Reproductive Responses Research Articles

Energy efficiency of grazing Hereford heifers classified by paternal residual feed intake

Abstract Residual feed intake (RFI) has become a widely spread index of feed efficiency. Although most of beef cattle systems in the world are pasture based, RFI evaluation and research is usually performed in confinement conditions. In this context, residual heat production (RHP) estimated as the difference between actual and expected heat production (HP), could allow to identify efficient animals. Thus, the aim of this work was to evaluate the relationship between paternal estimated breeding values (EBV) for RFI and beef heifer efficiency, measured as RHP, as well as its association with heifers’ productive and reproductive performance on grazing conditions. Seventy-one 25 ± 0.8-mo-old and seventy-four 24 ± 0.7-mo-old Hereford heifers were managed as contemporary groups in spring 2019 and 2020, respectively. Heifers were sired by 10 RFI-evaluated bulls and classified into three groups according to the paternal EBV for RFI: five bulls of low RFI (high efficiency, pHE), two bulls of medium RFI (medium efficiency), and three bulls of high RFI (low efficiency, pLE). The experimental period lasted 70 d prior to their first insemination where HP was determined by the heart rate-O2 pulse technique. In addition, reproductive performances during the first and second breeding and calving seasons were recorded. Heifers’ RHPs expressed as MJ/d and kJ/kg of body weight (BW)0.75/d were positively correlated with paternal RFI EBVs (P < 0.05; r > 0.60). Moreover, BW and average daily gain (ADG) were greater (P < 0.01) for pHE than pLE heifers while expressed as units of BW0.75/d, neither total HP nor metabolizable energy (ME) intake differed between groups, but pHE heifers had greater retained energy (RE; P < 0.01) and lower RHP (P < 0.05) than pLE ones. Gross energy efficiency (RE/ME intake) was greater (P < 0.001) for pHE than pLE heifers while the HP/ADG and RHP/ADG were reduced (P < 0.05) and feed-to-gain ratio (ADG/DM intake) tended to be greater (P = 0.07) for pHE than pLE heifers. In addition, during the first breeding and calving seasons, small but significant (P < 0.01) differences in reproductive responses between groups suggested an earlier pregnancy in pHE heifers than the pLE group, differences that disappeared during the second breeding and calving seasons. Thus, heifers sired by high-efficiency bulls measured as RFI were more efficient measured as RHP in grazing conditions, without significant differences in reproductive performance.

Follicular, luteal dynamics and effect of synchronization methods on reproductive responses in Cholistani cows.

The Cholistani cow is considered one of the milch breeds of Zebu cattle. Despite being heat and tick-resistant, it has not yet gained much popularity among Pakistan dairy farmers due to its long calving interval. This study aimed to understand the basic reproductive physiology of Cholistani cows using sonography and reproductive biotechnologies such as estrus synchronization and timed artificial insemination to improve reproductive efficiency. In experiment 1, six Cholistani cows with mixed parity 3.20 ± 1.30 and weighing 400 kg were selected to monitor ovarian dynamics on alternate days by the same sonographer from the onset of heat through ovulation until the next ovulation. Experiment 2 measured the effect of estrus synchronization methods, controlled internal drug release-gonadotropin-releasing hormone (CIDR-GnRH, n = 31) and Ovsynch (OVS, n = 32) on various reproductive parameters. The mean estrous cycle length was 19.81 ± 0.56 days with two follicular waves. The mean inter-ovulatory interval was 20.80 ± 0.52 days, with a preovulatory follicular size of 13.83 ± 2.37 mm. Estrus response was higher (p > 0.05) in controlled internal drug release (CIDR)-GnRH (93.54%) than in OVS (84.37%) cows. Similarly, ovulation and conception rates were higher in CIDR-GnRH (91.66% vs. 68.42%) than in OVS cows (41.37% vs. 33.33%), respectively. In conclusion, CIDR-GnRH results in a better estrus response, higher ovulation rate, and subsequently greater conception rate than OVS in Bos indicus dairy cows.

Role of chemokines in regulating luteal and uterine functions in pregnant cows.

Pregnancy is intricately regulated by the interactions between various bioactive substances secreted by the conceptus, uterus, and corpus luteum (CL). Interferon-τ, synthesized and secreted by the conceptus, plays a central role in the interaction mechanism of maternal recognition in cows. Chemokines, chemotaxis mediators that are primarily secreted by immune cells, regulate various reproductive responses in various species. Although there are scattered reports on the potential roles of chemokines in the bovine CL and the uterus during the estrous cycle, there is little information on chemokines in these organs during pregnancy. Therefore, in this review, we discuss the possible physiological roles of chemokines in the CL and uterus of pregnant cows, focusing on our recent findings on chemokines and changes in their receptor expression in the CL and endometrium of cows at some stages of pregnancy.

