Giant panda (Ailuropoda melanoleuca) spermatozoon decondensation in vitro is not compromised by cryopreservation

2006 ◽  
Vol 18 (7) ◽  
pp. 767 ◽  
Author(s):  
Rebecca E. Spindler ◽  
Huang Yan ◽  
JoGayle Howard ◽  
Wang PengYan ◽  
Zhang Hemin ◽  
...  
Keyword(s):  
Giant Panda ◽  
Acrosome Reaction ◽  
Ailuropoda Melanoleuca ◽  
Genetic Management ◽  
Giant Pandas ◽  
Before And After ◽  
In Captivity ◽  
Oocyte Cytoplasm ◽  
Natural Breeding

Natural breeding of giant pandas in captivity is compromised, making artificial insemination and spermatozoa cryopreservation essential for genetic management. This study examined the influence of freeze–thawing on traditional parameters such as motility and spermatozoon functionality, specifically decondensation in vitro. Giant panda spermatozoa were assessed before and after rapid cryopreservation (4°C to –130°C over 2 min) in liquid nitrogen vapour. Spermatozoa pre-incubated in medium for 6 h were co-incubated with cat zonae (2 zonae μL–1) for 30 min to effect capacitation and an acrosome reaction. Spermatozoa were then mixed with mature cat oocyte cytoplasm (2 cytoplasm μL–1) for 4 h and evaluated for decondensation. Frozen spermatozoa were less motile (P < 0.05) than fresh counterparts immediately post-thawing, but not after 6 h incubation. There were more (P < 0.05) spermatozoa with completely diffused chromatin post-thaw (10.4 ± 1.3%; mean ± s.e.m.) compared to fresh counterparts (5.1 ± 1.0%). However, there was no overall difference (P > 0.05) in the incidence of decondensation between fresh (4 h, 69.8 ± 5.9%) and thawed (4 h, 71.5 ± 4.9%) spermatozoa after exposure to cat oocyte cytoplasm. It is concluded that the ‘rapid’ method now used to cryopreserve giant panda spermatozoa has little impact on spermatozoon decondensation.

scholarly journals Acrosomal integrity and capacitation are not influenced by sperm cryopreservation in the giant panda

Reproduction ◽  
2004 ◽  
Vol 127 (5) ◽  
pp. 547-556 ◽  
Author(s):  
R E Spindler ◽  
Y Huang ◽  
J G Howard ◽  
P Wang ◽  
H Zhang ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Giant Panda ◽  
Calcium Ionophore ◽  
Sperm Cryopreservation ◽  
Management Tools ◽  
Giant Pandas ◽  
Before And After ◽  
Cryopreservation Protocol ◽  
Acrosomal Integrity

Sperm cryopreservation and artificial insemination are important management tools for giant panda breeding and the preservation of extant genetic diversity. This study examined the influence of freeze–thawing on sperm function, specifically capacitation. Sperm from nine giant pandas were assessed before and after rapid (− 40 and − 100 °C/min) cryopreservation by incubation in HEPES-buffered Ham’s F10 medium with and without the capacitation accelerators, 3-isobutyl-1-methylxanthine (IBMX) and dibutyryl cyclic AMP (dbcAMP). At 0, 3 and 6 h of exposure, aliquots were assessed for sperm motility traits and capacitation, defined as the proportion of sperm with intact acrosomes following exposure to solubilised zonae pellucidae (ursid or felid) or calcium ionophore subtracted from the proportion of sperm with intact acrosomes before exposure. Although mean±s.e.m. sperm motility post-thaw (56.1 ± 3.9% at 0 h) was less (P < 0.05) than pre-freeze (71.7 ± 6.0%), there was no difference (P > 0.05) in the proportion of acrosome-intact sperm (fresh, 93.0 ± 1.7% versus cryopreserved–thawed, 81.7 ± 4.7% at 0 h). Incidence of capacitation was greater (P < 0.05) in fresh sperm incubated with capacitation accelerators IBMX and dbcAMP (9 h: 50.9 ± 1.1) compared with fresh sperm incubated without accelerators (9 h: 41.2 ± 1.1%). Frozen–thawed sperm preincubated without accelerators underwent capacitation (49.6 ± 1.1%) to a greater extent (P < 0.05) compared with these fresh counterparts. Thawed samples with (9 h: 45.9 ± 1.4%) and without accelerators (9 h: 41.2 ± 1.1%) did not differ (P > 0.05) during the 9-h incubation. We conclude that giant panda spermatozoa (1) undergo capacitation in vitro with or without chemical accelerators and (2) withstand a rapid cryopreservation protocol, including retaining normal acrosomal integrity and functional capacitation ability.

