal pathogen that causes diseases in a wide range of insects including whiteflies. Methods and Results: Conidia of P. fumosoroseus were successfully transformed to hygromycin B resistance using the hph gene of Escherichia coli as the selectable marker. Transformation frequencies were 58·3 ± 18·5, 98·3 ± 24·8 and 169·7 ± 35·5 (±SEM) transformants per 105, 106 and 107 target conidia respectively. After confirmation by PCR, transformants were subjected to Southern analysis, and the results revealed that 45% (four of nine) of the transformants contained single-copy integration of the T-DNA. Conclusions: In our AMT system, we efficiently transformed conidia of P. fumosoroseus. The employment of this method circumvents time-consuming protoplast preparation and allows the isolation of transformants containing single-copy integration of the T-DNA. Significance and Impact of the Study: Considering the efficiency of Ag. tumefaciens-mediated transformation, this method represents a useful tool for insertional mutagenesis to characterize genes that are important for the pathogenicity of P. fumosoroseus. [ABSTRACT FROM AUTHOR]

The optimised use of mixed crude glycerin (MCG) and ractopamine hydrochloride (RH) in diets for finishing pigs requires a better understanding of glycerol metabolism. Therefore, the present study evaluated the performance, blood parameters, metabolite concentrations in the Longissimus lumborum muscle and the activity of enzymes in the liver, subcutaneous adipose tissue and kidney of pigs fed diets containing MCG and/or RH. According to the initial weight, 64 barrows were distributed in a 4 × 2 factorial design, corresponding to four concentrations of MCG (0, 100, 150, and 200 g/kg) in diets containing 0 or 10 mg/kg of RH. There was no MCG × RH interaction for any of the evaluated parameters (P > 0.05). Moreover, there was no isolated effect of dietary MCG or RH on the weight gain and the blood concentrations of glucose, insulin, triacylglycerols, total cholesterol, total protein, albumin, creatinine, cortisol and lactate (P > 0.05). There was a higher rate of phosphorylation of glycerol in the liver, stimulation of lipogenesis in adipose tissue and a likely increase in the utilisation of glycerol by muscle fibres in response to an increased concentration of circulating glycerol from the diet (P < 0.05). In contrast, when the diet contained up to 200 g/kg MCG and 10 mg/kg RH, there were indications of a lower utilisation of glycerol for the synthesis of triacylglycerols in muscle, liver and adipose tissue, in addition to increased oxidation of amino acids in the liver followed by improved feed conversion of pigs (P < 0.05). Crude glycerin is an important alternative energy ingredient in the diet of pigs and the ractopamine is a feed additive that has been associated with improvements in animal performance and meat quality. The present study contributes to the advancement of the animal science, because it provides information about a subject that remains still little explored, namely, the glycerol metabolism in pigs fed diets containing glycerin and ractopamine. Such understanding is necessary to contribute to the formulation of diets that are nutritionally more adequate. [ABSTRACT FROM AUTHOR]