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1.	Abraham, Maria Celina, et al. (författare) Comparison of three variants of colloid centrifugation for sperm selection: sperm quality and blastocyst development in vitro. 2016 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 169, s. 96-96 Konferensbidrag (refereegranskat)
2.	Al-Essawe, Essraa M, et al. (författare) Addition of seminal plasma to thawed stallion spermatozoa did not repair cryoinjuries 2018 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 196, s. 48-58 Tidskriftsartikel (refereegranskat)abstract Freezing and thawing processes induce structural and functional damage to sperm plasma membranes and internal organelles. Adding seminal plasma (SP) has been found to minimize or repair the cryoinjuries in some species. The objective of this study was to investigate whether adding SP from stallions of known freezability after thawing could repair cryoinjuries. Semen was collected from warmblood stallions (n = 8, three ejaculates/stallion) and processed by Single Layer Centrifugation (SLC) to remove SP prior to freezing. Pooled SP (5%) from bad freezer (BF) or good freezer (GF) stallions was added after thawing. Post-thaw sperm quality was assessed by flow cytometry in terms of chromatin integrity (ßI), membrane integrity, mitochondrial membrane potential (MMP), reactive oxygen species (ROS), and MitoSOX. Sperm kinematics were also assessed by computer-assisted sperm analysis. The ßI was lower in SLC control (C) than in BF or GF (P < 0.0001, P < 0.0003 respectively). The proportion of viable spermatozoa with intact cell membranes was higher in C than in SP treated groups (C vs. BF, P = 0.02; C vs GF, P = 0.05). There were fewer spermatozoa with low MMP and more with high MMP for C than GF (P = 0.006). The spermatozoa treated with SP from good freezers produced more ROS than when treated with SP from bad freezers (P = 0.007). Motility parameters were not affected by adding SP. In conclusion, adding SP after thawing does not have a beneficial effect on sperm quality, suggesting an inability to repair stallion sperm cryoinjuries, regardless of whether the SP originated from stallions semen, which has good or bad quality after thawing.
3.	Al-Essawe, Essraa M, et al. (författare) Extenders for alpaca epididymal spermatozoa: Comparison of INRA96 and andromed 2020 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 223 Tidskriftsartikel (refereegranskat)abstract Artificial insemination would be a useful technique for alpaca breeders to use as an aid to breeding to increase fleece quality. The technique, however, is not well developed in alpacas, partly because of the viscous nature of their seminal plasma. Castration conducted for husbandry purposes can provide a source of epididymal spermatozoa to test semen extenders or handling regimens, thus circumventing the problem of the viscous ejaculate. In this experiment, two semen extenders (Andromed and INRA96) developed for other species (bovine and equine, respectively) were tested with alpaca spermatozoa derived from the cauda epididymis. Sperm total motility (mean ± SEM A: 29.1 ± 4.8 % compared with I: 35.4 ± 4.8 %; NS), membrane integrity (A: 58 ± 9% compared with I: 56 ± 9%; NS) and acrosome integrity (A: 65 ± 7% compared with I: 54 ± 7%; NS) were not different between the two extenders. Progressive motility with use of INRA96 was greater after incubating for 30 min than after incubating for 10 min (35 ± 4% vs. 12 ± 4%, respectively; P = 0.03). In conclusion, viable epididymal spermatozoa could be extracted from the castrated organs after overnight transport. There were no differences in sperm quality between the two extenders; therefore, it appears that either extender could be used for alpaca spermatozoa. These results could help in the development of a technique for artificial insemination in alpacas.
4.	Al-Essawe, Essraa M, et al. (författare) Seminal plasma components differ between "good freezer" and "poor freezer" stallions 2016 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 169, s. 113-113 Konferensbidrag (refereegranskat)
5.	Al-Kass, Ziyad, et al. (författare) Deciphering sperm chromatin properties to predict stallion sperm fertility 2023 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 250 Tidskriftsartikel (refereegranskat)abstract Although previous studies have examined the relationship between the sperm DNA fragmentation index and fertility in stallions, other aspects of chromatin structure or packaging and fertility have not been explored. In the present study, relationships between fertility and DNA fragmentation index, protamine deficiency, total thiols, free thiols and disulfide bonds in stallion spermatozoa were investigated. Ejaculates (n = 36) were collected from 12 stallions and extended to prepare semen doses for insemination. One dose from each ejaculate was sent to the Swedish University of Agricultural Sciences. Aliquots of semen were stained for flow cytometry with acridine orange for the Sperm Chromatin Structure Assay (DNA fragmentation Index, %DFI), with chromomycin A3 (CMA) for protamine deficiency, and with monobromobimane (mBBr) for detection of total and free thiols and disulfide bonds. Per season pregnancy rates after insemination were obtained. Mixed linear models were used to analyze data. Negative correlations were found between pregnancy rate and %DFI (r = -0.35, P < 0.03) and pregnancy rate and free thiols (r = -0.60, P < 0.0001). Furthermore, there were positive correlations between total thiols and disulfide bonds (r = 0.95, P < 0.0001), and protamine and disulfide bonds (r = 0.4100, P < 0.01986). Since chromatin integrity, protamine deficiency and packaging were all associated with fertility, a combination of these factors could be used as a biomarker of fertility when assessing ejaculates.
6.	Al-Kass, Z., et al. (författare) Metagenomic analysis of bacteria in stallion semen 2020 Ingår i: Animal Reproduction Science. - : ELSEVIER. - 0378-4320 .- 1873-2232. ; 221 Tidskriftsartikel (refereegranskat)abstract Bacteria colonize stallion semen during collection and processing which may cause disease in inseminated females or negatively affect sperm quality during storage prior to insemination. Antibiotics are added to semen extenders to control the growth of these bacteria but may induce antimicrobial resistance. Research into alternatives to antibiotics for this purpose requires knowledge of which bacteria are present in semen. Not all bacteria in semen, however, can be identified by conventional microbiological techniques. The objectives of the study were to: i) determine which bacteria are present in stallion semen using metagenomics; and ii) investigate individual differences in bacterial content in semen from all stallions on one premises. Bacterial DNA was extracted from ejaculates from seven stallions (one ejaculate per stallion) and bacteria were identified using 16S sequencing. In total, 83 bacterial genera were identified, varying from 25 to 52 among different individuals. There was a negative correlation (r = -0.81212; P < 0.05) between the presence of Treponema spp. and Advenella spp. In conclusion, most of the bacteria present in stallion semen could be identified to genus level by 16S sequencing even when present at a low frequency. This method of identification may help to clarify individual variation in bacterial content and its potential effects on fertility.
7.	Alvarez-Rodriguez, Manuel, et al. (författare) Effect of length of time post-mortem on quality and freezing capacity of Cantabric chamois (Rupicapra pyrenaica parva) epididymal spermatozoa 2018 Ingår i: Animal Reproduction Science. - : ELSEVIER SCIENCE BV. - 0378-4320 .- 1873-2232. ; 198, s. 184-192 Tidskriftsartikel (refereegranskat)abstract Genome Resource Banks are keystones in the ex-situ conservation of wild species. Post-mortem (PM) collection of epididymal spermatozoa is an opportunistic and valuable source of germplasm, the time from the death of the animal limits its use. Seeking to improve germplasm preservation strategies for the chamois (Rupicapra sp.), the effect of PM time on epididymal sperm quality and freezability was studied using the Cantabrian chamois. Samples were classified according to PM collection time, up to 216 h (refrigerated), and cryopreserved (Tris-citric acid-fructose, 430 mOsm/kg, 15% egg yolk, 8% glycerol; freezing at - 20 degrees C/min). Sperm quality was assessed after recovery and post-thawing (motility by CASA, HOS test, abnormal forms, cytoplasmic droplets, and viability and acrosomal damage by flow cytometry). The sperm mass pH and osmolality showed a positive correlation with time. Total sperm motility dropped after 2 days PM, with progressivity and sperm velocities remained similar up to 3 days PM. Sperm freezability was acceptable, with the post-thawing HOST, motility, progressivity, VAP, VCL, VSL and BCF negatively correlating with PM time. Overall, chamois epidydimal samples were not adequate for preservation after 6 days PM. Freezability capacity could make these spermatozoa suitable for specific ART even if kept refrigerated for several days PM.
8.	Anel-López, Luis, et al. (författare) Reduced glutathione and Trolox (vitamin E) as extender supplements in cryopreservation of red deer epididymal spermatozoa 2012 Ingår i: Animal Reproduction Science. - : Elsevier. - 0378-4320 .- 1873-2232. ; 135:1-4, s. 37-46 Tidskriftsartikel (refereegranskat)abstract The use of assisted reproductive techniques in cervids is increasing as the commercial use of these species increase. We have tested the suitability of the antioxidants Trolox and reduced glutathione (GSH) for freezing red deer epididymal spermatozoa, aiming at improving post-thawing quality. Samples from 19 stags were frozen in a TES-Tris-fructose extender (20% egg yolk, 8% glycerol), with 1 or 5 mM of antioxidant. Motility (CASA), lipoperoxidation (malondialdehyde -MDA- production), membrane status, mitochondrial activity, acrosomal status (flow cytometry) and chromatin status (SCSA: %DFI and %HDS; flow cytometry) were assessed after thawing and after 6 h at 39°C. There were few differences between treatments after thawing, with Trolox reducing MDA production in a dose-response manner. After the incubation, sperm quality decreased and %DFI increased moderately, with no change for MDA. GSH improved motility, kinematic parameters and mitochondrial status, with a slight increase in %HDS. GSH 5 mM also increased moderately MDA production and %DFI, possibly due to enhanced metabolic activity and reducing power. Trolox maintained MDA low, but was detrimental to sperm quality. Trolox might not be appropriate for the cryopreservation of red deer epididymal spermatozoa, at least at the millimolar range. GSH results are promising, especially regarding motility improvement after the post-thawing incubation, and should be selected for future fertility trials.
