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Biology of Reproduction Jun 2024In cattle, the endometrium during diestrus and early pregnancy displays cellular responses that are consequences of prior, transient stimuli. Goal was to establish a...
In cattle, the endometrium during diestrus and early pregnancy displays cellular responses that are consequences of prior, transient stimuli. Goal was to establish a model to study cellular memory in the endometrium. The hypothesis is that stimuli given to endometrium in vivo are retained as a cellular memory that remains after bovine uterine epithelial cells (BUECs) are isolated, cultured, and further stimulated in vitro. Objectives were to measure BUEC proliferation/migration and responsiveness to recombinant bovine Interferon-tau (rbIFNT) in vitro: among cows that showed estrus (experiment 1 [Exp1]), cows that became or not pregnant to artificial insemination (Exp2), cows that received or not supplemental progesterone (P4; Exp3) and cows that received or not a COX-1/2 inhibitor (Exp4). Only cows that displayed estrus were included in studies. For all experiments endometrial cytology was collected 4 days after estrus, BUECs were cultured, propagated, and submitted to rbIFNT treatment and an in vitro scratch assay. In Exp1, different cows spontaneously grouped according to proliferative/migratory capacity and responsiveness to rbIFNT of their respective BUECs. In Exp2, BUECs from pregnant cows showed greater rbIFNT responsiveness and cellular proliferation. In Exp3, BUECs from cows supplemented with P4 presented inhibited proliferation and increased expression of RSAD2. In Exp4, Flunixin Meglumine modified rbIFNT responsiveness of BUECs in an IFN-signaling pathway-specific manner. In conclusion, physiological and pharmacological stimuli received by the endometrium in vivo were retained as cellular memory in BUECs, persisted in culture, and changed BUEC proliferation/migration and responsiveness to rbIFNT, which are characteristics associated with fertility in cattle.
PubMed: 38910516
DOI: 10.1093/biolre/ioae093 -
Animal Reproduction Science Jun 2024The objective of this study was to evaluate the impact of early progesterone removal on pregnancy rates to fixed-time artificial insemination (FTAI) in presynchronized...
The objective of this study was to evaluate the impact of early progesterone removal on pregnancy rates to fixed-time artificial insemination (FTAI) in presynchronized beef cows. Postpartum beef cows (n = 882) were randomly assigned to 1 of 2 treatments: 1) 7&7 Synch: cows received a controlled internal drug release insert (CIDR) and a 25-mg injection of prostaglandin F on day 0, 100 μg of GnRH on day 7, a second injection of prostaglandin F (PG2) at CIDR removal on day 14, and a second injection of GnRH at FTAI 60-66 h after PG2 (day 17); 2) 7&6 Synch: cows received the same treatment as 7&7 Synch; however, CIDR removal occurred in conjunction with PG2 on day 13, while FTAI remained at 60-66 h after CIDR removal (day 16). Ovarian ultrasonography was performed to determine follicle diameter at PG2 and FTAI in a subset of cows (n = 40). Cows exposed to the 7&7 Synch tended to have larger follicle diameter at PG2 compared with 7&6 Synch cows (P = 0.09); however, there were no differences in follicle diameter at FTAI. No differences were determined between treatments for the expression of estrus (7&7 Synch: 61.6 ± 5.30; 7&6 Synch: 54.1 ± 5.45; P = 0.31) or pregnancy rates to FTAI (7&7 Synch: 60.8 ± 3.83; 7&6 Synch: 57.0 ± 3.84; P = 0.42). In conclusion, early removal of progesterone did not impact pregnancy rates in presynchronized beef cows.
PubMed: 38909485
DOI: 10.1016/j.anireprosci.2024.107541 -
Theriogenology Jun 2024Single Layer Centrifugation (SLC) through a low density colloid offers an alternative solution to antibiotic use in boar semen extenders, with lower costs compared to...
