In vivo validation of in vitro quality tests for cryopreserved honey bee semen

Wegener, Jakob; May, Tanja; Knollmann, U.; Kamp, Günter; Müller, Karin; Bienefeld, Kaspar

doi:10.1016/j.cryobiol.2012.04.010

Cited by 29 publications

(41 citation statements)

References 27 publications

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“…However, the morphological changes observed in Figure B, as well as the greatly increased intensity of PI staining in LN‐killed cells (compared to dead cells in the sample, or heat killed cells) has not been reported in other species, and is not apparent in the images of freeze–thaw killed turkey and bull sperm reported previously . Researchers involved in developing cryopreservation techniques for honey bee sperm have struggled to maintain viability following the freeze–thaw process . Peng et al also found that honey bee sperm incur extensive cellular injuries from freeze–thawing.…”

Section: Discussionmentioning

confidence: 85%

Flow cytometry as a rapid and reliable method to quantify sperm viability in the honeybee Apis mellifera

et al. 2014

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An important measure of male quality is sperm viability; i.e., the percentage of live sperm within an ejaculate, as this provides an accurate measure of the number of sperm potentially available for egg fertilization. Sperm viability is often determined by fluorescence microscopy using dyes that differentially stain viable and nonviable sperm, but the technique has a number of limitations. Here, a flow cytometry (FCM) method was developed, which allows the rapid determination of honeybee sperm viability, facilitating high throughput analyses. Using samples with known sperm viabilities, it was found that data obtained from FCM were more accurate and less variable compared with data obtained for the same samples using fluorescence microscopy. It was also found that a previously reported additional population of honeybee sperm found in datasets using FCM is caused by freeze-thawing samples. In conclusion, the method described here allows to quantify sperm viability of honeybees quickly and with high accuracy. This will be of great value for future scientific research and could also be of value to guide future bee breeding programs, given the agricultural importance of honeybees as pollinators. V C 2014 International Society for Advancement of Cytometry Key terms honeybee; sperm; flow cytometry; microscopy; viability; SYBR 14; propidium iodide; liquid nitrogen EJACULATE quality is a fundamentally important reproductive male trait as the number of live sperm available for egg fertilization after being transferred to the female's reproductive tract is a key determinant of male competitiveness, reproductive success, and fitness (1-4). A number of methods have been developed in the past to quantify semen quality, including parameters such as sperm viability, motility, morphology, and total number (5-7). Such inspections are often done microscopically or using machinery in combination with software, capable of simultaneously tracking large numbers of sperm cells within a sample (8). Sperm viability, in particular, has become a key measure for semen quality in recent years, and often refers to the proportion of live sperm in an ejaculate. It is generally quantified by measuring the plasma membrane integrity, as the plasma membrane becomes more permeable in dead sperm and coincides with the loss of motility (9,10) and therefore fertilization capacity. Consequently, assessing sperm viability within ejaculates has become of major interest for the study of reproductive biology as well as for medical research and practices, for example to understand and treat male infertility (11).To determine the proportion of live and dead sperm in a semen sample, a number of dyes have been developed, which allow relatively easy quantification of sperm viability. This approach has been used for a number of different species and for the study of a wide range of different questions (6,(12)(13)(14). SYBR 14 and propidium

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Section: Discussionmentioning

confidence: 85%

Flow cytometry as a rapid and reliable method to quantify sperm viability in the honeybee Apis mellifera

et al. 2014

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“…It is estimated that only 2.5 % of inseminated spermatozoa actively migrate into long-term storage (Baer 2005). Wegener et al (2012) found a strong correlation between sperm motility and indicators of sperm performance in inseminated queens. In our study, we noticed a significant decline in sperm motility only for drones exposed to an extremely high concentration of IMD (200 ppb).…”

Section: Discussionmentioning

confidence: 97%

“…Live video pictures were taken using an Olympus BX40 microscope (Olympus Optical, Tokyo, Japan) with a 10× negative phase objective and a Sony CCD black and white camera (SPT-M108CE). Spermatozoa were rated as motile if active Sperm quality after exposure to IMD 213 movement (mostly circular with sperm head and tail overlapping) (Wegener et al 2012) was presented. For each experimental treatment, semen was obtained from 180 males (three IMD concentrations for six colonies, 10 males per dose at each colony).…”

Section: Sperm Motilitymentioning

confidence: 99%

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Sperm parameters of honeybee drones exposed to imidacloprid

et al. 2016

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-The objective of this study was to evaluate the effects of chronical exposure of honeybee drones to environmental (5 ppb) and non-environmental concentration (200 ppb) of imidacloprid (IMD) on sperm concentration, motility, viability, and mitochondrial membrane potential measured in semen obtained from 180 drones originating from 18 colonies. The results demonstrate that IMD exposure did not affect sperm concentration; however, there were significant differences in concentration within colonies. IMD exposure was associated with reductions in sperm motility, which also varied within colonies. Statistically significant interactions between IMD exposure and colony were found for active mitochondria and sperm viability. Our results strongly suggest that neonicotinoids can negatively affect honeybee drone sperm quality. It is important to emphasize that IMD actions can be strongly modulated according to the colony.Apis mellifera / imidacloprid / spermatozoa / motility / viability

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“…Superoxide dismutase is one of the antioxidant enzymes involved in the survival of drone spermatozoa (Wegener et al 2012). SOD level has an essential role in maintaining the balance between reactive oxygen species (ROS) generation and degradation (Gavella et al 1996).…”

Section: Discussionmentioning

confidence: 99%

Semen quality of honey bee drones maintained from emergence to sexual maturity under laboratory, semi-field and field conditions

Abdelkader

Kairo²,

Tchamitchian³

et al. 2013

Apidologie

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International audienceIn order to evaluate the semen quality among honey bee populations, emergent honey bee drones were maintained to sexual maturity for 20 days under laboratory, semi-field, and field conditions. The drones were successfully maintained in laboratory conditions. Drones under laboratory and field conditions presented a lower spermatozoa concentration and lower protein content than those under semi-field conditions. The viability of spermatozoa was higher under laboratory conditions, and the ATP content and the superoxide dismutase activity were higher both under laboratory and field conditions compared to drones kept under semi-field conditions. Hence, our data indicate that the semen quality was similar in drones maintained under laboratory and field conditions

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In vivo validation of in vitro quality tests for cryopreserved honey bee semen

Cited by 29 publications

References 27 publications

Flow cytometry as a rapid and reliable method to quantify sperm viability in the honeybee Apis mellifera

Flow cytometry as a rapid and reliable method to quantify sperm viability in the honeybee Apis mellifera

Sperm parameters of honeybee drones exposed to imidacloprid

Semen quality of honey bee drones maintained from emergence to sexual maturity under laboratory, semi-field and field conditions

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