9.	Axner, Eva, et al. (författare) Characteristics of reproductive organs and reproductive potential in Scandinavian female grey wolves (Canis lupus). 2023 Ingår i: Animal Reproduction Science. - 0378-4320 .- 1873-2232. ; 255 Tidskriftsartikel (refereegranskat)abstract The Swedish wolf population is closely monitored and managed to keep the population at a sustainable level while avoiding conflicts. Detailed knowledge about reproduction is crucial for estimates of population size and the reproductive potential of a population. Post-mortem evaluation of reproductive organs can be used as a complementary tool to field monitoring for evaluation of cyclicity and previous pregnancy, including litter size. Therefore, we evaluated reproductive organs from 154 female wolves that were necropsied during the period 2007-2018. The reproductive organs were weighed, measured, and inspected according to a standardised protocol. Presence of placental scars was evaluated for estimates of previous pregnancy and litter size. Data about individual wolves were also obtained from national carnivore databases. Body weight increased during the first year of life before levelling out. There was evidence of cyclicity the first season after birth in 16.3 % of the 1-year-old females. No females < 2 years had evidence of a previous pregnancy. Pregnancy rates were significantly lower in 2- and 3-year old females than in older females. Mean uterine litter size was 4.9 & PLUSMN; 2.3, and did not differ significantly between age groups. Our data supports earlier field data that female wolves usually start to reproduce at the earliest at 2-years of age but that they occasionally start to cycle one season earlier. All females & GE; 4 years of age had reproduced. Pathological findings of the reproductive organs were rare, indicating that reproductive health of female wolves is not a limiting factor for population growth.
10.	Balao da Silva, C, et al. (författare) Effect of Hoechst 33342 on stallion spermatozoa incubated in KMT or Tyrodes modified INRA96 2012 Ingår i: Animal Reproduction Science. - : Elsevier Masson. - 0378-4320 .- 1873-2232. ; 131:3-4, s. 165-171 Tidskriftsartikel (refereegranskat)abstract The only known means of effectively separating populations of X and Y bearing sperms is the Beltsville sexing technology. The technology implies that each individual sperm is interrogated for DNA content, measuring the intensity of the fluorescence after staining the spermatozoa with Hoechst 33342. Because there are no data regarding the effect of the staining on stallion sperm, ejaculates were incubated up to 90 min in presence of 0, 4.5, 9, 22.5, 31.5, 45, 54, 67.5, 76.5 and 90 mu M of Hoechst 33342, in two media, KMT or INRA-Tyrodes. After 40 and 90 min of incubation, motility (CASA) and membrane integrity (flow cytometry after YoPro-1/Eth staining) were evaluated. In KMT extender sperm motility significantly decreased after 45 min of incubation when sperm were incubated in the presence of concentrations of Hoechst of 45 mu M or greater (P andlt; 0.05). When incubated in modified INRA96, stallion spermatozoa tolerated greater concentrations of Hoechst, because sperm motility only decreased when incubated in presence of 90 mu M (P andlt; 0.05) and membrane integrity was not affected. After 90 min of incubation the same effect was observed, but in this case at concentrations over 45 mu M the percentage of total motile sperm was also reduced although only in samples incubated in KMT. To produce this effect in samples incubated in Tyrodes modified INRA 96, Hoechst had to be present at concentrations over 67.5 mu M. Apparently, the detrimental effect of Hoechst to stallion spermatozoa varies depending on the media, and INRA modified extender may be an alternative to KMT.
11.	Balao da Silva, C M., et al. (författare) Sex sorting increases the permeability of the membrane of stallion spermatozoa 2013 Ingår i: Animal Reproduction Science. - : Elsevier Masson. - 0378-4320 .- 1873-2232. ; 138:3-4, s. 241-251 Tidskriftsartikel (refereegranskat)abstract At present, the only repeatable means of selecting the sex of offspring is the Beltsville semen sorting technology using flow cytometry (FC). This technology has reached commercial status in the bovine industry and substantial advances have occurred recently in swine and ovine species. In the equine species, however, the technology is not as well developed. To better understand the changes induced in stallion spermatozoa during the sorting procedure, pooled sperm samples were sorted: sperm motility and kinematics were assessed using computer assisted sperm analysis, sperm membrane integrity was assessed using the YoPro-1 assay, while plasmalemmal stability and lipid architecture were assessed using Merocyanine 540/SYTOX green and Annexin-V, respectively. Lipid peroxidation was also investigated with the probe Bodipy(581/591)-C11. All assays were performed shortly after collection, after incubation and after sex sorting using FC. In order to characterize potential molecular mechanisms implicated in sperm damage, an apoptosis protein antibody dot plot array analysis was performed before and after sorting. While the percentage of total motile sperm remained unchanged, sex sorting reduced the percentages of progressive motile spermatozoa and of rapid spermatozoa as well as curvilinear velocity (VCL). Sperm membranes responded to sorting with an increase in the percentage of YoPro-1 positive cells, suggesting the sorted spermatozoa had a reduced energy status that was confirmed by measuring intracellular ATP content.
12.	Bergqvist, AS, et al. (författare) Detection of Fas ligand in the bovine oviduct 2005 Ingår i: Animal Reproduction Science. - : Elsevier Masson. - 0378-4320 .- 1873-2232. ; 86:1-2, s. 71-88 Tidskriftsartikel (refereegranskat)abstract Presence of a Fas-Fas ligand (FasL) system defines the immune-privileged status of certain tissues such as placenta. This study examined the fluids and tissue(s) of the bovine oviduct, where both spermatozoa and early embryos escape elimination by the female immune system, for the presence and the distribution of Fas and FasL, which might provide an explanation for the immune-privilegded site of this organ. In the present study, the immunolocalisation of FasL and Fas, as well as the gene expression of FasL, were determined in the uterotubal junction (UTJ), isthmic (I) and ampullar (A) segments of the oviduct during oestrus and the luteal phase of the oestrous cycle. The degree of apoptosis of oviductal epithelium was examined by the TUNEL method. Oviductal fluid (01317), collected chronically via indwelling catheters from the I or A segments during both non-luteal and luteal phases of the cycle, was analysed for the presence of FasL. The Fas immunostaining was scattered along the epithelium of all regions of the oviduct and cycle stages investigated, whereas FasL immunolabelling was more conspicuous in oestrous samples. This staining disappeared during the luteal phase, which was particularly evident in the sperm reservoir (UTJ and I). There were fewer TUNEL-positive cells than Fas- or FasL-positive cells in the oviductal epithelium, suggesting that tubal Fas and FasL are not directly involved in epithelia apoptosis. Western blot analyses detected FasL in ODF collected from both I and A, most conspicuously as a 24-27 kDa band but also at a 40-45 kDa band level. FasL mRNA was expressed in the epithelial cells from the sperm reservoir and A during both non-luteal and luteal phases. However, the level of expression differed significantly between segments during the luteal phase. The results provide novel evidence that the Fas-FasL system is present in the bovine oviduct and could be involved in mediating survival of spermatozoa and early embryos. (c) 2004 Elsevier B.V. All rights reserved.
13.	Bergqvist, AS, et al. (författare) Sulphated glycosaminoglycans (S-GAGs) and syndecans in the bovine oviduct 2006 Ingår i: Animal Reproduction Science. - : Elsevier Masson. - 0378-4320 .- 1873-2232. ; 93:1-2, s. 46-60 Tidskriftsartikel (refereegranskat)abstract In vivo, bull sperm capacitation seems to occur mainly in the oviduct. Capacitation of bull spermatozoa can be triggered in vitro by exposure to heparin, a heavily sulphated glycosaminoglycan (S-GAG). We determined the concentration of S-GAGs in oviductal fluid from dairy heifers, collected over the course of several oestrous cycles via surgically implanted intraluminal catheters. We also investigated the presence of syndecans, i.e. heparan sulphate proteoglycans, in the bovine oviductal epithelium of Swedish dairy cattle during standing oestrus and the luteal phase of the oestrous cycle, using immunohistochemistry for three different polyclonal antibodies raised against human syndecan-2 and rat syndecan-1 and syndecan-2, respectively. The concentration of S-GAGs in oviductal fluid obtained from the ampullar segment of the oviduct was significantly higher (P = 0.0026) than it was in fluid from the isthmic segment during the functional period, i.e. from prooestrus to metaoestrus (73.5 +/- 10.49 mg/L in ampullar ODF, compared to 43.2 +/- 10.74 mg/L in isthmic ODF); least square mean (L.S.M.) standard error of the mean (S.E.M.). There was also a sianificantly higher concentration of S-GAGs in the fluid from the oviduct ipsilateral to the ovulation side 73.5 +/- 10.54 mg/L on the ovulation side, compared to 43.1 +/- 10.71 mg/L in the oviduct on the contralateral side (L.S.M. +/- S.E.M., P = 0.0026) during this period. Both syndecan-1 and syndecan-2 were present in the epithelial cells lining all studied segments of the bovine oviduct, i.e. the UTJ, isthmus and ampulla, during both standing oestrus and dioestrus. The syndecans and S-GAGs found may influence the gametes. while they reside in the oviduct; the amounts of S-GAGs found in the bovine oviduct seem sufficient to act as capacitating factors in vivo. (c) 2005 Elsevier B.V. All rights reserved.