Single Layer Centrifugation (SLC) through a low density colloid offers an alternative solution to antibiotic use in boar semen extenders, with lower costs compared to high density colloids. The aim of this study was to explore the reproductive performance of sows when using SLC-prepared semen doses without antibiotics, employing low density Porcicoll to prepare semen doses for artificial insemination in a commercial swine herd in Thailand. Ejaculates were divided into two equal parts to create insemination doses, with each dose containing 3000 × 10 sperm/80 ml for intra-uterine insemination in individual sows. The sows were inseminated twice, with the interval between the two inseminations ranging from 8 to 16 h. The CONTROL group consisted of 206 semen doses treated with antibiotics, prepared for insemination in 103 sows, while the SLC group comprised 194 SLC-prepared semen doses without antibiotics for inseminating 97 sows. Fertility and fecundity traits, including non-return rate, conception rate, farrowing rate, and litter traits (i.e., the total number of piglets born per litter, number of piglets born alive per litter, number of stillborn piglets, and number of mummified fetuses), were compared between groups. Furthermore, data on piglet characteristics, including live-born and stillborn piglets (i.e., the prevalence of stillbirth (yes, no), birth weight, crown-rump length, body mass index (BMI), and ponderal index (PI)), were determined. No significant differences in non-return rate (75.7 % vs. 77.3 %), conception rate (73.8 % vs. 73.2 %), and farrowing rate (71.8 % vs. 73.2 %) were observed between the CONTROL and SLC groups, respectively (P > 0.05). Nevertheless, the total number of piglets born per litter in the SLC group was higher than in the CONTROL group (14.6 ± 0.9 vs. 12.3 ± 0.6, respectively, P = 0.049). Interestingly, the prevalence of stillbirth in the SLC group was lower than in the CONTROL group (6.2 % vs. 11.6 %, respectively, P < 0.001). Moreover, the newborn piglets in the SLC group exhibited higher birth weight and BMI compared to those in the CONTROL group (1.36 ± 0.03 vs. 1.26 ± 0.02 kg, P = 0.005, and 18.3 ± 0.3 vs. 17.3 ± 0.2 kg/m, P = 0.003). In conclusion, employing sperm doses after SLC through a low density colloid in artificial insemination within a commercial breeding operation did not have a detrimental impact on either fertility or fecundity traits but showed potential benefits in increasing the total number of piglets born per litter. Moreover, improvements were observed in the birth weight and body indexes of piglets, and the percentage of stillbirths was reduced. Our findings introduce new possibilities for antibiotic alternatives in semen extenders to reduce the risk of antimicrobial resistance in the swine industry. Additionally, they provide compelling reproductive outcomes supporting the integration of SLC-prepared semen doses into artificial insemination practices.
PubMed: 38909434
DOI: 10.1016/j.theriogenology.2024.06.016 -
Animal : An International Journal of... May 2024Small ruminant farming is of socio-economic and environmental importance to many rural communities around the world. The SMARTER H2020 project aims to redefine genetic...
Small ruminant farming is of socio-economic and environmental importance to many rural communities around the world. The SMARTER H2020 project aims to redefine genetic selection criteria to increase the sustainability of the sector. The objective of this study was to analyse the selection and breeding management practices of small ruminant producers and breeders, linked with socio-technical elements that shape them. The study is based on farm surveys using semi-structured interviews conducted in five countries (France, Spain, Italy, Greece, and Uruguay) across 272 producers and breeders of 13 sheep and goat breeds, and 15 breed × system combinations. The information was collected in four sections. The first and second sections dealt with general elements of structure and management of the system and the flock/herd. The third section focused on selection and breeding management practices: criteria for culling and replacement of females, selection criteria for males, use of estimated breeding values and global indexes, and preferences for indexing new traits to increase the sustainability of their system. The fourth section aimed to collect socio-technical information. We used a data abstraction method to standardise the representation of these data. A mixed data factor analysis followed by a hierarchical ascending classification allowed the characterisation of three profiles of selection and breeding management: (1) a profile of producers (n = 93) of small flocks/herds, with little knowledge or use of genetic selection and improvement tools (selection index, artificial insemination, performance recording); these farmers do not feel that new traits are needed to improve the sustainability of their system. (2) a profile of producers (n = 34) of multibreed flocks/herds that rely significantly on grazing; they are familiar with genetic tools, they currently use AI; they would like the indexes to include more health and robustness characteristics, to make their animals more resistant and to increase the sustainability of their system. And (3) a profile of producers or breeders (n = 145) of large flocks/herds, with specific culling criteria; these farmers are satisfied with the current indexes to maintain the sustainability of their system. These results are elements that can be used by private breeding companies and associations to support the evolution of selection objectives to increase the resilience of animals and to improve the sustainability of the small ruminant breeding systems.