Determination of reproductive response to different synchronization methods and profitability during the transition to anoestrus in multiparous Kangal sheep

Determination of reproductive response to different synchronization methods and profitability during the transition to anoestrus in multiparous Kangal sheep

Settling, Oviposition and Reproduction Response of Peach Fruit Fly, Bactrocera zonata (Saunders) to the Plant Extracts of Native Species of Kalat Division in Balochistan

Settling, Oviposition and Reproduction Response of Peach Fruit Fly, Bactrocera zonata (Saunders) to the Plant Extracts of Native Species of Kalat Division in Balochistan

Combining two injectable progesterone formulas for estrous synchronization in ewes.

The study aimed to evaluate the effectiveness of combining two injectable progesterone (iP4) formulas for estrous synchronization in ewes and to compare it with traditional intravaginal progesterone devices. Additionally, the study assessed whether the inclusion of GnRH enhances the reproductive outcomes of the iP4 treatment. Two experiments were conducted. In the first experiment, 20 Santa Inês ewes were divided into two groups: one group received intravaginal progesterone devices, and the other received combined long-acting and short-acting iP4. In the second experiment, 30 Corriedale ewes were divided into two groups: one received the combined iP4 with GnRH, and the other without GnRH. Estrous, ovulation, follicular populations, and progesterone concentrations were monitored. The combined iP4 treatment induced an artificial luteal phase and produced reproductive responses similar to those obtained with intravaginal devices. In the first experiment, the iP4 treatment tended to result in more synchronized ovulation compared to the control (P=0.095). In the second experiment, adding GnRH enhanced the quality of the corpus luteum, as indicated by increased diameter and vascularization on Day 23 (P=0.047 and P=0.02, respectively). The combined administration of long-acting and short-acting iP4 effectively synchronized estrous in ewes and showed similar efficacy to traditional intravaginal devices. The inclusion of GnRH improved luteal quality, suggesting potential benefits for reproductive management in ewes. Further studies are needed to evaluate the fertility outcomes of these protocols under field conditions.

Parasitoid cues modulate Drosophila germline development and stem cell proliferation

Environmental factors influence an organism's reproductive ability by regulating germline development and physiology. While the reproductive adaptations in response to extrinsic stress cues offer fitness and survival advantages to individuals, the mechanistic understanding of these modifications remains unclear. Here, we find that parasitoid wasps' stress signaling regulates Drosophila melanogaster oogenesis. We show that fruit flies dwelling in the wasp-infested area elevate their fecundity, and the observed reproductive response is specific to Pachycrepoideus sp., a pupal parasitoid wasp. Pachycrepoideus-specific olfactory and visual cues recruit the signaling pathways that promote germline stem cell proliferation and accelerate follicle development, increasing egg production in Drosophila females. Downregulation of signaling engaged in oocyte development by shifting flies to a non-wasp-infested environment increases apoptosis of the developing follicles. Thus, this study establishes host germline responsiveness to parasitoid-specific signals and supports a predator strategy to increase hosts for infection.

Metabolomic-based analysis reveals bile acid-mediated ovarian failure induced by low temperature in zebrafish.

As ectotherms, fish are highly sensitive to temperature fluctuations, which can profoundly impact their reproductive cycles. In this study, we investigated the fertility and histological characteristics of zebrafish ( Danio rerio) ovaries exposed to a temperature gradient ranging from the thermopreferendum temperature of the species, 27°C, to lower temperatures of 22°C, 20°C, and 13°C over a period of two weeks. Comparative metabolomic (six biological replicates for each temperature) and transcriptomic (four biological replicates for each temperature) analyses were conducted under the four temperature conditions. Results indicated that lower temperatures inhibited oocyte development and differential metabolites were involved in steroid hormone production, antioxidant function, and lipid and protein catabolism. Disrupted reproductive hormones, increased proteolysis, and lipid degradation significantly impeded oocyte development and egg maturation. Notably, a significant increase in bile acid content was noted in the ovaries of the cold-treated fish, indicating that bile acids play a critical role in ovarian failure. Overall, these findings provide valuable insights into the mechanisms governing the reproductive response of fish to cold stress.

