Assessment of Different Functional Parameters of Frozen–Thawed Buffalo Spermatozoa by Using Cytofluorimetric Determinations

Flow cytometry is a useful tool that provides an accurate, objective and rapid evaluation of semen quality. The use of this technique could significantly improve the quality of buffalo semen samples used in artificial insemination. This study was carried out to evaluate, by flow cytometry, frozen–thawed buffalo spermatozoa quality parameters such as sperm viability by SYBR‐14/propidium iodide staining; mitochondrial function by JC‐1 potentiometric probe; sperm chromatin stability (SCSA) by acridine orange; and acrosome reaction (AR) by FITC‐PNA staining. Semen samples from five Italian Mediterranean buffalo bulls were used. Sperm viability was not different between bulls and ranged from 33.4% to 43.6%. A consistent rate (55.1 ± 10.8%) of sperm cells showed high mitochondrial membrane potential (Δψ^high), with no significant differences between subjects. Sperm chromatin structure assay differed significantly between the five buffalo bulls; moreover, data showed high stability within each buffalo. DNA fragmentation indexes (DFI), such as %‐DFI, ‐DFI, SD‐DFI, were 11.2 ± 8.6, 153.3 ± 24.6 and 81.6 ± 21.2, respectively. Regarding AR, the percentage of acrosome‐reacted live (ARL) and acrosome‐reacted dead (ARD) spermatozoa was 0.3 ± 0.2 and 15.3 ± 5.5, respectively. This functional parameter differed significantly between buffalo bulls and showed high stability. Following to Ca²⁺ ionophore A23187 for 3 h, AR significantly differed between subjects and was characterized by an increase in both ARL (10.8%) and ARD population (22.0%). This study indicates that flow cytometry could be a useful tool for a quick multiparametric evaluation of sperm quality in buffalo. In particular, SCSA and AR resulted in sperm functional parameters sensitive enough for the diagnosis of frozen‐thawed semen fertilizing potential.     

Ungulate Herbivory Is Correlated with High Aspen Suckering Density but Reductions in Aspen Growth Rates and Recruitment

Aspen is a foundation species that provides vital habitat for hundreds of plant and animal species. However, changing ungulate herbivory regimes may be altering recruitment success and resilience of aspen forests. The objective of this observational study was to quantify the impacts of ungulate herbivory on aspen recruitment potential. We sampled 36 aspen dominant stands on the Fishlake National Forest in Utah for browse of apical meristems, suckering density (< 180 cm in height), and recruitment density (≥ 180 cm). Our analysis indicated that for each 10% of apical meristems browsed, annual vertical and radial growth rates and recruitment density were reduced by 1.2 cm, 0.02 mm, and 17 ha^? 1, respectively. In contrast, aspen suckering density increased by 1 230 suckers ha^? 1, with each 10% browse of apical meristems. This suggests that ungulates contribute to aspen suckers persisting in the understory for longer periods and may alter recruitment success of aspen stands. Aspen that were under high ungulate browse pressure were associated with lower growth rates and persisted for twice as long in the understory. For example, a 5-yr-old sucker was predicted to be 115 cm without browse of apical meristems and 60 cm with 100% browse of apical meristems. Further, aspens of the same height with 100% browse of apical meristems were nearly 4 yr older on average than aspens with no evidence of browse of apical meristems. Approximately 34% of aspen suckers had persisted in the understory < 100 cm for ≥ 6 yr, and 7% of suckers had persisted in the understory for ≥ 10 yr. Our results suggest that high rates of meristem removal are correlated with increased aspen suckering yet reduced aspen growth and recruitment.