Molecular characterization of gliotoxin-producing Aspergillus fumigatus in dairy cattle feed

Background and Aim: Several strains of Aspergillus fumigatus produce mycotoxins that affect the health and productivity of dairy cattle, and their presence in dairy cattle feed is a serious concern. This study aimed to determine the densities of A. fumigatus and gliotoxin in commercial dairy feed. Materials and Methods: More than 60 dairy feed samples were examined for fungal contamination, specifically for A. fumigatus, using phenotypic approaches and DNA sequencing of the internal transcribed spacer (ITS) and β-tubulin regions. Thin-layer chromatography and high-performance liquid chromatography (HPLC) were used to assess gliotoxin production in A. fumigatus. Real-time polymerase chain reaction (RT-PCR) was used to investigate the expression of gliZ, which was responsible for gliotoxin production. High-performance liquid chromatography was used to detect gliotoxin in feed samples. Results: Aspergillus was the most commonly identified genus (68.3%). Aspergillus fumigatus was isolated from 18.3% of dairy feed samples. Only four of the 11 A. fumigatus isolates yielded detectable gliotoxins by HPLC. In total, 7/11 (43.7%) feed samples tested had gliotoxin contamination above the threshold known to induce immunosuppressive and apoptotic effects in vitro. The HPLC-based classification of isolates as high, moderate, or non-producers of gliotoxin was confirmed by RT-PCR, and the evaluation of gliZ expression levels corroborated this classification. Conclusion: The identification of A. fumigatus from animal feed greatly depended on ITS and β-tubulin sequencing. Significant concentrations of gliotoxin were found in dairy cattle feed, and its presence may affect dairy cow productivity and health. Furthermore, workers face contamination risks when handling and storing animal feed

New FTY720-docetaxel nanoparticle therapy overcomes FTY720-induced lymphopenia and inhibits metastatic breast tumour growth

Purpose: Combining molecular therapies with chemotherapy may offer an improved clinical outcome for chemoresistant tumours. Sphingosine-1-phosphate (S1P) receptor antagonist and sphingosine kinase 1 (SK1) inhibitor FTY720 (FTY) has promising anticancer properties, however, it causes systemic lymphopenia which impairs its use in cancer patients. In this study, we developed a nanoparticle (NP) combining docetaxel (DTX) and FTY for enhanced anticancer effect, targeted tumour delivery and reduced systemic toxicity. Methods: Docetaxel, FTY and glucosamine were covalently conjugated to poly(lactic-co-glycolic acid) (PLGA). NPs were characterised by dynamic light scattering and electron microscopy. The cellular uptake, cytotoxicity and in vivo antitumor efficacy of CNPs were evaluated. Results: We show for the first time that in triple negative breast cancer cells FTY provides chemosensitisation to DTX, allowing a four-fold reduction in the effective dose. We have encapsulated both drugs in PLGA complex NPs (CNPs), with narrow size distribution of ~ 100 nm and excellent cancer cell uptake providing sequential, sustained release of FTY and DTX. In triple negative breast cancer cells and mouse breast cancer models, CNPs had similar efficacy to systemic free therapies, but allowed an effective drug dose reduction. Application of CNPs has significantly reversed chemotherapy side effects such as weight loss, liver toxicity and, most notably, lymphopenia. Conclusions: We show for the first time the DTX chemosensitising effects of FTY in triple negative breast cancer. We further demonstrate that encapsulation of free drugs in CNPs can improve targeting, provide low off-target toxicity and most importantly reduce FTY-induced lymphopenia, offering potential therapeutic use of FTY in clinical cancer treatment