1. Introduction
As the global population increases [
1], beef production is faced with challenges related to the changing climate and use of limited resources [
2,
3]. It is imperative that efficiency of cattle production is improved to increase both environmental and economic sustainability aspects of beef production. One method to improve sustainability is through the use of anabolic implants. Anabolic implants decrease greenhouse gas emissions by 8.9%, and overall land use by 9.1% [
2], effectively decreasing the environmental impact of beef production.
Anabolic implants contain steroid hormones to increase the efficiency and growth rate of cattle [
4,
5,
6], and have been routinely used in cattle production since the late 1950s in the U.S. [
7]. Although not all countries allow the use of anabolic implants in cattle production, there are currently over 40 commercially available implants approved by the Food and Drug Administration in the United States. These anabolic implants are approved for all stages of beef production; from suckling calves to finishing cattle [
5]. Roughly 90% of all cattle on feed in the U.S. receive at least one anabolic implant during production, with 80% receiving two or more [
8]. Anabolic implants can typically be classified as estrogenic, typically containing estradiol (E2), androgenic, typically containing the synthetic testosterone analogue trenbolone acetate (TBA), or as combined, being composed of both estrogenic and androgenic hormones [
7].
Anabolic implants increase economically viable traits such as average daily gain (ADG), feed intake, feed efficiency (FE), hot carcass weight (HCW), and ribeye area (REA) [
4,
7]. However, the exact physiological and molecular mechanisms by which anabolic implants operate to increase skeletal muscle growth in cattle remains elusive [
9,
10,
11,
12]. Furthermore, the increase in growth caused by anabolic implants, may increase trace mineral requirements to support skeletal muscle growth [
13], as lambs implanted with zeranol tended to retain greater amounts of Zn, and lose less Cu and Mn in feces and urine [
14]. When trace minerals are supplemented above national research council recommendations at feedlot consultant recommended concentrations, cattle receiving anabolic implants increase growth even further [
13]. Our hypothesis is that varying hormone type and concentration will alter economically viable traits and mineral status of Angus sired steers. Due to the complex nature of anabolic implants, and the unknowns in their mechanisms of operation to increase skeletal muscle growth, the purpose of this research was to investigate feedlot performance of steers receiving an estradiol only implant (E2), a trenbolone acetate only implant (TBA), or a combined estradiol and trenbolone acetate implant (ETBA) compared to non-implanted steers (CON). Additionally, due to the importance of trace minerals in growth, liver and plasma mineral concentrations were evaluated to better understand the relationship between anabolic implants and trace minerals in beef cattle.
4. Discussion
In the U.S. over 90% of cattle receive at least one anabolic implant at some point during production [
8], as anabolic implants increase overall performance and efficiency of beef cattle [
4]. Implants have the added benefit of also increasing both the environmental sustainability of the industry [
2] and economic return to producers [
15]. However, despite the clear benefits of implanting, the exact physiological and molecular mechanism that anabolic implants operate through to increase overall growth and efficiency remains elusive [
9,
12]. Furthermore, when trace minerals are supplemented at higher concentrations recommended by feedlot consultants, rather than national research council recommendations, an increase in growth is observed [
13]. This increase in growth is even further exacerbated when the animals receive anabolic implants, demonstrating that increased mineral concentrations may be required to support increased growth rates [
13]. Therefore, the purpose of this research was to examine varying hormone type and concentration, estradiol only, trenbolone acetate only, or a combined estradiol and trenbolone acetate implant, on performance in the feedlot, feeding behavior, and concentrations of trace minerals in the plasma and liver of Angus sired steers, to help improve our understanding of anabolic implants. The brief findings of this study were that anabolic implants containing TBA improved growth, and altered trace mineral concentrations, while an E2 only implant altered steer feeding behavior.
Current research suggests anabolic implants decrease land usage by 7.8–9.1% [
2,
19], and greenhouse gas emissions by 5.1% to 8.9% [
2,
19], creating a more environmentally sustainable end-product. This is through increasing ADG and G:F [
4]. In a review published by Duckett and Pratt, the authors state that anabolic implants increase ADG by 18%, feed efficiency by 6%, and feed intake by 6% [
4]. We found that a single anabolic implant containing 120 mg TBA and 24 mg E2 increased overall ADG by 25%, G:F by 14% and DMI by 7%. More recent research completed examining various implant protocols has found that anabolic implants can increase DMI from 5% to 12% [
20]. One likely reason steers used in this trial had increased performance compared to the numbers reported by the review, is that there are variable responses to implants when used in different stages of production [
21]. Specifically, cattle need adequate nutrition before implants can positively influence G:F and gain [
21]. Additionally, the number of implants and type of implants given [
20], the breed of cattle [
15], and sex of cattle [
22] can all influence how cattle respond to anabolic implants. The Duckett and Pratt review published an average of several studies [
4], taken together with the multitude of factors influencing response to anabolic implants, this could explain the increase in performance that was observed in this trial.
