| 1. |
- Al-Essawe, Essraa M, et al.
(författare)
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Seminal plasma influences the fertilizing potential of cryopreserved stallion sperm
- 2018
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 115, s. 99-107
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Tidskriftsartikel (refereegranskat)abstract
- Seminal plasma (SP) contains proteins that may influence cryosurvival and prevent capacitation-like changes due to freezing and thawing. The objective of this study was to investigate the effect of adding pooled SP from "good" (GF) or "bad" (BF) freezer stallions on sperm cells' fertilizing ability. "Good freezers" refers to stallions that usually produce ejaculates which can withstand cryopreservation, whilst "bad freezer" stallions produce ejaculates which cannot tolerate the freezing process. A heterologous zona binding assay with in vitro matured bovine oocytes was used to assess the binding ability of equine sperm cells as a possible alternative to artificial insemination trials. The effect of adding SP i) prior to cryopreservation; ii) after thawing of sperm cells selected by single layer centrifugation (SLC); iii) to capacitation medium, was evaluated. Adding SP from GE stallions prior to cryopreservation reduced the mean number of sperm cells bound to the zona pellucida (ZP) compared to control (P = 0.0003), SP-free sperm cells and group received SP from BF stallions (P < 0.0001 for both). After thawing SLC-selected sperm cells treated with 5% SP showed a decrease in binding ability compared with SP-free sperm cells (P < 0.0001). The binding affinity of sperm cells was higher in the group treated with SP from GF than with SP from BF stallions (P < 0.05). Prolonged exposure to SP impaired the ability of stallion sperm cells to undergo capacitation and bind to ZP, regardless of the source of SP (P < 0.0001). The response of equine sperm cells to SP is influenced by the ability of the sperm cells to withstand cryopreservation and is affected by the timing of exposure and the origin of SP. Customization of the protocol for individual stallions is recommended to optimize the effect. (C) 2018 Elsevier Inc. All rights reserved.
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| 2. |
- Alvarez-Rodriguez, Manuel, et al.
(författare)
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Effect of colloid (Androcoll-Bear, Percoll, and PureSperm) selection on the freezability of brown bear (Ursus arctos) sperm
- 2016
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Ingår i: Theriogenology. - : ELSEVIER SCIENCE INC. - 0093-691X .- 1879-3231. ; 85:6, s. 1097-1105
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Tidskriftsartikel (refereegranskat)abstract
- The development of a species-specific conservation protocol that involves artificial insemination with frozen semen needs to validate an effective methodology for freezing semen. Colloid centrifugation has been suggested and widely applied as an effective tool for selecting animal spermatozoa for artificial breeding. The objective of the present study was to compare different methods of centrifugation, single layer using Androcoll-Bear and Percoll and double layer using PureSperm 100 (in two different discontinuous gradients 40%-80% and 45%-90%), for the selection of fresh brown bear sperm samples. In the before freezing group, all selected samples showed a higher progressive motility and viability (except Percoll for motility 43.0 +/- 5.3 [P < 0.051); all colloids except PureSperm 45/90% rendered samples with fewer damaged acrosomes. In the after thawing group, all tested centrifugation colloids showed a good capacity to decrease the number of damaged acrosomes. Furthermore, PureSperm treatment (45/90%) resulted in an increase in apoptotic-like changes not only immediately after thawing but also after the incubation test, leading us to suggest that this gradient could induce some kind of deleterious effects on the sperm samples. On the other hand, PureSperm treatment (40/80%) yielded a quality preservation capacity similar to Androcoll-Bear in number of damaged acrosomes, different relative to the control (control, 5.3 +/- 0.6; PureSperm 80, 2.0 +/- 0.3; Androcoll, 2.1 +/- 0.9 [P < 0.051) but a decrease in the number of viable spermatozoa recovered after thawing relative to the control (control, 21.2 +/- 3.1; PureSperm 80, 13.7 +/- 2.7 [P < 0.051). In conclusion, Androcoll-Bear constitutes a useful tool for handling of brown bear ejaculates owing to its simple handling and procedure with a reliable sperm selection and freezability. This colloid yielded an improvement in several sperm parameters in brown bear frozen-thawed semen; the selected spermatozoa of fresh samples with this colloid showed a better resistance to freezing compared with the control sample not only for motility but also for viability. (C) 2016 Elsevier Inc. All rights reserved.
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| 3. |
- Alvarez-Rodriguez, Manuel, et al.
(författare)
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Exosomes in specific fractions of the boar ejaculate contain CD44: A marker for epididymosomes?
- 2019
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Ingår i: Theriogenology. - : ELSEVIER SCIENCE INC. - 0093-691X .- 1879-3231. ; 140, s. 143-152
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Tidskriftsartikel (refereegranskat)abstract
- Seminal plasma (SP) is a complex fluid containing proteins, peptides, enzymes, hormones as well as extracellular vesicles (EVs). The SP interacts with spermatozoa and the inner cell lining of the female genital tract, adsorbing proteins and exosomes that modulate sperm functions and female immune responsiveness. In the present study, boar sperm-free SP was studied using flow cytometry (FC) after membrane tetraspanins (CD9, CD63 and CD81) and membrane receptor CD44 marking of non-enriched (whole SP) or gradient fractions enriched through two-step discontinuous KBr-density-gradient ultracentrifugation, in whole ejaculate or in selected ejaculate fractions. The results, evaluated by transmission electron microscopy, confirmed the presence of exosomes in all fractions of the pig SP. Noteworthy, these pig SP-exosomes were CD44-bearing when analysed by FC, with bands detected by western blotting (WB) at the expected 85 kD size. The two-step discontinuous KBr-density-gradient ultracentrifugation enriched the population of exosomes in two specific gradient fractions, indicating exosomes (either prostasomes or epididymosomes) could be separated from low-density lipoprotein (LDL) but they co-sediment with the high-density lipoprotein (HDL)-bearing fraction. The findings pave for the selective isolation of exosomes in functional studies of their function when interacting with spermatozoa, the oocyte and/or the female genitalia, including hyaluronan-CD44 interplay. (C) 2019 Elsevier Inc. All rights reserved.