PubMed: 38905776
DOI: 10.1016/j.animal.2024.101208 -
Frontiers in Microbiology 2024The concept of a sterile uterus was challenged by recent studies that have described the microbiome of the virgin and pregnant uterus for species including humans and...
INTRODUCTION
The concept of a sterile uterus was challenged by recent studies that have described the microbiome of the virgin and pregnant uterus for species including humans and cattle. We designed two studies that tested whether the microbiome is introduced into the uterus when the virgin heifer is first inseminated and whether the origin of the microbiome is the vagina/cervix.
METHODS
The uterine microbiome was measured immediately before and after an artificial insemination (AI; Study 1; = 7 AI and = 6 control) and 14 d after insemination (Study 2; = 12 AI and = 12 control) in AI and non-AI (control) Holstein heifers. A third study (Study 3; = 5 Holstein heifers) that included additional negative controls was subsequently conducted to support the presence of a unique microbiome within the uterus despite the low microbial biomass and regardless of insemination. Traditional bacteriological culture was performed in addition to 16S rRNA gene sequencing on the same samples to determine whether there were viable organisms in addition to those detected based on DNA sequencing (16S rRNA gene sequence).
RESULTS AND DISCUSSION
Inseminating a heifer did not lead to a large change in the microbiome when assessed by traditional methods of bacterial culture or metataxonomic (16S rRNA gene) sequencing (results of Studies 1 and 2). Very few bacteria were cultured from the body or horn of the uterus regardless of whether an AI was or was not (negative control) performed. The cultured bacterial genera (e.g., , and ) were typical of those found in the soil, environment, skin, mucous membranes, and urogenital tract of animals. Metataxonomic sequencing of 16S rRNA gene generated a large number of amplicon sequence variants (ASV), but these larger datasets that were based on DNA sequencing did not consistently demonstrate an effect of AI on the abundance of ASVs across all uterine locations compared with the external surface of the tract (e.g., perimetrium; positive control samples for environment contamination during slaughter and collection). Major genera identified by 16S rRNA gene sequencing overlapped with those identified with bacterial culture and included , and .
PubMed: 38903779
DOI: 10.3389/fmicb.2024.1385505 -
Animal Reproduction Science Jun 2024The commercial swine industry utilizes artificial insemination (AI) in their breeding programs. With this assisted reproductive technology, the process starts by... (Review)
Review
The commercial swine industry utilizes artificial insemination (AI) in their breeding programs. With this assisted reproductive technology, the process starts by obtaining fresh ejaculates from desirable boars who are housed in a dedicated facility (i.e., stud) that also contains a clean-room laboratory where semen quality is assessed and then ejaculates processed into AI doses. In concert with AI adoption, disruptions in sow herd reproductive performance have been traced back to contributions made from the boar stud. Through field investigations and research, several extrinsic contaminants have been identified that impact semen quality either at the boar or AI-dose level. These contaminants can be categorized as either biological or chemical in origin, eliciting reprotoxic outcomes at the boar level and/or spermatotoxicity at the AI-dose level. Biological contaminants include multiple genera of primarily opportunistic microbes (i.e., bacteria, fungi), along with their secondary metabolites (e.g., endotoxins, exotoxins, mycotoxins). Chemical contaminants appear to originate from products used at the stud, and include cleaning agent/disinfectant residues, leachates from gloves and plastics, semen extender impurities, purified and drinking water impurities, and pesticides (i.e., biocides, fungicides, herbicides, insecticides, wood preservatives). In conclusion, contaminants are a real and constant threat to the health and productivity of a stud, and have caused significant reproductive and economic losses in the swine industry. The knowledge gained in recognizing the types and sources of contaminants provides a solid foundation for the development and implementation of pro-active strategies that mitigate risk to the industry.