Interestingly in our trial, E2 steers did not have altered performance compared to CON steers. In a compilation of implant trials published, animals receiving either a single mild estrogen (around 20 mg estrogen) implant or a single strong estrogen implant (around 200 mg estrogen) had increased ADG and DMI compared to steers that never received an anabolic implant [
23]. The payout period of anabolic implants is the effective period of the implant, which typically varies from 90–120 days [
24], with the payout period being impacted by the hormone concentration [
24]. The steers in the E2 group were implanted with Compudose, an implant containing 25.7 mg estradiol (Elanco Animal Health) with a 200 day payout, and according to the manufacturer, the payout occurs equally over the 200 days. The steers in this trial were harvested at 129 days, which may be part of the reason why E2 did not improve performance of the steers compared to CON.
Red Angus heifers categorized as having high ADG have longer FB durations than those heifers with a low ADG [
17]. We have previously found that Angus sired steers have numerically greater feedlot performance and tended to have longer feed bouts and longer bunk visits than Santa Gertrudis sired steers [
15]. In the current study, steers in the E2 group had shorter FB and BV, and spent less time with their heads down per BV and FB than CON steers. This is interesting, as there was no difference in performance between the E2 and CON steers. This suggests feeding behavior is not always related to feedlot performance, although more research needs to be done to determine the impacts of anabolic implants on feeding behavior.
Although the relationship between trace mineral nutrition and anabolic implants is not well understood, trace minerals can be linked to many aspects of growth. A clear connection between Zn and skeletal muscle protein synthesis has been observed using rodents to assess growth in response to Zn and protein supplementation [
25]. Zinc is vital to cellular proliferation [
26] and is a cofactor to metalloproteinases 2 and 9 [
27], both of which are associated with increased proliferation rates [
11], and protein turnover [
10] in bovine satellite cells. Satellite cells are essentially muscle precursor cells [
28] and are required to support skeletal muscle hypertrophy. Increasing satellite cell numbers allows for an increased capacity for skeletal muscle growth to occur [
29]. Furthermore, the Cu dependent enzyme, lysyl oxidase, is responsible for maintaining the structural integrity of the extracellular matrix [
30], a key component to proper muscle development. Given the strong molecular relationship between trace minerals and pathways associated with skeletal muscle growth, it is important that research is conducted to determine how different anabolic implants impact serum and liver concentrations of trace minerals.
Interestingly, both day 2 liver Cu and Zn were lesser for TBA than CON while day 2 liver Zn was lesser for ETBA than CON, suggesting the androgenic component of these treatments is influencing liver Cu and Zn more so than the estrogenic component. We have previously observed a decrease in liver Cu concentrations of implanted steers 14 days after a combination implant was administered, while liver Zn was greater for implanted steers than non-implanted at harvest [
13]. In agreement with the current work, a decrease in liver Zn concentrations 14 days post-implant administration was observed, coinciding with a decrease in plasma Zn concentrations of implanted steers compared to non-implanted controls on day 13 that remained through day 73 [
31]. The current study design was imperative to finding these TBA driven effects on liver Cu and Zn concentrations, as Niedermayer et al. [
13] and Messersmith [
30] both utilized combination implants that limited data interpretation to the effects of anabolic implant use rather than hormone type. Together, these data indicate trace mineral concentrations are influenced by hormone administration and hormone type. Additionally, it appears that trace minerals such as Cu and Zn, known for roles within many growth processes may be in greater demand by implanted cattle.
Peak hormonal payout of implants has been observed within the first 40 days post-implant administration [
24], indicating this time period should experience the greatest growth rates and subsequently, the greatest need for trace minerals to accommodate that growth. In the present study, the greatest differences in growth occurred within the first 28 days of implanting, interestingly, coinciding with many changes in trace mineral concentrations immediately following implant administration. These data emphasize the importance of trace mineral nutrition, especially Zn, during periods of high growth rates. Although liver Zn was lesser for TBA and ETBA than CON and liver Cu lesser for TBA than CON on day 2, no differences were observed by day 10. However, Niedermayer et al. [
13] and Messersmith [
31] still observed differences in liver trace mineral concentrations 14 days post administration of a combination implant. This difference may be due to the implant potencies used. Both previously mentioned studies [
13,
31] utilized aggressive combination implants (Component TE-200; 200 mg trenbolone acetate and 20 mg estradiol; Elanco Animal Health) compared to the less aggressive estrogen or trenbolone acetate only or combination implant (Revalor-S; 120 mg trenbolone acetate and 24 mg estradiol; Merck Animal Health) used in the current study.