14.	Bonato, Maud, et al. (författare) Predicting ejaculate quality and libido in male ostriches: Effect of season and age. 2014 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 1873-2232 .- 0378-4320. ; 151:1-2, s. 49-55 Tidskriftsartikel (refereegranskat)abstract The success of artificial breeding program depends largely on the reproductive performance of males. Male performance can vary with season and age impacting on quality and quantity of semen collected for artificial insemination purposes and therefore fertility of inseminated females. We examined variation in semen output and male libido of seven male ostriches (aged 2-5 years) over a period of 24 months. We collected ejaculates using a dummy female and measured semen characteristics (ejaculate volume, sperm concentration, number of spermatozoa per ejaculate, sperm motility and morphology) and male libido (willingness to mount the dummy). A total of 1006 ejaculates were collected. Across months, the volume of semen (mean±SEM) ranged from 1.03±0.12mL to 1.85±0.07mL, the sperm concentration from 3.21±0.12×10(9)/mL to 4.16±0.74×10(9)/mL, and the number of spermatozoa from 3.42±0.28×10(9) to 7.66±0.47×10(9). The largest volume of ejaculates and the highest number of sperm were collected in spring. Ejaculates with higher number of normal sperm were also collected in spring-early summer, whereas ejaculates with higher numbers of live abnormal and dead sperm were collected in winter. Sperm motility was relatively constant over months, despite a reduction in summer (January-February), while male libido peaked in winter (June-July) and spring (October-November). Furthermore, we observed high individual variation between males for all variables tested, except for motility. These results indicate that collections conducted in spring yield higher number of spermatozoa, when the libido of males is also at a maximum. Therefore in this species seasonal variation in semen quality should be considered in breeding programmes by artificial insemination to maximise fertility.
15.	Bonato, Maud, et al. (författare) The effect of temperature and pH on the motility and viability of ostrich sperm 2012 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 1873-2232 .- 0378-4320. ; 133:1-2, s. 123-128 Tidskriftsartikel (refereegranskat)abstract As the chemical environment of semen diluents can have a profound effect on sperm quality, we examined the effect of temperature and pH on the motility and viability of sperm in the ostrich. Semen was collected from four males, each male being replicated three times. Ejaculates were diluted and incubated for 10 min at 20 degrees C and 40 degrees C in four different buffers, temperature adjusted at pH 6, 7, 8 and 9 respectively. Average path velocity (VAP), curvilinear velocity (VCL), straight-line velocity (VSL), linearity (LIN), beat cross frequency (BCF) and amplitude of lateral displacement (ALH) were then recorded for each sample using CASA. The viability of sperm was assessed using nigrosin-eosin staining. Sperm incubated at 40 degrees C had higher motility parameters, except for ALH. At 40 degrees C, VAP, VSL and LIN increased with pH while VCL, BCF and ALH were higher for lower pHs. The viability of sperm was not affected by temperature but decreased at pH values > 7. A pH in the neutral range appeared to yield higher quality sperm after in vitro storage at 20 degrees C. However, the effect of different pH levels and temperatures on sperm longevity needs to be investigated further to develop viable ostrich specific diluents. (C) 2012 Elsevier B.V. All rights reserved.
16.	Brandt, Y., et al. (författare) Effects of continuous elevated cortisol concentrations during oestrus on concentrations and patterns of progesterone, oestradiol and LH in the sow 2009 Ingår i: Animal Reproduction Science. - : Elsevier Masson. - 0378-4320 .- 1873-2232. ; 110:1-2, s. 172-185 Tidskriftsartikel (refereegranskat)abstract This study investigated the effect of continuous elevated cortisol concentrations during standing oestrus on time of ovulation and patterns Of progesterone. oestradiol and luteinising, hormone (LH) in sows. The elevation of cortisol concentrations was achieved through repeated intravenous injections of synthetic adrenocorticotropic hormone (ACTH) every 2 It for approximately 48 h, from the onset of the second standing oestrus alter weaning. Treatment was terminated when ovulation was detected (monitored by transrectal ultrasonography every 4h) or when (lie sow had received a maximum of 24 injections. The close of ACTH (2.5 mu g/kg) was chosen to mimic the cortisol concentrations seen during mixing of unfamiliar SOWS. The sows (n = 14) Were surgically fitted with jugular vein catheters and randomly divided into a control (C group) where only NaCl solution were injected) or an ACTH group. Blood samples were collected every 2 h. In parallel with the blood sampling, saliva samples for cortisol analyses were taken from eight sows before onset of treatment and from four of the sows during treatment. There was no difference in time from onset of standing, oestrus to ovulation between the two groups. The interval between the peaks of oestradiol and LH to ovulation was prolonged in the ACTH group compared to the C group (p less than 0.05). with a tendency towards all earlier decline of oestradiol in the ACTH group. Cortisol and progesterone Concentrations were significantly elevated during treatment in the ACTH group (p less than 0.001). with cortisol peak concentrations occurring between 40 and 80 min after each ACTH injection. Cortisol concentrations in saliva and Plasma were highly correlated (p less than 0.001). In conclusion, elevated cortisol concentrations from the onset of standing oestrus increase progesterone concentrations and prolong the interval between oestradiol and LH peaks to ovulation, the latter possible due to an early decline in oestradiol concentrations and a change of the LH peak outline. the effect these hormonal changes have on reproductive performance need to be further investigated. (C) 2008 Elsevier B.V. All rights reserved.
17.	Brandt, Y, et al. (författare) Impact of ACTH administration on the oviductal sperm reservoir in sows: The local endocrine environment and distribution of spermatozoa 2006 Ingår i: Animal Reproduction Science. - : Elsevier Masson. - 0378-4320 .- 1873-2232. ; 92:1-2, s. 107-122 Tidskriftsartikel (refereegranskat)abstract The objective of the study was to investigate if short-term stress in sows (simulated by injections of synthetic adrenocorticotrophic hormone (ACTH)) during standing oestrus had a negative effect on the local environment in the utero-tubal junction (UTJ) and isthmus and the distribution of spermatozoa in these segments. Fourteen sows were monitored for ovulation using ultrasonography in two consecutive oestruses. The sows were fitted with jugular catheters and, from onset of the second oestrus, blood samples were collected every second hour. In the 2nd oestrus, seven sows were given ACTH every second hour, from the onset of standing oestrus until the sow ovulated (ACTH-group), whereas the other seven sows remained as controls (C-group) and were given NaCl solution. The sows were artificially inseminated 16-18 h before expected ovulation. Six hours after ovulation the sows were anaesthetised, and blood samples were repeatedly taken from veins draining the uterus and the UTJ-isthmus, respectively. This oviduct was thereafter removed and divided in four adjacent sections consisting of: (i) the UTJ, (ii) the first, and (iii) the second isthmus segment prior to (iv), the ampullary-isthmic junction (AIJ) and the ampulla. The three first-mentioned segments were flushed to retrieve spermatozoa, whereas the last one was flushed to collect oocytes/ova. The number of spermatozoa attached to the zona pellucida was counted. The concentrations of cortisol in jugular blood of the ACTH-group sows during the time of ACTH-injections were significantly higher than of the C-group sows (p less than 0.05), as were the levels of progesterone (p less than 0.001). Progesterone and cortisol concentrations measured in the blood samples draining the UTJ-isthmic region 6 h after ovulation did not significantly differ between the groups, but the C-group displayed significantly higher concentrations of progesterone in the UTJ-isthmic region compared with the levels measured in parallel samples taken of jugular blood (P less than 0.01). The C-group, but not the ACTH-group, also displayed a significant elevation in progesterone concentration 6 h after ovulation compared with the basal levels before ovulation (p less than 0.01). Numbers of retrieved spermatozoa were not significantly different between the C-group and the ACTH-group. However, there was a tendency for a larger number of spermatozoa among sows in the ACTH-group, especially in the isthmic segment adjacent to the AIJ. In conclusion, simulated stress induced by injections of ACTH during standing oestrus results in elevated concentrations of progesterone before ovulation and may interfere with the rise of progesterone after ovulation. However, ACTH-injections appeared to augment transport of spermatozoa through the female genital tract of pigs. (c) 2005 Elsevier B.V. All rights reserved.
18.	Brandt, Y, et al. (författare) Impact of ACTH during oestrus on the ultrastructure of the spermatozoa and their environment in the tubal reservoir of the postovulatory sow 2006 Ingår i: Animal Reproduction Science. - : Elsevier Masson. - 0378-4320 .- 1873-2232. ; 93:3-4, s. 231-245 Tidskriftsartikel (refereegranskat)abstract This study investigated whether injections of synthetic ACTH (simulating short-term stress) in sows during standing oestrus have a negative effect on spermatozoa and the local intraluminal environment in the utero-tubal junction (UTJ) and isthmus. Seven of the 14 sows were given ACTH through a jugular catheter every 2 h from the onset of standing oestrus until the sow ovulated (ACTH-group), while the other seven sows were given NaCl solution (C-group). All sows were artificially inseminated before ovulation. Six hours after ovulation (detected with transrectal ultrasonography) the sows were anaesthetised, the right oviduct was fixed in toto by vascular perfusion with glutaraldehyde, and the UTJ and specimens from the isthmus were prepared for scanning electron microscopy (SEM). SEM revealed that a seemingly viable population of spermatozoa remained in the UTJ 6 It after ovulation. A majority of sows in the ACTH-group had moderately to exaggerated amounts of mucus in the intraluminal environment of the sperm reservoir. In conclusion, stress simulated by exogenous ACTH in sows may alter the intraluminal environment of the sperm reservoir. (c) 2005 Elsevier B.V. All rights reserved.