PubMed: 38897823
DOI: 10.1016/j.anireprosci.2024.107519 -
Journal of Reproductive Immunology Jun 2024Many endocrine or non-endocrine factors are involved in sperm production. Although reproductive hormones are very important for the initiation and maintenance of...
Many endocrine or non-endocrine factors are involved in sperm production. Although reproductive hormones are very important for the initiation and maintenance of spermatogenesis, other factors, such as inflammation and oxidative stress, affect spermatogenesis. The aim of this study is to evaluate the relationships between sperm parameters and hormones, oxidative stress, and inflammation status. We conducted this study on 40 rats. Sperm parameters (motility, abnormal sperm rate, and dead sperm rate), oxidative stress (malondialdehyde, glutathione, glutathione peroxidase, and catalase), inflammation (NF-κβ, TNF-α, IL-1β, IL-6, and IL-10), and hormone parameters (follicle-stimulating hormone, luteinizing hormone, testosterone, melatonin, and corticosterone) were determined. Relationships between mentioned parameters were investigated by canonical correlation analysis. Canonical correlation coefficients for these data sets (sperm-oxidative stress, sperm-inflammation, and sperm-hormone parameters) were found to be strongly significant (r= 0.875, p<0.001; r= 0.868, p<0.001; r= 0.886, p<0.001, respectively). The rate of explanation of oxidative stress, inflammation parameters and hormones by sperm parameters was 61.80 %, 56.10 % and 63.90 %, respectively. Canonical correlation analysis results have revealed that dead sperm rate is mostly related to nuclear factor-kappa beta (NF-κβ), catalase, and corticosterone. CCA, which has taken into account the multiple relationships, has revealed that multidimensional evaluation of data sets can provide important and innovative information to researchers for the assessment of relationships between sperm, oxidative stress, inflammation, and hormone parameters.
PubMed: 38896933
DOI: 10.1016/j.jri.2024.104276 -
Veterinary Medicine and Science Jul 2024Anti-Müllerian hormone (AMH) holds potential as a biomarker for assessing the superovulation (SO) response in cattle. Nonetheless, there exists scant information...
BACKGROUND
Anti-Müllerian hormone (AMH) holds potential as a biomarker for assessing the superovulation (SO) response in cattle. Nonetheless, there exists scant information regarding this aspect in the literature concerning dairy heifers. Given this gap, our objective is to explore the viability of AMH as an indicator for gauging the SO response specifically in Holstein heifers. Furthermore, our aim encompasses examining the variations in AMH levels within the same individuals before and after undergoing SO.
METHODS
The study included 41 Holstein heifers. All heifers were superovulated and blood samples were taken both before and after the SO protocol.
RESULTS
The findings revealed that the mean values of serum AMH concentrations before and after SO were 0.122 ng/mL (0.093-0.248 ng/mL) and 0.119 ng/mL (0.084-0.170 ng/mL), respectively. AMH concentrations in heifers were stratified into low (<0.106 ng/mL), medium (0.107-0.126 ng/mL) and high (>0.127 ng/mL) categories both before and after SO.
CONCLUSIONS
There was no significant correlation between AMH levels in the heifers both before and after SO treatment with the number of follicles, corpora lutea, total embryos collected or embryos transferred (p > 0.05). Furthermore, this study showed that serum AMH concentrations in Holstein heifers did not change after SO treatment. In this study, as AMH levels in Holstein heifers were in a narrow range, a relationship between AMH and SO response could not be determined. In future studies, we believe that it would be more useful to plan more studies in Holstein donor heifers, taking into account the number of animals and AMH levels.