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| 4. |
- Axner, Eva, et al.
(författare)
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Concentrations of anti-Müllerian hormone in the domestic cat. Relation with spay or neuter status and serum estradiol
- 2015
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 83, s. 817-821
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Tidskriftsartikel (refereegranskat)abstract
- Female cats with unknown history can be diagnosed as spayed or intact with a GnRH-stimulation test or an LH test independent of the stage in the estrous cycle. However, although most females are correctly diagnosed with the LH test, the sensitivity and specificity are not 100%. The GnRH-stimulation test, although reliable, requires an injection of buserelin 2 hours before the blood sample is collected. Granulosa cells are the only cell type that produces anti-Mullerian hormone (AMH) in females, whereas Sertoli cells produce AMH in males. Anti-Mullerian hormone has been linked to spay status in dogs and cats and to ovarian and testicular pathology and fertility in different species. Our aim was to evaluate serum AMH concentrations in spayed female cats and in intact female cats of known age and reproductive stage (inactive ovaries or luteal phase). In addition, our aim was to compare serum AMH concentrations in intact and neutered male cats. We analyzed serum AMH concentrations in 15 spayed and 16 intact females and in 15 intact and 12 neutered male cats. Serum AMH was below the lowest standard point (<0.14 ng/mL) in all spayed females and neutered males, ranged between 1.3 and 19.0 ng/mL in the intact females and between 4.8 and 813 ng/mL in intact males. Thus, the AMH test had 100% sensitivity and specificity to diagnose the presence or absence of ovaries and testes in this study. In addition, in contrast to serum estradiol, serum AMH was not affected by buserelin stimulation (P = 0.459). Serum AMH was not correlated with serum estradiol before (r(s) = -0.188, P = 0.519) or after (r(s) = 0.335, P = 0.242) buserelin stimulation in the intact females. Four 6-month-old intact cats (two females and two males) had the highest AMH concentrations which in the females might represent a prepubertal peak previously described in other species and in males is likely due to high concentrations before puberty. In conclusion, we found that the AMH Gen II ELISA is reliable for diagnosing spay and neuter status of cats and that the domestic cat might be an interesting model for studies on AMH dynamics. (C) 2015 Elsevier Inc. All rights reserved.
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| 5. |
- Axner, Eva, et al.
(författare)
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Macroscopic and microscopic evaluation of Eurasian lynx (Lynx lynx) female tubular reproductive organs in relation to ovarian structures
- 2015
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 84, s. 710-715
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Tidskriftsartikel (refereegranskat)abstract
- Although monitoring wild animals in the field is essential for estimations of population size and development, there are pitfalls associated with field monitoring. In addition, some detailed data about reproductive physiology can be difficult to obtain in wild live animals. Studying reproductive organs from the Eurasian lynx killed at hunting or found dead could be used as a valuable addition to other field data. We evaluated reproductive organs from 39 Eurasian lynx females (Lynx lynx) killed in Sweden during the hunting seasons in 2009, 2010, and 2011. According to notes on ovarian structures, the animals were categorized as being in one of four different reproductive stages: juvenile (n = 10), follicular stage (n = 8), luteal stage (n = 11), and anestrus (n = 10). Corpora lutea were classified as fresh CL from the present season or as luteal bodies from previous cycles. Microscopic evaluations were blindly coded while the outer measurements of the vagina and uterus were taken at the time of organ retrieval. The width of the endometrium, myometrium, outer width of the uterine horns, and the diameter of the vagina differed significantly with the reproductive stage (P < 0.001) and were largest in the follicular and luteal phases. The number of endometrial glands evaluated blindly coded on a subjective scale was significantly associated with the reproductive stage (P < 0.0001) and was significantly higher in the luteal phase than that in any other reproductive stages (P < 0.05). Cornification of the vaginal epithelium was only observed in females in the follicular stage or in females with signs of a recent ovulation. In conclusion, both macroscopic and histologic measurements are useful for a correct classification of the reproductive stage when evaluating reproductive organs in the Eurasian lynx killed during the hunting season. Routine evaluation of reproductive organs has a potential to be a useful additional tool to field studies of live lynx to monitor their reproduction. (C) 2015 Elsevier Inc. All rights reserved.
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| 6. |
- Barranco, Isabel, et al.
(författare)
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Levels of activity of superoxide dismutase in seminal plasma do not predict fertility of pig AI-semen doses
- 2019
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Ingår i: Theriogenology. - : ELSEVIER SCIENCE INC. - 0093-691X .- 1879-3231. ; 140, s. 18-24
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Tidskriftsartikel (refereegranskat)abstract
- Superoxide dismutase (SOD) is a major antioxidant enzyme in boar seminal plasma (SP). This study evaluated how SP-SOD affected sperm attributes when semen of boars of various breeds, included in commercial artificial insemination (Al)-programs, was extended and liquid-stored at 17 degrees C for AI; as well as their in vivo fertility (farrowing rate and litter size of 10,952 AI-sows). SP-SOD-activity was assessed in 311 ejaculates (100 boars) while sperm motility (by CASA), viability and intracellular H2O2 generation in viable spermatozoa (by flow cytometry) were measured at 0 and 72 h of liquid storage. SP-SOD activity was not affected by breed but differed (P amp;lt; 0.001) between boars (n = 50), ranging from 1.16 +/- 0.11 to 7.02 +/- 0.75 IU/mL. Semen Al-doses (n =44) hierarchically grouped (P amp;lt; 0.001) with low SP-SOD activity showed lower (P amp;lt; 0.05) sperm motility and intracellular H2O2 at 72 h of liquid storage. Fertility did not differ between AI-boars (n = 39) hierarchically grouped (P amp;lt; 0.001) with high or low SP-SOD activity. In conclusion, SP-SOD activity is boar dependent and positively related with sperm functionality of liquid stored semen AI-doses. However, this positive effect is not reflected on in vivo fertility post-AI. (C) 2019 Elsevier Inc. All rights reserved.
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| 7. |
- Chatdarong, Kaywalee, et al.