19.	Chanrot, Metasu, et al. (författare) Dose related effects of LPS on endometrial epithelial cell populations from dioestrus cows 2017 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 177, s. 12-24 Tidskriftsartikel (refereegranskat)abstract Lipopolysaccharides (LPS) from Gram negative bacteria are involved in the pathogeny of uterine diseases in cows. This study aimed to investigate LPS effects on the growth of bovine endometrial epithelial cells (bEEC) and relationships between LPS response and tissue characteristics. Uteri from 35 females were characterized for parity and stage of oestrous cycle. Densities of glandular tissue (dGT), CD11b+ cells and Ki67+ cells were measured in the endometrial tissue. Cells from 13 dioestrus cows were exposed to 0, 2, 4, 8, 12, 16 or 24 mu g/mL LPS. Effects of parity and stage of the oestrous cycle on tissue characteristics and effects of LPS dosage, cow and tissue characteristics on changes in cell numbers were analyzed by ANOVA. The dGT was higher in metoestrus and dioestrus samples than in pro-oestrus ones whereas densities of CD11b+ and Ki67+ cells were higher at pro-oestrus (p < 0.05-p < 0.01). LPS influenced bEEC populations in a dose related manner. An increase in number of live cells was observed for dosages ranging from 2 to 12 mu g/mL LPS (p < 0.0001 vs controls). No effect was found on numbers and frequencies of dead cells. With higher dosages, the numbers of live cells did not increase but the numbers of dead did increase. No relationships were observed between cow or tissue characteristics and growth patterns or frequencies of viable bEEC in controls nor in the response to LPS. To conclude this model is suitable for further studies on dysregulations induced by LPS in endometrial tissue. (C) 2016 The Authors. Published by Elsevier B.V.
20.	Crespo-Felez, I., et al. (författare) Effect of Single Layer Centrifugation Porcicoll (70%, 80% and 90%) or supplementation with reduced glutathione, seminal plasma and bovine serum albumin on frozen-thawed boar sperm 2017 Ingår i: Animal Reproduction Science. - : ELSEVIER SCIENCE BV. - 0378-4320 .- 1873-2232. ; 187, s. 167-173 Tidskriftsartikel (refereegranskat)abstract Selecting the optimal sperm population is essential for success with reproductive techniques. Porcicoll (formerly Androcoll-P) is a colloid formulation for selection of high-quality boar spermatozoa by single layer centrifugation (SLC). To date, most studies have been carried out with fresh semen and large volumes. We carried out 2 experiments to test the use of Porcicoll for thawed boar semen in small volumes. In Experiment 1, cryopreserved semen doses were thawed, split in 200-pL aliquots and layered on 1 mL of Porcicoll 70%, 80% or 90%, or buffer without colloid. We assessed sperm recovery (the proportion of the loading dose that appeared in the pellet, %), and the physiology of the selected spermatozoa (flow cytometry: Viability, apoptotic changes, capacitation, mitochondrial activity, intracellular reactive oxygen species). The most suitable proportion was Porcicoll 80%, allowing acceptable sperm recovery (16.9 4.2%, compared to 70% (35.4% 3.0, p amp;lt; 0.001) and 90% (8.2% 3.0, P = 0.001), and improved quality (mitochondrial activity: Porcicoll 80%: 77.7 1% vs Control: 60.3 0.7%, P amp;lt; 0.05). In Experiment 2, we compared 3 supplements to Porcicoll 80%: 500 mM reduced glutathione (GSH), 20% seminal plasma (SP) and 0.5% bovine serum albumin (BSA). Supplementation with GSH or BSA did not cause relevant changes relative to Control. In contrast, SP induced membrane and acrosomal changes resembling capacitation, which might preclude its use in some applications, and decreased recovery (5.5% 1.9 vs. 24.3% 1.2 Control; P amp;lt; 0.001). However, it could be useful prior to other applications such as in vitro fertilisation. Overall, Porcicoll is an effective colloid for isolating a high-quality population from thawed boar sperm, 80% being a balanced option for good recovery and high quality. Supplements could be useful depending on the proposed use of the spermatozoa.
21.	Dalin, Anne-Marie (författare) Short communication: Concentration of TGF-beta 1, IL-10 and IL-6 in boar seminal plasma and TGF-beta 1 level in different fractions of ejaculates 2012 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 131, s. 194-198 Tidskriftsartikel (refereegranskat)abstract To conclude: Boar seminal plasma contained TGF-beta 1 and IL-10 but with high individual variation. IL-6 was low or undetectable. The TGF-beta 1 level was highest in the first 10 mL of the sperm-rich fraction of the ejaculate. Further studies are needed on the role of different levels of cytokine in boar semen on porcine female reproductive tissue, especially for TGF-beta 1. (C) 2012 Elsevier B.V. All rights reserved.
22.	Deori, Sourabh, et al. (författare) Seminal plasma protein profiles in young bulls at different ages 2020 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 220, s. 23-24 Konferensbidrag (refereegranskat)
23.	Einarsson, Stig, et al. (författare) Effects of early vaccination with Improvac (R) on the development and function of reproductive organs of male pigs 2011 Ingår i: Animal Reproduction Science. - : Elsevier Masson. - 0378-4320 .- 1873-2232. ; 127:1-2, s. 50-55 Tidskriftsartikel (refereegranskat)abstract Gonadotropin-releasing hormone (GnRH) vaccine (Improvac (R)) is effective at diminishing boar taint by interfering with testis function. Early pre-pubertal vaccination at 10 and 14 weeks-of-age could be desirable if sufficient and sustained effects could be achieved. Crossbred male pigs (n = 24) were randomly assigned to three groups each with eight individuals: an unvaccinated control group, one group vaccinated with Improvac (R) early at ages 10 and 14 weeks, and a third group vaccinated with Improvac at the standard ages of 16 and 20 weeks. The average age at slaughter was 25 weeks. At slaughter, reductions in testes weight and bulbourethral gland length of vaccinated pigs compared with controls were observed (P andlt; 0.001), accompanied by lowered testosterone concentrations in peripheral blood (P andlt; 0.001). The diameter of tubuli seminiferi was affected: being 18% smaller in standard and 38% smaller in early vaccinated males, compared with controls (P andlt; 0.01). Leydig cells in vaccinated pigs became pycnotic, and their number decreased in early vaccinated pigs. Spermatogenesis was disrupted, evidenced by spermatocyte loss among standard vaccinated pigs to severe spermatogenic arrest among early vaccinated pigs. This histological picture was reflected in the absence of epididymal spermatozoa in 5 of 8 early vaccinated pigs and a dramatic reduction in the remaining 3 early vaccinated pigs. Among standard vaccinated pigs, 5% of the spermatozoa were morphologically normal (andgt;70% in controls, P andlt; 0.01). Early vaccination caused a more severe disruption of testicular structure and function than standard vaccination, thus providing an alternative for immunocastration of male pigs.
24.	Einarsson, Stig (författare) Influence of the uterine inflammatory response after insemination with frozen-thawed semen on serum concentrations of acute phase proteins in mares 2014 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 146, s. 182-186 Tidskriftsartikel (refereegranskat)abstract The aim of this study was to investigate the clinical relevance of measuring blood concentrations of serum amyloid A (SAA), haptoglobin (Hp) and fibrinogen (Fib) in horse reproductive management, and changes in response to artificial insemination (Al) with frozen-thawed semen. Standardbred mares (n = 18) with different reproductive status (eight healthy mares in first postpartum oestrus, five healthy barren mares and five mares with endometritis) were inseminated with frozen-thawed semen. Endometritis was evaluated during oestrus by bacteriological culture, cytology and presence of ultrasonically visible intrauterine fluid during oestrus. Concentrations of SAA, Hp and Fib were analysed in the blood in every 48 h during oestrus and until 5, 6 or 7 days after Al. The day of sampling and number of blood samples varied between mares because of length of the oestrus and time of Al. Changes in concentrations of SAA, Hp and Fib were evaluated based on the day of sampling regard to Al and classification of the mares. There were no differences in SAA, Hp and Fib concentrations over time before or after Al or between the groups of mares. The insemination of mares with frozen-thawed semen did not increase the plasma concentrations of SAA, Hp and Fib above clinical threshold concentration and there were no differences between susceptible or healthy mares. (C) 2014 Published by Elsevier B.V.
25.	Grundin, Johanna, et al. (författare) Individual male dependent improvement in post-thaw dromedary camel sperm quality after addition of catalase 2019 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 209 Tidskriftsartikel (refereegranskat)abstract Cryopreservation is stressful to sperm cells inducing an increase in the production of reactive oxygen species and subsequently reducing post-thaw sperm quality. With the present study, there was evaluation of the protective effects of two antioxidants, epigallocatechin (1 mM) and catalase (500 IU/ml), added at thawing, as well as inter-individual variation on quality of cryopreserved dromedary camel spermatozoa. Semen was collected from six males and sperm, selected using single layer centrifugation, were cryopreserved. Post-thaw sperm quality was evaluated by assessing motility variables, viability and acrosome integrity then sperm were co-incubated with or without antioxidant (control) and further assessed at 1.5 and 3 h of the incubation period. Oxidative damage was measured colorimetrically for malondialdehyde production at 3 h of the incubation period. With the use of epigallocatechin there were not promising results, however, with use of catalase there were greater total and progressive motility, and values for some kinematic variables (P<0.05) at both incubation time points, although there were some differences among males. There was no overall effect of antioxidant based on production of malonaldehyde. The capacity of thawed sperm to fertilize, with and without addition of catalase at thawing, was studied using artificial insemination (n=10 per treatment) with no differences between treatments (10% for both). It is concluded that catalase supplementations to semen extender prolong sperm survival, however, there is no improvement of in vivo fertilization as a result of this supplementation. There was an obvious male effect, necessitating further studies to understand the mechanisms of action of catalase.