scholarly journals Immune profiles of male giant panda (Ailuropoda melanoleuca) during breeding season

2021 ◽  
Author(s):  
haibo shen ◽  
Caiwu Li ◽  
Ming He ◽  
Yan Huang ◽  
Jing Wang ◽  
...  
Keyword(s):  
Pattern Recognition ◽  
Breeding Season ◽  
Giant Panda ◽  
Expression Profiles ◽  
Killer Cell ◽  
Pattern Recognition Receptors ◽  
Ailuropoda Melanoleuca ◽  
Giant Pandas ◽  
Altered Expression ◽  
Immune Related Genes

Abstract Background: The giant panda (Ailuropoda melanoleuca) is a threatened endemic Chinese species and a flagship species of national and global conservation concern. Life history theory proposes that reproduction and immunity can be mutually constraining and interrelated. Knowledge of immunity changes of male giant pandas during the breeding season is limited.Results: Here, we researched peripheral blood gene expression profiles associated with immunity. Thirteen captive giant pandas, ranging from 9 to 11 years old, were divided into two groups based on their reproductive status. We identified 318 up-regulated DEGs and 43 down-regulated DEGs, which were enriched in 87 GO terms and 6 KEGG pathways. Additionally, we obtained 45 immune-related genes with altered expression, mostly up-regulated, and identified four hub genes HSPA4, SUGT1, SOD1, and IL1B in PPI analysis. These 45 genes were related to pattern recognition receptors, autophagy, peroxisome, proteasome, natural killer cell, antigen processing and presentation. SUGT1 and IL1B were related to pattern recognition receptors. HSP90AA1 was the most up-regulated gene and is a member of heat shock protein 90 family. HSP90 contributes to the translocation of extracellular antigen. KLRD1 encodes CD94, whose complex is an inhibitor of the cytotoxic activity of NK cells, was down-regulated. IGIP, which has the capability of inducing IgA production by B cells, was down-regulated, suggesting low concentration of IgA in male giant pandas. Our results suggest that most immune-related genes were up-regulated and more related to innate immune than adaptive immune. Conclusions: Our results indicated that breeding male giant pandas presented an immunoenhancement in innate immunity, enhanced antigen presentation and processing in cellular immunity compared to non-breeding males. The humoral immunity of male giant pandas may show a tendency to decrease during the breeding season. This study will provide a foundation for further studies of immunity and reproduction in male giant pandas.

Anogenital gland secretions code for sex and age in the giant panda, Ailuropoda melanoleuca

2004 ◽  
Vol 82 (10) ◽  
pp. 1596-1604 ◽  
Author(s):  
Hong Yuan ◽  
Dingzhen Liu ◽  
Lixing Sun ◽  
Rongping Wei ◽  
Guiquan Zhang ◽  
...  
Keyword(s):  
Giant Panda ◽  
Fatty Acid Esters ◽  
Ailuropoda Melanoleuca ◽  
Giant Pandas ◽  
Analog Form ◽  
Males And Females ◽  
Relative Abundances ◽  
Anogenital Gland Secretions ◽  
Acid Esters ◽  
Form Information

Anogenital gland secretions play a major role in chemical communication by giant pandas, Ailuropoda melanoleuca (David, 1869). We analyzed 45 samples of anogenital gland secretions collected from 24 captive pandas (5 male adults, 6 female adults, 6 male subadults, and 7 female subadults) by gas chromatography and mass spectrometry. The secretions contained over 95 compounds. Based on 56 common compounds (relative abundances >0.1%) shared by more than three individuals, we identified steroids, long-chain fatty acids, fatty-acid esters, aldehydes, alkanes, alkenes, amines, terpenes, and furans. The chemical composition of each secretion was individual-specific. Although none of these individual compounds was age- or sex-specific, the relative abundances of several compounds differed between males and females and between adults and subadults. This result shows that information about sex and age could be coded in analog form. Information about age but not gender could also be digitally coded by the presence or absence of some of the 56 compounds, in addition to the analog coding.