(författare)
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The development of cat testicular sperm cryopreservation protocols: Effects of tissue fragments or sperm cell suspension
- 2016
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 85, s. 200-206
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Tidskriftsartikel (refereegranskat)abstract
- In endangered animals that have been found dead or sterilized for medical reasons, testis isthe ultimate source of haploid DNA or sperm. Thus, preservation of testicular sperm maybe performed to rescue their genetics. The aim of this study was to evaluate protocols fortesticular sperm freezing: as tissue fragments or cell suspension in domestic cats as amodel. A pair of testes from each cat (n ¼ 9) were cut into eight equal pieces. Fourrandomly selected pieces were cryopreserved as: (1) tissue pieces using two-step freezing;(2) tissue pieces using a slow passive cooling device (CoolCell); (3) sperm suspension aftersingle-layer centrifugation (SLC) through colloids; and (4) sperm suspension without beingprocessed through SLC. A testicular piece from each cat served as fresh control. Testicularsperm membrane and DNA integrity were evaluated before, and after, the cryopreservationprocess. In addition, spermatogenic cell types (testicular sperm, spermatogonia, spermatocyte,and spermatid) present in the suspension samples were counted before andafter SLC. The results found that testicular sperm membrane integrity in the suspensionafter SLC process was higher than that in the fragment form neither using the two-step norCoolCell freezing, both before and after freezing (before freezing: 92.3 3.4 vs. 81 4.5and 80.0 7.0; after freezing: 84.5 4.6 vs. 71.2 12 and 76.2 4.6; P 0.05). Testicularsperm DNA integrity was, however, not different among groups. Furthermore, the samplesprocessed through the SLC had higher ration of sperm cells: other spermatogenic cells thanthose were not processed through the SLC (88.9 3.8 vs. 30 7.9; P 0.05). In summary,testicular sperm cryopreserved as a minced suspension is considered suitable in terms ofpreventing sperm membrane integrity, and SLC is considered a selection tool for enrichinghaploid sperm cells from castrated or postmortem cats.
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| 8. |
- Einarsson, Stig, et al.
(författare)
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Occurrence of bacteria and polymorphonuclear leukocytes in fetal compartments at parturition; relationships with foal and mare health in the peripartum period
- 2015
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 84, s. 163-169
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Tidskriftsartikel (refereegranskat)abstract
- This study investigated the relationship of the health of the newborn foal and (1) number of polymorphonuclear leukocytes (PMNLs) in the amniotic fluid, (2) bacteria present in the amniotic fluid and the venous umbilical blood, and (3) bacteria present in the uterus of the newly foaled mare. A further aim was to investigate relationships between the bacteriologic findings in the amniotic fluid, umbilical blood, and uterus postpartum. Samples were taken from 50 Standardbred trotter foaling mares from a well-managed stud in Sweden. Parturition was spontaneous in all cases. Length of pregnancy, parturition and postpartum complications, health status of the foal, the time between foaling and the expulsion of the placenta, and the number of postfoaling mares becoming pregnant after insemination were recorded. Amniotic fluid was collected when the amniotic vesicle was clearly visible; it was analyzed for bacteriology and occurrence of PMNLs. Umbilical blood was analyzed for the presence of bacteria and the concentration of serum amyloid A. The uterus of themarewas swabbed for bacteriology 6 to 17 hours postpartum. A blood samplewas taken from the foal before administering plasma. The foals were divided into two groups: group 1 required up to 2 hours to rise after birth (2 hours; 31 foals) and group 2 requiredmore than two hours (>2 hours; 19 foals). The length of gestation varied between 332 and 356 days; there was no significant difference in gestation length between the two foal groups. Partus and postpartum complications occurred in a significantly higher proportion of mares giving birth to group 2 foals than group 1 foals (P ¼ 0.02), although uterine culture postpartum and the subsequent pregnancy rate per season were not different between the groups. Compromised health status was significantly higher among foals belonging to group 2 than group 1 (P ¼ 0.001). Most of the amniotic samples contained 5% or less PMNLs. Only three samples contained more than 30% PMNLs; group 2 foals had the highest percentage of PMNLs. Bacterial growth was found in both amniotic fluid (57%) and umbilical blood (35%) in mares irrespective of whether their foals were healthy or compromised. Coagulase-negative staphylococci were the most frequent bacteria. There were no differences in bacterial occurrence in amniotic fluid or in umbilical blood between the two foal groups
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| 9. |
- Elwing, Bodil, et al.
(författare)
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Effect of different freezing rates and thawing temperatures on cryosurvival of dromedary camel spermatozoa
- 2019
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 125, s. 43-48
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Tidskriftsartikel (refereegranskat)abstract
- The objective of this study was to evaluate the effect of different freezing rates and thawing temperatures on the post-thaw quality of camel spermatozoa. Ten ejaculates from five male camels were frozen at five different freezing rates, achieved by placing the straws at specific heights above the surface of liquid nitrogen for different lengths of time (4 cm for 15 min; 1 cm for 15 min; 7 cm for 15 min; 7 cm for 5 min + 4 cm for 3 min; 4 cm for 5 min + 1 cm for 3 min) followed by storage in liquid nitrogen. Two thawing temperatures (37 degrees for 30 s and 60 degrees C for 10 s) were subsequently tested. Post-thawing, the samples were evaluated for total and progressive motility, kinematics, membrane and acrosome integrity, and membrane functionality (hypoosmotic swelling test) at zero and 1 h post thawing. Total and progressive motility were significantly higher for the fastest freezing rate (at 1 cm) at 0 h (p < 0.01 for both), as were VCL (p < 0.01), VSL (p < 0.05) and STR (p < 0.05). Freezing at 4 cm produced the lowest values of STR compared to other treatments (p < 0.05). At 1 h, no differences in total motility were observed between freezing at 4 cm and 1 cm, both being significantly better than freezing rate 7 cm + 4 cm (p < 0.01). For progressive motility and VSL, only freezing at 1 cm was superior to the 7 cm + 4 cm combination (p < 0.001 and p < 0.05 respectively). Membrane integrity at 1 h was higher for freezing at 7 cm than at 1 cm (p < 0.01). For thawing temperatures, total motility and progressive motility at 0 h and 1 h (p < 0.001), and acrosome integrity at 1 h (p < 0.01) were higher for 60 degrees C thawing temperature than 37 degrees C. The kinematics VCL (p < 0.001), VSL and STR (p < 0.01), and VAP (p < 0.05) showed higher values for 60 degrees C thawing temperature than 37 degrees C at 0 h. After 1 h, higher values for VSL, VCL and VAP (p < 0.05) were observed for 60 degrees C than for 37 degrees C. In conclusion, a fast freezing rate would probably be beneficial for camel semen, and thawing should be conducted at 60 degrees C. (C) 2018 Published by Elsevier Inc.