26.	Gustafsson, Hans (författare) Uterine bacterial flora in postpartum Danish Holstein dairy cows determined using DNA-based fingerprinting: Correlation to uterine condition and calving management 2013 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 138, s. 39-48 Tidskriftsartikel (refereegranskat)abstract The overall aim of this study was to describe uterine bacterial flora during the postpartum period in Danish Holstein cows using the Terminal Restriction Fragment Length Polymorphism (T-RFLP) method. This method produces a pattern of nucleic acid fragments from the microorganisms present, reflecting the "fingerprint" of the actual microbial flora. As well as characterizing changes in flora with time from calving and between herds, data were examined for strong relations between uterine bacterial flora, calving management and uterine condition. In total 125 Holstein cows from five herds were included, and for each cow calving management was recorded. Cows were clinically examined on average 8 (range 0-19) and 28 (range 22-38) days after calving, and a uterine sample was taken for bacterial identification using T-RFLP. Milk samples were taken weekly for progesterone analysis. Bacteria were found in all cows at both examinations, and the flora was composed of many species, including species not traditionally reported to be present in the bovine uterus. The bacterial composition differed according to days from calving and herd. In all five herds Fusobacterium necrophorum, Pseudomonas/Acinetobacter and Bacteroides/Sphingobacterium/Prevotellaceae were among the most common at both examinations. In four herds there was a percentage decrease of F. necrophorum from first to second examination, and in all herds there was a percentage increase of Pseudomonas/Acinetobacter from first to second examination. No differences in bacterial flora were found between cows with different uterine scores, which were influenced by herd, calving difficulty and retained placenta. (C) 2013 Elsevier B.V. All rights reserved.
27.	Hagos, Lidya, et al. (författare) Effect of rosmarinic acid on bull sperm quality and bacteria in semen 2020 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 220 Konferensbidrag (övrigt vetenskapligt/konstnärligt)
28.	Hurri, Emma, et al. (författare) Semen from young bulls: how soon are the ejaculates acceptable for freezing? 2020 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 220, s. 11-12 Konferensbidrag (refereegranskat)
29.	Jiwakanon, Jatesada, et al. (författare) Cytokine expression in the gilt oviduct: Effects of seminal plasma, spermatozoa and extender after insemination 2010 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 119, s. 244-257 Tidskriftsartikel (refereegranskat)abstract Effects of semen components [fresh semen in extender, spermatozoa in extender (Spz), seminal plasma (SP)], or extender alone (Beltsville thawing solution, BTS) on the expression of selected cytokines [interleukin (IL)-1 beta, IL-6, IL-10 and transforming growth factor (TGF)-beta 1)] as well as the presence of cells positive for CD8 or CD25 were studied in the pig oviduct. In addition, cytokines in SP and oviductal flushings were analyzed.In experiment (Exp) I, groups of gilts were sampled at 5-6 h after insemination with SP, Spz, fresh semen in BTS or only BTS (control). In Exp II, gilts were sampled 35-40 h after insemination with SP, Spz, BTS or only catheter insertion (control).Most oviductal flushing samples were positive (detectable limits) for IL-10 and TGF-beta 1 but only few for IL-6. The IHC-labelling of IL-6. IL-10 and TGF-beta 1 was evident, especially in the epithelial cells of the isthmus and infundibulum as well as in the cells of the regional (mesometrial) lymph node. Cilia of the epithelium were positive for IL-6 (strongest in the infundibulum) and TGF-beta 1 (strongest in the isthmus) but negative for IL-10. There were no consistent differences in IHC-labelling of the cytokines in relation to different treatments, except at 35-40 h after insemination (Exp II), when IL-6 was slightly higher in epithelium of the SP group and IL-10 in the infundibular connective tissue was higher in the SP and Spz groups.In the isthmus and infundibulum, there were no differences between animals inseminated with BTS (control) and the semen components for any of the cytokine mRNAs at 5-6 h after insemination (Exp I). However, later (35-40 h, Exp II), insemination with SP, Spz and BTS alone appeared to up-regulate TGF-beta 1 mRNA expression compared with the control group (without any fluid infused). In all treatment groups, the mRNA level for TGF-beta 1 was higher than for IL-1 beta, IL-6 and IL-10. Higher mRNA levels of all cytokines were found in the isthmus compared with the infundibulum.Numbers of CD8-positive cells (both in epithelium and connective tissue) appeared higher in the infundibulum compared with the isthmus and were mostly higher shortly (Exp I) after treatment with SP, SPZ and BTS than later (Exp II) in both segments. CD25-positive cells were few and found solely in the sub-epithelial connective tissue.The results indicate that in the porcine oviduct, IL-6. IL-10 and TGF-beta 1 are endogenous produced and that TGF-beta 1 may have a more important role for immunomodulation than the other cytokines, especially in isthmus. Differences between isthmus and infundibulum in cytokine mRNA expression and in presence of CD8-positive cells indicate different patterns of immune reactivity in the upper and lower parts of the oviduct. (C) 2010 Elsevier B.V. All rights reserved.
30.	Jiwakanon, Jatesada, et al. (författare) Influence of seminal plasma, spermatozoa and semen extender on cytokine expression in the porcine endometrium after insemination 2011 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 123, s. 210-220 Tidskriftsartikel (refereegranskat)abstract The effects of semen components or extender alone on the expression of selected cytokines [interleukine (IL)-1β, IL-6, granulocyte-macrophage colony stimulating factor (GM-CSF), IL-10 and transforming growth factor (TGF)-β1] on the porcine endometrium were studied, as well as the presence of polymorphonuclear neutrophilic granulocytes (PMNs). In experiment (Exp) I, groups of gilts were sampled at 5-6h after insemination with fresh semen in extender (Beltsville thawing solution, BTS), spermatozoa in extender (Spz), seminal plasma (SP), or only BTS (control). In Exp II, gilts were sampled 35-40h after insemination with Spz, SP, BTS or only catheter inserted (as control). Immunohistochemical (IHC) labelling of IL-6, IL-10 and TGF-β1 was evident, especially in surface and glandular epithelia of the porcine endometrium. There were no consistent differences in IHC-labelling of the cytokines in relation to different treatments. However, the scores for IL-6 and IL-10 in surface epithelium and sub-epithelial connective tissue compartments were higher at 35-40h than shortly (5-6h) after treatment. Cytoplasmic labelling in the sub-epithelial connective tissue was observed in scattered individual cells but not in PMNs. Shortly (5-6h) after insemination, there were no differences between animals inseminated with BTS (control) and the semen components for any of the cytokine mRNAs. Later however, at 35-40h, lower endometrial expression of TGF-β1 mRNA was observed in the Spz and BTS groups compared with the control (catheter only). The same pattern was found for IL-10 (NS). The mRNA expression of IL-6 in the BTS inseminated group was higher compared to the control group. Insemination with SP resulted in significantly lower PMN cell infiltration in the sub-epithelial connective tissue compared with Spz or BTS groups shortly (5-6h) after insemination. Later (35-40h), a significant difference was found between SP (lower) and the control group (only catheter). To conclude, our results show that insemination and/or inseminated components modulated cytokine expression in the gilt endometrium. The semen extender BTS stimulated immune reactivity, as shown by down-regulation of the suppressive cytokine TGF-β1. Insemination with solely SP clearly decreased PMN cell infiltration of the gilt endometrium. However, no clear relation between the cytokines studied and PMN cell presence was found
31.	Johannisson, Anders, et al. (författare) Androcoll-E selects a subset of live stallion spermatozoa capable of producing ROS. 2012 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 132, s. 74-82 Tidskriftsartikel (refereegranskat)
32.	Johannisson, Anders, et al. (författare) Colloidal centrifugation with Androcoll-E (TM) prolongs stallion sperm motility, viability and chromatin integrity 2009 Ingår i: Animal Reproduction Science. - : Elsevier Masson. - 0378-4320 .- 1873-2232. ; 116:1-2, s. 119-128 Forskningsöversikt (refereegranskat)abstract The objective was to investigate the changes in stallion sperm quality (sperm motility, viability, membrane integrity and chromatin integrity) occurring during cool storage, and to study the effect of sperm selection by single layer colloidal centrifugation on these parameters of sperm quality. Spermatozoa from 3 stallions (10 ejaculates, 3-4 per stallion) were selected by centrifugation through a single layer of colloid (SLC). The resulting sperm preparations and the control samples (extended but unselected semen samples) were stored at 5 degrees C for 48 h. Assessments of sperm quality, such as sperm motility, viability (SYBR-14/PI staining), membrane stability (Annexin-V/PI staining) and chromatin integrity, were performed on aliquots of the selected sperm preparations and unselected samples on the day of collection (3 h) and after 24 and 48 h of storage. In the SLC-selected sperm samples, sperm motility, sperm viability, proportions of spermatozoa with normal morphology and with intact chromatin were significantly better than in unselected samples (motility: 77 +/- 4% vs. 64 +/- 8% at 3 h; P less than 0.001; viability: 79.5 +/- 9% vs. 64.7 +/- 9%, P less than 0.001: normal morphology 89 +/- 6% vs. 69 9%; chromatin integrity DFI 11.3 +/- 5% vs. 22.1 +/- 10%). Membrane stability, however, was not different in the SLC-selected and unselected samples (74.6 +/- 8% vs. 69.3 +/- 8%). The deterioration seen in sperm quality in the unselected samples was prevented by SLC, so that sperm viability, membrane stability and chromatin integrity were unchanged in the selected samples by 48 h compared to 3 h (Pless than0.001), whereas the unselected samples were significantly worse by 48 h (Pless than0.001). Furthermore, it should be possible to send an aliquot of a normal insemination dose (i.e. unselected spermatozoa) overnight to a reference laboratory for analysis of both plasma membrane and chromatin integrity. In conclusion, centrifugation of stallion spermatozoa through a single layer of colloid is a useful technique for selecting the best spermatozoa from an ejaculate and, moreover, sperm quality is maintained during storage. (C) 2009 Elsevier B.V. All rights reserved.