92 EVALUATION OF BOAR SPERM FUNCTIONALITY AFTER A CUSHIONED CENTRIFUGATION TECHNIQUE

2006 ◽  
Vol 18 (2) ◽  
pp. 154
Author(s):  
J. Gadea ◽  
S. Martínez-Miró ◽  
G. Decuadro-Hansen ◽  
C. Matás
Keyword(s):  
Seminal Plasma ◽  
Acrosome Reaction ◽  
Sperm Viability ◽  
Lipid Packing ◽  
Lipid Disorder ◽  
Merocyanine 540 ◽  
Centrifuge Tube ◽  
Before And After ◽  
Centrifugation Technique

Separation of sperm from seminal plasma is required in most semen freezing procedures. Semen is typically subjected to centrifugation to concentrate sperm into a pellet and allow removal of the seminal plasma prior to dilution in freezing extender. Centrifugation is a relatively effective method to recover sperm, however, the process also causes considerable sperm damage. The use of a dense, inert, and isotonic solution as a cushion in the bottom of the centrifuge tube allows a greater centrifugation speed to be applied and results in greater sperm recovery. The aim of the present work was to evaluate the effects of this cushioned centrifugation technique on in vitro sperm viability and functionality. Sperm-rich fractions from 16 fertile boars were diluted and cooled to 15�C; then subsamples were centrifuged by one of two different techniques. A standard method (SM), 800 g for 10 min in 50-mL tubes (Westendorf et al. 1975 Dtsch. Tier�rztl. Wschr. 82, 261-267) and a cushioned method (CM), 1000 g for 20 min using 45 mL of diluted semen on 5 mL of an isotonic iodixanol solution (60% w/v gradient) were performed. Sperm samples were stained with merocyanine 540 (M540) and Yo-Pro 1 (Harrison et al. 1996 Mol. Rep. Dev. 45, 378-391) to detect changes in lipid packing disorder of the plasma membrane. Another set of sperm samples was incubated in the presence of (0.7 �M) 22,72-dichlorodihydrofluorescein diacetate (Gadea et al. 2005 J. Androl. 26, 396-404) to estimate production of reactive oxygen species (ROS). A final set of sperm samples was stained with peanut aggultinin-fluorscein isothiocyanate (PNA-FITC) and propidium iodide to evaluate the acrosome reaction. All of these parameters were evaluated by flow cytometry before and after centrifugation. ANOVA analysis revealed that centrifugation altered lipid packing disorder and viability. Raw semen (RS) had a larger number of viable low lipid disorder sperm than centrifuged semen (RS = 86.9a vs. SM = 81.64b vs. CM = 80.6b, P < 0.01) and a decreased number of dead sperm cells (RS = 9.5a vs. SM = 15.0b vs. CM = 16.3b, P < 0.01). However, the cushioned and standard centrifugation methods yielded similar results for all the parameters measured. No significant differences were found for generation of ROS or in the number of sperm exhibiting the acrosome reaction. In conclusion, compared to the standard centrifugation method, this simple cushioned modification is a more efficient means of processing boar semen for freezing because significantly less sample losses are detected; also, it provides similar levels of sperm viability and functionality, and consequently a higher number of doses per ejaculation can be produced.

scholarly journals Topographic habitat features preferred by the Endangered giant panda Ailuropoda melanoleuca: implications for reserve design and management

Oryx ◽  
2011 ◽  
Vol 45 (2) ◽  
pp. 252-257 ◽  
Author(s):  
Ming-Hao Gong ◽  
Yan-Ling Song
Keyword(s):  
Geographical Information Systems ◽  
Giant Panda ◽  
Decisive Role ◽  
Reserve Design ◽  
Geographical Information ◽  
Limiting Factors ◽  
Ailuropoda Melanoleuca ◽  
High Quality ◽  
Giant Pandas ◽  
Habitat Features