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| 10. |
- Gonzalez-Arto, Marta, et al.
(författare)
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Melatonin receptors MT1 and MT2 are expressed in spermatozoa from several seasonal and nonseasonal breeder species
- 2016
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Ingår i: Theriogenology. - : ELSEVIER SCIENCE INC. - 0093-691X .- 1879-3231. ; 86:8, s. 1958-1968
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Tidskriftsartikel (refereegranskat)abstract
- Melatonin is a ubiquitous and multipurpose molecule, and one of its roles is to regulate reproduction in some seasonal mammals. Our group has previously reported the variation in the melatonin levels in ram seminal plasma along the year and identified MT1 and MT2 receptors in ram spermatozoa. The objective of this study was to elucidate whether the presence of melatonin receptors (MT1 and MT2) in the sperm plasma membrane, and melatonin in the seminal plasma is related to seasonal breeding. For this purpose, the presence of melatonin receptors and tseasonal breeder (epididymal spermatozoa); bull as a conventional nonseasonhe levels of melatonin in seminal plasma have been examined in several species: donkey and stallion as long-day breeders; red deer as a wild, short-day, highly al breeder; boar as a seasonal breeder, under management techniques; and dog as possible a seasonal breeder not regulated by melatonin. We have detected measurable levels of melatonin in the seminal plasma of all ejaculated semen samples (from donkey, stallion, boar, bull, and dog). Also, and for the first time, we have demonstrated the presence of MT1 and MT2 melatonin receptors in the spermatozoa of all these species, regardless their type of reproduction or sperm source (ejaculated or epididymal), using indirect immunofluorescence techniques and Western blotting. Our findings suggest that melatonin and melatonin receptors may be universally distributed in the reproductive system of mammals and that the sperm melatonin receptors cells may not be necessarily related with seasonal reproduction. Furthermore, the presence of MT1 at the cytoplasmic droplet in immature ejaculated stallion spermatozoa found in one sample and epididymal red deer spermatozoa suggests that melatonin may be involved in specific functions during spermatogenesis and sperm maturation, like protecting spermatozoa from oxidative damage, this activity being mediated through these receptors. (C) 2016 Elsevier Inc. All rights reserved.
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| 11. |
- Gonzalez Herrero, Raquel, et al.
(författare)
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Blood plasma collected after adrenocorticotropic hormone administration during the preovulatory period in the sow negatively affects in vitro fertilization by disturbing spermatozoa function
- 2015
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 83, s. 1128-1139
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Tidskriftsartikel (refereegranskat)abstract
- Successful fertilization is essential for reproduction and might be negatively affected by stressful events, which could alter the environment where fertilization occurs. The aim of the study was to determine whether an altered hormonal profile in blood plasma caused by adrenocorticotropic hormone (ACTH) administration could affect in vitro fertilization in the pig model. In experiment 1, gametes were exposed for 24 hours to plasma from ACTHtreated, non-ACTH-treated sows, or medium with BSA. Fertilization, cleavage, and blastocyst rates were lower in the ACTH group compared with the no ACTH or BSA control groups (P < 0.01). In experiment 2, the exposure of matured oocytes for 1 hour before fertilization to the same treatments did not have an impact on their ability to undergo fertilization or on embryo development. In experiment 3, spermatozoa were incubated for 0, 1, 4, and 24 hours under the same conditions. There was no effect of treatment on sperm viability. The percentage of acrosome-reacted spermatozoa remained higher in the ACTH group compared with the non-ACTH-treated group through the incubation period (P < 0.001). Protein tyrosine phosphorylation (PTP) patterns were also affected by treatment (P < 0.001). The presence of an atypical PTP pattern was higher in the ACTH group at all the analyzed time points compared with the BSA and no ACTH groups (P < 0.001). In conclusion, this altered environment may not affect oocyte competence but might affect the sperm fertilizing ability through alterations in the acrosome reaction and correct sequence of PTP patterns. (C) 2015 Elsevier Inc. All rights reserved.
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| 12. |
- Hagman, Ragnvi
(författare)
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Serum tryptophan and its metabolites in female dogs undergoing ovariohysterectomy as treatment of pyometra or as elective spay surgery
- 2015
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 83, s. 1279-1286
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Tidskriftsartikel (refereegranskat)abstract
- This study compares serum concentrations of tryptophan (TRP), kynurenine (KYN), kynurenic acid (KYNA), and indoleamine 2,3-dioxygenase (IDO) activity in healthy bitches and bitches with bacterial uterine infection (pyometra). The effects of surgery were also assessed by measuring these variables in both groups of dogs before and after ovariohysterectomy. Presurgery, mean (+/- standard deviation) TRP, KYN, and KYNA concentrations and IDO activity were 68.44 +/- 1.77, 2.00 +/- 0.33, 112.11 +/- 111.91 mu mol/L, and 29.22 +/- 10.10, respectively, in the healthy dogs; and 40.16 +/- 12.11, 8.27 +/- 3.94, 411.11 +/- 199.60 mu mol/L, and 205.92 +/- 154.20, respectively, in the dogs with pyometra. Tryptophan and KYN levels had normalized on suture removal (10 days after surgery) though IDO activity and KYNA concentrations remained elevated during the postoperative period compared with presurgery values in both study groups. Our results suggest that KYNA concentrations and IDO activity could be useful indicators of the inflammation induced by pyometra and could be also used to monitor recovery following ovariohysterectomy in both healthy dogs and dogs with pyometra. (C) 2015 Elsevier Inc. All rights reserved.