33.	Johannisson, Anders, et al. (författare) Simultaneous evaluation of superoxide content and mitochondrial membrane potential in stallion semen samples provides additional information about sperm quality 2018 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 192, s. 290-297 Tidskriftsartikel (refereegranskat)abstract An improved fertility prediction for stallions is of importance for equine breeding. Here, we investigate the potential of a combined staining of stallion spermatozoa for superoxide and mitochondrial membrane potential (MMP) for this purpose. Semen samples were analysed immediately after arrival at the laboratory, as well as after 24 h. Superoxide was measured by MitoSOXRed, while MMP was measured with JC-1. Menadione was used to stimulate superoxide production. In addition, other parameters of sperm quality, namely motility, membrane integrity, chromatin integrity, sperm kinematics and Hoechst 33258 exclusion were measured and correlated to superoxide production and MMP. Both bivariate correlations between measured parameters as well as multivariate analysis were performed. Measured values in the superoxide/MMP assay did not correlate with other parameters. However, there was a strong negative correlation (r = 0.96 after 0 h, r = 0.95 after 24 h) between membrane integrity and chromatin integrity. Moderate positive correlations were found between motility parameters and membrane integrity, as well as moderate negative correlations between motility parameters and chromatin integrity. The multivariate analysis revealed that membrane integrity, chromatin integrity and motility contributed to the first principal component, while the second was influenced by superoxide/MMP parameters as well as sperm kinematics. Storage of samples for 24 h decreased motility, chromatin integrity and membrane integrity. In conclusion, combined measurement of superoxide and MMP provides additional information not obtained by other assays of sperm quality.
34.	Johannisson, Anders, et al. (författare) Spermatozoa in the sperm-peak-fraction of the boar ejaculate show a lower flow of Ca2+ under capacitation conditions post-thaw which might account for their higher membrane stability after cryopreservation 2011 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 128, s. 37-44 Tidskriftsartikel (refereegranskat)abstract Boar spermatozoa collected in the ejaculate sperm peak-portion (P1, first 10 mL of the sperm-rich fraction, SRF), had shown a higher resilience to freezing and thawing compared to spermatozoa from the rest of the ejaculate (2nd portion of the SRF plus the post-sperm-rich fraction, PSRF), even when using a simplified freezing technique, as long as spermatozoa were incubated in their own seminal plasma (SP). This experiment studied the stability of P1- and SRF-P1 boar spermatozoa frozen in MiniFlatPacks (MFP), post-thaw, using flow cytometry. Since spermatozoa from either portion showed similar cryosurvival and low proportions of unstable membranes (<3%, annexin-V/propidium iodide staining), and only a tendency for SRF-P1 live spermatozoa to depict acrosome exocytosis (FITC-PNA/PI/H33342); they were explored for Ca2+ contents using a Fluo-4 probe under in vitro capacitating conditions (mBO+ medium), as well they were tested for their ability to sustain a short Ca2+-ionophore (A23187) in vitro challenge. The proportions of live spermatozoa depicting high Ca2+-levels were initially <2% but increased over incubation time, particularly in SRF-P1(P < 0.05), while proportions of live spermatozoa with low Ca2+-levels were basically constant over incubation time (similar to 11-14%), for either portion. Incubation in capacitation medium did not modify the proportions of low-Ca2+ but dramatically increased the proportions of high-Ca2+ spermatozoa (P < 0.001) already after 15 min exposure, highest for SRF-P1 spermatozoa. While the proportion of live spermatozoa with intact acrosome was significantly decreased among SRF-P1 (P < 0.001), that of P1-spermatozoa remained unchanged, probably owing to the lowest relative content of cytosolic Ca2+. The results suggest that spermatozoa in the P1-portion are more resilient to express acrosome exocytosis post-thaw compared to those bathing in the rest of the SRF-fraction when cryopreserved using a simplified technique, in MFPs. (C) 2011 Elsevier B.V. All rights reserved.
35.	Kunkitti, Panisara, et al. (författare) Osmotic tolerance of feline epididymal spermatozoa 2017 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 185, s. 148-153 Tidskriftsartikel (refereegranskat)abstract During the cryopreservation process, spermatozoa are exposed to hypertonic solutions contributed by the high concentration of cryoprotectant. During addition and removal of cryoprotectant the spermatozoa are subjected to a substantial osmotic stress. Spermatozoa of different I species and different stages of maturation may have different susceptibility to osmotic stress depending on the biology of the cell membrane and this will affect their tolerance to the freezing thawing stress. The aims of this study were to determine the osmotic tolerance limits for motility, membrane integrity and mitochondrial membrane potential of feline epididymal spermatozoa and to study the effect of osmotic stress on the feline spermatozoa of different epididymal regions. Epididymal spermatozoa from three regions (caput, corpus and cauda) were pre-exposed to various osmolalities (75, 300, 600, 900, 1200 mOsm) in a single step for 10 min and returned to 300 mOsm afterward. Percentage of motile spermatozoa was measured subjectively and membrane integrity (SYBR-14 positive cells) was evaluated prior to and after exposure to different osmolalities. The mitochondrial membrane potential (JC1) of spermatozoa were evaluated using flow cytometer and compared between epididymal regions (caput, corpus and cauda). All the parameters were compared using a mixed procedure. The, percentage of motile epididymal spermatozoa decreased significantly when spermatozoa were exposed to 75 mOsm and 600 mOsm. Epididymal spermatozoa showed signs of damage when pre-exposed to 900 and 1200 mOsm and returned to isotonic condition as significant reduction of membrane integrity and mitochondrial membrane potential were observed (P < 0.05). The plasma membrane of spermatozoa from the cauda epididymal region showed higher susceptibility to osmotic stress than the other regions as demonstrated by a significant difference between regions after return to isotonicity from 900 mOsm (P > 0.01) and a difference between caput and corpus after return from 1200 mOsm (P < 0.05). The corpus and cauda epididymal spermatozoa had higher percentage of spermatozoa with high mitochondrial membrane potential than those from caput when exposed to 75, 300 and 600 mOsm (P < 0.05). In conclusion, a single step exposure to hypertonic solution of greater than 600 mOsm prior to return to isotonic condition can cause severe damage to sperm membrane and mitochondrial membrane potential compared to non returning (exposure to various osmolality but not returned to isotonic condition). Changes in osmolality impacted mostly on sperm motility. Spermatozoa from cauda epididymis were more susceptible to osmotic stress compared to those from corpus and caput indicating that the maturation changes in the sperm membrane during passage through the epididymis increase susceptibility to the osmotic changes that may occur during sperm cryopreservation.
36.	Kunkitti, Panisara, et al. (författare) The ability of feline spermatozoa in different epididymal regions to undergo capacitation and acrosome reaction 2015 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 161, s. 64-74 Tidskriftsartikel (refereegranskat)abstract The sperm maturation process that occurs in the epididymis is a necessary process for spermatozoa to acquire motility and the ability to undergo capacitation, which is an important key for fertilization. The aim of this study was to evaluate the ability of feline spermatozoa from different regions of the epididymis to undergo capacitation and acrosome reaction. Experiment I: epididymal spermatozoa from caput, corpus and cauda regions were placed in phosphate buffered saline (control medium) and in vitro fertilization medium (capacitating conditions). Sperm motility, motility patterns, plasma membrane integrity and tyrosine phosphorylation were evaluated at time 0 and 60 min after incubation. Experiment II: spermatozoa were treated with 2 mu M of calcium ionophore (A23187) to induce the acrosome reaction and acrosome reaction was evaluated. The results showed a significant effect of region with a higher percentage of tyrosine phosphorylation in spermatozoa from the cauda than in the caput or corpus regions (P=0.0061; P=0.0088). Spermatozoa from corpus and cauda showed higher values in the majority of the measured motility parameters than spermatozoa from the caput (P<0.0001). Spermatozoa from all epididymal regions can undergo the acrosome reaction in vitro in response to induction by calcium ionophore with no difference between regions (P>0.05). Spermatozoa from all epididymal regions were able to undergo capacitation. Higher percentage of tyrosine phosphorylation in spermatozoa from the cauda reflect that they more easily underwent capacitation compared to spermatozoa from caput and corpus which required more time of incubation for capacitation. In conclusion feline epididymal spermatozoa from all regions can undergo capacitation and acrosome reaction in vitro and do not require incubation under capacitating conditions. (C) 2015 Elsevier B.V. All rights reserved.
