Investigations on different Semen Extenders for Cockatiel Semen

Abstract

The aim of the present study was the modification and evaluation of three different semen extenders for cockatiel semen in order to achieve a long survival time for transport, examination purpose and for potential cryopreservation, respectively. Therefore, individual and pooled semen samples of 30 cockatiels (Nymphicus hollandicus) were investigated for pH and osmolality values and subsequently pH and osmolality values of the semen extenders were adjusted to those values in the semen. Pooled semen samples were then partitioned into four equal parts and diluted with the three different semen extenders in 1:4 and 1:8 dilution. 1 % glucose-Ringer’s solution was used as control, respectively. A total of 64 incremental diluted semen samples were obtained for investigation. Each dilution was investigated regarding sperm motility immediately after dilution and another four times every 30 minutes. Sperm viability was evaluated 0 and 120 minutes after dilution via eosin B-stain on the diluted semen samples and in pure semen samples. Additionally, the fluorescence stain SYBR® Green/propidium iodide was used to assess sperm viability. The results indicate that cockatiel spermatozoa are highly sensitive to variations in pH and osmolality, requiring adjustment of commercial diluents to pH = 7.42 and osmolality = 300 mOsm/kg. Modified Lake diluent maintained higher viability and motility than other diluents tested. Sperm morphology was indicated to be least adversely affected by modified Lake diluent in 1:4 concentration compared to other semen extenders and concentrations used.

Viability assessment of spermatozoa in large falcons (Falco spp.) using various staining protocols

“…Using eosin blue, the differentiation of dead from alive sperm was effectively possible, as reported from viability analysis in other avian species (Behncke, 2002;Fischer, Neumann, Purchase, et al, 2014;Lierz et al, 2013;Neumann et al, 2013;Schneider et al, 2018Schneider et al, , 2019Stelzer et al, 2005Stelzer et al, , 2009 (Schneider et al, 2017(Schneider et al, , 2018.…”

“…For this reason, we intended the evaluation of conventional live/dead stains in the first part of the study and the comparison with a modern fluorescence dye in the second part of the study; both backed up by a correlation with sperm motility results of the same samples. In particular, eosin blue, a single colorimetric stain that has been successfully used in sharp-tailed grouse (Schneider et al, 2019) and various psittacine species (Behncke, 2002;Fischer, Neumann, Purchase, et al, 2014;Lierz et al, 2013;Schneider et al, 2017Schneider et al, , 2018Stelzer et al, 2005Stelzer et al, , 2009 was evaluated in semen samples of falcons for the first time. Furthermore, eosin blue was compared in two different preparations (EB and EB2) to various established staining combinations of eosin yellow and eosin blue with different proportions of the background stains nigrosin or aniline (EYN5, EBN5 EYN10, EYA, EBA), respectively, to bromophenol blue-nigrosin (BBN) and the fluorescence stain SYBR ® Green-propidium iodide (SYBR-PI).…”

“…Likewise, in turkey semen the combination of SYBR with PI has been demonstrated as one of two most effective live/ dead stains of the various stains tested in the panel of different dilutions (Donoghue et al, 1995;Friars & Chatterjee, 1969). In semen of cockatiels, SYBR-PI enabled an efficient viability assessment as well as the conventional stain eosin blue 2% (Schneider et al, 2017(Schneider et al, , 2018, but detailed correlations to sperm motility have not been investigated in this species.…”

Sperm morphology and evidence for sperm competition among parrots

“…Sperm samples were taken directly from the cloaca using scaled glass capillaries (Wiretrol II, 1–5 µl; Drummond Scientific Company). From the samples collected in 2012–2013 and the Kākāpō samples (the latter were previously diluted, one with NaCl and the other with the semen extender Blanco, Schneider et al, ), smears were made onto microscope slides, stained with Eosin B2% and covered with a mounting medium (Entellan New, 107961; Merck KGaA), whereas the ones from 2018 were fixed in 50–100 µl of 5% formalin solution. From the samples fixed in formalin, we pipetted a 10 µl aliquot onto a microscope slide, spread it with the side of the pipette tip and allowed it to air‐dry.…”

Modification and Clinical Application of the Inner Perivitelline Membrane Test in Different Avian Species

“…Cock spermatozoa were diluted 1:2, and falcon spermatozoa were diluted at a 1:1 ratio with a commercially available diluent (6-Hour SemAid, PHL Associates, Inc., Davis, CA, USA) at 4 °C. The cockatiel spermatozoa were diluted at a ratio of 1:1, up to 1:3 depending on the concentration with a specific diluent for cockatiels [18]. Diluted samples were stored in the refrigerator (4 °C) before use.…”