Role of Electromagnetic Activity in Cellular Mechanics

The cell is the building block of life since it is the fundamental unit of all living things. Organelles, the cytoplasm, the nucleus, and other structures all have specific roles in cells. Genetic makeup, which controls cell growth, reproduction, and environmental response, determines the structure and function of cells. It is commonly known that cells communicate chemically and electrically. Cellular interactions via other physical fields are exactly the opposite. The electromagnetic field (EMF) is the most likely option for a different type of cellular interaction. This review delves into the role of electromagnetic fields in cells and their consequential impact on cell behavior. This gives a broad perspective on the basic construction and operation of cells, presents the notion of cellular mechanics, and deepens the understanding of the pivotal function of microtubules and centrosomes in cellular division. It also explores their interplay with the electromagnetic field. The discussion extends to how EMF influences ion channels on the cell membrane, a critical component in cellular communication, and the electrical properties of neurons. This review concludes by identifying the current limitations and gaps in EMF research in cells, such as constraints in experimental methods and technology, and the need for more comprehensive theories and models to fully understand these complex processes.

Maternal or Paternal Antibiotics? Intergenerational Transmission and Reproductive Toxicity in Zebrafish.

Despite the known direct toxicity of various antibiotics to aquatic organisms, the potential chronic impact through intergenerational transmission on reproduction remains elusive. Here, we exposed zebrafish to a mixture of 15 commonly consumed antibiotics at environmentally relevant concentrations (1 and 100 μg L-1) with a cross-mating design. A high accumulation of antibiotics was detected in the ovary (up to 904.58 ng g-1) and testis (up to 1704.49 ng g-1) of F0 fish. The transmission of antibiotics from the F0 generation to the subsequent generation (F1 offspring) was confirmed with a transmission rate (ki) ranging from 0.11 to 2.32. The maternal transfer of antibiotics was significantly higher, relative to paternal transfer, due to a greater role of transmission through ovarian enrichment and oviposition compared to testis enrichment. There were similar impairments in reproductive and developmental indexes on F1 eggs found following both female and male parental exposure. Almost all antibiotics were eliminated in F2 eggs in comparison to F1 eggs. However, there were still reproductive and developmental toxic responses observed in F2 fish, suggesting that antibiotic concentration levels were not the only criterion for evaluating the toxic effects for each generation. These findings unveil the intergenerational transmission mechanism of antibiotics in fish models and underscore their potential and lasting impact in aquatic environments.

Study on epigenotoxicity, sex hormone synthesis, and DNA damage of benzo[a]pyrene in the testis of male Ruditapes philippinarum

Research on the mechanisms of reproductive toxicity caused by persistent organic pollutants (POPs) in marine animals has received significant attention. One group of typical POPs, called polycyclic aromatic hydrocarbons (PAHs), has been found to cause various reproductive toxicities in aquatic organisms, including epigenotoxicity, reproductive endocrine disruption, DNA damage effects and other reproductive toxicity, thereby affecting gonadal development. Interestingly, male aquatic animals are more susceptible to the disturbance and toxicity of environmental pollutants. However, current studies primarily focus on vertebrates, leaving a large gap in our understanding of the reproductive toxicity and mechanisms of PAHs interference in marine invertebrates. In this study, male Ruditapes philippinarum was used as an experimental subject to investigate reproduction-related indexes in clams under the stress of benzo[a]pyrene (B[a]P) at different concentrations (0, 0.8, 4 and 20 μg/L) during the proliferative, growth, maturity, and spawning period. We analyzed the molecular mechanisms of reproductive toxicity caused by PAHs in marine bivalves, specifically epigenotoxicity, reproductive endocrine disruption, and gonadal damage-apoptotic effect. The results suggest that DNA methylation plays a crucial role in mediating B[a]P-induced reproductive toxicity in male R. philippinarum. B[a]P may affect sex hormone levels, impede spermatogenesis and testis development in clams, by inhibiting the steroid hormone synthesis pathway and downregulating genes critical for cell proliferation, testis development, and spermatid expulsion. Moreover, the spermatids of male R. philippinarum were severely impaired under the B[a]P stress, leading to reduced reproductive performance in the clams. These findings contribute to a better understanding of the reproductive toxicity response of male marine invertebrates to POPs stress.