AbstractThe effect of landscape on populations is of great importance, especially given the number of species inhabiting patchy landscapes. Developments in geographical information systems are facilitating a greater application of spatial analyses to threatened species, such as the Endangered giant panda Ailuropoda melanoleuca, for which habitat patchiness and quality are key limiting factors. Of all factors that influence the suitability of habitat for the giant panda, topography is not subject to change. Here, we report a spatial and statistical analysis of the high quality topographic habitat preferred by the giant panda across its stronghold in the Qinling Mountains, China. High quality topographic habitat, as indicated by a combination of elevation, slope and aspect, covers 92,788 ha, accounting for 15% of the current range of the species. The distribution of the giant panda closely follows patterns of topography and areas with patches of high quality topographic habitat are strongly associated with areas supporting greater numbers of giant pandas. However, comparisons between our model and the existing reserve system reveals a number of inadequacies. Some of the reserves contain little high quality topographic habitat and many patches of high quality topographic habitat are unprotected. Given the importance of topography and the decisive role it plays in shaping habitat, landscapes containing high quality topographic features must be a critical consideration in the design of reserves for the giant panda. The existing system of nature reserves is heavily weighted towards judicial and administrative boundaries, to the detriment of other factors such as topography.

scholarly journals Testosterone Changes in Female Pandas in Estrus

2019 ◽  
Author(s):  
Xuefeng Liu ◽  
Lei Yuan ◽  
Wei Wang ◽  
Yuyan You ◽  
Yanhui Liu ◽  
...  
Keyword(s):  
Artificial Insemination ◽  
Giant Panda ◽  
Estradiol Level ◽  
Giant Pandas ◽  
Testosterone Levels ◽  
Natural Breeding ◽  
And Behavior ◽  
Estradiol Levels ◽  
Human Primate

AbstractCurrently, the majority of giant panda breeding is carried out by cage-mating or artificial insemination based on estrogen levels and behavior in female pandas. However, studies have shown that testosterone levels both in women and in non-human primate females have a significant effect on the desire to mate. In this study, we wanted to explain how testosterone levels of female giant pandas would change during estrous. In this study, 23 accounts of rutting were recorded in 10 female pandas from 2009 to 2012. Changes in urinal testosterone levels were monitored and compared with estradiol values. Our data showed that, for female pandas in estrus, testosterone levels after the estradiol peak was significantly higher than before, and the testosterone peak occurred 4 days after the estradiol peak. Furthermore, testosterone and estradiol level were only significantly correlated after peak estradiol levels peaked, and not before. Finally, out findings suggest that testosterone could help us better understand hormone variation during panda estrus, as well as help aid in the natural breeding of pandas.

scholarly journals Contrasting Vaginal Bacterial Communities Between Estrus and Non-estrus of Giant Pandas (Ailuropoda melanoleuca)

2021 ◽  
Vol 12 ◽  
Author(s):  
Chanjuan Yue ◽  
Xue Luo ◽  
Xiaoping Ma ◽  
Dongsheng Zhang ◽  
Xia Yan ◽  
...  
Keyword(s):  
Bacterial Communities ◽  
Giant Panda ◽  
16S Rrna Gene Sequencing ◽  
Abundant Species ◽  
Rrna Gene ◽  
Ailuropoda Melanoleuca ◽  
Sequencing Analysis ◽  
Vaginal Microbiome ◽  
Giant Pandas ◽  
Vaginal Secretions