Topics: Animals; Cattle; Anti-Mullerian Hormone; Superovulation; Female; Biomarkers
PubMed: 38895914
DOI: 10.1002/vms3.1507 -
Reproduction in Domestic Animals =... Jun 2024This study evaluated the effect of bovine somatotropin (bST) on pregnancy rate (PR) and size of the dominant follicle (DF) on the day of intravaginal progesterone (P4)...
This study evaluated the effect of bovine somatotropin (bST) on pregnancy rate (PR) and size of the dominant follicle (DF) on the day of intravaginal progesterone (P4) removal in protocols for fixed-time artificial insemination (FTAI). Bos indicus (Nellore) females (n = 392) were distributed into three groups. The control group (CG; n = 92) received an intravaginal P4 device + estradiol benzoate on day (d)0; prostaglandin F2α on d7 (first application); removal of P4 + estradiol cypionate (EC) + PGF2α (second application) + ultrasound (US) of the DF on d9; the FTAI was performed on d11; and pregnancy diagnosis (PD) was performed on d45. The bST group (bSTG; n = 142) underwent the same protocol as the CG, except that the animals received 125 mg of bST on d7. The equine chorionic gonadotropin (eCG) group (eCGG; n = 158) underwent the same protocol as the CG, except that the animals received 300 IU of eCG on d9. The PRs of the bSTG, eCGG, and CG were 48%, 48%, and 35%, respectively (p < .05); the bSTG and eCGG showed greater PRs, with follicles 6-7.9 mm (p < .05) and 8-8.9 mm in diameter, respectively. The bSTG exhibited a greater dimension of the DF on d9 of the protocol (p < .05). The eCGG had higher PRs with a body condition score (BCS) of 2.5, and the bSTG had a BCS of 3.0 (p < .05). It was concluded that bST increased PR, bST showed better performance in smaller DF and larger follicular diameter on d9 of the protocol, eCG acted better on animals with lower BCSs, and bST can be used in FTAI.
Topics: Animals; Female; Insemination, Artificial; Pregnancy; Cattle; Growth Hormone; Progesterone; Pregnancy Rate; Estradiol; Ovarian Follicle; Dinoprost; Estrus Synchronization; Administration, Intravaginal
PubMed: 38894646
DOI: 10.1111/rda.14642 -
Animals : An Open Access Journal From... May 2024Circulating microRNAs (miRNAs) were investigated as biomarkers for the diagnosis of early pregnancy in cattle. The levels of prospective miRNA biomarkers and the...
Circulating microRNAs (miRNAs) were investigated as biomarkers for the diagnosis of early pregnancy in cattle. The levels of prospective miRNA biomarkers and the features of extracellular vesicles (EVs) in the blood were evaluated. In Study 1, plasma samples from cows 21 days after artificial insemination (AI) were examined using RT-qPCR to determine the levels of seven circulating miRNAs. Only the levels of miR-126-3p were significantly lower in the pregnant group than in the non-pregnant group. In Study 2, among individuals not pregnant at the first AI, the miRNA levels were compared between the individuals pregnant at the second AI and those who remained non-pregnant. The miR-25 levels were significantly higher in the pregnant group at the second AI than in the pregnant group at the first AI; miR-19b, miR-27b, and miR-29a levels were also high. In the non-pregnant group, changes were absent in the miRNA levels in the same individual between the first and second AIs. In Study 3, Western blotting and RT-qPCR showed the presence of miRNAs in EVs and their levels were lower than in plasma. Thus, circulating miR-126-3p may serve as a biomarker for the diagnosis of early pregnancy in cattle. In addition, the expression of some miRNAs tended to be higher during pregnancy than during non-pregnancy in the same individual, suggesting their potential as an index to determine pregnancy and non-pregnancy rates using a comparative method.
PubMed: 38891639
DOI: 10.3390/ani14111592