Effect of Short-Term Glycerin Supplementation on Follicle Dynamics and Pregnancy Rate in Goats

We investigated the effects of short-term glycerin supplementation on follicular dynamics and pregnancy rates. Twenty-five goats with synchronized estrus and follicular waves with three injections of a prostaglandin analog every 7 days were used. Two days after the second injection, 13 goats were randomly chosen to receive an oral drench of 200 mL of glycerin (glycerin group [GG], n = 13) for 6 days, whereas the remaining 12 animals received an oral drench of saline (control group [CG], n = 12). At 24 and 48 h after the third injection, the goats mated. The animals were kept in a collective stall and received the same diet. The GG had higher blood glucose levels during the supplementation period than the CG (76.4 ± 1.9 vs. 50.3 ± 0.7 mg/dL; p < 0.01). The glycemic peak was recorded 4 h after the glycerin administration (102.3 ± 5.1 mg/dL) and remained higher than that in the CG 8 and 12 h later. The GG goats had a higher rectal temperature, heart rate, and respiratory rate than the CG goats and showed an increase in these parameters 4, 8, and 12 h after glycerin drenching. The GG animals also exhibited increased stress, urination, and drinking behaviors and reduced rumination. The ultrasonographic analysis showed a higher number of follicles with a diameter >4 mm (p < 0.05) and a greater follicular diameter (p < 0.01) in the waves before and after ovulation induction. The pregnancy and twinning rates and litter size at parturition were not different between the groups. Short-term supplementation with glycerin positively affects ovarian stimulation but has no effect on the reproductive response after mating.

Maternal effects of human-mediated disturbances on the growth and reproductive traits of different ecotypes of an annual invasive plant Erigeron annuus

A multifactorial combination of human-mediated disturbances is now recognized as a major driver of the global rise and spread of invasive alien plants. Among the numerous invasive alien plants, annuals have fast adaptability and evolution via maternal effect in human-mediated disturbance environments, such as chemical and physical controls. The characteristic enables them to quickly disperse and establish new habitats. Thus, understanding the adaptability and evolution of invasive plants in response to multifactorial human-mediated disturbances is crucial for developing effective management strategies to control their spread and minimize their impacts on ecosystems and human activities. In order to test the growth and reproductive responses in both the parent and offspring generations, two ecotypes (clump and scatter) of an annual invasive plant, Erigeron annuus, were subjected to simulated trampling, defoliation (simulated herbivory), and herbicide disturbances. We found that disturbances negatively affected the growth and reproduction of E. annuus in parent and offspring generations. In comparison to the scatter ecotype plants, the clump ecotype plants had greater growth and reproductive production, as well as a reduced maternal interference effect to cope with human-mediated disturbances. It enhanced the adaptability of their offspring to disturbances. The findings imply that the evolutionary advantage of clump ecotype plants may explain why they can thrive in disturbed habitats and become invasive in some regions.

Beta-Carotene Supplementation for Sows: Reproductive Parameters and Productive Performance of Piglets

The rapid fetal development and the increased demand for milk result in a catabolic state and oxidative stress in hyperprolific sows. Despite animal defense mechanisms, the dietary supplementation of antioxidants is being evaluated to reduce the impacts of excess free radicals. The aim of this study was to evaluate the short- and long-term effects of beta-carotene supplementation for sows on the reproductive response and performance of suckling piglets. A total of 120 sows were distributed in a 3 × 4 factorial arrangement of three supplementation levels (B0–no supplementation, B200–200 mg beta-carotene/day and B400–400 mg beta-carotene/day) and four groups of parity order (1st, 2nd, 3rd, above 4th). Beta-carotene supplementation during lactation resulted in a higher litter weight at weaning. A higher average weight of piglets and litter weight at birth were observed, as well as a greater number of piglets weaned and litter weight at weaning in sows supplemented with 400 mg of beta-carotene during gestation and lactation periods. Supplementation with 200 and 400 mg of beta-carotene resulted in a greater weight at weaning and daily weight gain in piglets. Daily supplementation with 400 mg of beta-carotene in the prepartum and lactation phases provides a greater litter weight at weaning and, when supplemented in the pre-gestation and gestation periods, results in a greater litter weight at birth and at weaning.

CDC Division of Reproductive Health's Emergency Preparedness Resources and Activities for Radiation Emergencies: Public Health Considerations for Women's Reproductive Health.

Pregnant, postpartum, and lactating people, and infants have unique needs during public health emergencies, including nuclear and radiological incidents. This report provides information on the CDC Division of Reproductive Health's emergency preparedness and response activities to address the needs of women of reproductive age (aged 15-49 years), people who are pregnant, postpartum, or lactating, and infants during a radiation emergency. Highlighted preparedness activities include: (1) development of a quick reference guide to inform key questions about pregnant, postpartum, and lactating people, and infants during radiation emergencies; and (2) exercising the role of reproductive health experts during nuclear and radiological incident preparedness activities.

