A comparison of alternative assays to measure DNA damage in stallion spermatozoa: TUNEL test versus ‘Nicoletti assay’

The aberrations of sperm DNA may cause various problems and have negative consequences on fertility. These influence embryonic development or might lead to early embryo loss. Sperm Chromatin Structure Assay (SCSA) is the flow cytometric method most often used for the detection of DNA lesions; however, some studies using that method reached confusing conclusions. The aim of this pilot study was to adjust and compare two alternative tests, namely the TUNEL test and the Nicoletti assay. The above-mentioned two flow cytometric methods capable of detecting the fragmented DNA of sperm were tested on 12 frozen-thawed stallion semen samples. The TUNEL test demonstrated much higher DNA fragmentation ratio than the Nicoletti assay (mean ± SD: 30.77 ± 13.03% vs. 1.93 ± 0.89%, respectively). A fluorescent microscopic check of the samples showed that TUNEL labelled the plasma membrane and the mitochondria in a nonspecific way, rather than detecting only the fragmented DNA, thus eventually resulting in a false positive sign. The Nicoletti assay is simpler, quicker and does not detect nonspecific binding; however, further analyses are required to determine its diagnostic value

Soil seed bank of the invasive Robinia pseudoacacia in planted Pinus nigra stands

Pinus nigra and Robinia pseudoacacia are exotic trees used for afforestation in Hungary. Pinus nigra was non-invasive, however R. pseudoacacia escaped from cultivation and invaded several vegetation types including pine plantations. It has recently been planned to cut P. nigra plantations and replace them by native tree stands, especially in nature reserves. The scattered presence of R. pseudoacacia specimens in pine stands might place constraints on planned tree replacement because of their vegetative resprouting and recolonization from an established seed bank. The aim of this study was to investigate the soil seed bank under the canopy of solitary R. pseudoacacia specimens found in P. nigra plantations. Altogether 250 soil samples were collected from the 0–6 and 6–12 cm soil layers under solitary Robinia trees of varying ages (with basal areas between 62.4 and 1089.3 cm2). Seeds were separated by sieving then scarified and germinated. Seed bank density ranged between 640 and 2285 seedsm–2 with an average distribution of 82.7% and 17.3% in the upper and lower soil layer, respectively. Total density of the seed bank and also the seed bank ratio of the lower soil layer increased with tree age. The accumulated seed bank of R. pseudoacacia should be considered in the careful planning of tree replacement operations in Pinus nigra stands