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| 13. |
- Hessle, Anna
(författare)
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Automated activity monitoring and visual observation of estrus in a herd of loose housed Hereford cattle: Diagnostic accuracy and time to ovulation
- 2017
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 87, s. 205-211
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Tidskriftsartikel (refereegranskat)abstract
- A prospective cohort study was performed in the purebred Hereford herd at Gotala Beef and Lamb Research Centre, Sweden. The study's first objective was to assess the ability of an automatic activity monitoring system (AAMS) to detect estrus in beef suckler cows, and its second objective was to estimate the time from estrus to ovulation. The study sample (n = 38) consisted of 14 Hereford heifers and 24 Hereford cows. Standardized visual observation of estrus was performed for 20 minutes thrice daily, and animal activity was recorded with an AAMS system, Heatime (SCR Engineers Ltd., Israel). Cows in estrus underwent transrectal ultrasonography every 8 hours, to estimate the time of ovulation. Blood samples for progesterone analysis were collected thrice weekly throughout the study period. A cutoff value of 1-ng progesterone/mL of serum was used to define luteal activity. The AAMS had a 90% (95% confidence interval [CI] 77%-97%) sensitivity and 100% specificity (95% CI 94%-100%), and visual detection of estrus had a 77% sensitivity (95% CI 62%-88%) and a 89% specificity (95% CI 79%-95%) for identifying estrus when compared to the gold standard defined by temporal pattern of serum progesterone concentration. When both methods were used in parallel, the sensitivity increased to 96% (95% CI 86%-99%), and the specificity increased to 90% (95% CI 80%-96%). The time of ovulation after estrus was determined on 50 occasions. The median estrus (AAMS detected) to ovulation interval was 25 hours for heifers and 23 hours for cows (interquartile range 11-29 hours and 19-25 hours, respectively). The median estrus (visually detected) to ovulation interval was 28 hours for heifers and 21 hours for cows (interquartile range 13-29 hours for both categories). In conclusion, the AAMS had both a higher sensitivity and specificity for estrus detection than thrice-daily visual observation. The time from detection of estrus to ovulation observed in this study indicates that reproductive performance might be improved if Hereford cattle are inseminated sooner after detection of estrus than is currently recommended. (C) 2016 Elsevier Inc. All rights reserved.
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| 14. |
- Humblot, Patrice
(författare)
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Reproductive performance of Bos indicus beef cows treated with different doses of equine chorionic gonadotropin at the end of a progesterone-estrogen based protocol for fixed-time artificial insemination
- 2018
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 118, s. 150-156
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Tidskriftsartikel (refereegranskat)abstract
- Two experiments were performed to evaluate the reproductive performance of zebu beef cows treated with different doses of eCG at the end of a progesterone (P4)/estrogen based protocol for timed artificial insemination (TAI). In Experiment 1, suckling Bos indicus Nelore cows (n = 261) received, on day 0, a progesterone (P4) intravaginal device (PD) and an injection of 1 mg estradiol benzoate (EB). On day 8, the PD was removed, 500 pig of cloprostenol was injected, and cows were assigned to one of the following groups: Control (no treatment), 300 (300 IU of eCG), 600 (600 IU of eCG), and 900 (900 IU of eCG). On day 9, all cows received 1 mg EB and TAI performed 54-56 h after cloprostenol injection. A pregnancy diagnosis was done by ultrasound scanning 40 days after TAI, and the number of fetuses and calves was recorded at pregnancy diagnosis and at birth. More cows treated with eCG displayed estrus within 48 h after removal of the PD (42.3% vs. 11.6%, P < 0.01), and ovulated more than one follicle (42%, 58/138 vs. 1.8%, 1/54; P < 0.01). This effect on ovulation rate was dose dependent (P < 0.05). The pregnancy rate was affected only by cow parity (primiparous, 25.3% vs. multiparous, 48.9%; P < 0.01). Twin pregnancy was higher (P < 0.01) in cows treated with eCG (42%, 58/138) than controls (0%, 0/54). However, few cows (33.3%) were able to keep both fetuses intact until birth. For evaluation of ovarian characteristics by B mode and Doppler ultrasonography, 43 Nelore cows were submitted In Experiment 2 to the same four groups described in Experiment 1. Although no difference (P > 0.1) was observed for size and blood perfusion in the pre-ovulatory follicles, corpus luteum was larger and with greater blood perfusion (P <0.05) in eCG-treated cows. In conclusion, eCG increased the number of double/multiple ovulations in a dose-dependent manner, induced larger and more vascularized corpora lutea, but did not affect the fertility of cyclic or anestrous cows. Although eCG results in twin pregnancies, most of cows underwet embryo/fetus loss and birth a single calf. (C) 2018 Elsevier Inc. All rights reserved.
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| 15. |
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| 16. |
- Karlsson, Iulia, et al.
(författare)
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Pathogenic Escherichia coli and lipopolysaccharide enhance the expression of IL-8, CXCL5, and CXCL10 in canine endometrial stromal cells
- 2015
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 84, s. 34-42
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Tidskriftsartikel (refereegranskat)abstract
- Chemokines play a central role in cellular communication in response to bacterial infection. However, the knowledge of the chemokine responses to bacterial infections in dogs remains limited. Uterine bacterial infection (pyometra) is one of the most common bacterial diseases in dogs and causes sepsis in most of the cases. We have shown previously that dogs with pyometra have higher messenger RNA (mRNA) levels of chemokines in uterus. To assess whether the stromal part of the endometrium expresses chemokines in response to bacterial infection, we cultured endometrial stromal cells isolated from healthy dogs and exposed them to either live pathogenic Escherichia coli, isolated from the uterus of a dog with pyometra, or lipopolysaccharide. Changes in the mRNA expression of ELR+ CXC chemokines, IL-8, CXCL5, CXCL7, and ELR- CXC chemokine, CXCL10, were measured after 24 hours using quantitative real-time polymerase chain reaction. Levels of IL-8, CXCL5, and CXCL10 were upregulated in endometrial stromal cells exposed to E coli and lipopolysaccharide, whereas the level of CXCL7 was decreased or unaffected. In addition, levels of IL-8 and CXCL5, but not CXCL7 or CXCL10, were significantly higher in dogs with pyometra than those in healthy dogs. Our findings show that pathogenic uterine-derived E coli induces a CXC chemokine response both in cultured endometrial stromal cells within 24 hours and in pyometra-affected uteri from dogs. Stromal cells could therefore play an important role in early neutrophil and T cell recruitment to the site of inflammation during gram-negative bacterial infection of the uterus. Further studies are needed to clarify the role of chemokines in host response to bacterial infection in dogs and the possibility of using chemokines as diagnostic parameters for bacterial infection in this species. (C) 2015 Elsevier Inc. All rights reserved.