37.	Li, Junwei, et al. (författare) Is boar sperm freezability more intrinsically linked to spermatozoa than to the surrounding seminal plasma? 2018 Ingår i: Animal Reproduction Science. - : ELSEVIER SCIENCE BV. - 0378-4320 .- 1873-2232. ; 195, s. 30-37 Tidskriftsartikel (refereegranskat)abstract This study aimed to elucidate the effect of seminal plasma (SP) from post-SRF on boar sperm freezability and, in addition, to determine the relevance of sperm itself to sustain cryopreservation, regardless of the SP surrounding them. Twelve ejaculates from three boars were manually collected in fractions/portions, P1: the first 10 mL of the SRF, P2: the rest of the SRF and the post-SRF. Immediately, samples were centrifuged to separate spermatozoa from the surrounding SP. Spermatozoa from P1 and P2 were then incubated with its own SP or that from post-SRF, diluted in BTS (1:1, v/v) at 17 degrees C overnight before being frozen in 0.5 mL straws using a standard protocol. Sperm motility (total and progressive) deteriorated (P amp;lt; 0.05) when P1- or P2-sperm when incubated overnight in SP from post-SRF, while sperm viability differed between P1 and P2 (P amp;lt; 0.05) regardless of the SP they were incubated in. Post-thaw sperm quality and functionality differed between P1 and P2, regardless of the SP used for overnight pre-freezing incubation. Post-thaw motility (P amp;lt; 0.05) and viability (P amp;lt; 0.01), as well as plasma membrane fluidity (P amp;lt; 0.05) or lipid peroxidation values (P amp;lt; 0.01) were best in P1 sperm compared to those of P2. The protein profile of sperm from P1 and P2, analyzed by 2D-PAGE, showed qualitative differences, which suggest that sperm rather than SP would explain differences in sperm freezability between ejaculate fractions/portions. Use of P1 fraction spermatozoa seems thus optimal for cryopreservation.
38.	Macias Garcia, B., et al. (författare) Centrifugation on a single layer of colloid selects improved quality spermatozoa from frozen-thawed stallion semen 2009 Ingår i: Animal Reproduction Science. - : Elsevier Masson. - 0378-4320 .- 1873-2232. ; 114:1-3, s. 193-202 Tidskriftsartikel (refereegranskat)abstract The present study attempted to select the subpopulation of stallion spermatozoa that best survived a conventional freezing and thawing procedure, using centrifugation of post-thawed semen samples through a single layer of a glycidoxypropyltrimethoxysilane-coated silica colloid with a species-specific formulation (Androcoll-E (TM)). Sperm motility, sperm chromatin structure, membrane integrity and mitorchondrial membrane potential were studied in filtered and non-filtered spermatozoa. Single-layer centrifugation (SLC) using Androcoll-E (TM) significantly improved all the sperm parameters studied, implying SLC may be a simple approach to improve the quality of frozen-thawed (FT) spermatozoa for AI. (c) 2008 Elsevier B.V. All rights reserved.
39.	Malmsten, Jonas, et al. (författare) Characteristics of spermatozoa and reproductive organs in relation to age and body weight in Swedish moose (Alces alces) 2015 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 153, s. 76-86 Tidskriftsartikel (refereegranskat)abstract Knowledge of the reproductive biology of game species is vital for sustainable management. In moose (Alces alces), research in reproductive characteristics has focused on the female, whereas there are few studies in male moose. The aim of the present study was to investigate sperm morphology and chromatin integrity (SCSA), and their relationships with testicular and epididymal features, as well as temporal aspects with respect to the hunting season. In total, 143 male moose aged 1.5-11.5 years were sampled from 2008 to 2011. The proportion of normal spermatozoa (PNS) ranged from 1.5% to 82.0%, with a mean of 51%, and the %DFI (DNA fragmentation index) ranged from 2.5% to 36.7% (mean 9.5). PNS decreased temporally, and was positively associated with carcass and testes weight. Body weight and testes weight had positive effect on PNS regardless of age. No effect of any explanatory variables was observed on the DEI. The testis/body weight ratio of moose (0.033%) is among the lowest reported among mammals, indicating a less polygynous mating system than in roe deer and red deer. For reproduction success in moose, a high body weight in males is favorable, as is a balanced sex ratio. Thus, males should not be harvested prior to the time when the majority of females have passed their first oestrus of the season. (C) 2014 The Authors. Published by Elsevier B.V.
40.	Mata-Campuzano, María, et al. (författare) Refrigerated storage of ram sperm in presence of Trolox and GSH antioxidants : effect of temperature, extender and storage time 2014 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 151:3-4, s. 137-147 Tidskriftsartikel (refereegranskat)abstract Antioxidants have a potential to improve the quality and fertility of refrigerated-stored ram semen. Reduced glutathione (GSH) and Trolox (0.2, 1 and 5mM) were evaluated in ram semen preserved at 15 and 5°C up to 48 and 96h, respectively. Extenders were also evaluated (15°C: Tris-citrate-fructose, TCF, without lipids, and TES-Tris-fructose 10% egg yolk, TTF-EY; 5°C: TTF-EY and 3.5% soybean lecithin, TTF-SL; INRA96 at both temperatures). Storage at 5°C resulted in poorer quality than 15°C up to 48h, while allowing acceptable quality at 96h. Antioxidants had few effects on sperm quality, with use of Trolox resulting in reduced motility and viability in TCF. Storage at 15°C in the TCF extender resulted in decreased motility, viability and mitochondrial activity compared with use of TTF-EY. Sperm quality when storage was at 5°C was similar, but storage in TTF-SL resulted in decreased motility and mitochondrial activity. Acrosomal status was only slightly affected by extender and antioxidant. Mitochondrial activity was improved by antioxidants in TTF-SL, and GSH at 5mM when semen was stored at 5°C in TTF-EY. A preliminary artificial insemination trial indicated that supplementation with GSH has the potential for improving lambing (P<0.1). In conclusion, use of antioxidants resulted in lesser effects than extender composition or storage time on quality of ram semen. Use of Trolox negatively impacted sperm quality and GSH had some positive impacts. The use of soybean lecithin requires further research to assess its impact on mitochondria.
41.	Miguel-Jimenez, Sara, et al. (författare) In vitro assessment of egg yolk-, soya bean lecithin- and liposome-based extenders for cryopreservation of dairy bull semen 2020 Ingår i: Animal Reproduction Science. - : Elsevier. - 0378-4320 .- 1873-2232. ; 215 Tidskriftsartikel (refereegranskat)abstract The study was conducted to compare the effect of four commercially available extenders (Triladyl®- egg yolk-based; Andromed® and Bioxcell®-plant based and Optixcell®-liposome-based) on post-thaw sperm quality and functionality variables evaluated using computer-assisted sperm analysis and flow cytometry. A total of 30 ejaculates from five bulls were analysed. With use of Triladyl®, sperm had a greater post-thaw total motility than with use of Bioxell® and Optixcell® but there was no difference as compared with use of Andromed® with the greatest (P < 0.05) percentage of progressively motile cells. With use of Optixcell®, there was a greater (P < 0.05) percentage of sperm with an intact membrane than with use of Triladyl® and Bioxcell®, but values were similar with use of Andromed®. Acrosome damage in semen preserved with use of Optixcell® was less than with use of Bioxcell® and Andromed®. With use of Optixcell®, there was a greater percentage of viable spermatozoa with a lesser lipid disruption (P < 0.05) when compared with the other extenders. Production of peroxides was greater for sperm cryopreserved with use of Triladyl® and Optixcell® while less superoxide was produced in the samples cryopreserved with the egg yolk-based extender. Optixcell® appears to be a promising alternative to replace traditional egg yolk extenders. With use of Optixcell®, however, there were greater peroxide concentrations after thawing. With use of Andromed®, there were similar results as with use of Optixcell®, therefore, it could be an effective substitute for egg-yolk based media due to the greater proportion of highly and progressively motile spermatozoa at thawing.
42.	Morillo Rodriguez, A, et al. (författare) Freezing stallion semen with the new Caceres extender improves post thaw sperm quality and diminishes stallion-to-stallion variability 2011 Ingår i: Animal Reproduction Science. - : Elsevier Masson. - 0378-4320 .- 1873-2232. ; 127:1-2, s. 78-83 Tidskriftsartikel (refereegranskat)abstract Ejaculates from 7 stallions were split and simultaneously frozen in three different extenders, INRA 96 egg yolk glycerol, Ghent and the newly developed extender Caceres. After thawing, samples were evaluated for motility (CASA system) sperm membrane integrity and early membrane changes (YoPro-1/Eth staining), acrosome integrity (FICT-PNA), and mitochondrial membrane potential (JC-1) (flow cytometry). Samples frozen in Caceres extender consistently showed the best results in post-thaw motility (increases ranging from 11 to 17%, p andlt; 0.05) and velocity (p andlt; 0.05), membrane integrity (increases ranging from 11 to 14%, p andlt; 0.05) and mitochondrial membrane potential (p andlt; 0.05). It is concluded that this new extender should be included in a freezeability test to determine the best extender for each individual.
43.	Morrell, Jane (författare) An update on semen collection, preservation and artificial insemination in the dromedary camel (Camelus dromedaries) 2018 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 194, s. 11-18 Forskningsöversikt (refereegranskat)abstract Artificial insemination (AI) in domestic animals is an important tool to maximise the use of genetically superior males and thereby insure rapid genetic progress. However, the application of AI in camelids has been hindered by the difficulties involved in collecting, as well as handling the semen due to the viscous nature of the seminal plasma. This review describes the challenges of semen collection and discusses the role of seminal plasma as well as the reasons for the viscosity and how to liquefy it so that ejaculates can be more accurately evaluated. It also reports on the use of various extenders used for liquid storage of fresh and chilled semen and how pregnancy rates are affected by numbers of spermatozoa inseminated, site of insemination and timing of insemination in relation to GnRH injection given to induce ovulation. In addition, this paper reviews the latest research in cryopreservation of camel semen and addresses the various problems involved and possible improvements that can be made so that pregnancy rates can be increased with frozen semen.