Bacterial infection and imbalance of bacterial community in the genitourinary system of giant panda could affect the reproductive health. In severe cases, it can also lead to abortion. In this study, 13 of vaginal secretions in the estrue (E) group and seven of vaginal secretions in the non-estrue (NE) group were used to study the composition and diversity of vaginal bacterial communities between estrus and non-estrus by 16S rRNA gene sequencing analysis. The results showed that the vaginal microbiome in giant pandas shared the same top five abundant species between estrus and non-estrus at the phylum level. However, the vaginal microbiome changed significantly during estrus at the genus level. In top 10 genera, the abundance of Escherichia, Streptococcus, and Bacteroides in the E group was significantly higher than that in the NE group (p&lt;0.05); Azomonas, Porphyromonas, Prevotella, Campylobacter, and Peptoniphilus in the NE group was significantly higher than that in the E group (p&lt;0.05). The richness and diversity of vaginal microbiome in giant panda on estrus were significantly lower than those on non-estrus (p&lt;0.05). It is noteworthy that the abundance of Streptococcus, Escherichia, and Bacteroides of vagina in giant pandas maintained low abundance in the daily. Whereas, they increased significantly during estrus period, which may play an important role in female giant pandas during estrus period. It was hypothesized that hormones may be responsible for the changes in the vaginal microbiome of giant pandas between estrus and no-estrus stages.

scholarly journals Immune profiles of male giant panda (Ailuropoda melanoleuca) during breeding season

2020 ◽  
Author(s):  
haibo shen ◽  
Caiwu Li ◽  
Ming He ◽  
Yan Huang ◽  
Jing Wang ◽  
...  
Keyword(s):  
Breeding Season ◽  
Antigen Processing ◽  
Giant Panda ◽  
Life History Theory ◽  
Expression Profiles ◽  
Killer Cell ◽  
Ailuropoda Melanoleuca ◽  
Giant Pandas ◽  
Altered Expression ◽  
Immune Related Genes

Abstract Background The giant panda (Ailuropoda melanoleuca), regarded as the flagship species of conservation, is a most threatened species endemic to China. Life history theory proposes that reproduction and immunity can be mutually constraining and interrelated. The immunity changes of male giant pandas during reproduction is not clear. Results Here, we did research on peripheral blood gene expression profiles associated with immunity. 13 giant pandas with ages ranging from 9 to 11 years old were divided into two groups depending on whether or not they were in breeding season. We identified 318 up-regulated DEGs and 43 down-regulated DEGs, which were enriched in 87 GO terms and 6 KEGG pathways. What’s more, we obtained 45 immune-related genes with altered expression, mostly up-regulated, and identified four hub genes HSPA4, SUGT1, SOD1 and IL1B in PPI analysis. These genes were related to pattern recognition receptors, autophagy, peroxisome, proteasome, natural killer cell, antigen processing and presentation. Our results suggest that most immune-related genes were up-regulated and more related to innate immune than adaptive immune. Conclusions Male giant pandas presented an immunoenhancement in innate immunity, enhanced antigen presentation and processing in cellular immunity, and a low humoral immunity during breeding season. This study will provide a foundation for further studies of immunity and reproduction in giant pandas.

scholarly journals Diagnosing Zygosity in Giant Panda Twins Using Short Tandem Repeats

2018 ◽  
Vol 21 (6) ◽  
pp. 527-532
Author(s):  
Maiju Qiao ◽  
Yingmin Zhou ◽  
Thomas Connor ◽  
Rengui Li ◽  
Dan Tang ◽  
...  
Keyword(s):  
Giant Panda ◽  
Tandem Repeats ◽  
Monozygotic Twins ◽  
Giant Pandas ◽  
West China ◽  
Conservation Effort ◽  
In Captivity ◽  
Mz Twins ◽  
Insight Into ◽  
Short Tandem

The giant panda, native to mountains of south-west China, is one of the world's rarest bear species and is subject to considerable conservation effort. In captivity, the proportion of twins accounts for 54% of the total number of births. To date, little is known about zygosity in panda populations — specifically, the proportion of monozygotic and dizygotic twins. In this study, we used 10 microsatellite markers for reliable zygosity testing, and the probability of monozygotic twins was 99.963% when all 10 markers were concordant. Out of 43 studied twin pairs, no MZ twins were found, indicating that there may be no identical panda twins (or the incidence is very low). We speculate that the fertilized eggs of giant pandas do not have the capability to split into two identical embryos, or that this ability is very poor, which is likely due to delayed implantation that is common in bear species. The results of this study deepen our understanding of giant panda breeding, yield insight into panda twins’ likely mechanism of formation, and reduce the uncertainty of individual identity in wild population surveys.

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