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| 17. |
- Kumaresan, Arumugam, et al.
(författare)
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Effect of bovine oviductal fluid on motility, tyrosine phosphorylation, and acrosome reaction in cryopreserved bull spermatozoa
- 2019
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 124, s. 48-56
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Tidskriftsartikel (refereegranskat)abstract
- This study was conducted to investigate the complex interactions between oviducts and cryopreserved spermatozoa. Herein we report the dynamic changes in bull sperm functions during in vitro incubation with bovine estrus and luteal oviductal fluid. Frozen-thawed bull spermatozoa was incubated either in non-capacitating medium, capacitating medium, non-capacitating medium containing 20% v/v estrus oviductal fluid or non-capacitating medium containing 20% v/v luteal oviductal fluid for 6 h at 38 degrees C under 5% CO2. At hourly interval spermatozoa were evaluated for kinematics, tyrosine phosphorylation and acrosome reaction. The sperm velocity parameters were higher (P < 0.05) in capacitating medium compared to the other treatments. At 4 and 5 h of incubation, the proportion of live tyrosine phosphorylated spermatozoa was higher (P < 0.05) in estrus oviductal fluid compared to all other treatments. From 4 to 6 h of incubation the proportion of live acrosome reacted spermatozoa was higher (P < 0.05) in estrus oviductal fluid compared to the other treatments. We conclude that estrus oviductal fluid induced tyrosine phosphorylation and acrosome reaction in a higher proportion of frozen-thawed bull spermatozoa compared to luteal oviductal fluid, although sperm kinematics were not significantly influenced by oviductal during incubation. (C) 2018 Elsevier Inc. All rights reserved.
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| 18. |
- Kunkitti, Panisara, et al.
(författare)
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In vitro fertilization using frozen-thawed feline epididymal spermatozoa from corpus and cauda regions
- 2016
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 86, s. 1403-1408
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Tidskriftsartikel (refereegranskat)abstract
- Epididymal sperm preservation offers a potential for rescuing genetic material from endangered or valuable animals after injury or death. Spermatozoa from corpus, as well as from cauda, have the capability to be motile and to undergo capacitation and can thus potentially be preserved for assisted reproductive technologies. In the present study, feline frozen-thawed epididymal spermatozoa from corpus and cauda regions were investigated for their ability to fertilize homologous oocytes and further embryo development in vitro. Epididymal spermatozoa from corpus and cauda of seven cats were cryopreserved and used for IVF. Cumulus-oocyte complexes (n = 419) were obtained from female cats after routine spaying. Frozen-thawed corpus epididymal spermatozoa showed similar properties of acrosome integrity, membrane integrity, and chromatin integrity as frozen-thawed spermatozoa from cauda except corpus spermatozoa showed lower motility (P < 0.05). The fertilizing capacity of frozen-thawed corpus epididymal spermatozoa was confirmed by similar number of embryos developing to the two- and four-cell stages compared with sperm from cauda (32.03% vs. 33.33%). However, oocytes fertilized with corpus spermatozoa had lower potential to develop to the blastocyst stage (6.79%) and had lower cell numbers compared to oocytes fertilized with cauda spermatozoa (14.08%). In conclusion, spermatozoa from corpus epididymis had a similar capability to fertilize homologous oocytes in vitro as sperm from cauda but resulted in fewer embryos developing to the blastocyst stage compared to spermatozoa from the cauda. (C) 2016 Elsevier Inc. All rights reserved.
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| 19. |
- Kunkitti, Panisara, et al.
(författare)
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The tolerance of feline corpus and cauda spermatozoa to cryostress
- 2016
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 85, s. 502-508
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Tidskriftsartikel (refereegranskat)abstract
- Epididymal sperm preservation can be used to avoid the total loss of genetic material in threatened species. Spermatozoa from the corpus, as from the cauda, are motile and can undergo capacitation. Thus, they can potentially be preserved for assisted reproductive technologies. However, cryopreservation of spermatozoa has a direct detrimental effect on sperm quality. The aim of this study was to compare the chromatin stability and the survival rate of spermatozoa from the corpus and cauda epididymis after cryopreservation. Epididymal spermatozoa were collected and cryopreserved from the corpus and cauda of 12 domestic cats. Sperm motility, progressive motility, membrane integrity, acrosome integrity, and DNA integrity were evaluated before and after freezing thawing. The average total number of spermatozoa collected from the corpus was lower (10.2 x 10(6) +/- 7.4) than that from the cauda epididymis (24.9 x 10(6) +/- 14.4; P = 0.005). The percentage of spermatozoa with intact DNA did not differ significantly whether it was collected from the corpus or cauda regions and did not decrease after freezing thawing in either region. However, motility of spermatozoa from both regions was affected by the freezing thawing process with a significant decline in motility after thaw compared with fresh spermatozoa. A significant difference in the percentage of motile sperm between the corpus and cauda was observed after the freezing thawing process (P < 0.001). Although sperm motility was lower in postthaw spermatozoa from the corpus epididymidis than from the cauda, the rate of the reduction did not differ between regions. This study indicates that the cryopreservation process does not have a negative effect on chromatin stability of feline epididymal spermatozoa. Spermatozoa from the corpus region have a similar freezability as spermatozoa from the cauda region. Therefore, preservation of spermatozoa from the corpus and the cauda epididymidis might be of value in preserving genetic material from endangered or valuable felids. (c) 2016 Elsevier Inc. All rights reserved.
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| 20. |
- Laskowski, Denise, et al.