Indeed, implant hormone potency and type can influence mineral stores. The observed increase in day 10 liver Fe concentrations for E2 and ETBA treatments compared to CON indicates a role for E2 in Fe metabolism. Research has found that E2 impairs the transcription of the Fe exporter, ferroportin, through an E2 responsive element [
32]. Therefore, steers implanted with E2 appeared to have limited Fe export from the liver in the current study, however, no effects of E2 implant strategies were observed for plasma Fe concentrations. In addition to Fe metabolism, emerging research has found that heifers implanted with an aggressive two implant strategy (Revalor-200, Merck Animal Health; on days 0 and 91) had lesser liver Mn concentrations than heifers implanted with an extended-release implant (Revalor-XH, Merck Animal Health) on day 0 [
33]. Interestingly, both Niedermayer et al. [
13] and Messersmith [
31] observed decreases in liver Mn concentrations of aggressively implanted steers 14 days post implant administration. These data are in agreement with the decrease observed in liver Mn of TBA steers compared to CON on day 2 and 10. However, the lack of differences in liver Mn due to ETBA suggests either the lesser concentration of TBA in the ETBA implant did not as aggressively affect skeletal muscle protein degradation as the TBA implant alone, or that the addition of E2 in the combination implant supported more skeletal muscle net protein gain. Regardless, the decrease in liver Mn may be due to less skeletal muscle protein degradation in implanted cattle resulting in lesser demand for the urea cycle. Therefore, the Mn dependent terminal enzyme of the urea cycle, arginase [
34,
35], is likely down regulated leading to the decrease in liver Mn observed. However, more work is warranted to confirm how liver Mn is being utilized.
Serum urea nitrogen is a marker of lean tissue anabolism, as it inversely indicates increased N retention [
7]. Implant strategies have been shown to impact SUN concentrations [
20], as the hormones used in implants increase protein accumulation in vivo [
36] and protein synthesis rates in vitro [
10]. In the present study, SUN was investigated on days 0, 2, 10, 28 and 56 from CON, E2, TBA, and ETBA steers. Interestingly, no differences were observed in SUN for these different implant treatments. Research conducted using a different combination estradiol trenbolone acetate implant did not find a difference in SUN until day 213 [
20]. Another study that investigated the effects of anabolic implants on plasma urea nitrogen (PUN) found that steers that received a mild implant (14 mg E2 and 80 mg TBA) and were re-implanted 56 days later with a more aggressive implant (20 mg E2 and 200 mg TBA) had decreased PUN on day 70 of the trial [
13]. These findings taken together suggest that the implant protocol and strength of the implant influence SUN concentrations in steers.
As the use of anabolic implants increases, so does the concern with quality grade of the beef [
37,
38], which is a key component of the grid system used to determine payments to producers for producers in the United States [
39]. A combined high quality grade and low yield grade is optimal for producers paid on the grid system [
39]. Increased marbling increases quality grade, while increased subcutaneous fat undesirably increases yield grade [
40]. Estradiol and TBA have been shown to decrease both marbling [
6] and subcutaneous fat [
37]. In the current study, none of the implant treatments altered marbling, while TBA steers only tended to have improved yield grade compared to CON steers. This is most likely explained as the steers were finished at a group average of 7 mm of rib fat and implants were administered 129 days prior to harvest, both of which helped to minimize any negative effects of implants on carcass characteristics. Research has found that giving cattle implants earlier in the feeding period, rather than later, helps offset potential negative effects of implants on marbling [
41]. Additionally, as previously mentioned, peak payout of the implants typically occurs within the first 40 days post-implanting with most anabolic implants having an effective payout period of 90–120 days [
24]. As the steers were finished harvest at day 129 post-implanting, the negative effects that anabolic implants sometimes have on marbling were not observed in this trial. If the steers were finished to a set weight or were kept on feed longer to reach the U.S. industry standard of 12 mm of rib fat, differences may have been observed in both marbling and yield grade.
Steers in the ETBA group did have increased HCW when compared to the CON steers, with the ETBA implant increasing HCW by 8%. In Duckett and Pratt’s review, they found that on average implants increased carcass weight by 5% [
4]. Interestingly, TBA implants increased REA by 10.7% compared to CON steers, most likely due to increased skeletal muscle protein accretion and muscle growth. Additional emerging research has found that increasing the hormone concentration of anabolic implants leads to an increase in HCW and REA linearly in yearling beef steers [
42]. These findings, taken together, help confirm that anabolic implants increase HCW and REA of cattle.