44.	Morrell, Jane, et al. (författare) Bull breed affects which parameters of sperm quality are indicative of fertility 2016 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 169, s. 112-113 Tidskriftsartikel (övrigt vetenskapligt/konstnärligt)
45.	Morrell, Jane, et al. (författare) Colloid centrifugation of fresh semen improves post-thaw quality of cryopreserved dromedary camel spermatozoa 2018 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 192, s. 28-34 Tidskriftsartikel (refereegranskat)abstract Colloids have been successfully used in a number of species to improve sperm populations for IVF and for cryopreservation The usefulness of Single Layer Centrifugation (SLC) for freezing dromedary camel spermatozoa in two different extenders was evaluated by examining the motility, viability, acrosome status, DNA integrity, and ability of cryopreserved sperm to penetrate oocytes in vitro in a heterologus IVF system. Two ejaculates from each of five males were divided into four aliquots: two were processed by SLC (selected) while two were centrifuged without colloid (control). Pellets were cryopreserved in Green Buffer or INRA-960 degrees containing 3% glycerol and evaluated at 0 and 1 h post thawed. The SLC improved post-thaw total and progressive motility at 0 (both P < 0.0001) and 1 (P < 0.001; P < 0.01, respectively) h, and STR (both P < 0.05) and BCF (both P < 0.001) at 0 h. Sperm viability and acrosome integrity (both P < 0.001) were improved at both time points. Sperm frozen in Green Buffer had greater total and progressive motilities at 0 (both P < 0.001) and 1 (both P < 0.001) h than INFtA-965 samples. Spermatozoa in Green Buffer also had a greater VAP, VCL and VSL at 0 h and improved viability and acrosome integrity at Oh (P < 0.05; P = 0.001, respectively) and 1 h (P < 0.05; P < 0.001, respectively). Viability of SLC spermatozoa was improved in Green Buffer at 1 h (P < 0.05). Oocyte penetration (P < 0.05) and pronuclear formation (P < 0.01) were greater with SLC-selected spermatozoa than non-selected spermatozoa, regardless of extender. No difference was observed between treatments or extenders in the mean number of spermatozoa per oocyte penetrated. The SLC spermatozoa had less (P < 0.01) DNA fragmentation compared to controls. The DNA fragmentation was moderately and negatively correlated with penetration (r = -0.4162; P = 0.02) and pronuclear formation (r = -0.3390; P < 0.01). In conclusion, colloid centrifugation of spermatozoa and cryopreservation in Green Buffer improves post thaw motility variables and IVF performance of dromedary camel spermatozoa.
46.	Morrell, Jane (författare) Colloid centrifugation reduces bacterial load in chilled dog semen 2020 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 219 Tidskriftsartikel (refereegranskat)abstract Conventional semen extenders contain antibiotics to prevent bacterial growth. Finding alternatives would be beneficial to minimize the development of bacterial resistance mechanisms. The aim of this study was to determine the effect of Single Layer Centrifugation (SLC) with Canicoll of dog semen on microbial load and sperm quality during cooled storage. Twenty-four ejaculates were obtained from healthy dogs by digital manipulation. Samples were diluted in Tris-citratefructose extender without antibiotics and divided into two treatment groups: SLC-selected samples and unselected samples. Sperm motility (CASA), viability and acrosome integrity (PI/FITC-PNA) as well as bacterial load of each microorganism species (colony-forming units/mL) were assessed at 0 and 48 h of storage at 4 degrees C. Results indicate SLC-selected dog spermatozoa have greater percentages of motility, viability and acrosome integrity (P < 0.05). Bacterial growth in SLC sperm samples was less (P < 0.05) than unselected samples. Removal of individual bacterial species varied from 91 % to 98 % for Escherichia coll. (91.62 %), Streptococcus spp. (98.18 %), Staphylococcus spp.(95.33 %) and Pseudomonas spp. (92.50 %). In conclusion, the use of SLC with Canicoll has the potential to decrease bacterial load in chilled dog semen.
47.	Morrell, Jane, et al. (författare) Comparison of differents methods of sperm selection of llama raw semen 2016 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 173, s. 8-12 Tidskriftsartikel (refereegranskat)abstract The objective of this study was to compare the efficiency of different sperm selection methods applied to the same llama ejaculate. Four treatments were compared: two variants of the swim up technique (with and without seminal plasma), and two different colloids, Androcoll-E-Large and Percoll (R). Using electroejaculation, 21 semen samples were obtained from 7 llama males (n = 7, r=3). The ejaculates were incubated in a solution of 0.1% collagenase, to decrease thread formation, and then split into 4 aliquots: one aliquot was layered over a column of Androcoll-E-Large (SLC) and the second over a column of Percoll (45%). The third aliquot was deposited in a tube with culture medium and was incubated at a 45 degrees angle for 30 min at 37 degrees C (SU1). The last aliquot was centrifuged to separate the spermatozoa and seminal plasma. The sperm pellet obtained was resuspended, and transferred to a tube with culture medium which was incubated at an angle of 45 degrees for 30 min at 37 degrees C (SU2). Both aliquots SLC and P showed higher proportions of progressive motility and plasma membrane functionality (p <= 0.05) than raw semen. There were no significant differences (p > 0.05) in sperm viability and in normal spermatozoa between raw semen and treatments. Nevertheless, only SLC did not have a significant increase of bent tails. In conclusion SLC centrifugation would be the method of choice for selecting llama spermatozoa.
48.	Morrell, Jane, et al. (författare) Comparison of DNA fragmentation of frozen-thawed epididymal sperm of dogs using Sperm Chromatin Structure Analysis and Sperm Chromatin Dispersion test 2017 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 187, s. 74-78 Tidskriftsartikel (refereegranskat)abstract The aim of this study was to compare sperm DNA fragmentation of frozen-thawed epididymal sperm of dogs using the SCSA (Sperm Chromatin Structure Assay) and SCDt (Sperm Chromatin Dispersion test). For this purpose, epididymis from neutered dogs were minced and incubated in a Tris-based extender. The recovered sperm were frozen in a two-step cooling protocol with Tris-based, egg yolk extender and increasing glycerol concentrations, and stored in liquid nitrogen. After thawing, each replica was incubated at 38 degrees C for 24 h. Sperm DNA fragmentation index (sDFi) was assessed by SCSA and SCDt at 0, 3, 6 and 24 h of incubation and compared within treatments. The relationship and agreement between techniques were evaluated by Pearson's coefficient and Intraclass Correlation Coefficient (ICC). The results were expressed as mean standard error of the mean (SEM). Both techniques indicated there was a significant increase of DNA fragmentation after 24 h of incubation. Moderate correlation (r = 0.65; P < 0.01) but lack of agreement (ICC = 0.451; P > 0.05) was found between SCSA and SCDt. The lack of agreement indicates that SCSA and SCDt measure different aspects of DNA fragmentation. Four halo morphologies were detected after 24 h of incubation using the SCDt: un-fragmented DNA with a small halo, fragmented DNA with large halo and two new halo presentations never described before for dog sperm: receding sperm with a disappearing halo and "bald" sperm without chromatin dispersion halo around the core. Sperm without a halo of chromatin dispersion are not described by the manufacturer and are similar to un-fragmented sperm, which could lead to erroneous results when using the SCDt. Further studies with different incubation periods and including the new morphologies described in this study should be performed. In conclusion, although SCSA and SCDt can evaluate the changes in the sperm DNA fragmentation dynamics of frozen-thawed epididymal dog sperm, these provided different findings on sperm DNA fragmentation.
49.	Morrell, Jane (författare) Effect of cryopreservation and single layer centrifugation on canine sperm DNA fragmentation assessed by the sperm chromatin dispersion test 2013 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 143, s. 118-125 Tidskriftsartikel (refereegranskat)abstract The aims of this study were: 1) to assess the effect of freezing and thawing on dog sperm DNA fragmentation index (sDFI) using the sperm chromatin dispersion test (SCDt); and 2) to determine whether or not the sperm selection by single layer centrifugation (SLC) using Androcoll-C improves sperm DNA longevity in SLC-selected frozen-thawed dog semen samples. Semen samples were collected from 4 dogs using digital manipulation. After collection, ejaculates were pooled and cryopreserved following a standard protocol. Sperm motility and morphology were assessed before freezing and after thawing as a control for the cryopreservation method used. In experiment 1, sDFI was analyzed immediately before freezing and after thawing (baseline values), showing no significant differences between fresh and frozen-thawed semen samples. In experiment 2, frozen-thawed semen samples were processed or not by SLC using Androcoll-C and longevity of DNA were assessed in terms of sDFI after 24 h of in vitro incubation at physiological temperature (38 degrees C). The results showed low values of sDFI in SLC-selected semen in comparison to unselected samples. In conclusion, no effect of cryopreservation was observed on baseline values of dog sperm DNA fragmentation. Additionally, SLC-selection using Androcoll-C improved longevity of frozen-thawed sperm DNA assessed by the SCDt. (C) 2013 Elsevier B.V. All rights reserved.
50.	Morrell, Jane (författare) Effects of single layer centrifugation with Androcoll-P on boar sperm 2013 Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 138, s. 276-281 Tidskriftsartikel (refereegranskat)

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