(författare)
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Insulin during in vitro oocyte maturation has an impact on development, mitochondria, and cytoskeleton in bovine day 8 blastocysts
- 2017
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 101, s. 15-25
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Tidskriftsartikel (refereegranskat)abstract
- Insulin is a key metabolic hormone that controls energy homeostasis in the body, including playing a specific role in regulating reproductive functions. Conditions associated with hyperinsulinemia can lower developmental rates in bovine in vitro embryo production and are linked to decreased fertility in humans, as in cases of obesity or type 2 diabetes. Embryo quality is important for fertility outcome and it can be assessed by choosing scoring standards for various characteristics, such as developmental stage, quality grade, cell number, mitochondrial pattern or actin cytoskeleton structure. Changes in the embryo's gene expression can reflect environmental impacts during maturation and may explain morphological differences. Together with morphological evaluation, this could enable better assessment and possibly prediction of the developmental potential of the embryo. The aim of this study was to use a bovine model to identify potential gene signatures of insulin-induced changes in the embryo by combining gene expression data and confocal microscopy evaluation. Bovine embryos were derived from oocytes matured in two different insulin concentrations (10 mu g mL(-1) and 0.1 mu g mL(-1)), then stained to distinguish f-Actin, DNA and active mitochondria. The total cell number of the embryo, quality of the actin cytoskeleton and mitochondrial distribution were assessed and compared to an insulin-free control group. A microarray-based transcriptome analysis was used to investigate key genes involved in cell structure, mitochondrial function and cell division. Our results indicate that insulin supplementation during oocyte maturation leads to lower blastocyst rates and a different phenotype, characterised by an increased cell number and different actin and mitochondrial distribution patterns. These changes were reflected by an up-regulation of genes involved in cell division (MAP2K2; DHCR7), cell structure (LMNA; VIM; TUBB2B; TUBB3; TUBB4B) and mitochondrial activation (ATP5D; CYP11A1; NDUFB7; NDUFB10; NDUFS8). Taken together, we hypothesise that the increased proliferation in the insulin-treated groups might impair the developmental potential of the embryos by inducing metabolic stress on the molecular level, which could be detrimental for the survival of the embryo. (C) 2017 Elsevier Inc. All rights reserved.
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| 21. |
- Laskowski, Denise, et al.
(författare)
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The functional role of insulin in fertility and embryonic development-What can we learn from the bovine model?
- 2016
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 86, s. 457-464
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Forskningsöversikt (refereegranskat)abstract
- Insulin is a key metabolic hormone that plays a crucial role in regulating energy homeostasis in the body. In addition, insulin-dependent signaling has important functions in reproduction and early embryo development. As metabolism and reproduction are closely linked, metabolic challenges may be the source of reproductive disorders and decreased fertility. This is known for the dairy cow and for other species including the human. Although metabolic disorders in the dairy cow often derive from a failure to adapt to a high milk production, the situation in the human is often linked to emerging conditions and associated diseases in our modern society such as obesity and diabetes, where an excess energy intake causes decreased fertility in women. Both energy excess and energy deficit are associated with a deviation of insulin concentrations in serum and follicular fluid from normal levels. Although many studies have shown that extreme variation in energy supply can negatively influence early embryo development by inducing changes in circulating concentrations of several metabolites or hormones like insulin, several in vitro culture media are still supplemented with insulin in high concentrations. In this review, direct and indirect effects of insulin on fertility will be described. Differences between the in vivo and in vitro situations will also be discussed. (C) 2016 Elsevier Inc. All rights reserved.
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| 22. |
- Li, Junwei, et al.
(författare)
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Seminal plasma antioxidants are directly involved in boar sperm cryotolerance
- 2018
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Ingår i: Theriogenology. - : ELSEVIER SCIENCE INC. - 0093-691X .- 1879-3231. ; 107, s. 27-35
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Tidskriftsartikel (refereegranskat)abstract
- Boar ejaculates are ejected in fractions with a specific composition in terms of sperm numbers and seminal plasma (SP), which is reflected in the varying sperm cryotolerance observed among different fractions. As boar sperm are particularly sensitive to oxidative stress, this study evaluated the role of SP antioxidants in the observed differences in sperm cryotolerance among ejaculate fractions. Ten ejaculates from five boars were manually collected in fractions: the first 10 mL of the sperm-rich fraction (SRF), the rest of the SRF and the post-SRF. Semen samples comprising the entire ejaculate (EE) were created by proportionally mixing the three fractions described above. Each of the 40 resulting semen samples was split into two aliquots: one was used for sperm cryopreservation following a standard protocol utilizing 0.5-mL straws, and the other was used to collect SP for antioxidant assessment. Frozen-thawed (FT) sperm from the SRF (the first 10 mL of the SRF and the rest of the SRF) and those from post-SRF were of the highest and worst quality, respectively, which was measured in terms of total and objective progressive motility and viability (P amp;lt; 0.01). Viable FT sperm from the post-SRF generated more reactive oxygen species and experienced more lipid peroxidation than those from the SRF (both the first 10 mL and the rest of the SRF) (P amp;lt; 0.01). The percentage of FT sperm exhibiting fragmented nuclear DNA did not differ among ejaculate fractions and the EE. Catalase, glutathione peroxidase and glutathione peroxidase 5 (GPx-5) were lowest in SP from the first 10 mL of the SRF (P amp;lt; 0.001), whereas superoxide dismutase (SOD) and paraoxonase 1 (PON-1) were highest in SP of the SRF (both the first 10 mL and the rest of the SRF) (P amp;lt; 0.01). Trolox-equivalent antioxidant capacity (TEAC) and the ferric-reducing ability of plasma (FRAP) were highest in SP from the first 10 mL of the SRF and lowest in the post-SRF (P amp;lt; 0.001), whereas cupric-reducing antioxidant capacity was lowest (P amp;lt; 0.05) in SP from the first 10 mL of the SRF. Regression analyses indicated that certain SP antioxidants had good predictive value for post-thaw recovery rates of total motility (R-2 = 54.8%, P amp;lt; 0.001; including SOD, TEAC and FRAP) and viability (R-2 = 56.1%, P amp;lt; 0.001; including SOD, PON-1, GPx-5 and TEAC). These results demonstrated that certain SP antioxidants are positively involved in boar sperm cryotolerance, minimizing the oxidative stress imposed by cryogenic handling. (C) 2017 Elsevier Inc. All rights reserved.