Survival, Growth, and Reproduction Responses in a Three-Generation Exposure of the Zebrafish (Danio rerio) to Perfluorooctane Sulfonate.

A prior multigenerational perfluorooctane sulfonic acid (PFOS) exposure investigation in zebrafish reported adverse effects at 0.734 µg/L, among the lowest aquatic effect levels for PFOS reported to date. The present three-generation PFOS exposure quantified survival, growth, reproduction, and vitellogenin (VTG; egg yolk protein) responses in zebrafish, incorporating experimental design and procedural improvements relative to the earlier study. Exposures targeting 0.1, 0.6, 3.2, 20, and 100 µg/L in parental (P) and first filial (F1) generations lasted for 180 days post fertilization (dpf) and the second filial generation (F2) through 16 dpf. Survival decreased significantly in P and F2 generation exposures, but not in F1, at the highest PFOS treatment (100 µg/L nominal, 94-205 µg/L, measured). Significant adverse effects on body weight and length were infrequent, of low magnitude, and occurred predominantly at the highest exposure treatment. Finally, PFOS had no significant effects on P or F1 egg production and survival or whole-body VTG levels in P or F1 male fish. Overall, the predominance and magnitude of adverse PFOS effects at <1 µg/L reported in prior research were largely nonrepeatable in the present study. In contrast, the present study indicated a threshold for ecologically relevant adverse effects in zebrafish at 117 µg/L (SE 8 µg/L, n = 10) for survival and 47 µg/L (SE 11 µg/L, n = 19) for all statistically significant negative effects observed. Environ Toxicol Chem 2024;43:115-131. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Two distinct mechanisms lead to either oocyte or spermatocyte decrease in C. elegans after whole developmental exposure to γ-rays.

Wildlife is subject to various sources of pollution, including ionizing radiation. Adverse effects can impact the survival, growth, or reproduction of organisms, later affecting population dynamics. In invertebrates, reproduction, which directly impacts population dynamics, has been found to be the most radiosensitive endpoint. Understanding the underlying molecular pathways inducing this reproduction decrease can help to comprehend species-specific differences in radiosensitivity. From our previous studies, we found that decrease in reproduction is life stage dependent in the roundworm Caenorhabditis elegans, possibly resulting from an accumulation of damages during germ cell development and gamete differentiation. To go further, we used the same experimental design to assess more precisely the molecular determinants of reproductive toxicity, primarily decreases in gamete number. As before, worms were chronically exposed to 50 mGy·h-1 external gamma ionizing radiation throughout different developmental periods (namely embryogenesis, gametogenesis, and full development). To enable cross species extrapolation, conserved molecular pathways across invertebrates and vertebrates were analysed: apoptosis and MAP kinase Ras/ERK (MPK-1), both involved in reproduction and stress responses. Our results showed that these pathways are life-stage dependent, resulting from an accumulation of damages upon chronic exposure to IR throughout the life development. The Ras/ERK pathway was activated in our conditions in the pachytene region of the gonad where it regulates cell fate including apoptosis, but not in the ovulation zone, where it controls oocyte maturation and ovulation. Additionally, assessment of germ cell proliferation via Ras/ERK pathway showed no effect. Finally, a functional analysis of apoptosis revealed that while the decrease of the ovulation rate is caused by DNA-damaged induced apoptosis, this process does not occur in spermatocytes. Thus, sperm decrease seems to be mediated via another mechanism, probably a decrease in germ cell proliferation speed that needs further investigation to better characterize sex-specific responses to IR exposure. These results are of main importance to describe radio-induced reprotoxic effects and contribute as weight of evidence for the AOP #396 "Deposition of ionizing energy leads to population decline via impaired meiosis".

Profiling the Physiological Roles in Fish Primary Cell Culture.