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| 23. |
- Martinez, C. A., et al.
(författare)
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Peroxidized mineral oil increases the oxidant status of culture media and inhibits in vitro porcine embryo development
- 2017
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Ingår i: Theriogenology. - : ELSEVIER SCIENCE INC. - 0093-691X .- 1879-3231. ; 103, s. 17-23
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Tidskriftsartikel (refereegranskat)abstract
- The use of oils with undetected alterations is a long-recognized problem for in vitro embryo production systems. Since peroxides in oils have been associated with reduced embryo production outcomes, our goals were (1) to evaluate the effects of a batch of mineral oil (MO) that was suspected to be altered on the in vitro production of pig embryos and (2) to determine oil peroxide values throughout culture and the transfer of oxidant agents from oil to culture media. Sunflower oil, which has a completely different chemical composition than MO but a higher oxidative status, and unaltered MO were used as controls. Oocyte maturation, fertilization and embryo development were affected differently depending on the oil overlay used. While the suspected MO was not able to sustain in vitro maturation and fertilization, the oocytes incubated in the presence of sunflower oil were matured and fertilized similarly to those of the unaltered MO group. Moreover, the cleavage rate of presumed zygotes cultured under the suspected MO was severely reduced compared with those cultured under the other oils, and none of the cleaved embryos developed to the blastocyst stage. Although the cleavage rates in the sunflower oil and unaltered MO groups were similar, embryos cultured under sunflower oil also failed to develop to the blastocyst stage. Our results revealed that the suspected MO and sunflower oil had similar levels of peroxides and that these levels were much higher than those of the unaltered MO. The total oxidant status was higher in media incubated under peroxidized oils than in fresh media or media incubated without an oil overlay or under unaltered MO, indicating that oxidant agents were transferred to the incubation media. However, unlike the sunflower oil group, the culture media incubated under the suspected MO had high levels of total oxidant status and low levels of hydrogen peroxide and reactive oxygen species, suggesting the presence of other unknown oxidant agents in that oil. These results indicate that a peroxidized MO overlay dramatically decreases embryo production outcomes. This decrease could be associated with the higher peroxide values of the oil but cannot be explained by the levels of hydrogen peroxide and reactive oxygen species transferred from the oil to the culture media. It is likely that different oxidant agent(s) and/or other toxic compounds present in the peroxidized MO are responsible for its damaging effects on oocytes and embryos. (C) 2017 Elsevier Inc. All rights reserved.
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| 24. |
- Martinez, C. A., et al.
(författare)
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Porcine blastocyst viability and developmental potential is maintained for 48 h of liquid storage at 25 degrees C without CO2 gassing
- 2019
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Ingår i: Theriogenology. - : ELSEVIER SCIENCE INC. - 0093-691X .- 1879-3231. ; 135, s. 46-55
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Tidskriftsartikel (refereegranskat)abstract
- Short- and medium-term storage of pig embryos has become relevant for commercial application of nonsurgical deep uterine embryo transfer (NsDU-ET) in the light of the strict legal and administrative requirements posed by the International Association for Air Transport (IATA) to allow shipment of liquid nitrogen (LN2) containers and the technical drawbacks when using vitrified embryos. Therefore, this study developed an efficient method for the liquid storage of in vivo-derived porcine blastocysts for a moderate duration (48 h) without controlled CO2 gassing. We evaluated two storage temperatures (25 degrees C and 37 degrees C) and three HEPES-supplemented media: the chemically defined media TL-PVA and NCSU-PVA and the semi-defined medium NCSU-BSA. We observed no differences in survival, hatching rate or final developmental stage between the two temperatures, but storage at 25 degrees C was more efficient to preserve zona pellucida (ZP) integrity. Blastocysts were successfully stored for 24 h in a chemically defined medium. Yet, only 48 h storage in NCSU-BSA medium supported blastocyst development. Although all storage conditions resulted in an embryonic developmental delay, blastocysts stored in NCSU-BSA at either tested temperature could hatch and attain the same final developmental stage as control blastocysts when cultured under standard conditions after storage. Moreover, blastocysts stored at 25 degrees C for 48 h in NCSU-BSA medium could produce pregnancies after surgical transfer. In conclusion, porcine blastocysts maintain their viability and developmental potential after storage in the semi-defined medium NCSU-BSA for at least 48 hat 25 degrees C. (C) 2019 Elsevier Inc. All rights reserved.
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| 25. |
- Martinez, C. A., et al.
(författare)
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Simple storage (CO2-free) of porcine morulae for up to three days maintains the in vitro viability and developmental competence
- 2018
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Ingår i: Theriogenology. - : ELSEVIER SCIENCE INC. - 0093-691X .- 1879-3231. ; 108, s. 229-238
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Tidskriftsartikel (refereegranskat)abstract
- The advancement of porcine embryo transfer (ET) technology is constrained by regulatory hurdles (liquid nitrogen transportation) or, more importantly, the technical obstacles of using vitrified embryos in combination with nonsurgical deep uterine ET technology. Maintaining embryos in culture during transport and prior ET collides with the need of CO2 gassing and the best choice of culture medium. In this work, we describe storage conditions for short-term embryo CO2-free storage that allowed for a majority of in vivo-derived porcine morulae to survive after 3 days of storage in a liquid state, and to develop to the blastocyst stage unhatched, a sanitary prerequisite for ET. The storage conditions included NCSU-23 medium supplemented with bovine serum albumin, where bicarbonate was partially replaced by HEPES to avoid the need for CO2 gassing, and a temperature of 37 degrees C. These conditions were able to maintain the functionality of the stored embryos (hatching capacity after exposure to conventional culture conditions) and their developmental competence after ET (normal fetuses by day 38 of pregnancy). Use of this strategy of CO2-free storage should allow the shipment of fresh embryos worldwide in the absence of liquid nitrogen. (C) 2017 Elsevier Inc. All rights reserved.
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