Fish primary cell culture has emerged as a valuable tool for investigating the physiological roles and responses of various cell types found in fish species. This review aims to provide an overview of the advancements and applications of fish primary cell culture techniques, focusing on the profiling of physiological roles exhibited by fish cells in vitro. Fish primary cell culture involves the isolation and cultivation of cells directly derived from fish tissues, maintaining their functional characteristics and enabling researchers to study their behavior and responses under controlled conditions. Over the years, significant progress has been made in optimizing the culture conditions, establishing standardized protocols, and improving the characterization techniques for fish primary cell cultures. The review highlights the diverse cell types that have been successfully cultured from different fish species, including gonad cells, pituitary cells, muscle cells, hepatocytes, kidney and immune cells, adipocyte cells and myeloid cells, brain cells, primary fin cells, gill cells, and other cells. Each cell type exhibits distinct physiological functions, contributing to vital processes such as metabolism, tissue regeneration, immune response, and toxin metabolism. Furthermore, this paper explores the pivotal role of fish primary cell culture in elucidating the mechanisms underlying various physiological processes. Researchers have utilized fish primary cell cultures to study the effects of environmental factors, toxins, pathogens, and pharmaceutical compounds on cellular functions, providing valuable insights into fish health, disease pathogenesis, and drug development. The paper also discusses the application of fish primary cell cultures in aquaculture research, particularly in investigating fish growth, nutrition, reproduction, and stress responses. By mimicking the in vivo conditions in vitro, primary cell culture has proven instrumental in identifying key factors influencing fish health and performance, thereby contributing to the development of sustainable aquaculture practices.

The effect of stress on rates of asexual reproduction in an invasive planarian

Animal reproduction under stressful conditions is often reduced, with current survival and future reproduction being generally traded off against current reproductive activity. This study examines the impacts of physical and chemical stressors on the rates of asexual reproduction of the invasive planarian Girardia tigrina. 320 wild-caught planaria (mixed size class) were kept individually in Petri dishes such that their individual rates of fission through fragmentation could be easily monitored. Four treatment groups were compared, one chemical (5 mg/L ammonia) and one physical (decapitation), in comparison to a negative control (animals were starved of food) and a positive control where the animals were given an abundance of food. The two treatment groups immediately began reproducing asexually and accumulated the highest number of fissions over the course of the 12-day investigation period, while the positive control only began to fission after 7 days. We propose that the reproductive response observed here is an adaptive one to stressful conditions, whereby the likelihood of survival through numerical abundance is enhanced, although the size and vulnerability of resulting fragments may impose a balancing cost. The response may play a role in the invasiveness of G. tigrina by making it able to colonize environments where adverse conditions prevail.

181 Effects of Moderate Exercise Regimen on Reproductive Development of Replacement Beef Heifers Reared in Drylots at a High Stocking Density

Abstract This experiment evaluated the physiological and reproductive responses of beef heifers reared at a high stocking density and provided with access to an exercise area. Ninety Angus × Brahman heifers were ranked by age (257 ± 2 days), body weight (BW; 216 ± 2 kg), and temperament score on day 0 and assigned to a) 1 of 6 drylot pens (10x14 m pens; 10 heifers/pen) resulting in a stocking density of 14 m2/heifer (DENS), or b) 1 of 3 paddocks (2-ha paddocks; 10 heifers/paddock) resulting in a stocking density of 2,000 m2/heifer (CON). On day 15, 3 DENS pens were randomly selected and given access to an exercise area (DENS-EX) for 1 hour three times weekly. Negligible forage was available for CON heifers, whereas all heifers received the same limit fed diet which averaged 13 kg/heifer daily of a corn-based diet. Heifer shrunk BW was recorded after 16 h of feed and water withdrawal on days -3 and 225 for average daily gain calculation. On day 0, heifers were fitted with a pedometer behind their shoulder and an ear tag (CowManager, Select Sires, Plain City, OH) to record behavioral responses. Each week, pedometer results were recorded, and blood samples were collected for puberty evaluation via plasma progesterone. Heifers from CON had more (P &amp;lt; 0.05) steps per week compared with DENS and DENS-EX, and DENS-EX heifers had more (P &amp;lt; 0.05) steps per week compared with DENS. Heifers from CON spent more (P &amp;lt; 0.05) time ruminating and eating compared with DENS and DENS-EX. No treatment effects (P ≥ 0.12) were detected for heifer BW gain or temperament variables. Heifers from DENS experienced delayed puberty attainment given a greater (P &amp;lt; 0.05) proportion of CON heifers were pubertal by day 98 compared with DENS and DENS-EX, whereas a greater (P &amp;lt; 0.05) proportion of DENS-EX heifers were pubertal by day 154 compared with DENS heifers (treatment × day interaction, P &amp;lt; 0.01). Collectively, results from this experiment demonstrate that providing heifers reared at a high stocking density access to an exercise area does not result in similar reproductive responses as those reared on pasture at a low stocking density. This work is supported by the USDA National Institute of Food and Agriculture (#2021-67015-34083).